KR100479198B1 - Anti-inflammatory and immunosuppressive active compounds - Google Patents

Abstract

The compound of the following Chemical Formula 1 provided in the present invention has anti-inflammatory and immunosuppressive activity.

[Formula 1]

Wherein R 'is hydrogen or alkyl; A is substituted with adamantyl or optionally unsaturated mono-, bi- or tricyclic moieties, heterocyclic moieties and / or hydroxy, alkanoyl oxy, amino, aminoalkyl, halogen, alkyl, trialkylamnoniumalkyl Heterocyclic moiety; (a) optionally comprises a double bond or NR ′ as a chain having 1 to 5 carbon atoms; R is hydrogen or phenyl; X is 0 or NR ′ or absent; r and m are each 0, 1 or 2; B is phenylene or cyclohexylene; Y is hydroxy or aminoalkyl, optionally with an oxygen atom in between.

Description

Anti-inflammatory and immunosuppressive active compounds

The present invention relates to novel compounds and their use as anti-inflammatory and immunosuppressive agents.

The role of cytokines (IL-1β, IL-1a, TNFα and IL-6) in the progression of inflammatory reactions is well known (Dinarello CA and Wolff SM, New Eng. J. Med. 328 (2): 106113, 1993 : Tracey KJ and Cerami A., Crit. Care Med. 21: S415m 1993; Melli M. and Parente L., Cytokines and Lipocortin in Inflammation and Cell Differentiation, Wiley-Liss.New York 1990: Dawson MM Lymphoka Causal Interleukin, CRC Publisher, Boca Raton, Florida, 1991). Much research has been done to find compounds called Cytokine Suppressive Anti-Inflammatory Drugs that exert inhibitory effects on pro-inflammatory cytokines, especially IL-1β and TNFα (Lee JC et al., Nature 372: 739, 1994; Davidsen SK and Summers JB, Exp. Opin. Ther. Patents. 5 (10): 1087, 1995), and recently a wide range of agents, also referred to as non-traditional non-steroidal anti-inflammatory drugs (Chiou GCY and Liu SXL, Exp. Opin. Ther. Patents. 6 (1): 41, 1996).

Tanaka et al. (Tanaka et al., Chem. Pharm. Bull., 31 (8), 2810-2819 (1983)) reported the importance of the hydroxamine group in anti-inflammatory activity: Samine groups play a major role in the colorlessness of other parts of the molecule, where groups such as methoxy increase inflammatory efficacy, while groups such as acet amido reduce inflammatory activity.

Surprisingly, the inventors have found that hydroxamic acid derivatives containing amidobenzoic acid components have an excellent anti-inflammatory action with immunosuppressive activity, as reported in the prior literature.

Accordingly, the present invention relates to compounds of the following [Formula 1]:

[Formula 1]

Wherein R 'is hydrogen or (Ci _-4 ) alkyl; A is a mono-, bi- or tricyclic moiety that is adamantyl or optionally partially or wholly unsaturated, and may include one or more heteroatoms selected from the group consisting of N, S or 0, optionally hydroxy , Alkanoyloxy, primary-, secondary- or tertiary amino, amino (C _1-4 ) alkyl, mono- or di (C _1-4 ) alkyl-amino (C _1-4 ) alkyl, halogen, ( C _1-4 ) alkyl, tri (C _1-4 ) alkylammonium (C _1-4 ) alkyl;

The wavy line Is a chain of 1 to 5 carbon atoms optionally containing a double bond or NR 'wherein R' is as defined above;

R is hydrogen or phenyl;

X is an oxygen atom or NR 'and R' is as defined above, or X is absent:

r and m are 0, 1 or 2 respectively:

B is a phenylene or cyclohexylene ring;

Y is a hydroxy or amino (C _1-4 ) alkyl chain, optionally with an oxygen atom in between; only,

The tricyclic group defined in A is fluorenyl, wherein X is not O and at the same time Y is not unless the fluorenyl is substituted with a tri (C _1-4 ) alkylammonium (C _1-4 ) alkyl group It is conditioned only if it is not a hydroxy group.

As described below, the alkyl groups defined above are, for example, methyl, ethyl, 2-methylethyl, 1,3-propyl, 1,4-butyl, 2-ethylethyl, 3-methylpropyl, 1,5-pentyl , 2-ethylpropyl, 2-methylbutyl and analogs thereof, while the mono-, bi- or tricyclic groups defined above are phenyl, cyclohexyl, pyridyl, piperidyl, pyrimidyl, pyridazyl, naphthyl, Nil, anthranyl, phenanthryl, fluorenyl, furanyl, pyranyl, benzofuranyl, chromenyl, xanthyl, isothiazolyl, isoxazolyl, phenothiazyl, phenoxazyl, morpholyl, thiophenyl, benzo Thiophenyl and the like. The halogen atom may be chlorine, bromine or fluorine. Finally, alkanoyloxy groups mean acetyloxy, propionyloxy, isopropionyloxy, butanoyloxy and the like.

The first group of preferred compounds of Formula 1 is wherein R 'is hydrogen; A is phenyl, 1- or 2-naphthyl, cyclohexyl, 1- or 2- 1,2,3,4-tetrahydronaphthyl, adamantyl, quinolinyl, isoquinolinyl, 1- or 2 -Selected from indenyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, optionally substituted as described above; Wavy lines C ₁ -C ₅ , preferably a chain of C ₁ -C ₃ , for example a methylene, ethylene, propylene or propenyl group; Y is OH and R, B, m and r are the compounds as defined above.

A second preferred group is that R 'is hydrogen; A is optionally substituted phenyl or 1- or 2-naphthyl, more preferably 1- or 2-naphthyl; R is phenyl when A is phenyl or hydrogen when A is 1- or 2-naphthyl; R, B, m and r are as defined above; Y is OH and the C ₁ to C ₃ alkylene chain is a compound as defined in the first group of the above preferred compounds.

The A group is preferably decided to be substituted by an alkyl group, _{- (C 1 - C 4)} - alkylamino (C ₁ ~ C _4).

Another object of the present invention relates to the use of the compound as an anti-inflammatory and immunosuppressive agent and to mixing the compound in a pharmaceutical composition containing a pharmaceutically acceptable excipient.

Compounds of the present invention are prepared according to methods known to those skilled in the art. The starting materials used in the preparation methods can be purchased commercially or prepared according to the methods described in the literature. When X is present, the compound of [Formula 1] is prepared using the compound of the following [Formula 2] as a starting material.

[Formula 2]

Here, R and A are as defined above, X 'is an oxygen atom or NR' group, R 'is as defined above. The compounds and carbonic acid reactive derivatives such as, for example, disuccinimidyl carbonate or carbonyl diimidazole (CDI), may be used in combination with triethylamine in an inert solvent such as acetonitrile, tetrahydrofuran (THF), dioxane, or chlorinated solvent. In the presence of a quaternary amine or an aromatic amine such as pyridine, the reaction is carried out at room temperature to the flux temperature of the solvent for about 1 hour to about 48 hours.

As a result, the compound of the following [Formula 3],

[Formula 3]

Wherein R, A and X 'are as defined above and R ₁ is an imidazolyl or hydroxysuccinimidyl group;

In a mixture of organic solvents which can be mixed with water, such as tetrahydrofuran, acetonitrile or alcohol, together with the desired amino acid and water, alkali metals such as sodium hydroxide, alkali or alkaline earth metal carbonates such as sodium carbonate or In the presence of bicarbonate, the reaction is carried out at room temperature for about 1 to about 48 hours.

As a result, the resulting compound of [Formula 4],

[Formula 4]

Wherein R, A, X ', B, m and r are as previously defined;

Treatment with thionyl chloride in a chlorinated solvent or thionyl chloride as a solvent is made to react with acyl chloride by reaction for from about 1 hour to about 12 hours at room temperature to the reflux temperature of the solvent.

Next, when the compound of [Formula 1] in which Y is hydroxy is desired, the acyl chloride and hydroxylamine-HC1 are reacted, and in other cases, the acyl chloride is reacted with an appropriate alkylenediamine. The reaction is carried out for about 1 to about 48 hours at room temperature, in the presence of the inorganic salt described in the previous step, in a mixture of water and an organic solvent that can be mixed with water such as tetrahydrofuran and acetonitrile. It is carried out to generate the compound of [Formula 1].

Alternatively, the acid compound of formula [4] and hydroxylamine HCl or alkylenediamine may be used in an inert solvent such as acetonitrile, tetrahydrofuran, dioxane, or chlorinated solvent, such as a condensing agent such as CDI and triethyleneamine. In the presence of a quaternary amine, the reaction is carried out at room temperature for about 1 hour to about 24 hours. The final product is purified according to conventional chromatography or crystallization methods.

The compound of [Formula 1] in which X is not present is prepared by using the compound of the following Formula [5] as a starting material.

[Formula 5]

Here, R and A are as defined above. The compound is treated with thionyl chloride in a chlorinated solvent or reacted for about 1 to about 12 hours at room temperature to the reflux temperature of the solvent using thionyl chloride as a solvent to activate acylchloride. Alkali or alkaline earth metal carbonates or bicarbonates, such as alkali metal hydroxides such as sodium hydroxide or sodium carbonate, in a mixed solution of the acyl chloride with the desired amino acid in water, an organic solvent such as THF or acetonitrile, and water Under reaction at room temperature for about 1 hour to about 12 hours.

As a result, the compound of the following [Formula 6],

[Formula 6]

Wherein R, A, B, m and r are as defined above;

It is converted to the compound of Formula 1 according to the method already described above.

In the following, some examples of preparing some representative compounds of the present invention are described. Unless otherwise indicated, ¹ H-NMR spectra were recorded in dimethyl sulfoxide (DMSO) using a spectrometer (VARIAN GEMINI 200).

Example 1

4- (5-phenylpentaneamido) benzohydroxyxamic acid

A. Thionyl chloride (2.5 mL, 34 mmol) was added to a 5-phenylpentanone (5 g, 28 mmol) solution dissolved in chloroform (100 mL). The reaction mixture was refluxed for 4 hours and then evaporated to dryness. The residue was redissolved in chloroform and evaporated to dryness three times. The resulting 5-phenylpentanoyl chloride was dissolved in THF (50 mL), and the solution was slowly added to a solution of aminomethylbenzoic acid (3.8 g, 28 mmol) in 1 N sodium hydroxide (56 mL). The reaction mixture was stirred at room temperature for 3 hours, then THF was evaporated under cooling and the aqueous solution was acidified with hydrochloric acid. The precipitate formed was filtered, redissolved in THF, dried over anhydrous calcium chloride and the solvent was evaporated to give 6.7 g of 4- (5-phenylpentaneamido) benzoic acid (yield 81%): m.p. = 227-230 ° C.

¹ H-NMR d 12.5 (s, 1H, exchanged with D ₂ O), 10.3 (s, 1H), 7.92 (d, 2H), 7.75 (d, 2H), 7.25 (m, 5H), 2.63 (t, 2H), 2.41 (t, 2H), 1.65 (m, 4H).

B. Thionyl chloride (3.3 mL, 45 mmol) and 3 drops of pyridine were added to a solution in which the compound (6.7 g, 22 mmol) obtained in step A was dissolved in chloroform (100 mL). The reaction mixture was stirred at rt for 5 h and then evaporated to dryness. The residue was redissolved in chloroform and evaporated to dryness three times. The resulting 4- (5-phenylpentaneamido) benzoyl chloride was dissolved in THF (50 mL), and hydroxylamine-HCl (1.9 g, 27 mmol) and bicarbonate in 1 N sodium hydroxide (27 mL, 27 mmol). To the solution in which sodium (1.9 g, 22 mmol) was dissolved, the solution was slowly added. The reaction mixture was stirred at rt for 3 h, then made acidic with 1N hydrochloric acid and the THF was evaporated under cooling. The aqueous phase was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate and the solvent was evaporated. The crude product was treated with lukewarm ethyl ether and filtered to give 4.8 g of the title compound. (Yield 69%); m.p. = 189-191 ° C

¹ H-NMR d 11.12 (s, 1H), 10.13 (s, 1H), 8.97 (s, 1H), 7.691 (m, 4H), 7.33 ÷ 7.13 (m, 5H), 2.61 (m, 2H), 2.38 (m, 2H), 1.65 ÷ 58 (m, 4H).

Example 2

4- [2,2- (diphenyl) ethoxycarbamoylmethyl] benzohydroxyxamic acid

A. A solution of 2,2-diphenylethanol (6 g, 30 mmol) and CDI (4.9 g, 30 mmol) dissolved in THF (30 mL) was stirred at room temperature for 2 hours, followed by 1 N sodium hydroxide (30 A solution of 4-aminomethylbenzoic acid (4.6 g, 30 mmol) dissolved in mL) was added. The reaction mixture was stirred at room temperature for 3 hours, then acidified with hydrochloric acid, THF was evaporated under cooling, and the aqueous phase was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and the solvent was evaporated. The crude product was treated with lukewarm n-hexane and filtered to give 10 g of 4- [2,2- (diphenyl) ethoxycarbamoyl-methyl] benzoic acid (yield 90%); m.p. = 102-105 ° C.

¹ H-NMR d 12.7 (s, 1H, exchanged for D ₂ 0), 7.91 (d, 2H), 7.78 (t, 1H), 7.50-7.20 (m, 12H), 4.61 (d, 2H), 4.38 ( t, 1H), 4.23 (d, 2H).

B. Thionyl chloride (1.3 mL, 8 mmol) and 3 drops of pyridine were added to a solution of the compound (4.5 g, 1 mmol) obtained in step A in chloroform (50 mL). The reaction mixture was stirred at rt for 3 h and then evaporated to dryness. The residue was redissolved in chloroform and evaporated to dryness three times. The resulting 4- [2,2 (diphenyl) ethoxycarbamoylmethyl] -benzoyl chloride was dissolved in THF (50 mL), 1N sodium hydroxide (14.4 mL, 14.4 mmol) and THF (20 mL). To the solution dissolved hydroxylamine HCl (1 g, 14.4 mmol) and sodium bicarbonate (1 g, l2 mmol) was slowly added. The reaction mixture was stirred at room temperature for 3 hours, then acidified with 1N hydrochloric acid and THF was evaporated under cooling. The aqueous phase was extracted with ethyl acetate, the organic phase was dried together under anhydrous sodium sulfate and the solvent was evaporated. The resulting crude product was purified by column chromatography using a silica gel column (eluent: methylene chloride / methanol 15: 1), and the resulting product was treated with lukewarm ethyl ether and filtered to yield 1.5 g of the title compound. Obtained (yield 32%): mp = 164-166 ° C.

¹ H-NMR d 11.21 (s, 1H), 9.04 (s, 1H), 7.74 (t, 1H), 7.72 (m, 2H), 7.38 ÷ 7.20 (m, 12H), 4.60 (d, 2H), 4.38 (t, 1 H), 4.20 (d, 2 H).

Example 3

4- [2- (adamant-1-yl) -ethoxycarbamoyl] benzohydroxyxamic acid

A. Dissolves 2- (adamant-1-yl) ethanol (2.1 g, l2 mmol), dissuccinimidyl carbotate (3.3 g, l3 mmol) and pyridine (0.86 mL, 10 mmol) in acetonitrile (50 mL). The mixed solution was stirred at room temperature for 24 hours, then the solvent was evaporated under cooling, and the residue was dissolved in methylene chloride (100 mL). The solution was washed with water (50 mL) and 1N hydrochloric acid (50 mL) and again with water (50 mL). The organic phase was dried over anhydrous sodium sulfate and the solvent was evaporated to afford 3 g (9 mmol) of 2- (adamant-1-yl) ethyl-succinimidyl carbonate crude product. The resulting crude product was dissolved again in THF (30 mL), and the resulting solution was dissolved in a solution of 4-aminobenzoic acid (1.3 g, 9 mmol) and sodium carbonate (0.98 g, 9 mmol) in water (30 mL). After addition, it was stirred overnight at room temperature and acidified with hydrochloric acid. THF was evaporated under cooling and the aqueous phase was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate to evaporate the solvent. The resulting crude product was treated with lukewarm n-hexane and filtered to afford 3.2 g of 4- [2- (adamant-1-yl) ethoxycarbamoyl] benzoic acid (yield 77%); m.p. = 197-200 ° C.

¹ H-NMR d 12.6 (s, 1H, exchanged with D ₂ 0), 10.0 (s, 1H), 7.89 (d, 2H), 7.60 (d, 2H), 4.19 (t, 2H), 1.93 (m, 5H), 1.75 ÷ 1.40 (m, 12H).

B. Thionyl chloride (1.2 mL, 17.4 mmol) and 3 drops of pyridine were added to the solution obtained by dissolving the compound (3 g, 8.7 mmol) obtained in step A in chloroform (50 mL). The reaction mixture was refluxed for 5 hours and then evaporated to dryness. The residue was redissolved in chloroform and evaporated to dryness three times. The resulting 4- [2- (adamant-1-yl) ethoxycarbamoyl] -benzoyl chloride was dissolved in THF (50 mL), 1N sodium hydroxide (10.4 mL, 10.4 mmol) and THF ( 20 ml) was added slowly to a solution of hydroxylamine HCl (0.7 g, 10.4 mmol) and sodium bicarbonate (0.7 g, 8.7 mmol), stirred at room temperature for 8 hours, and then acidified with 1 N hydrochloric acid. And THF were evaporated under cooling. The aqueous phase was extracted with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate to evaporate the solvent. The resulting crude product was treated with lukewarm ethyl ether and filtered to give 1.5 g of the title compound (yield 48%); m.p. = 198-200 ° C.

¹ H-NMR d 11.08 (s, 1H), 9.84 (s, 1H), 8.94 (s, 1H), 7.70 (m, 2H), 7.53 (m, 2H), 4.17 (m, 2H), 1.94 (m , 3H) 1.73 ÷ 1.37 (m, 14H).

Example 4

4-[(naphthyl-1-yl) methoxycarbamoyl] benzohydroxyxamic acid

A. 7 g of pure 4-[(naph-1-yl) methoxycarbamoyl] benzoic acid according to the method described in Example 3A with naphthyl-1-yl-methanol (5 g, 31 mmol) as starting material. Was obtained (yield 81%).

¹ H-NMR d 12.7 (s, 1H, exchanged with D ₂ O), 10.2 (s, 1H), 8.18 (d, 1H), 7.97 (m, 4H), 7.61 (m, 6H), 5.69 (s, 2H).

B. Using the compound prepared in Step A (6 g, 19 mmol) as a starting material, 1.86 g of the title compound were obtained according to the method described in Example 3B (yield 30%); m.p. = 192-194 ° C.

¹ H-NMR d 11.12 (s, 1H), 10.06 (s, 1H), 8.96 (s, 1H), 8.14 (m, 1H), 8.03 ÷ 7.96 (m, 2H), 7.76 ÷ 7.50 (m, 8H) , 5.67 (s, 2 H).

Example 5

4-[(1,2,3,4-tetrahydronaphth-2-yl) methoxycarbamoyl] -benzohydroxamic acid

A. According to the method described in Example 3A, starting with (1,2,3,4-tetrahydronaphth-2-yl) methanol (4 g, 24 mmol), 5.2 g of pure 4-[(1 , 2,3,4-tetrahydronaphth-2-yl) methoxycarbamoyl] benzoic acid was obtained (yield 67%): mp = 203-206 ° C.

¹ H-NMR d 12.6 (s, 1H, exchanged with D ₂ O), 10.1 (s, 1H), 7.92 (d, 2H), 7.63 (d, 2H), 7.1 (m, 4H), 4.14 (d, 2H), 2.88 (m, 3H), 2.54 (m, 1H), 2.25-1.90 (m, 2H), 2.49 (m, 1H).

B. Using the compound obtained in Step A (5.2 g, 16 mmol) as starting material, 4.1 g of the title compound were obtained according to the method described in Example 3B (yield 75%); m.p. = 184-186 ° C.

¹ H-NMR d 11.10 (s, 1H), 9.97 (s, 1H), 8.98 (s, 1H), 7.72 (m, 2H), 7.54 (m, 2H), 7.09 (s, 4H), 4.11 (d , 2H), 2.92 ÷ 2.75 (m, 3H), 2.61 ÷ 2.47 (m, 1H), 2.17 ÷ 1.93 (m, 2H), 1.56 ÷ 1.35 (m, 1H).

Example 6

4-[(3-phenylpropoxy) carbamoyl] benzohydroxyxamic acid

A. 7.7 g of pure 4-[(3-phenylpropoxy) carbamoyl] benzoic acid was obtained according to the method described in Example 3A using 3-phenylpropanol (5 g, 36 mmol) as a starting material (yield 7I%). mp; 171-173 ° C.

¹ H-NMR d 12.6 (s, 1H, exchanged with D ₂ O), 10.1 (s, H), 7.90 (d, 2H), 7.61 (d, 2H), 7.28 (m, 5H), 4.13 (t, 2H), 2.72 (t, 2H), 1.98 (m, 2H).

B. Using the compound obtained in Step A (7 g, 23 mmol) as a starting material, 5.8 g of the title compound were obtained according to the method described in Example 3B (yield 80%); m.p. = 179-181 ° C.

¹ H-NMR d 11.09 (s, 1H), 9.94 (s, 1H), 8.95 (s, 1H), 7.71 (m, 2H), 7.54 (m, 2H), 7.36 ÷ 7.15 (m, 5H), 4.10 (t, 2H), 2.70 (m, 2H), 1.95 (m, 2H).

Example 7

4-[(naphth-2-yl_methoxycarbamoyl] benzohydroxyxamic acid

A. 6.6 g of pure 4-[(naph-2-yl) methoxycarbamoyl] according to the method described in Example 3A with naphth-2-yl-methanol (5 g, 31 mmol) as starting material Benzoic acid was obtained (yield 68%); mp = 241-243 ° C.

¹ H-NMR d 12.7 (s, 1H, exchanged for D ₂ O), 10.2 (s, 1H), 7.95 (m, 6H), 7.60 (m, 5H), 5.39 (s, 2H).

B. 4.5 g of the title compound were obtained according to the method described in Example 3B using the compound (6 g, 18.6 mmol) obtained in step A as a starting material (yield 72%); m.p. = 220-222 ° C.

¹ H-NMR d 11.10 (s, 1H), 11.01 (s, 1H), 8.95 (s, 1H), 7.99 ÷ 7.90 (m, 4H), 7.73 (m, 2H), 7.61 ÷ 7.50 (m, 5H) , 5.36 (s, 2 H).

Example 8

(E) -4-[(3-phenyl-prop-2-enyl) carbamoyl] benzohydroxyxamic acid

A. 1.75 g (13 mmol) of trans-cinnamol was dissolved in l30 ml of acetonitrile under argon atmosphere, and then 5 g (l 9 mmol) of N, N'-disuccinimidylcarbonate were added under heating to completely dissolve. The mixture was cooled to 20 ° C. and pyridine (0.65 mL, 1 g, 12 mmol) was added. The reaction mixture was covered with aluminum and stirred at room temperature for 30 hours. The solvent was evaporated under cooling, the residue was dissolved in ethyl acetate and washed repeatedly with 0.1 N hydrochloric acid and finally with water. The solution was dried and evaporated to dryness in a cold state to obtain a crude product, which was dissolved in 26 mL of dioxane and p-aminobenzoic acid (1.79 g) in water (26 mL) with 1.38 g (13 mmol) of sodium carbonate. 13 mmol) was added to the dissolved solution. The solution was stirred for 60 hours at room temperature, then THF and brine were added. The organic phase was separated, washed with 0.1 N hydrochloric acid (twice) and brine was added again, followed by drying to evaporate the solvent. The resulting crude product was dissolved in isopropyl ether and filtered to give 0.7 g of (E) -4-[(3-phenyl-prop-2-enyl) carbamoyl] benzoic acid (yield 18 ° C.); m.p. = 176-178 ° C.

¹ H-NMR d 12.70 (s, 1H, exchanged with D ₂ O), 10.20 (s, 1H), 7.93 (d, 2H), 7.61 (d, 2H), 7.55 ÷ 7.25 (m, 5H), 6.78 ( d, 1H), 6.46 (dt, 1H), 4.87 (d, 2H).

B. The compound of step A (0.7 g, 2.3 mmol) was dissolved in 8 mL of anhydrous THF and CDI (0.46 g, 2.8 mmol) was added at 0 ° C. The mixture was stirred overnight at room temperature and then hydroxylamine HCl (0.2 g, 2.3 mmol) was added thereto and then stirring continued for another 60 hours at room temperature. The precipitated product was filtered off, suspended in 1N hydrochloric acid and stirred overnight. Filtration and vacuum drying gave 400 mg of the title compound (yield 55.6%): m. P. = 194-195 ° C.

¹ H-NMR d 11.14 (exchanged with s, 1H, D ₂ O), 10.07 (s, 1H), 8.97 (exchanged with s, 1H, D ₂ O), 7.75 (d, 2H), 7.65 ÷ 7.25 (m , 7H), 6.79 (d, 1H), 6.48 (dt, 1H), 4.85 (d, 2H).

Example 9

(Z) -4-[(3-phenyl-prop-2-enyl) carbamoyl] benzohydroxyxamic acid

A. 2.4 g of (Z) -4-[(3-phenyl-prop-2-enyl) carba according to the method described in Example 8A with cis-cinnamol (2.82 g, 20 mmol) as starting material Moyl] benzoic acid was obtained (yield 40%).

¹ H-NMR d 12.72 (s, 1H, exchanged with D ₂ O), 10.20 (s, 1H), 7.92 (d, 2H), 7.60 (d, 2H), 7.55 ÷ 7.25 (m, 5H), 6.73 ( d, 1H), 5.91 (dt, 1H), 4.96 (d, 2H).

B. 400 mg of the title compound were obtained according to the method described in Example 8B using the step A product (0.7 g, 2.3 mmol) as a starting material (yield 55.6%); m.p. = 169-170 ° C.

¹ H-NMR d 11.13 (s, 1H, exchanged with D ₂ O), 10.05 (s, 1H), 8.98 (s, 1H, exchanged with D ₂ O), 7.74 (d, 2H), 7.55 (d, 2H ), 7.50 ÷ 7.25 (m, 5H), 6.77 (d, 1H), 5.90 (dt, 1H), 4.94 (d, 2H).

Example 10

4- [3,3- (diphenyl) propoxycarbamoyl] benzohydroxyxamic acid

A. 4.2 g of 4- [3,3- (diphenyl) propoxycarbamoyl] benzoic acid was obtained according to the method described in Example 3A using 3,3-diphenylpropanol (3 g, 14 mmol) as a starting material. (Yield 80%); m.p. = 156-159 ° C.

¹ H-NMR d 12.6 (s, 1H, exchanged with D ₂ O), 10.1 (s, 1H), 7.90 (d, 2H), 7.60 (d, 2H), 7.40 ÷ 7.15 (m, 10H), 4.17 ( t, 1H), 4.03 (t, 2H), 2.46 (m, 2H).

B. Using the compound of step A (4.0 g, 10 mmol) as a starting material, 1 g of the title compound was obtained according to the method described in Example 3B (yield 26%), m. P. = 196-197 ° C.

¹ H-NMR d 11.08 (s, 1H), 9.93 (s, 1H), 8.93 (s, 1H), 7.70 (m, 2H), 7.54 (m, 2H), 7.39 ÷ 7.15 (m, 10H), 4.15 (t, 1 H), 3.99 (t, 2H), 2.44 (q, 2H).

Example 11

N- (2-aminoethyl-4-[(fluoren-9-yl) methoxy-carbamoylmethyl] benzamide HCl

A. A solution of 9-fluorenylmethyl chloroformate (5 g, 19 mmol) dissolved in THF (50 mL) was slowly added to 4-aminomethyl benzoic acid solution (2.9 g, 19 mmol). The reaction mixture was stirred at room temperature for 3 hours and then acidified with 1N hydrochloric acid and THF was evaporated under cooling. The aqueous phase was extracted with ethyl acetate and the organic phases were combined and dried over anhydrous sodium sulfate to evaporate the solvent. The resulting crude product was treated with lukewarm ethyl ether and filtered to give 7 g of 4-[(fluorene-9-yl) methoxy-carbamoylmethyl] benzoic acid (yield 98%); m.p. = 232-235 ° C.

¹ H-NMR d 12.7 (s, 1H, exchanged for D ₂ O), 7.94 (m, 5H), 7.77 (d, 2H), 7.58 + 7.30 (m, 6H), 4.42 (d, 2H), 4.30 ( m, 3H).

B. Thionyl chloride (1.46 mL, 20 mmol) was added to a solution of compound A (3.7 g, 10 mmol) in chloroform (100 mL), and the reaction mixture was refluxed for 2 hours and then evaporated to dryness. . The residue was dissolved in chloroform and evaporated three times. The resulting 4-[(fluorene-9-yl) methoxy-carbamoylmethyl] benzoyl chloride was dissolved in THF (30 mL) and 2- (t in water (20 mL) and THF (30 mL). To this solution was slowly added butyloxyimino) ethylamine (1.6 g, 10 mmol) and sodium bicarbonate (0.8 g, 10 mmol). The reaction mixture was stirred at rt for 12 h and then evaporated THF under cooling, acidified the aqueous phase with 1N hydrochloric acid and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate to evaporate the solvent. The resulting crude product was treated with lukewarm ethyl ether and filtered to give 3.85 g of N- (2-t-butyloxycarbamoylmethyl) -4-[(fluoren-9-yl) methoxy carbamoylmethyl] benza Mid was obtained (yield 75%): mp = 167-169 ° C.

¹ H-NMR d 8.45 (t, 1H), 7.94 ÷ 7.70 (m, 7H), 7.54 ÷ 7.25 (m, 6H), 6.96 (t, 1H), 4.42 (d, 2H), 4.27 (m, 3H) , 3.32 (q, 2H), 3.13 (q, 2H), 1.41 (s, 9H).

C. The product of step B (3.6 g, 7 mmol) was added in small portions to trifluoroacetic acid (25 mL) at 0 ° C. The mixture was stirred at rt for 4 h, then the acid was evaporated and the residue dissolved in lukewarm ethyl ether and filtered to give the product. The product was dissolved in THF and methanol and hydrochloric acid ether solution was added thereto. The solvent was evaporated and the process repeated four times. The resulting crude product was dissolved in some water at 0 ° C. and filtered to give 2 g of the title compound (yield 65%); m.p. = 182-184 ° C.

¹ H-NMR d 8.79 (t, 1H), 8.17 (s, 3H), 7.96 ÷ 7.86 (m, 5H), 7.72 (m, 2H), 7.48 ÷ 7.29 (m, 6H), 4.38 (m, 2H) , 4.25 (m, 3H), 3.55 (q, 2H), 3.00 (t, 2H).

Example 12

4- [6- (diethylaminomethyl) naphth-2-ylmethyloxycarbamoyl] benzohydroxamic acid HCl

A. 1- (3-dimethylaminopropyl) in a solution in which 2,6-naphthalenedicarboxylic acid (25 g, 115 mmol) and hydroxybenzotriazole (15.6 g, 115 mmol) were dissolved in dimethylformamide (l800 mL). ) -3-ethylcarbodiimide HCl (EDCI) (22.2 g, 115 mmol) was added and the mixture was stirred at rt for 2 h. Diethyl amine (34.3 mL, 345 mmol) was added and the solution stirred overnight at room temperature. The solvent was evaporated under reduced pressure and the resulting crude product was treated with 1N hydrochloric acid (500 mL) and ethyl acetate (500 mL), then the insoluble compound was filtered and layered. The organic phase was extracted with 5% sodium carbonate (3 x 200 mL) and the aqueous solution was combined, acidified with concentrated hydrochloric acid and extracted with ethyl acetate (3 x 200 mL). The organic phase was washed with 1N hydrochloric acid (6x100 mL), dried under anhydrous sodium sulfate and the solvent removed under reduced pressure to yield 18.5 g (60% yield) of pure 6- (diethylaminocarbonyl) -2-naphthalenecarboxylic acid. Obtained: mp = 122-124 ° C.

¹ H-NMR d 8.67 (s, 1 H), 8.25-8.00 (m, 4 H), 7.56 (d, 1 H), 3.60-3.20 (m, 4 H), 1.30-1.00 (m, 6H).

B. 6- (diethylaminocarbonyl) -2-naphthalenecar dissolved in THF (200 mL) in a reflux suspension suspended in lithium aluminum hydroxide (7.5 g, 199 mmol) in THF (500 mL). Acid (18 g, 66 mmol) was added slowly. The mixture was refluxed for one hour and then cooled to room temperature, treated with THF (25 mL) and water (3.5 mL), treated with 20% sodium hydroxide (8.5 mL) and finally water (33 mL). ). The white solid was filtered off and the solvent was removed under reduced pressure. The crude product was dissolved in diethyl ether (200 mL) and extracted with 1N hydrochloric acid (3 × 100 mL). The aqueous phase was treated with 32% sodium hydroxide and extracted with diethyl ether (3x100 mL). The organic phase was dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure to give 12.7 g (yield 79%) of pure 6- (diethylaminomethyl) -2-naphthalenemethanol as a thick oil.

¹ H-NMR d 7.90-7.74 (m, 4H), 7.49 (m, 2H), 5.32 (t, 1H, exchanged with D ₂ O), 4.68 (d, 2H), 3.69 (s, 2H), 2.52 ( S, 4H), 1.01 (t, 6H).

C. A solution of 6- (diethylaminomethyl) -2-naphthalenemethanol (12.5g, 51mmol) and N, N'-disuccinimidyl carbonate (13.2g, 51mmol) in acetonitrile (250ml) was dissolved. After stirring for 3 hours at room temperature, the solvent was removed and the crude product was dissolved in THF (110 mL). The solution was added to a solution of 4-aminobenzoic acid (7.1 g, 51 mmol) and sodium carbonate (5.5 g, 51 mmol) dissolved in water (200 mL) and THF (100 mL). The mixture was stirred overnight at room temperature, then THF was removed under reduced pressure and the solution was treated with 1N hydrochloric acid (102 mL, 102 mmol). The resulting precipitate was filtered, dried under reduced pressure, triturated in diethyl ether and filtered to give 13.2 g (64% yield) of pure 4- [6- (diethylaminomethyl) naphth-2-ylmethyloxycarbamoyl [- Benzoic acid was obtained; m.p. = 201-205 ° C. (dec).

¹ H-NMR d 10.26 (s, 1H), 8.13 (s, 1H), 8.05-7.75 (m, 6H), 7.63 (m, 3H), 5.40 (s, 2H), 4.32 (s, 2H), 2.98 (q, 4H), 1. 24 (t, 6H).

D. 4- [6- (diethylaminomethyl) naphth-2-ylmethyloxycarbamoyl] benzoic acid (13.1 g, 32 mmol) and thionyl chloride (7 mL, 96 mmol) dissolved in chloroform (300 mL) The solution was refluxed for 4 hours and then the solvent and thionyl chloride were evaporated. The crude product was dissolved in chloroform (100 mL) and evaporated to dryness three times. Solid crude product was dissolved in water (150 mL) and THF (50 mL) with hydroxylamine HCl (2.7 g, 39 mmol), sodium bicarbonate (5.4 g, 64 mmol) and 1N sodium hydroxide (39 mL, 39 mmol). To the prepared solution. The mixture was stirred overnight at room temperature, then THF was removed under reduced pressure and the aqueous phase was extracted with ethyl acetate (3 × 100 mL). The organic phases were combined and dried under anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude product was dissolved in THF and treated with 1.5 N hydrochloric acid ether solution. The solid product was filtered and dried to give 6 g (41% yield) of pure 4- [6- (diethylaminomethyl) naphth-2-yl-methyloxycarbamoyl] benzohydroxyxamic acid HCl as a white solid; m.p. = 162-165 ° C.

¹ H-NMR d 11.24 (exchanged with s, 1H, D ₂ O), 10.88 (exchanged with s, 1H, D ₂ O), 10.16 (s, 1H), 8.98 (exchanged with bs, 1H, D ₂ O) , 8.21 (s, 1H), 8.10-7.97 (m, 3H), 7.89 (d, 1H), 7.80-7.55 (m, 5H) 5.39 (s, 2H), 4.48 (d, 2H), 3.09 (m, 4H), 1.30 (t, 6H).

Example 13

4- [6- (dipropylaminomethyl) naphth-2-ylmethyloxycarbamoyl] benzohydroxyxamic acid HCl

1 g of pure 4- [6- (dipropylamino) according to the same method as described in Example 12, using 2,6-naphthalenedicarboxylic acid (5 g) and dipropyl amine (9.6 ml) as starting materials. Methyl) naphth-2-ylmethyloxycarbamoyl] benzohydroxamic acid HCl as a white solid; m.p. = 140-142 ° C.

¹ H-NMR d 11.15 (s, 1H, swapped with D ₂ O), 10.95 (s, 1H, swapped with D ₂ O), 10.16 (s, 1H), 8.20 (s, 1H), 8.10-7.97 (m , 3H), 7.89 (d, 1H), 7.80-7.55 (m, 5H) 5.40 (s, 2H), 4.50 (d, 2H), 2.98 (m, 4H), 1.79 (m, 4H), 0.88 (t , 6H).

Example 14

4- [6- (dibutylaminomethyl) naphth-2-ylmethyloxycarbamoyl] benzohydroxyxamic acid HCl

1.2 g of pure 4- [6- (dibutyl) was prepared according to the same method as described in Example 12, using 2,6-naphthalenedicarboxylic acid (5 g) and dibutyl amine (11.8 ml) as starting materials. Aminomethyl) naphth-2-ylmethyloxycarbamoyl] benzohydroxyxamic acid HCl as a white solid; m.p. = 137-141 ° C.

¹ H-NMR d 11.19 (exchange with s, 1H, D ₂ O), 10.91 (exchange with s, 1H, D ₂ O), 10.16 (s, 1H), 8.96 (exchange with bs, 1H, D ₂ O) , 10.16 (s, 1H), 8.96 (bs, 1H, exchanged with D ₂ O), 8.21 (s, 1H), 8.10-7.98 (m, 3H), 7.87 (d, 1H), 7.80-7.55 (m, 5H) 5.38 (s, 2H), 4.52 (d, 2H), 3.02 (m, 4H), 1.77 (m, 4H), 1.30 (m, 4H), 0.89 (t, 6H).

Example 15

4- [4- (diethylaminomethyl) naphth-1-ylmethyloxycarbamoyl] benzohydroxamic acid HCl

1.1 g of pure 4- [4- (diethyl) according to the same method as described in Example 12, using 1,4-naphthalenedicarboxylic acid (5 g) and diethyl amine (7.3 ml) as starting materials. Aminomethyl) naphth-2-ylmethyloxycarbamoyl] benzohydroxyxamic acid HCl as a white solid; m.p. = 162-165 ° C.

¹ H-NMR d 11.24 (s, 1H, exchanged for D ₂ O), 10.48 (s, 1H, exchanged for D ₂ O), 10.13 (s, 1H), 8.43 (m, 1H), 8.26 (m, 1H ), 8.04 (d, 1H), 7.82-7.70 (m, 5H) 7.58 (d, 2H), 5.73 (s, 2H), 4.84 (d, 2H), 3.17 (m, 4H), 1.32 (t, 6H ).

Example 16

4- [6- (diethylaminomethyl) naphth-1-ylmethyloxycarbamoyl] benzohydroxamic acid HCl

A. 6- (diethylaminocarbamoyl) -2-naphthalenecarboxylic acid (prepared according to the method of Example 12A) (14 g, 52 mmol) and thionyl chloride (3.8 mL, 52 mmol) in chloroform (200 mL) The dissolved solution was refluxed for 3 hours and then the solvent and thionyl chloride were evaporated. The crude product was dissolved in chloroform (100 mL) and evaporated to dryness three times. The crude product was dissolved in THF (50 mL) and added at 0 ° C. to 32% ammonium hydroxide (10 mL) solution dissolved in water (50 mL) and THF (50 mL). The mixture was stirred at rt overnight, then THF was removed under reduced pressure and the solid was filtered dried to afford 11.6 g (yield 81%) of 6- (diethylaminocarbamoyl) -2-naphthalenecarboxamide. This product was used for the next step without further purification.

B. A solution of 6- (diethylaminocarbonyl) -2-naphthalenecarboxamide (11.6 g, 42 mmol) in THF (100 mL) was dissolved in lithium aluminum hydride (4.9 g, THF (100 mL)). 128 mmol) was added slowly to the suspended suspension. The mixture was refluxed for 2 hours and then cooled to room temperature, treated with a mixture of THF (16 ml) and water (2.2 ml), then treated with 20% sodium hydroxide (5.5 ml) and finally water (22 ml). ). The white solid was filtered off and the solvent was removed under reduced pressure. The crude product was purified by chromatography using silica gel (eluent chloroform-methanol-ammonium hydroxide 15: 1: 0.1) to give 8.1 g (yield 80%) of pure 6- (diethylaminomethyl) -2- Naphthylmethylamine was obtained as a wax solid.

¹ H-NMR d 7.78 (m, 4H), 7.49 (m, 2H), 3.89 (s, 2H), 3.67 (s, 2H), 2.50 (q, 4H), 1.00 (t, 6H).

C. A solution of 6- (diethylaminomethyl) -2-naphthylmethylamine (6 g, 24 mmol) and N, N'-disuccinimidyl carbonate (6.3 g, 24 mmol) in acetonitrile (200 mL) After stirring for 3 hours at room temperature, the solution was dissolved in a solution of 4-amizobenzoic acid (3.4 g, 24 mmol) and sodium carbonate (2.6 g, 24 mmol) in water (100 mL) and THF (100 mL). Added. The mixture was stirred at rt for 48 h, then 1N hydrochloric acid (48 mL, 48 mmol) was added and the solvent was removed under reduced pressure. The crude product was purified by chromatography using silica gel (eluent chloroform-methanol-ammonium hydroxide 7: 3: 0.5) to give 3.5 g (yield 36% pure 4- [6- (diethylaminomethyl) naphth- 2-ylmethylaminocarbamoyl] benzoic acid was obtained mp = 179-183 ° C.

¹ H-NMR d 9.58 (s, 1H), 7.95-7.78 (m, 6H), 7.65-7.45 (m, 4H), 7.29 (t, 1H), 4.51 (d, 2H), 3.81 (s, 2H) , 2.62 (q, 4 H), 1.07 (t, 6 H).

D. 4- [6- (diethylaminomethyl) naphth-2-ylmethylaminocarbamoyl] benzoic acid (3.1 g, 7.6 mmol) and thionyl chloride (1.1 mL, 15.2 mmol) in dimethylformamide (30 mL) ) Was stirred overnight at room temperature, then the suspension was diluted with diethyl ether and the solid was filtered-dried to give 3.2 g of 4- [6- (diethylaminomethyl) naphth-2-ylmethylamino Carbamoyl] benzoyl chloride crude product. This compound was dissolved in water (30 mL) and THF (40 mL) with hydroxylamine HCl (0.6 g, 8.5 mmol), sodium bicarbonate (1.2 g, 14 mmol) and 1N sodium hydroxide (8.5 mL, 8.5 mmol). To the prepared solution, in solid state. The mixture was stirred at rt for 2 h, then the mixture was saturated with sodium chloride, the organic phase was separated and the aqueous phase was extracted with THF. The organic phases were combined and dried under anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude product was dissolved in hot THF (150 mL) and the insoluble material was filtered off. The solution was cooled to room temperature and treated with 1.5 N hydrochloric acid ether solution. The solid product was filtered to dryness to obtain 1.2 g (yield 38%) of pure 4- [6- (diethylaminomethyl) naphth-2-ylmethylaminocarbamoyl] -benzohydroxamic acid HCl as a white solid; m.p. = 167-168 ° C.

¹ H-NMR d 11.07 (exchanged with s, 1H, D ₂ O), 10.49 (exchanged with bs, 1H, D ₂ O), 9.49 (s, 1H), 8.93 (exchanged with s, 1H, D ₂ O) , 8.15 (s, 1H), 8.05-7.87 (m, 3H), 7.81 (d, 1H), 7.72 (d, 2H), 7.65-7.50 (m, 3H) 7.27 (t, 1H), 4.54 (d, 2H), 4.48 (s, 2H), 3.11 (m, 4H), 1.30 (t, 6H).

Example 17

4- [N-isopropyl-1,2,3,4-tetrahydroisoquinol-3-yl) methyloxycarbamoylmethyl] benzohydroxyxamic acid HCl

A. 1,2,3,4-tetrahydro-3-isoquinoline-carboxylic acid (9 g, 42 mmol), 2-bromopropane (8 mL, 84 mmol) and 1 N sodium hydroxide in ethanol (l70 mL) 168 mmol) dissolved solution was refluxed for 5 hours, then 2-bromopropane (8 mL, 84 mmol) and 1N sodium hydroxide (168 mL, 168 mmol) were added and the mixture was refluxed for 5 hours. . Ethanol was removed and the aqueous solution was treated with 6N hydrochloric acid to pH = 7. The unreacted starting material was recovered by filtration and the solvent was removed under reduced pressure. The crude product was dissolved in ethanol, the inorganic salts were filtered off and the organic phase was evaporated under reduced pressure. This process was repeated three times to obtain 8.1 g (87% yield) of 1,2,3,4-tetrahydro-3-isoquinolinecarboxylic acid, which was used in the next step without further purification.

¹ H-NMR δ 7.13 (m, 4H), 4.16 (d, 1H), 3.89 (d, 1H), 3.58 (t, 1H), 3.43 (m, 1H), 3.13 (dd, 1H), 2.94 (dd) , 1H), 1.19 (d, 3H), 1.11 (d, 3H).

B. A solution of N-isopropyl-1,2,3,4-tetrahydro-3-isoquinolinecarboxylic acid (8.0 g, 36 mmol) suspended in THF (100 mL) was suspended in THF (100 mL). It was added slowly to the lithium aluminum hydride (2.1 g, 54 mmol) reflux suspension. The mixture was refluxed for 2 hours and then cooled to room temperature, treated with a mixture of THF (7 mL) and water (0.9 mL), followed by 20% sodium hydroxide (2.3 mL) and finally water (9.2 mL). ). The white solid was filtered and the solvent was removed under reduced pressure to give 5.0 g (68% yield) of pure N-isopropyl-1,2,3,4-tetrahydroisoquinol-3-ylmethanol as a thick oil.

¹ H-NMR δ 7.12 (m, 4H), 4.54 (bs, 1H, exchanged for D ₂ O), 3.68 (s, 2H), 3.55-3.35 (m, 2H), 3.20-2.90 (m, 2H), 2.79 (d, 2H), 1.10 (d, 3H), 1.01 (d, 3H).

C. N-isopropyl-1,2,3,4-tetrahydroisoquinoline-3-ylmethanol (4.6 g, 22.4 mmol) and 1,1'-carbonyldiimidazole (3.63 g) in THF (50 mL) , 22.4 mmol) was dissolved at room temperature for 3 hours. The stirred solution was added to a solution of 4-aminomethylbenzoic acid (3.38 g, 22.4 mmol) and 1 N sodium hydroxide (22.4 mL, 22.4 mmol) in water (20 mL). The solution was stirred overnight at room temperature, then 1N hydrochloric acid (22.4 mL, 22.4 mmol) was added and the solvent was removed under reduced pressure. The crude product was purified by chromatography on silica gel (eluent chloroform-methanol-ammonium hydroxide 8: 2: 0.5) to give 3.1 g (yield 35%) of pure 4-[(N-isopropyl-1,2, 3,4-tetrahydroisoquinol-3-yl) -methyloxycarbamoyl] benzoic acid was obtained as a white solid. m.p. = 93-95 ° C.

¹ H-NMR δ 7.92 (d, 2H), 7.86 (t, 1H), 7.36 (t, 2H), 7.14 (m, 4H), 4.26 (d, 2H), 4.07 (dd, 1H), 3.73 (s , 2H), 3.68 (dd, 1H), 3.25 (m, 1H), 3.02 (m, 1H), 2.89 (dd, 1H), 2.72 (dd, 1H), 1.10 (d, 3H), 1.04 (d, 3H).

D. 4-[(N-isopropyl-1,2,3,4-tetrahydroisoquinol-3-yl) methyloxycarbamoyl] benzoic acid (3.0 g, 7.8 mmol) and thionyl in chloroform (100 mL) The solution in which chloride (1.7 mL, 23 mmol) was dissolved was refluxed for 3 hours, and then the solvent and thionyl chloride were evaporated. The crude product was dissolved in chloroform (100 mL) and evaporated to dryness three times. The crude product was dissolved in THF (50 mL) and dissolved in water (30 mL) and THF (20 mL) with hydroxylamine HCl (0.65 g, 9.4 mmol), sodium bicarbonate (1.3 g, 15.7 mmol) and 1N sodium hydroxide. (9.4 mL, 9.6 mmol) was added to the dissolved solution. The mixture was stirred at rt for 1 h, then THF was removed under reduced pressure and the aqueous phase was extracted with ethyl acetate (3 × 100 mL). The organic phases were combined and dried under anhydrous sodium sulfate and the solvent was removed under reduced pressure. The crude product was dissolved in THF and treated with 1.5 N hydrochloric acid ether solution. The solid product was filtered and dried to yield 2.1 g (yield 62%) of pure 4-[(N-isopropyl-1,2,3,4-tetrahydroisoquinol-3-yl) methyloxycarbamoyl] benzohydroxyxamin Acid HCl was obtained as a white solid. m.p. = 154-157 ° C.

¹ H-NMR δ 11.24 (s, 1H), 10.75 (s, 1H), 9.07 (bs, 1H), 8.01 (t, 1H), 7.70 (d, 2H), 7.34 (d, 2H), 7.32 (m , 4H), 4.38 (d, 2H), 4.26 (m, 4H) 4.02 (m, 1H), 3.78 (m, 1H), 3.15 (d, 2H), 1.42 (d, 3H), 1.30 (d, 3H) ).

The following compounds can be prepared according to methods analogous to those described above:

4-[(4-dimethylaminomethyl-naphth-2-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(4-Diethylaminoethyl-naphth-2-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(4-dimethylaminoethyl-naphth-2-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(6-dimethylaminomethyl-naphth-2-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(6-di-isopropylaminomethyl-naphth-2-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(4-dimethylaminomethyl-naphth-2-yl) methoxycarbamoyl] methylbenzohydroxyxamic acid

4-[(4-dimethylaminomethyl-naphth-2-yl) ethoxycarbamoyl] benzohydroxyxamic acid

4-[(5,6,7,8-tetrahydronaphth-2-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(1,2,3,4-tetrahydronaphth-2-yl) glycineamido] benzohydroxyxamic acid

4-[(4-Diethylaminomethyl-naphth-2-yl) ethoxycarbamoyl] benzohydroxyxamic acid

4-[(6-dimethylaminomethyl-naphth-2-yl) ethoxycarbamoyl] benzohydroxyxamic acid

4-[(6-diethylaminomethyl-naphth-2-yl) ethoxycarbamoyl] benzohydroxyxamic acid

4-[(1,2,3,4-tetrahydronaphth-2-yl) methoxycarbamoyl] benzohydroxamic acid

4-[(4-Dimethylaminomethyl-naphth-1-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(4-dimethylaminoethyl-naphth-1-yl) methoxycarbamoyl] benzohydroxylamino acid

4-[(5-Dimethylaminomethyl-naphth-1-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(5-Diethylaminomethyl-naphth-1-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(5-Di-n-propylaminomethyl-naphth-1-yl) methoxycarbamoyl] benzohydroxylamino acid

4-[(5-Di-isopropylaminomethyl-naphth-1-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(5-Di-n-butylaminomethyl-naphth-l-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(6-dimethylaminomethyl-naphth-1-yl) methoxycarbamoyl] benzohydroxylamino acid

4-[(6-diethylaminomethyl-naphth-1-yl) methoxycarbamoyl] benzohydroxylamino acid

4-[(6-di-n-propylaminomethyl-naphth-1-yl) methoxycarbamoyl] benzohydroxylamino acid

4-[(6-Di-isopropylaminomethyl-naphth-1-yl) methoxycarbamoyl] benzohydroxylamino acid

4-[(6-di-n-butylaminomethyl-naphth-1-yl) methoxycarbamoyl] benzohydroxylamine acid

4-[(4-dimethylaminomethyl-naphth-1-yl) methoxycarbamoyl] methylbenzohydroxyxamic acid

4-[(4-Dimethylaminomethyl-naphth-1-yl) ethoxycarbamoyl] benzohydroxyxamic acid

4-[(4-Diethylaminomethyl-naphth-l-yl) ethoxycarbamoyl] benzohydroxyxamic acid

4-[(5-Dimethylaminomethyl-naphth-1-yl) ethoxycarbamoyl] benzohydroxyxamic acid

4-[(5-Diethylaminomethyl-naphth-1-yl) ethoxycarbamoyl] benzohydroxylamino acid

4-[(6-Dimethylaminomethyl-naphth-1-yl) ethoxycarbamoyl] benzohydroxyxamic acid

4-[(6-diethylaminomethyl-naphth-1-yl) ethoxycarbamoyl] benzohydroxyxamic acid

4- [N- (naphth-1-yl-methyl) glycineamido] benzohydroxyxamic acid

4- [N- (naphth-2-yl-methyl) glycineamido] benzohydroxyxamic acid

4-[(N-methyl-1,2,3,4-tetrahydroisoquinol-5-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(N-ethyl-1,2,3,4-tetrahydroisoquinol-5-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(isoquinol-5-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(N-methyl-1,2,3,4-tetrahydroisoquinol-6-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(N-ethyl-1,2,3,4-tetrahydroisoquinol-6-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(isoquinol-6-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(N-methyl-1,2,3,4-tetrahydroisoquinol-1-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(N-ethyl-1,2,3,4-tetrahydroisoquinol-1-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(isoquinol-1-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(N-methyl-1,2,3,4-tetrahydroisoquinol-3-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(N-ethyl-1,2,3,4-tetrahydroisoquinol-3-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(isoquinol-3-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(N-methyl-1,2,3,4-tetrahydroisoquinol-4-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(N-ethyl-1,2,3,4-tetrahydroisoquinol-4-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(isoquinol-4-yl) methoxycarbamoyl] benzohydroxyxamic acid

4- [3- (1,2,3,4-tetrahydroisoquinol = 2-yl) propionamido] benzohydroxyxamic acid

4-[(benzothiophen-4-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(benzothiophen-5-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(benzofuran-4-yl) methoxycarbamoyl] benzohydroxyxamic acid

4-[(benzofuran-5-yl) methoxycarbamoyl] benzohydroxyxamic acid

4- [4- (diethylaminopropyl) naphth-1-ylmethyloxycarbamoyl] benzohydroxamic acid HCl

4- [3- (diethylaminomethyl) naphth-1-ylmethyloxycarbamoyl] benzohydroxyxamic acid HCl

4- [3- (diethylaminoethyl) naphth-1-ylcarbamoyl] benzohydroxamic acid HCl

4- [3- (diethylaminopropyl) naphth-1-ylmethyloxycarbamoyl] benzohydroxyxamic acid HCl

4- [4- (diethylaminopropyl) naphth-1-ylmethylaminocarbamoyl] benzohydroxamic acid HCl

4- [3- (diethylaminomethyl) naphth-1-ylmethylaminocarbamoyl] benzohydroxamic acid HCl

4-3- (diethylaminoethyl) naphth-1-ylmethylaminocarbamoyl] benzohydroxamic acid HCl

4- [3- (diethylaminopropyl) naphth-1-ylmethylaminocarbamoyl] benzohydroxamic acid HCl

4- [6- (dipropylaminomethyl) naphth-2-ylmethylaminocarbamoyl] benzohydroxamic acid HCl

4- [6- (dibutylaminomethyl) naphth-2-ylmethylaminocarbamoyl] benzohydroxamic acid HCl

4- [4- (diethylaminomethyl) naphth-1-ylmethylaminocarbamoyl] benzohydroxamic acid HCl

4- [4- (dipropylaminomethyl) naphth-1-ylmethylaminocarbamoyl] benzohydroxamic acid HCl

4- [4- (diethylaminoethyl) naphth-1-ylmethylaminocarbamoyl] benzohydroxamic acid HCl

Pharmacological activity

The anti-inflammatory and immunosuppressive activity of the compounds of the invention was tested through in vitro and in vivo experiments.

1. In vitro cytokine production test

The compounds of the invention were tested for their activity, particularly on IL-1β production in humans. Mononuclear cells (PBMCs) were prepared from peripheral blood ("Buffie coat") obtained from healthy subjects using a density gradient centrifuge (Ficoll-Hypaque, Biochrom KG, Berlin, Germany). Under humid air containing 37 ° C., 5% carbon dioxide, cells (2.5 × 10 ⁶ per ml) were cultured in 96-well plates (Nunc) with a final volume of 200 μl. The cell culture medium was RPMI 1640 N-acetyl-L-alanyl-L-glutamine (“low endotoxin”; Biochrom KG) with low endotoxin content, with 1% fetal bovine serum (Hyclone Laboratories Inc., Logan, Utah), 100 Ul / ml penicillin and 100 μg / ml streptomycin were added. The production of cytokines was induced by stimulating cells with 10 ng / ml LPS (lipopolysaccharide) obtained from E. coli serotype 055: B5 (Sigma Chemical Co., St. Louis, MT). The cells were pretreated for 60 minutes with compounds of the invention (final concentration: 0.05%) dissolved in DMSO. The compound of the present invention and LPS were present for a total of 20 hours of incubation time. Unstimulated cells were used as negative controls. At the end of the test, the supernatants were harvested and tested for production of IL-1β using specific ELISA assays (“sandwich antigen capture”, R & D system, Minneapolis, Minnesota). To cover 96-well microtiter plates (Nunc), goat's polyclonal serum specific for human IL-1β was used and purified using affinity. After washing, the wells were incubated for 2 hours with 3% bovine serum albumin (BSA) in buffered saline. Recombinant human IL-1β was used to obtain a standard graph for each test, and the superior samples were diluted l / 20 and l / 80 with PBS + 0.1% BSA. The plates were then incubated at 4 ° C. After washing, a secondary antibody, ie, mouse anti-human IL-1β monoclonal antibody, was added. After incubation and washing, peroxidase-conjugated goat anti-Ig-G polyclonal antibody (Zymed) was added followed by the pigmented substrate tetramethylbenzidine dichloride (from Sigma). The reaction was stopped using 4N sulfuric acid and the absorbance at 450 nm was measured using an automatic spectrometer (Perkin-Elmer). Inhibitory activity of the compounds of the present invention was expressed as the concentration of compounds that inhibit the production of IL-1β by 50% compared to IC _50, the control.

2. In Vivo LPS-Induced TNFα Production Test

Male mouse BALB / c (18-2Og) was purchased from Harlan-Nossan (Correza, Italy); Lipoplissaccharide (LPS) was obtained from Streptococcus enterotidis (code L-6011) purchased from Sigma (St. Louis, Montana); Dexamethasone (Soldsam) was purchased from Lavoratorio Pharmacologico Milanes (Caronno P., Italy).

LPS ( S. enteritidis , 7.5 mg / kg) was intraperitoneally administered to mice. Dexamethasone was intraperitoneally administered 30 minutes prior to LPS administration. Compounds of the invention were orally administered 90 minutes prior to LPS administration. Two hours after administration of LPS, anesthetized experimental animals were regenerated, blood was collected through cardiac puncture, and the blood was allowed to coagulate. Serum TNF α was measured by ELISA using mouse recombinant TNF α as a standard and using a mouse reagent set (Genzyme, Cambridge, Mass.) According to the manufacturer's instructions.

From the above test results, it was demonstrated that the test compound is as effective or more effective in inhibiting IL-1β and TNF α production as dexamethasone, control compound and well-known anti-inflammatory agents.

The present invention also relates to methods of using the compounds of Formula 1 as anti-inflammatory and immunosuppressive agents, i.e., to the use as well as to any industrially applicable method in this regard, including mixing the compounds into pharmaceutical compositions. will be. Examples of such pharmaceutical compositions include tablets, dragee creams, ointments and vials, of which the vials are suitable for both oral and intramuscular or intravenous administration. Such compositions comprise the active ingredient alone or in admixture with conventional pharmaceutically acceptable excipients and carriers.

The dosage of the active ingredient can be varied within a wide range depending on the type of compound used, and can be administered once a day or several times a day depending on the therapeutic conditions.

Claims (6)

The compound of Formula 1; [Formula 1] Wherein R 'is hydrogen or (C _1-4 ) alkyl; A is a mono-, bi- or tricyclic moiety that is adamantyl or optionally partially or wholly unsaturated, and may include one or more heteroatoms selected from the group consisting of N, S or 0, optionally hydroxy , Alkanoyloxy, primary-, secondary- or tertiary amino, amino (C _1-4 ) alkyl, mono- or di (C _1-4 ) alkyl-amino (C _1-4 ) alkyl, halogen, ( C _1-4 ) alkyl, tri (C _1-4 ) alkylammonium (C _1-4 ) alkyl; The wavy line Is a chain of 1-5 carbon atoms, optionally including a double bond, or NR ', wherein R' is as defined above: R is hydrogen or phenyl; X is an oxygen atom or NR 'and R' is as defined opposite, or X is absent; r and m are each 0, 1 or 2; B is a phenylene or cyclohexylene ring; Y is a hydroxy or amino (C _1-4 ) alkyl chain optionally with an oxygen atom in between; only, The tricyclic group defined in A is fluorenyl, wherein X is not 0 and at the same time Y is not unless the fluorenyl is substituted with a tri (C _1-4 ) alkylammonium (C _1-4 ) alkyl group It is conditioned only if it is not a hydroxy group. The compound of claim 1, wherein R ′ is hydrogen; A is phenyl, 1- or 2-naphthyl, cyclohexyl, 1- or 2-1,2,3,4-tetrahydronaphthyl, adamantyl, quinolinyl, isoquinolinyl, 1- or Selected from the group consisting of 2-indenyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, optionally substituted as defined in claim 1, and a wavy line Is a C ₁ to C ₅ chain, Y is OH and R, B, m and r are as defined in claim 1. The compound of claim 1, wherein R ′ is hydrogen; A is optionally substituted phenyl or 1- or 2-naphthyl; R is phenyl when A is phenyl and hydrogen when A is 1- or 2-naphthyl; R, B, m and r are as defined in claim 1; Y is OH and the C ₁ to C ₅ alkylene chain is as defined in claim 2. The compound of claim 1, which is used to prepare a medicament suitable as an anti-inflammatory agent. The compound of claim 1, which is used to prepare a medicament suitable as an immunosuppressive agent. A pharmaceutical composition comprising at least one compound of claim 1 as an active ingredient with a pharmaceutically acceptable excipient.