CHIR-090

CHIR-090 is an L-threonine derivative obtained by formal condensation of the carboxy group of 4-({4-[(morpholin-4-yl)methyl]phenyl}ethynyl)benzoic acid with the amino group of N-hydroxy-L-threoninamide. It has a role as an antimicrobial agent, a lipopolysaccharide biosynthesis inhibitor and an EC 3.5.1.108 (UDP-3-O-acyl-N-acetylglucosamine deacetylase) inhibitor. It is an acetylenic compound, a member of morpholines, a member of benzamides, a L-threonine derivative and a hydroxamic acid.

Sodium methoxide (25 wtpercent in MeOH, 1.860 g, 8.60 mmol) was added to a stirred solution of hydroxylaminehydrochloride (400 mg, 5.76 mmol) in anhydrous MeOH (5 mL) at 0 °C under N2 atmosphere. After stirring 20 min, asolution of methyl ester 4 (829 mg, 1.90 mmol) in 1:1 MeOH/THF (6 mL) was added and the reaction mixture stirred at0 °C for 1 h and at room temperature for 4 h. The reaction was quenched with 1.0 M HCl (6 mL), concentrated by rotaryevaporation to remove organic solvents, and diluted with DMSO (4 mL). Analytical RP-HPLC (C18 column, CH3CNgradient 5-35percent, 0.1percent TFA, UV analysis 300 nm, 16 min) indicated a purity of 85percent for the crude product mixture.Purification by preparative RP-HPLC and lyophilization of the collected fractions gave 701 mg (81percent) of 5 as a fluffywhite solid. LRMS (ES+) m/z 438.1 (C24H27N3O5 + H requires 438.20); RP-HPLC (300 nm, 16 min run) 8.7 min.Preparation of (2S, 3R)-N-(2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide (5). Sodium methoxide (25 wt percent in MeOH, 1.860 g, 8.60 mmol) was added to a stirred solution of hydroxylamine hydrochloride (400 mg, 5.76 mmol) in anhydrous MeOH (5 mL) at 0° C. under NTo a solution of compound 6 (7.8 g, 17.9 mmol) in methanol/THF (1:1) was added aqueous 50percent hydroxylamine (15 mL) followed by catalytic amount of KCN (10 mg) and the resulting solution was stirred at room temperature for 18 h. The reaction mass was acidified to pH ~6 with aqueous 10percent citric acid solution (20 mL) and diluted with water (100 mL). The solution was extracted with 10percent methanol in chloroform (150 mL x 2).The combined organic layer was washed with brine solution (100 mL), dried over anhydrous MgSO4 and evaporated under reduced pressure to get crude compound. The crude product was purified by column chromatography (silica gel, 100-200 mesh) using 1.5percent methanol in chloroform as mobile phase to yield pure product (4.5 g, 57percent). LC-MS 438.6 m/z (M+l). MW = 437.2 amu 'H NMR (400 MHz, DMSO-d6) 10.69(1H, NH, 1H), 8.87(s, OH, 1H), 8.14-8.17(d, NH, 1H), 7.94-7.96(d, 2H), 7.64-7.67(d, 2H), 7.54-7.56(d, 2H), 7.38-7.40(d, 2H), 4.90-4.91(s, NH, 1H), 4.26-4.28(dd, 1H), 4.01-5.05(dd, 1H), 3.57-3.59(br s, 4H), 3.50(s, 2H), 2.36-2.49(s, 4H), 1.09-1.10(s, 3H) ppmGeneral procedure: A solution of the amine (1.5 mmol) and trimethyl orthoformate (180 muL, 1.625 mmol) in THF (20 mL) was added to the aldehyde-containing resin (105 mg, 0.25 mmol). An atmosphere of nitrogen was established and after mixing for 5 minutes, acetic acid (180 muL, 3.08 mmol) followed by a solution of NaBH3CN (71 mg, 1.125 mmol) in methanol (1 mL) was added. Mixing was continued for 44 hours after which the resin was filtered, drained and washed with DMF (2 x 10 mL) and methanol (3 x 10 mL), drained again and dried in vacuo. Cleavage from the resin was achieved by treatment with trifluoroacetic acid (10 mL) for 30 minutes. The solution was collected and concentrated to dryness to give a crude residue which was purified by preparative HPLC using a e.g. Gilson GX-281 semi-preparative HPLC system equipped with a Luna 200 x 25 mm (C18, 10 mu) or a Gemini 150 x 30 mm (C18, 5 mu) column applying a gradient consisting of 0.1% TFA/water and acetonitrile.To a solution of compound 6 (7.8 g, 17.9 mmol) in methanol/THF (1:1) was added aqueous 50% hydroxylamine (15 mL) followed by catalytic amount of KCN (10 mg) and the resulting solution was stirred at room temperature for 18 h. The reaction mass was acidified to pH ~6 with aqueous 10% citric acid solution (20 mL) and diluted with water (100 mL). The solution was extracted with 10% methanol in chloroform (150 mL x 2).The combined organic layer was washed with brine solution (100 mL), dried over anhydrous MgSO4 and evaporated under reduced pressure to get crude compound. The crude product was purified by column chromatography (silica gel, 100-200 mesh) using 1.5% methanol in chloroform as mobile phase to yield pure product (4.5 g, 57%). LC-MS 438.6 m/z (M+l). MW = 437.2 amu 'H NMR (400 MHz, DMSO-d6) 10.69(1H, NH, 1H), 8.87(s, OH, 1H), 8.14-8.17(d, NH, 1H), 7.94-7.96(d, 2H), 7.64-7.67(d, 2H), 7.54-7.56(d, 2H), 7.38-7.40(d, 2H), 4.90-4.91(s, NH, 1H), 4.26-4.28(dd, 1H), 4.01-5.05(dd, 1H), 3.57-3.59(br s, 4H), 3.50(s, 2H), 2.36-2.49(s, 4H), 1.09-1.10(s, 3H) ppmSodium methoxide (25 wt% in MeOH, 1.860 g, 8.60 mmol) was added to a stirred solution of hydroxylaminehydrochloride (400 mg, 5.76 mmol) in anhydrous MeOH (5 mL) at 0 C under N2 atmosphere. After stirring 20 min, asolution of methyl ester 4 (829 mg, 1.90 mmol) in 1:1 MeOH/THF (6 mL) was added and the reaction mixture stirred at0 C for 1 h and at room temperature for 4 h. The reaction was quenched with 1.0 M HCl (6 mL), concentrated by rotaryevaporation to remove organic solvents, and diluted with DMSO (4 mL). Analytical RP-HPLC (C18 column, CH3CNgradient 5-35%, 0.1% TFA, UV analysis 300 nm, 16 min) indicated a purity of 85% for the crude product mixture.Purification by preparative RP-HPLC and lyophilization of the collected fractions gave 701 mg (81%) of 5 as a fluffywhite solid. LRMS (ES+) m/z 438.1 (C24H27N3O5 + H requires 438.20); RP-HPLC (300 nm, 16 min run) 8.7 min.Preparation of (2S, 3R)-N-(2-Hydroxy-1-hydroxycarbamoyl-propyl)-4-(4-morpholin-4-ylmethyl-phenylethynyl)-benzamide (5). Sodium methoxide (25 wt % in MeOH, 1.860 g, 8.60 mmol) was added to a stirred solution of hydroxylamine hydrochloride (400 mg, 5.76 mmol) in anhydrous MeOH (5 mL) at 0 C. under N2 atmosphere. After stirring 20 min, a solution of methyl ester (4) (829 mg, 1.90 mmol) in 1 : 1 MeOH/THF (6 mL) was added and the reaction mixture stirred at 0 C. for 1 h and at room temperature for 4 h. The reaction was quenched with 1.0 M HCl (6 mL), concentrated by rotary evaporation to remove organic solvents, and diluted with DMSO (4 mL). Analytical RP-HPLC (C18 column, CH3CN gradient 5-35%, 0.1% TFA, UV analysis 300 ran, 16 min) indicated a purity of 85% for the crude product mixture. Purification by preparative RP-HPLC and lyophilization of the collected fractions gave 701 mg (8 l%) of(5) as a fluffy white solid. LRMS (ES+) m/z 438.1 (C24H27N3O5+H requires 438.20); RP-HPLC (300 ran, 16 min run) 8.7 min.

Molecular Weight:437.5
XLogP3:1.3
Hydrogen Bond Donor Count:4
Hydrogen Bond Acceptor Count:6
Rotatable Bond Count:8
Exact Mass:437.19507097
Monoisotopic Mass:437.19507097
Topological Polar Surface Area:111
Heavy Atom Count:32
Complexity:680
Defined Atom Stereocenter Count:2
Covalently-Bonded Unit Count:1
Compound Is Canonicalized:Yes