4-(dimethylamino)butanoic acid;hydrochloride

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

Procedure: The above-titled compound was prepared according to the method of Example 8 by reacting equimolar quantities of {[10-hydroxy-5,5-dimethyl-8-(1,2-dimethylheptyl)-1,2,3,4-tetrahydro-5H-[1]benzopyrano[4,3-c]pyridin-2-yl]acetyl}urea and 4-(dimethylamino)butyric acid hydrochloride in the presence of dicyclohexylcarbodiimide.

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

Procedure: BOP reagent (27 mg, 0.060 mmol) was added to a solution of (+)-7-amino-4-methyl-5-(3 -hydroxyphenyl)-2-(3-phenylpropyl)-2-azabicyclo[3.3.1]nonane (23, 20 mg, 0.055 mmol), 4-(dimethylamino)butyric acid hydrochloride (18 mg, 0.11 mmol) and triethylamine (0.038 mL, 0.27 mmol) in dry THF (5 mL). The reaction mixture was stirred under N2 at room temperature for 4 h. The mixture was diluted with Et2O (20 mL), washed with saturated NaHCO3, followed by water, organic layer collected, dried (Na2SO4) and solvent removed under reduced pressure yielding crude product. This was purified by flash chromatography (33% (80% CHCl3:18% CH3OH:2% NH4OH) in CHCl3) to afford 4-(dimethylamino)-N-[(1R,4S,5S,7R)-5-(3-hydroxyphenyl)-4-methyl-2-(3-phenylpropyl)-2-azabicyclo[3.3.1]non-7-yl]butanamide (13) (23 mg, 89%) as an off-white foam. 1H-NMR (CDCl3) δ7.28-7.09 (m, 6H), 6.89 (d, 1H, J=7.0 Hz), 6.64-6.59 (m, 3H), 6.31 (m, 1H), 4.65 (br., 1H), 3.16 (br., 1H), 3.02 (d, 1H, J=7.8 Hz), 2.69-2.14 (m, 16H), 1.91 (m, 1H), 1.86-1.75 (m, 4H), 1.58 (m 1H), 1.36-0.83 (m, 3H), 0.71 (d, 3H, J=6.5 Hz); LRMS (ES) m/z 478.7 (M+H)+. washed with saturated NaHCO3 collected dried (Na2SO4) and solvent removed under reduced pressure yielding crude product This was purified by flash chromatography (33% (80% CHCl3:18% CH3OH:2% NH4OH) in CHCl3)

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

Procedure: In 6 ml of methylene chloride-N,N-dimethylformamide (1:2) was dissolved 120 mg (0.20 mmol) of 1,2-O-dioleylglycerol, followed by addition of 168 mg (1 mmol) of 4-dimethylaminobutyric acid hydrochloride. Then, 206 mg (1 mmol) of N,N'-dicyclohexylcarbodiimide (DCC) and 25 mg (0.2 mmol) of 4-dimethylaminopyridine were further added and the reaction was conducted at ambient temperature overnight. The precipitated byproduct urea was filtered off using a glass filter and the filtrate was concentrated to dryness under reduced pressure and treated with methylene chloride-saturated aqueous sodium hydrogen carbonate solution. After phase separation, the methylene chloride layer was dried over sodium sulfate and the solvent was distilled off under reduced pressure. The residue was subjected to column chromatography (silica gel/methylene chloride-methanol) to provide 123 mg (87%) of the title compound of the invention. The precipitated byproduct urea was filtered off filter the filtrate was concentrated to dryness under reduced pressure treated with methylene chloride-saturated aqueous sodium hydrogen carbonate solution After phase separation the methylene chloride layer was dried over sodium sulfate the solvent was distilled off under reduced pressure

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

Procedure: 10 g (60 mmol) of 4-dimethylaminobutyric acid hydrochloride (Janssen; Bruggen/Germany) are heated for approximately 3 h at 65° C. in 30 ml of SOCl2. SOCl2 is evaporated off and the residue is stirred to yield the title compound: TLC of a sample dissolved in methanol, Rf (U)=0.67; TLC of 4-dimethylaminobutyric acid hydrochloride, Rf (U)=0.50. SOCl2 is evaporated off

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

Procedure: 10 g (60 mmol) of 4-dimethylaminobutyric acid hydrochloride (Janssen; Bruggen/Germany) are heated for approximately 3 h at 65° C. in 30 ml of SOCl2. SOCl2 is evaporated off and the residue is stirred to yield the title compound: TLC of a sample dissolved in methanol, Rf (U)=0.67; TLC of 4-dimethylaminobutyric acid hydrochloride, Rf (U)=0.50. SOCl2 is evaporated off

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

Procedure: Grams 10 4-dimethylaminobutyric acid hydrochloride are heated up till melting together with 6.45 g o-phenylendiamine then it is allowed to cool and 150 ml water are added thereto. The solution is washed with dichloromethane, made alkaline and extracted with dichloromethane. The extracts are collected together, filtered on charcoal, made anhydrous on sodium sulphate and evaporated to dryness. A thick oily reddish residue is obtained, which is triturated with 200 ml diethyl ether to give 5.8 g 2-(3-dimethylaminopropyl)benzimidazole melting at 117°-119° C. to cool The solution is washed with dichloromethane extracted with dichloromethane The extracts are collected together filtered on charcoal evaporated to dryness A thick oily reddish residue is obtained which is triturated with 200 ml diethyl ether

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

Procedure: From 4-(dimethylamino)butyric acid hydrochloride, HATU and N-ethyldiisopropylamine in THF, then treatment with (+)-7-[1,4]dioxan-2-yl-4-methoxy-benzothiazol-2-ylamine. [α]D20=+24.2° (c=0.53, CHCl3), ES-MS m/e (%): 380 (M+H+, 100).

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

Procedure: In 6 ml of methylene chloride-N,N-dimethylformamide (1:2) was dissolved 120 mg (0.20 mmol) of 1,2-O-dioleylglycerol, followed by addition of 168 mg (1 mmol) of 4-dimethylaminobutyric acid hydrochloride. Then, 206 mg (1 mmol) of N,N'-dicyclohexylcarbodiimide (DCC) and 25 mg (0.2 mmol) of 4-dimethylaminopyridine were further added and the reaction was conducted at ambient temperature overnight. The precipitated byproduct urea was filtered off using a glass filter and the filtrate was concentrated to dryness under reduced pressure and treated with methylene chloride-saturated aqueous sodium hydrogen carbonate solution. After phase separation, the methylene chloride layer was dried over sodium sulfate and the solvent was distilled off under reduced pressure. The residue was subjected to column chromatography (silica gel/methylene chloride-methanol) to provide 123 mg (87%) of the title compound. The precipitated byproduct urea was filtered off filter the filtrate was concentrated to dryness under reduced pressure treated with methylene chloride-saturated aqueous sodium hydrogen carbonate solution After phase separation the methylene chloride layer was dried over sodium sulfate the solvent was distilled off under reduced pressure

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

Procedure: In 6 ml of methylene chloride-N,N-dimethylformamide (1:2) was dissolved 120 mg (0.20 mmol) of 1,2-O-dioleylglycerol, followed by addition of 168 mg (1 mmol) of 4-dimethylaminobutyric acid hydrochloride. Then, 206 mg (1 mmol) of N,N'-dicyclohexylcarbodiimide (DCC) and 25 mg (0.2 mmol) of 4-dimethylaminopyridine were further added and the reaction was conducted at ambient temperature overnight. The precipitated byproduct urea was filtered off using a glass filter and the filtrate was concentrated to dryness under reduced pressure and treated with methylene chloride-saturated aqueous sodium hydrogen carbonate solution. After phase separation, the methylene chloride layer was dried over sodium sulfate and the solvent was distilled off under reduced pressure. The residue was subjected to column chromatography (silica gel/methylene chloride-methanol) to provide 123 mg (87%) of the title compound. The precipitated byproduct urea was filtered off filter the filtrate was concentrated to dryness under reduced pressure treated with methylene chloride-saturated aqueous sodium hydrogen carbonate solution After phase separation the methylene chloride layer was dried over sodium sulfate the solvent was distilled off under reduced pressure

Conditions: temp:0.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: DIPEA (0.253 ml, 1.452 mmol) was added at 0° C. to a stirred suspension of 3-(1-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-4-(1,2,3,6-tetrahydropyridin-4-yl)-1H-isochromen-1-one hydrochloride (Intermediate D23, 0.200 g, 0.363 mmol), HATU (0.166 g, 0.436 mmol) and 4-dimethylaminobutyric acid hydrochloride (0.073 g, 0.436 mmol) in THF/DMF 5:1 (6 mL). The mixture was stirred at 0° C. for 15 min, then at rt for further 45 min. The reaction mixture was poured into saturated aqueous sodium bicarbonate and extracted with DCM. The collected organic phases were concentrated under reduced pressure and the crude purified by RP-flash chromatography (Biotage 30 g C18 column, gradient elution from 100:0 to 70:30 A/B in 15 CV; A: water/MeCN 95/5+0.01% HCOOH, B: water/MeCN 5/95+0.01% HCOOH) to give title compound (0.153 g, 0.227 mmol, 62.6% yield) as a white powder. at rt for further 45 min extracted with DCM The collected organic phases were concentrated under reduced pressure the crude purified by RP-flash chromatography (Biotage 30 g C18 column, gradient elution from 100:0 to 70:30 A/B in 15 CV

Conditions: temp:0.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: DIPEA (0.253 ml, 1.452 mmol) was added at 0° C. to a stirred suspension of 3-(1-(4-amino-3-iodo-1H-pyrazolo[3,4-d]pyrimidin-1-yl)ethyl)-4-(1,2,3,6-tetrahydropyridin-4-yl)-1H-isochromen-1-one hydrochloride (Intermediate D23, 0.200 g, 0.363 mmol), HATU (0.166 g, 0.436 mmol) and 4-dimethylaminobutyric acid hydrochloride (0.073 g, 0.436 mmol) in THF/DMF 5:1 (6 mL). The mixture was stirred at 0° C. for 15 min, then at rt for further 45 min. The reaction mixture was poured into saturated aqueous sodium bicarbonate and extracted with DCM. The collected organic phases were concentrated under reduced pressure and the crude purified by RP-flash chromatography (Biotage 30 g C18 column, gradient elution from 100:0 to 70:30 A/B in 15 CV; A: water/MeCN 95/5+0.01% HCOOH, B: water/MeCN 5/95+0.01% HCOOH) to give title compound (0.153 g, 0.227 mmol, 62.6% yield) as a white powder. at rt for further 45 min extracted with DCM The collected organic phases were concentrated under reduced pressure the crude purified by RP-flash chromatography (Biotage 30 g C18 column, gradient elution from 100:0 to 70:30 A/B in 15 CV

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

Procedure: A mixture of 3-(2,3-dihydro-1H-indol-5-yl)-5-vinyl-3H-imidazo[4,5-b]pyridine (48.0 mg, 0.14 mmol) (Intermediate 10.1), 4-(dimethylamino)butyric acid hydrochloride (36.3 mg, 0.21 mmol), N-ethyl-N′-(3-dimethylaminopropyl)carbodiimide hydrochloride (54.9 mg, 0.29 mmol), 4-dimethylaminopyridine (122.5 mg, 1.00 mmol), and dichloromethane (8 ml) is stirred for 24 h at ambient temperature. The mixture is successively extracted with saturated aqueous ammonium chloride and saturated aqueous sodium bicarbonate. The organic layer is dried over sodium sulfate and concentrated in vacuo to render 50.4 mg (94%) of the respective amide. HPLC (over 10 min 10-85% MeCN/100 mM aq. NaOAc): 92% purity, tR 6.27 min (tR(SM) 6.61 min). The mixture is successively extracted with saturated aqueous ammonium chloride and saturated aqueous sodium bicarbonate The organic layer is dried over sodium sulfate concentrated in vacuo HPLC (over 10 min 10-85% MeCN/100 mM aq. NaOAc) 92% purity, tR 6.27 min (tR(SM) 6.61 min)