GSK1324726A (I-BET726)

Procedure 24-{(2S.4/?)-1 -Acetyl-4-r(4-chlorophenyl)aminol-2-methyl-1.2.3.4-tetrahvdro-6- quinolinvDbenzoic acidEthyl 4-{(2S, 4R)-1 -acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1 , 2, 3, 4-tetrahydro-6- quinolinyl}benzoate (for a preparation see Example 158) (5.41 g, 1 1.69 mmol) was dissolved in ethanol (100 mL) and the solution was treated with 2M NaOH aqueous solution (50 mL, 100 mmol). The resulting mixture was stirred at room temperature (air atmosphere) for approximately 2 h then most of the ethanol was removed in vacuo. The resulting yellow solution was diluted with water (resulting in the formation of an oily yellow precipitate). The aqueous phase was washed twice with DCM (which didn't dissolve the precipitate previously formed) then was acidified with a 2N hydrochloric acid aqueous solution to pH 1 and extracted twice with AcOEt. The combined AcOEt phases were washed with brine, dried using a hydrophobic frit and concentrated in vacuo. The residual yellow foam was triturated with EtEthyl 4-{(2S, 4R)-1 -acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1 , 2, 3, 4-tetrahydro-6- quinolinyl}benzoate (for a preparation see Intermediate 17) (5.41 g, 1 1.69 mmol) was dissolved in ethanol (100 mL) and the solution was treated with aqueous NaOH solution (2M, 50 mL, 100 mmol). The resulting mixture was stirred at room temperature (air atmosphere) for approximately 2 h then most of the ethanol was removed in vacuo. The resulting yellow solution was diluted with water (resulting in the formation of an oily yellow precipitate). The aqueous phase was washed twice with DCM (which didn't dissolve the precipitate previously formed) then was acidified with hydrochloric acid (2N) to pH 1 and extracted with EtOAc (x2). The combined EtOAc phases were washed with brine, dried (hydrophobic frit) and concentrated in vacuo. The residual yellow foam was triturated with EtEthyl 4-{(2S, 4R)-1 -acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1 , 2, 3, 4-tetrahydro-6- quinolinyl}benzoate (for a preparation see Intermediate 17) (5.41 g, 1 1.69 mmol) was dissolved in ethanol (100 mL) and the solution was treated with aqueous NaOH solution (2M, 50 mL, 100 mmol). The resulting mixture was stirred at room temperature (air atmosphere) for approximately 2 h then most of the ethanol was removed in vacuo. The resulting yellow solution was diluted with water (resulting in the formation of an oily yellow precipitate). The aqueous phase was washed twice with DCM (which didn't dissolve the precipitate previously formed) then was acidified with hydrochloric acid (2N) to pH 1 and extracted with EtOAc (x2). The combined EtOAc phases were washed with brine, dried (hydrophobic frit) and concentrated in vacuo. The residual yellow foam was triturated with EtIntermediate 18 4-{(2S, 4R)-1-Acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1, 2, 3, 4-tetrahydro-6-quinolinyl}benzoic acid [0199] [0200] Ethyl 4-{(2S, 4R)-1-acetyl-4-[(4-chlorophenyl)amino]-2-methyl-1, 2, 3, 4-tetrahydro-6-quinolinyl}benzoate (for a preparation see Intermediate 17) (5.41 g, 11.69 mmol) was dissolved in ethanol (100 mL) and the solution was treated with aqueous NaOH solution (2M, 50 mL, 100 mmol). The resulting mixture was stirred at room temperature (air atmosphere) for approximately 2 h then most of the ethanol was removed in vacuo. The resulting yellow solution was diluted with water (resulting in the formation of an oily yellow precipitate). The aqueous phase was washed twice with DCM (which didn't dissolve the precipitate previously formed) then was acidified with hydrochloric acid (2N) to pH 1 and extracted with EtOAc (×2). The combined EtOAc phases were washed with brine, dried (hydrophobic frit) and concentrated in vacuo. The residual yellow foam was triturated with Et[0125] A solution of Intermediate 8 (320mg, 0.73 mmol) in EtOH (10ml) and 1N NaOH (1.5ml, 1.5mmol) was heatedto reflux. After 1 hour a tlc monitoring indicated the completion of the reaction. The crude mixture was evaporated todryness and the residue taken up in water (10mL). Acidification of the mixture at pH=3 was carried out by addition of a1N HCl solution. The organic materials were extracted with EtOAc (3x 25 mL) and the organic phase combined andwashed with brine and dried over Na2SO4. After concentration under vacuo the residue was taken up in a DCM/ hexanemixture to give a red solid after filtration. The compound was recrystallised in EtOAC, filtered and washed with i-Pr2O.The resulting white powder was solubilised in MeOH/H2O, concentrated to dryness and taken up with H2O. Finallyfiltration of the precipitate afforded the title compound as a white powder (147mg), mp: 275°C.HRMS calculated for C25H23N2O3Cl (M-H)- 433.1319, found : 433.1299. Rt: 2.21 min.1 H NMR (300 MHz, CHLOROFORM-d) δ ppm 1.2 (d, 3 H) 1.35 (m, 1 H) 2.3 (s, 3 H) 2.7 (m, 1 H) 4.25 (dd, 1 H) 4.95(m, 1 H) 6.65 (d, 2 H) 7.15 (d, 2 H) 7.25 (s, 1 H) 7.55 (m, 4 H), 8.15(d, 2H)[α]D = + 395 (c= 0.96 g/cl, EtOH) measured at the EtOAc recrystallisation stage.The title compound eluted at 4.51 min by HPLC as the first peak using a Chiralpak IA (250x4.6 mm 5mm) column withtert-butyl methyl oxide (MTBE) +0.1percent TFA /Ethanol :90/10 as the mobile phase. A 1 ml/mn flow rate was applied and10mL of sample prepared with the dilution of 1 mg of the title compound in 1ml of eluent was injected. Detection of thecompound was carried out with both 210 and 254 nM UV wavelengths. The other enantiomer came off at 5.92 min.Pd/C (22.1 mg, 10percent, 0.208 mmol, containing -50percent water) was added to a solution of 4- ((2S, 4R)-1-acetyl-4-((4-chlorophenyl)amino)-2-methyl-1 , 2, 3, 4-tetrahydroquinolin-6- yl)benzoic acid (for a preparation see Intermediate 18)(369 mg, 0.848 mmol) and ammonium formate (134 mg, 2.121 mmol) in ethanol (5 ml) and DMF (1 ml) and reaction mixture heated at reflux. After 1.5h the reaction mixture was cooled to r.t.. The reaction mixture was filtered through Celite.(TM). and filter cake washed with EtOH. The filtrate was concentrated to give a light brown oil (251 mg). The filter cake was washed with a further portion of 1 :1 EtOH:DMF (50ml) and combined with the previous material and concentrated to give 4-((2S, 4R)-1 -acetyl-2-methyl-4-(phenylamino)-1 , 2, 3, 4- tetrahydroquinolin-6-yl)benzoic acid (306mg, 0.764mmol, 90percent yield) as a yellow solid. LCMS (Formate, 2min), Rt=1.01 min, MH+ = 401.

Molecular Weight:434.9
XLogP3:5.1
Hydrogen Bond Donor Count:2
Hydrogen Bond Acceptor Count:4
Rotatable Bond Count:4
Exact Mass:434.1397203
Monoisotopic Mass:434.1397203
Topological Polar Surface Area:69.6
Heavy Atom Count:31
Complexity:643
Defined Atom Stereocenter Count:2
Covalently-Bonded Unit Count:1
Compound Is Canonicalized:Yes