diethyl (2S)-2-aminopentanedioate

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of L-glutamic acid diethyl ester (0.81 g, 4.0 mmoles) and 4-(((3-amino-1,2-dihydro-1-oxobenzo[f]quinazolin-9-yl)amino)sulfonyl)benzoic acid (0.43 g, 0.9 mmoles) in dry dimethylformamide (20 ml) was added 1-hydroxybenzotriazole monohydrate (0.43 g, 3.2 mmoles) and dicyclohexylcarbodiimide (0.66 g, 3.2 mmoles). The solution was stirred at room temperature under a nitrogen atmosphere for 20 hrs, after which time the solvent was removed under reduced pressure. The residue was recrystallized successively from methylene chloride:methanol/9:1 and methanol to give diethyl N-(4-(((3-amino-1,2-dihydro-1-oxobenzo[f]quinazolin-9-yl)amino)sulfonyl)benzoyl)-L-glutamate as a white solid. (0.215 9, 36%). 1HNMR(DMSO-d6, 300 MHz) δ: 1.12(t, J=7 Hz, 3H, ester-CH3); 1.15(t, J=7 Hz, 3H, ester-CH3); 1.89-2.13(m, 2H, glu-CH2); 2.40(t, J=7.6 Hz, 2H, glu-CH2); 4.00(q, J=7 Hz, 2H, ester-CH2); 4.08(q, J=7 Hz, 2H, ester-CH2); 6.55(br s, 2H, NH2); 7.16(d, J=8.9 Hz, 1H, Ar); 7.23(dd, J=8.7, 2 Hz, 1H, Ar); 7.73(d, J=8.7 Hz, 1H, Ar); 7.89(d, J=8.9 Hz, 1H, Ar); 7.96(m, 4H, Ar); 8.89(d, J=7.4 Hz, 1H, glu-NH); 9.48(d, J=2 Hz, 1H, Ar); 10.67(s, 1H, NH); 11.08(s, 1H, NH). Anal. Calculated for C28H29N5O8S: C, 56.46; H, 4.91; N, 11.76; S. 5.38. Found: C, 56.45; H, 4.94; N, 11.75; S, 5.43. after which time the solvent was removed under reduced pressure The residue was recrystallized successively from methylene chloride

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of 0.2 g of 2-acetamido-4-hydroxy-6-[2-(4-carboxyphenyl)prop-1-enyl]pyrido[2,3-d]pyrimidine in 50 mL of N-methylpyrrolidinone containing 0.18 g of N-methylmorpholine was added 0.22 g of phenyl N-phenylphosphoramidochloridate in a single portion. After stirring the mixture at room temperature for 1 hour, 0.20 g of diethyl L-glutamate was added. The reaction mixture was stirred overnight, the solvent was removed under reduced pressure and the residue was triturated with chloroform. The mixture was filtered and the filtrate was evaporated under reduced pressure. The residue was subjected to preparative thin layer chromatography on silica gel using a 5% methanol in chloroform mixture as the eluent. This gave 74.6 mg (25%) of the title compound as a pale yellowish solid; NMR (CDCl3, 250 MHz) d 1.22 (t, 3H, J=7.1 Hz), 1.30 (t, 3H J=7.1 Hz), 2.11-2.57 (m, 10H), 4.14 (q, 2H, J=7.1 Hz), 4.24 (q, 2H, J=7.1 Hz), 4.75-4.83 (m, 1H), 6.86 (brs, 1H), 7.18 (brs, 1 H), 7.57 (d, 2H, J=8.42 Hz), 7.84 (d, 2H, J=8.42 Hz), 8.50 (d, 1H, J=2.01 Hz), 8.96 (brs, 1H), 10.34 (brs, 1H). The reaction mixture was stirred overnight the solvent was removed under reduced pressure the residue was triturated with chloroform The mixture was filtered the filtrate was evaporated under reduced pressure

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a 100 mL 14/20 round bottom flask under a nitrogen atmosphere was charged 1.0 g (4.81 mmmol) of 2-bromo-4-thiazolecarboxylic acid in 10 mL of benzene, followed by the addition of 1.4 mL (19 mmol) of thionyl chloride, and a catalytic amount of dimethylformamide. The reaction was heated to reflux for 2 hours The volatiles were removed in vacuo, and this residue was then dissolved in 15 mL of methylene chloride and added dropwise to an ice-bath cooled mixture of 1.21 g (5.05 mmol) of L-glutamic acid diethyl ester, 1.41 mL (10.1 mmol) of triethylamine, and 5 mg of dimethylaminopyridine in 15 mL methylene chloride. After the addition, the ice bath was removed and the reaction was stirred at room temperature for 2 hours The reaction was diluted with methylene chloride, washed with 0.1 N hydrochloric acid, water, 5% sodium bicarbonate, water, dried over sodium sulfate, and removed in vacuo. The crude residue was purified using silica gel flash chromatography eluting with a gradient of 1:2 ethyl acetate/hexanes to 1:1 ethyl acetate/hexanes to give 0.9 g (48%) of N-[(2-bromo-5-thiazolyl)carbonyl]-L-glutamic acid diethyl ester as a yellow oil. Rf =0.30 (1:2 ethyl acetate/hexanes), 1H NMR (300 MHz, CDCl3) δ 1.24-1.34 (m, 6H), 2.15-2.32 (m, 2H), 2.47-2.55 (m, 2H), 4.12-4.30 (m, 4H), 4.64-4.71 (m, 1H), 7.97 (s, 1H). The reaction was heated to reflux for 2 hours The volatiles were removed in vacuo this residue was then dissolved in 15 mL of methylene chloride added dropwise to an ice-bath cooled After the addition the ice bath was removed was diluted with methylene chloride washed with 0.1 N hydrochloric acid, water, 5% sodium bicarbonate, water dried over sodium sulfate removed in vacuo The crude residue was purified eluting with a gradient of 1:2 ethyl acetate/hexanes to 1:1 ethyl acetate/hexanes

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a 100 mL 14/20 round bottom flask under a nitrogen atmosphere was charged 1.0 g (4.81 mmmol) of 2-bromo-4-thiazolecarboxylic acid in 10 mL of benzene, followed by the addition of 1.4 mL (19 mmol) of thionyl chloride, and a catalytic amount of dimethylformamide. The reaction was heated to reflux for 2 hours The volatiles were removed in vacuo, and this residue was then dissolved in 15 mL of methylene chloride and added dropwise to an ice-bath cooled mixture of 1.21 g (5.05 mmol) of L-glutamic acid diethyl ester, 1.41 mL (10.1 mmol) of triethylamine, and 5 mg of dimethylaminopyridine in 15 mL methylene chloride. After the addition, the ice bath was removed and the reaction was stirred at room temperature for 2 hours The reaction was diluted with methylene chloride, washed with 0.1 N hydrochloric acid, water, 5% sodium bicarbonate, water, dried over sodium sulfate, and removed in vacuo. The crude residue was purified using silica gel flash chromatography eluting with a gradient of 1:2 ethyl acetate/hexanes to 1:1 ethyl acetate/hexanes to give 0.9 g (48%) of N-[(2-bromo-5-thiazolyl)carbonyl]-L-glutamic acid diethyl ester as a yellow oil. Rf =0.30 (1:2 ethyl acetate/hexanes), 1H NMR (300 MHz, CDCl3) δ 1.24-1.34 (m, 6H), 2.15-2.32 (m, 2H), 2.47-2.55 (m, 2H), 4.12-4.30 (m, 4H), 4.64-4.71 (m, 1H), 7.97 (s, 1H). The reaction was heated to reflux for 2 hours The volatiles were removed in vacuo this residue was then dissolved in 15 mL of methylene chloride added dropwise to an ice-bath cooled After the addition the ice bath was removed was diluted with methylene chloride washed with 0.1 N hydrochloric acid, water, 5% sodium bicarbonate, water dried over sodium sulfate removed in vacuo The crude residue was purified eluting with a gradient of 1:2 ethyl acetate/hexanes to 1:1 ethyl acetate/hexanes

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a 100 mL 14/20 round bottom flask under a nitrogen atmosphere was charged 1.7 g (8.17 mmmol) of 2-bromo-4-thiazolecarboxylic acid in 17 mL of benzene, followed by the addition of 2.4 mL (33 mmol) of thionyl chloride, and a catalytic amount of dimethylformamide. The reaction was heated to reflux for 2 hours The volatiles were removed in vacuo, and this residue was then dissolved in 20 mL of methylene chloride and added dropwise to an ice-bath cooled mixture of 2.06 g (8.58 mmol) of L-glutamic acid diethyl ester, 2.39 mL (10.1 mmol) of triethylamine, and 10 mg of dimethylaminopyridine in 30 mL of methylene chloride. After the addition, the ice bath was removed and the reaction was stirred at room temperature for 2 hours The reaction was diluted with methylene chloride, washed with 0.5 N hydrochloric acid, water, 5% sodium bicarbonate, water, dried over sodium sulfate, and removed in vacuo. The crude residue was purified using silica gel flash chromatography eluting with 3:1 chloroform/ether to give 2.7 g (84%) of diethyl N-(2-bromo-4-thiazolylcarbonyl)-L-glutamate as a yellow oil. Rf =0.43 (3:1 chloroform/ether). 1H NMR (300 MHz, DMSO-d6) δ 1.14 (q, J=7.1 Hz, 6H), 1.98-2.18 (m, 2H), 2.35 (t, J=7.3 Hz, 2H), 3.97-4.11 (m, 4H), 4.37-4.50 (m, 1H), 8.28 (d, J=5.9 Hz, 1H), 8.73 (d, J=7.7 Hz, 1H) The reaction was heated to reflux for 2 hours The volatiles were removed in vacuo this residue was then dissolved in 20 mL of methylene chloride added dropwise to an ice-bath cooled After the addition the ice bath was removed was diluted with methylene chloride washed with 0.5 N hydrochloric acid, water, 5% sodium bicarbonate, water dried over sodium sulfate removed in vacuo The crude residue was purified eluting with 3:1 chloroform/ether

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Following the procedures outlined in Example 34, substituting 3-(6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)aniline (intermediate 31.5) for 4-(6,8-dichloro-2-methyl-1,2,3,4-tetrahydroisoquinolin-4-yl)aniline and (S)-diethyl 2-aminopentanedioate for (S)-dimethyl 2-aminosuccinate gave, after purification by preparative HPLC, the title compound as a TFA salt. 1H-NMR (300 MHz, DMSO-d6, ppm): δ 8.74 (s, 1H), 7.67 (s, 1H), 7.42 (m, 1H), 7.27-7.25 (m, 2H), 6.79 (m, 2H), 6.52-6.49 (m, 1H), 4.63-4.58 (m, 1H), 4.44 (m, 2H), 4.20-4.16 (m, 1H), 3.72-3.64 (m, 2H), 2.99 (s, 3H), 2.34-2.27 (m, 2H), 2.01-1.97 (m, 2H), 1.82-1.77 (m, 2H). MS 480 [M+H]+. gave after purification by preparative HPLC

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Diethyl N-(4-iodobenzoyl)-L-glutamate is prepared from 4-iodobenzoyl chloride and diethyl L-glutamate in 56% yield by the same method. m.p. 105°-106° C.; NMR (CDCl3, 300 MHz) delta 1.26 (t, 3H, J=7.2 Hz), 1.33 (t, 3H, J=7.2 Hz), 2.11-2.60 (m, 4H), 4.14 (q, 2H, J=7.2 Hz), 4.78 (m, 1H), 7.15 (d, 1H, J=7.2 Hz), 7.58 (d, 2H, J=8.4 Hz), 7.83 (d, 2H, J=8.4 Hz).

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a mixture of 0.92 g of 4-bromobenzoyl chloride and 1.0 g of diethyl L-glutamate in 50 ml to dry methylene chloride is added 1.16 ml of triethylamine. The reaction mixture is stirred overnight under nitrogen. After a standard workup, the methylene chloride solution is concentrated under reduced pressure, and the residue recrystallized from hexanes to give 0.68 g (46%) of analytically pure diethyl N-(4-bromobenzoyl)-L-glutamate as a white solid: m.p. 82.5°-83.5° C.; NMR (CDCl3, 300 MHz) delta 1.26 (t, 3H, J=6.9 Hz), 1.33 (t, 3H, J=6.9 Hz), 2.11-2.60 (m, 4H), 4.14 (q, 2H, J=6.9 Hz), 4.27 (q, 2H, J=6.9 Hz) 4.78 (m, 1H), 7.14 (d, 1H, J=7.2 Hz), 7.61 (d, 2H, J=8.4 Hz), 7.73 (d, 2H, J=8.4 Hz); IR (KBr) 3320, 2980, 1745, 1720, 1635, 1520, 1375, 1300, and 1200 cm-1. Anal. Calcd. for C16H20BrNO5 : C, 49.75; H, 5.22; N, 3.63; Br, 20.69. Found: C, 49.70; H, 5.15; N, 3.65; Br, 20.90. After a standard workup, the methylene chloride solution is concentrated under reduced pressure the residue recrystallized from hexanes

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of 0.55 g of 4-ethynylbenzoic acid (obtained from tert.-butyl 4-ethynylbenzoate in 84% yield by hydrolysis with trifluoroacetic acid) in 50 ml of anhydrous ether and 25 ml of anhydrous tetrahydrofuran is added 1.58 ml of triethylamine. This is followed by 1.00 g of phenyl N-phenylphosphoraminochloridate. After stirring the reaction mixture at room temperature under nitrogen for 0.5 hour, 0.90 g of diethyl L-glutamate is added in one portion. The mixture is allowed to stir for another 8 hrs. After a workup, the residue is subjected to column chromatography using a 1% methanol:methylene chloride mixture as the eluent. The major fraction isolated from the column contained 0.68 g (54%) of diethyl N-(ethynylbenzoyl)-L-glutamate as an oil which slowly solidified: NMR (CDCl3, 300 MHz) delta 1.25 (t, 3H, J=6.9 Hz), 1.33 (t, 3H, J=6.9 Hz), 2.11-2.60 (m, 4H), 3.23 (s, 1H), 4.09 (q, 2H, J=6.9 Hz), 4.27 (l, 2H, J=6.9 Hz), 4.80 (m, 1H), 7.12 (d, 1H, J=7.2 Hz), 7.59 (d, 2H, J=8.4 Hz), 7.81 (d, 2H, J=8.4 Hz); IR (KBr) 3330, 3280, 2990, 1735, 1640, 1520, 1380, 1200, 1105, 1020, 855, and 770 cm- 1. to stir for another 8 hrs The major fraction isolated from the column

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Diethyl N-(4-iodobenzoyl)-L-glutamate is prepared from 4-iodobenzoyl chloride and diethyl L-glutamate in 56% yield by the same method. m.p. 105°-106° C.; NMR (CDCl3, 300 MHz) delta 1.26 (t, 3H, J=7.2 Hz), 1.33 (t, 3H, J=7.2 Hz), 2.11-2.60 (m, 4H), 4.14 (q, 2H, J=7.2 Hz), 4.78 (m, 1H), 7.15 (d, 1H, J=7.2 Hz), 7.58 (d, 2H, J=8.4 Hz), 7.83 (d, 2H, J=8.4 Hz)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Diethyl N-(4-iodobenzoyl)-L-glutamate is prepared from 4-iodobenzoyl chloride and diethyl L-glutamate in 56% yield by the same method. m.p. 105°-106° C.; NMR (CDCl3, 300 MHz) delta 1.26 (t, 3H, J=7.2 Hz), 1.33 (t, 3H, J=7.2 Hz), 2.11-2.60 (m, 4H), 4.14 (q, 2H, J=7.2 Hz), 4.78 (m, 1H), 7.15 (d, 1H, J=7.2 Hz), 7.58 (d, 2H, J=8.4 Hz), 7.83 (d, 2H, J=8.4 Hz)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a mixture of 0.92 g of of 4-bromobenzoyl chloride and 1.0 g of diethyl L-glutamate in 50 ml to dry methylene chloride is added 1.16 ml of triethylamine. The reaction mixture is stirred overnight under nitrogen. After a standard workup, the methylene chloride solution is concentrated under reduced pressure, and the residue recrystallized from hexanes to give 0.68 g (46%) of analytically pure diethyl N-(4-bromobenzoyl)-L-glutamate as a white solid: m.p. 82.5°-83.5° C.; NMR (CDCl3, 300 MHz) delta 1.26 (t, 3H, J=6.9 Hz), 1.33 (t, 3H, J=6.9 Hz), 2.11°-2.60 (m, 4H), 4.14 (q, 2H, J=6.9 Hz), 4.27 (q, 2H, J=6.9 Hz) 4.78 (m, 1H), 7.14 (d, 1H, J=7.2 Hz), 7.61 (d, 2H, J=8.4 Hz), 7.73 (d, 2H, J= 8.4 Hz); IR (KBr) 3320, 2980, 1745, 1720, 1635, 1520, 1375, 1300, and 1200 cm-1. After a standard workup, the methylene chloride solution is concentrated under reduced pressure the residue recrystallized from hexanes