Kappa opioid agonists and uses thereof
09815824 · 2017-11-14
Assignee
Inventors
- Neel K. Anand (San Mateo, CA)
- Franco J. Duarte (Millbrae, CA, US)
- Wen Zhang (San Ramon, CA)
- Zhongxu Ren (Foster City, CA)
Cpc classification
C07D409/12
CHEMISTRY; METALLURGY
C07D241/04
CHEMISTRY; METALLURGY
C07C233/40
CHEMISTRY; METALLURGY
C07D207/09
CHEMISTRY; METALLURGY
C07D401/12
CHEMISTRY; METALLURGY
A61P1/06
HUMAN NECESSITIES
C07C233/05
CHEMISTRY; METALLURGY
C07D205/04
CHEMISTRY; METALLURGY
C07D207/14
CHEMISTRY; METALLURGY
International classification
C07D409/12
CHEMISTRY; METALLURGY
C07C233/40
CHEMISTRY; METALLURGY
C07D401/12
CHEMISTRY; METALLURGY
C07D241/04
CHEMISTRY; METALLURGY
C07D207/14
CHEMISTRY; METALLURGY
C07D205/04
CHEMISTRY; METALLURGY
C07C233/05
CHEMISTRY; METALLURGY
C07C333/00
CHEMISTRY; METALLURGY
C07D295/125
CHEMISTRY; METALLURGY
C07D207/09
CHEMISTRY; METALLURGY
Abstract
Provided are compounds of Formula I; and pharmaceutically acceptable salts and solvates thereof. ##STR00001## The present disclosure relates to novel compounds and to their use as agonists of the kappa opioid receptor. The disclosure also relates to methods for preparation of the compounds and to pharmaceutical compositions containing such compounds. Kappa opioid agonists that exhibit full agonist properties at the kappa opioid receptor have been widely shown to be efficacious in preclinical models of pain, particularly visceral pain.
Claims
1. A compound having the structure: ##STR00286## wherein: R.sup.1 is ##STR00287## R.sup.2 is hydrogen; R.sup.3 is hydrogen; R.sup.4 is —X-POLY; R.sup.5 is hydrogen; R.sup.6 is hydrogen; X is —O— or —OC(O)NH—; POLY is —(CH.sub.2CH.sub.2O).sub.n—Y; n is 1 to 30; and Y is hydrogen or substituted or unsubstituted lower alkyl; and pharmaceutically acceptable salts and solvates thereof.
2. A compound selected from: 2-(3, 4-dichlorophenyl)-N-{(1S)-2-[(3S)-3-(2-methoxyethoxy)pyrrolidin-1-yl]-1-phenylethyl}-N-methylacetamide; 2-(3, 4-Dichlorophenyl)-N-{(1S)-2-[(3S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl]-1-phenylethyl}-N-methylacetamide; (3S)-1-[(2S)-2-{[(3,4-Dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]pyrrolidin-3-yl{2-[2-(2-methoxyethoxy)ethoxy]ethyl}carbamate; (3S)-1-[(2S)-2-{[(3,4-Dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]pyrrolidin-3-yl 2,5,8,11,14-pentaoxahexadecan-16-ylcarbamate; and 2-(3,4-Dichlorophenyl)-N-methyl-N-{(1S)-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethyl}acetamide; and pharmaceutically acceptable salts thereof.
3. A pharmaceutical composition comprising a compound of claim 1 and at least one pharmaceutically acceptable excipient.
4. A method of treating pain or inflammation comprising administering a compound of claim 1 to a patient in need thereof.
5. The compound of claim 1, having the structure: ##STR00288## wherein: n is 1 to 30; R.sup.10 is hydrogen; and R.sup.11 is —H, —CH.sub.3, or —CF.sub.3; and pharmaceutically acceptable salts and solvates thereof.
Description
EXAMPLES
(1) All chemical reagents referred to in the appended examples are commercially available unless otherwise indicated. The preparation of PEG-mers is described in, for example, U.S. Patent Application Publication No. 2005/0136031.
Example 1
Preparation of 2-(3,4-dichlorophenyl)-N-{(1S)-2-[(3S)-3-(2-methoxyethoxy)pyrrolidin-1-yl]-1-phenylethyl}-N-methylacetamide, hydrochloride salt (1)
(2) ##STR00065##
(3) 2-(3,4-dichlorophenyl)-N-{(1S)-2-[(3S)-3-(2-methoxyethoxy)pyrrolidin-1-yl]-1-phenylethyl}-N-methylacetamide, hydrochloride salt (1) was prepared according to the following steps.
Step 1: Preparation of 2-(3,4-Dichlorophenyl)-N-{(1S)-2-((3S)-3-hydroxypyrrolidin-1-yl)-1-phenylethyl}-N-methylacetamide, hydrochloride salt (3)
(4) ##STR00066##
(5) (3S)-1-[(2S)-2-(Methylamino)-2-phenylethyl]pyrrolidin-3-ol (2) (Ghosh, A. et. al., Chemical Communications (2002), #15, 1644) (0.70 g, 2.39 mmol), 2-(3,4-dichlorophenyl)acetic acid (0.45 g, 2.17 mmol), and N,N-diisopropylethylamine (0.56 g, 4.34 mmol) were dissolved in 20 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU) (0.84 g, 2.61 mmol) was added into the solution. The reaction mixture was stirred for 16 hours and then was concentrated. The residue was dissolved in ethyl acetate (50 mL) and was washed with saturated sodium bicarbonate (2×30 mL), brine (30 mL) and was dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded 2-(3,4-dichlorophenyl)-N-{(1S)-2-((3S)-3-hydroxypyrrolidin-1-yl)-1-phenylethyl}-N-methylacetamide (3) (0.86 g, 97% yield). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.45-7.25 (m, 7H), 7.15 (m, 1H), 6.10 (m, 0.85H), 5.05 (m, 0.15H), 4.30 (m, 1H), 3.75 (m, 2H), 3.20 (t, 1H), 3.05 (m, 1H), 2.80 (m, 2H), 2.62 (s, 3H), 2.64 (m, 1H), 2.38 (m, 1H), 2.15 (m, 1H), 1.73 (m, 1H); MS (EI) for C.sub.21H.sub.24Cl.sub.2N.sub.2O.sub.2: 407 (MH.sup.+). The free base (20 mg) was dissolved in 1 mL of acetonitrile. To the solution was added 0.2 mL of 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt.
Step 2: Preparation of 2-(3,4-Dichlorophenyl)-N-{(1S)-2-[(3S)-3-(2-methoxyethoxy)pyrrolidin-1-yl]-1-phenylethyl}-N-methylacetamide, hydrochloride salt (1)
(6) ##STR00067##
(7) 2-(3,4-Dichlorophenyl)-N-{(1S)-2-((3S)-3-hydroxypyrrolidin-1-yl)-1-phenylethyl}-N-methylacetamide (0.060 g, 0.15 mmol) was dissolved in 5 mL of N,N-dimethylformamide and sodium hydride (0.017 g of 60% in mineral oil, 0.44 mmol) was added into the solution. 1-bromo-2-methoxyethane (0.061 g, 0.44 mmol) in 1 mL of N,N-dimethylformamide was added under stirring. The reaction mixture was stirred at room temperature for 3 hours. 150 mL of dichloromethane was added into the reaction mixture. The solution was washed with water (100 mL×3) and was dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded 2-(3,4-dichlorophenyl)-N-{(1S)-2-[(3S)-3-(2-methoxyethoxy)pyrrolidin-1-yl]-1-phenylethyl}-N-methylacetamide (1) (0.020 g, 29% yield). The free base (18 mg) was dissolved in 1 mL of acetonitrile. To the solution was added 0.2 mL of 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (3). .sup.1H NMR (500 MHz, HCl salt in MeOD): δ 7.50 (m, 2H), 7.31 (m, 3H), 7.26 (m, 1H), 7.21 (m, 2H), 6.35 (d, 1H), 4.62 (d, 1H), 4.25 (m, 2H), 4.10-3.95 (m, 1H), 3.85 (m, 2H), 3.70-3.50 (m, 1H), 3.45 (m, 2H), 3.35 (m, 1H), 3.32 (s, 3H), 3.25 (m, 1H), 2.60 (d, 3H), 2.45-2.20 (m, 2H), 2.10 (m, 1H), 1.96 (m, 1H); MS (EI) for C.sub.24H.sub.30Cl.sub.2N.sub.2O.sub.3: 465 (MH.sup.+).
(8) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 2
Preparation of 2-(3,4-Dichlorophenyl)-N-{(1S)-2-[(3S)-3-(2-(2-(2-methoxyethoxy)ethoxy]ethoxy)pyrrolidin-1-yl)-1-phenylethyl}-N-methylacetamide, hydrochloride salt (4)
(9) ##STR00068##
Step 1: Preparation of (S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidine
(10) ##STR00069##
(11) (S)-tert-Butyl-3-hydroxypyrrolidine-1-carboxylate (6.0 g, 32.04 mmol) was dissolved in THF (120 mL). The mixture was cooled to 0° C., NaH (1.54 g, 60% in mineral oil, 38.45 mmol) was added and reaction mixture was stirred for 30 min. 1-Bromo-2-(2-(2-methoxyethoxy)ethoxy)ethane (10.91 g, 48.06 mmol) dissolved in THF (25 mL) was added to the above mixture while maintaining the temperature of reaction at 0° C. The reaction mixture was stirred for additional 1 hour at 0° C. and allowed to warm to 22-25° C. and stirred for 18 hours at that temperature. The above mixture was concentrated under reduced pressure. The resulting residue was dissolved in DCM (120 mL) and the resulting solution was washed with water (50 mL×2). The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum to get a thick oil which was dissolved in DCM/TFA (2:1) (25 mL) and stirred for 4 hours at 22-25° C. and then concentrated under vacuum. The residue was dissolved in water (35 mL) and pH of mixture was adjusted to 9 by adding sodium carbonate. The solution was saturated with sodium chloride and extracted with DCM (50 mL×3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure and the crude product was purified by column chromatography yielded (S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidine (2.2 g, 29.5% yield) as an oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 4.05 (m, 1H), 3.70-3.6 (m, 8H), 3.55 (m, 4H), 3.38 (s, 3H), 3.10 (m, 2H), 2.75 (m, 2H), 1.90 (m, 1H); MS (EI) for C.sub.11H.sub.23NO.sub.4: 234 (MH.sup.+).
Step 2: Preparation of benzyl((S)-2-(S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl)-2-oxo-1-phenylethyl)carbamate
(12) ##STR00070##
(13) (S)-3-(2-(2-(2-Methoxyethoxy)ethoxy)ethoxy)pyrrolidine (2.0 g, 8.57 mmol), (S)-2-(((benzyloxy)carbonyl)amino)-2-phenylacetic acid (2.44 g, 8.57 mmol) and DIPEA (3.32 g, 25.71 mmol) were dissolved in acetonitrile (40 mL). The above mixture was stirred for 15 min at 22-25° C. and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU) (3.30 g, 10.28 mmol) was added into the solution. The reaction mixture was stirred for 1 h at 0° C. and then 4 hours at room temperature. The reaction mixture was concentrated under reduced pressure. The resulting residue was dissolved in DCM (50 mL) and washed with brine (25 mL×2). The solution was dried over anhydrous sodium sulfate and concentrated. The obtained residue was then purified by column chromatography to yield benzyl((S)-2-((S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl)-2-oxo-1-phenylethyl)carbamate (3.27 g, 76% yield). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.43 (m, 2H), 7.34 (m, 8H), 6.40-6.25 (dd, 1H), 5.40 (m, 1H), 5.23 (d, 1H), 5.04 (t, 1H), 4.08 (m, 1H), 3.70-3.50 (m, 12H), 3.42 (m, 2H), 3.41 (s, 3H), 3.21 (m. 2H), 2.12-1.80 (m, 2H); MS (EI) for C.sub.27H.sub.36N.sub.2O.sub.7: 501 (MH.sup.+).
Step 3: Preparation of (S)-2-((S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl)-N-methyl-1-phenylethanamine
(14) ##STR00071##
(15) Benzyl((S)-2-((S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl)-2-oxo-1-phenylethyl)carbamate (4.0 g, 8.0 mmol) was dissolved in THF (40 mL). The mixture was cooled to 0° C. LAH tablet (1.52 g, 40 mmol) was added to the above mixture and the mixture was stirred for 15 min at 0° C. and then heated to 65° C. and maintained at that temperature for four hours. A 3N aq. sodium carbonate solution was added cautiously until effervescence ceased. The solid was removed by filtration and was washed with DCM (100 mL). The filtrate was concentrated and the residue was dissolved in DCM (150 mL). The solution was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum to obtain the crude product which was purified by column chromatography to yield (S)-2-((S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl)-N-methyl-1-phenylethanamine (1.35 g, 46% yield) as a thick oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.35 (m, 4H), 7.25 (m, 1H), 4.05 (m, 1H), 3.67 (m, 8H), 3.55 (m, 5H), 3.40 (s, 3H), 2.85 (m, 1H), 2.80 (m, 2H), 2.55 (m, 1H), 2.48 (m, 1H), 2.28 (s, 3H), 2.05 (m, 1H), 1.80 (m, 2H); MS (EI) for C.sub.20H.sub.34N.sub.2O.sub.4: 367 (MH.sup.+).
Step 4: Preparation of 2-(3,4-dichlorophenyl)-N—((S)-2-((S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl)-1-phenylethyl)-N-methylacetamide, hydrochloride salt (4)
(16) ##STR00072##
(17) (S)-2-((S)-3-(2-(2-(2-Methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl)-N-methyl-1-phenylethanamine (3) (1.3 g, 3.54 mmol), 2-(3,4-dichlorophenyl)acetic acid (0.725 g, 3.54 mmol), and N,N-diisopropylethylamine (0.915 g, 7.08 mmol) were dissolved in acetonitrile (13 mL). The resulting mixture was stirred for 15 min at 22-25° C. and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU) (1.36 g, 4.25 mmol) was added into the solution. The reaction mixture was stirred for four hours at 22-25° C. and concentrated. The residue was dissolved in DCM (100 mL) and washed with water and dried over sodium sulfate. Evaporation of solvent and purification of the resulting residue by flash chromatography yielded 2-(3,4-dichlorophenyl)-N—((S)-2-((S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl)-1-phenylethyl)-N-methylacetamide (0.78 g, 40% yield). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.40 (m, 2H), 7.32 (m, 2H), 7.25 (m, 2H), 7.20-7.10 (m, 2H), 6.10 (m, 1H), 4.05 (m, 1H), 3.78 (d, 1H), 3.65 (m, 8H), 3.55 (m, 4H), 3.40 (s, 3H), 3.15 (t, 1H), 3.05 (m, 1H), 2.85-2.70 (m, 2H), 2.70 (br, s, 3H), 2.50 (m, 2H), 2.05 (m, 1H), 1.80 (m, 1H); MS (EI) for C.sub.28H.sub.38Cl.sub.2N.sub.2O.sub.5: 554 (MH.sup.+). The free base was dissolved in 4M hydrochloride in 2-propanol. The mixture was concentrated to afford product as hydrochloride salt (4). The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 3
Preparation of (3S)-1-[(2S)-2-{[(3,4-Dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]pyrrolidin-3-yl{2-[2-(2-methoxyethoxy)ethoxy]ethyl}carbamate, hydrochloride salt (5)
(18) ##STR00073##
(19) 2-(3,4-Dichlorophenyl)-N-{(1S)-2-((3S)-3-hydroxypyrrolidin-1-yl)-1-phenylethyl}-N-methylacetamide (3) (0.030 g, 0.074 mmol) and 4-nitrophenyl carbonochloride (0.017 g, 0.081 mmol) were dissolved in 3 mL of dichloromethane. N,N-diisopropylethylamine (0.018 g, 0.15 mmol) was added under stirring. The reaction mixture was stirred at room temperature for two hours. 2-(2-(2-methoxyethoxy)ethoxy)ethanamine (0.033 g, 0.20 mmol) was added. The reaction mixture was stirred at room temperature for 6 hours. 100 mL of dichloromethane was added into the reaction mixture. The resultant solution was washed with saturated sodium chloride (60 mL×3) and was dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded (3S)-1-[(2S)-2-{[(3,4-Dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]pyrrolidin-3-yl {2-[2-(2-methoxyethoxy)ethoxy]ethyl}carbamate (0.023 g, 58% yield). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.45-7.26 (m, 7H), 7.16 (m, 1H), 6.10 (m, 0.85H), 5.20 (m, 2H), 5.05 (m, 0.15H), 3.85-3.62 (m, 10H), 3.57 (m, 4H), 3.38 (s, 3H), 3.15 (t, 1H), 2.98 (m, 2H), 2.75 (m, 4H), 2.40 (m, 1H), 2.20 (m, 1H), 1.82 (m, 2H); MS (EI) for C.sub.29H.sub.39Cl.sub.2N.sub.3O.sub.6: 596 (Mil).
(20) The free base (20 mg) was dissolved in 1 mL of acetonitrile. To the solution was added 0.2 mL of 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (5).
(21) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 4
(3S)-1-[(2S)-2-{[(3,4-Dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]pyrrolidin-3-yl 2,5,8,11,14-pentaoxahexadecan-16-ylcarbamate, hydrochloride salt (6)
(22) ##STR00074##
(23) 2-(3,4-Dichlorophenyl)-N-{(1S)-2-((3S)-3-hydroxypyrrolidin-1-yl)-1-phenylethyl}-N-methylacetamide (3) (0.035 g, 0.086 mmol) and 4-nitrophenyl carbonochloride (0.020 g, 0.10 mmol) were dissolved in 3 mL of dichloromethane. N,N-diisopropylethylamine (0.022 g, 0.172 mmol) was added under stirring. The reaction mixture was stirred at room temperature for 2 hours. 2,5,8,11,14-Pentaoxahexadecan-16-amine (0.060 g, 0.24 mmol) was added. The reaction mixture was stirred at room temperature overnight. Dichloromethane (100 mL) was added into the reaction mixture. The solution was washed with saturated sodium chloride (100 mL×3) and was dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded (3S)-1-[(2S)-2-{[(3,4-dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]pyrrolidin-3-yl 2,5,8,11,14-pentaoxahexadecan-16-ylcarbamate (0.025 g, 47% yield). .sup.1H NMR (500 MHz, DMSO-d6): δ 7.56 (m, 1H), 7.50 (s, 1H), 7.40-7.20 (m, 6H), 7.12 (m, 1H), 5.82 (m, 0.8H), 5.20 (m, 0.2H), 4.95 (m, 1H), 3.92-3.65 (m, 2H), 3.50 (m, 16H), 3.48 (m, 4H), 3.24 (s, 3H), 3.10 (m, 3H), 2.90 (m, 1H), 2.75 (m, 4H), 2.42 (m, 1H), 2.20 (m, 1H), 1.65 (m, 1H); MS (EI) for C.sub.33H.sub.47Cl.sub.2N.sub.3O.sub.8: 684 (MH.sup.+).
(24) The free base (22 mg) was dissolved in 1 mL of acetonitrile. To the solution was added 0.5 mL of 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (6).
(25) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 5
2-(3,4-Dichlorophenyl)-N-methyl-N-{(1S)-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethyl}acetamide, hydrochloride salt (7)
(26) ##STR00075##
(27) 2-(3,4-Dichlorophenyl)-N-methyl-N-{(1S)-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethyl}acetamide, hydrochloride salt (7) was prepared according to the following steps.
Step 1: Preparation of (3S)-3-(2,5,8,11,14-Pentaoxahexadecan-16-yloxy)pyrrolidine
(28) ##STR00076##
(29) (S)—N-Boc-3-pyrrolidinol (1.0 g, 5.34 mmol) was dissolved in 10 mL of tetrahydrofuran. The resultant solution was cooled in an ice-bath and then sodium hydride (0.257 g of 60% in mineral oil, 6.41 mmol) was added. The mixture was stirred for five minutes before 16-bromo-2,5,8,11,14-pentaoxahexadecane (2.02 g, 6.41 mmol) in 5 mL of tetrahydrofuran was added. The reaction mixture was stirred at 0° C. for one hour and then overnight at room temperature. 150 mL of dichloromethane was added into the reaction mixture. The solution was washed with water (100 mL×3) and dried over sodium sulfate. An oil was obtained after removing solvent, which was dissolved in 10 mL of trifluoroacetic acid/dichloromethane (1:2). The mixture was stirred for 4 hours and then was concentrated. 5 mL of water was added into the mixture. The pH of the mixture was adjusted to 9 by adding sodium carbonate. The solution was saturated with sodium chloride and was extracted with dichloromethane (150 mL×3). The organic phase was dried over sodium sulfate. After removing solvent, an oil was obtained, which was purified by flash chromatography to afford (3S)-3-(2,5,8,11,14-Pentaoxahexadecan-16-yloxy)pyrrolidine (0.944 g, 56% yield). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 4.24 (m, 1H), 3.69 (m, 15H), 3.66 (m, 6H), 3.52 (m, 1H), 3.42 (m, 1H), 3.40 (s, 3H), 3.30 (m, 1H), 2.15 (m, 2H); MS (EI) for C.sub.15H.sub.31NO.sub.6: 322 (MH.sup.+).
Step 2: Preparation of Benzyl{(1S)-2-oxo-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethyl}carbamate
(30) ##STR00077##
(31) Z-Phg-OH ((2S)-{[(Benzyloxy)carbonyl]amino}(phenyl)acetic acid) (1.0 g, 3.49 mmol), (3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidine (from step 1) (1.02 g, 3.17 mmol), and N,N-diisopropylethylamine (1.22 g, 9.52 mmol) were dissolved in 20 mL of acetonitrile. The mixture was stirred for 10 minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU) (1.22 g, 3.81 mmol) was added into the solution. The reaction mixture was stirred for 1 hour at 0° C. and then overnight at room temperature. 100 mL of dichloromethane was added into the reaction mixture, and the resultant solution was washed with 5% of sodium chloride (100 mL×3). The solution was dried over sodium sulfate and was concentrated. The purification of the residue by flash chromatography yielded benzyl {(1S)-2-oxo-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethyl}carbamate (1.1 g, 59% yield). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.43 (m, 2H), 7.34 (m, 8H), 6.40-6.30 (dd, 1H), 5.40 (m, 1H), 5.23 (d, 1H), 5.04 (t, 1H), 4.08 (m, 1H), 3.70-3.50 (m, 20H), 3.42 (m, 2H), 3.41 (s, 3H), 3.21 (m. 2H), 2.12-1.80 (m, 2H); MS (EI) for C.sub.31H.sub.44N.sub.2O.sub.9: 589 (MH.sup.+).
Step 3: Preparation of (1S)—N-Methyl-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethanamine
(32) ##STR00078##
(33) A solution of benzyl {(1S)-2-oxo-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethyl}carbamate (from step 2) (0.95 g, 1.61 mmol) in tetrahydrofuran was added dropwise to a stirred 2.0 M solution of lithium aluminum hydride (4.0 mL, 8.0 mmol) at room temperature. The mixture was stirred for 30 minutes at room temperature and then was heated to 65° C. for four hours. After cooling down to room temperature, a 3N sodium carbonate solution was added into the reaction mixture carefully until effervescence ceased. The solid was filtered out and was washed with dichloromethane (100 mL). The filtrate was concentrated and the residue was dissolved in 150 mL of dichloromethane. The solution was washed with saturated sodium chloride solution and was dried over sodium sulfate. (1S)—N-Methyl-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethanamine (0.60 g, 82% yield) was obtained after removing solvent. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.39 (m, 2H), 7.35 (m, 3H), 4.08 (m, 1H), 3.67 (m, 16H), 3.18 (m, 5H), 3.40 (s, 3H), 2.90 (m, 1H), 2.80 (m, 2H), 2.55 (m, 1H), 2.48 (m, 1H), 2.30 (s. 3H), 2.10 (m, 3H), 1.82 (m, 1H); MS (EI) for C.sub.24H.sub.42N.sub.2O.sub.6: 455 (MH.sup.+).
Step 4: Preparation of 2-(3,4-Dichlorophenyl)-N-methyl-N-{(1S)-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethyl}acetamide, hydrochloride salt (7)
(34) ##STR00079##
(35) (1S)—N-Methyl-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethanamine (from step 3) (0.100 g, 0.22 mmol), 2-(3,4-dichlorophenyl)acetic acid (0.045 g, 0.22 mmol), and N,N-diisopropylethylamine (0.056 g, 0.44 mmol) were dissolved in 3 mL of acetonitrile. The mixture was stirred for 10 minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU) (0.085 g, 0.26 mmol) was added into the solution. The reaction mixture was stirred for 4 hours at room temperature and was concentrated. The residue was dissolved in dichloromethane (100 mL), was washed with water and was dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded 2-(3,4-dichlorophenyl)-N-methyl-N-{(1S)-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethyl}acetamide (0.090 g, 64% yield). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.40 (m, 2H), 7.32 (m, 2H), 7.30 (m, 3H), 7.15 (m, 1H), 6.10 (m, 0.83H), 5.02 (m, 0.17H), 4.05 (m, 1H), 3.78 (d, 1H), 3.65 (m, 17H), 3.55 (m, 4H), 3.39 (s, 3H), 3.16 (t, 1H), 3.05 (m, 1H), 2.85 (m, 1H), 2.70 (m, 4H), 2.50 (m, 2H), 2.05 (m, 1H), 1.80 (m, 1H); MS (EI) for C.sub.32H.sub.46Cl.sub.2N.sub.2O.sub.7: 641 (MH.sup.+).
(36) The free base (85 mg) was dissolved in 1 mL of acetonitrile. To the solution was added 0.5 mL of 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (7).
(37) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 6
Preparation of N-Methyl-N-{(1S)-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethyl}-N2-phenylglycinamide, hydrochloride salt (8)
(38) ##STR00080##
(39) (1S)—N-Methyl-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethanamine (from example 5, step 3) (0.070 g, 0.154 mmol), 2-(phenylamino)acetic acid (0.023 g, 0.154 mmol), and N,N-diisopropylethylamine (0.038 g, 0.308 mmol) were dissolved in 3 mL of acetonitrile. The mixture was stirred for 10 minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU) (0.059 g, 0.185 mmol) was added into the solution. The reaction mixture was stirred for 4 hours at room temperature and was concentrated. The residue was dissolved in dichloromethane (100 mL), was washed with water and was dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded N-methyl-N-{(1S)-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethyl}-N.sup.2-phenylglycinamide (0.050 g, 55% yield). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.34 (m, 5H), 7.20 (m, 2H), 6.75 (m, 1H), 6.65 (m, 2H), 6.09 (m, 1H), 5.02 (m, 1H), 4.05 (m, 1H), 3.90 (m, 2H), 3.60 (m, 20H), 3.40 (s, 3H), 3.20 (m, 1H), 3.05 (m, 1H), 2.85 (m, 2H), 2.78 (s, 3H), 2.55 (m, 1H), 2.15 (br., 1H), 2.05 (m, 1H), 1.80 (m, 1H); MS (EI) for C.sub.32H.sub.49N.sub.3O.sub.7: 588 (MH.sup.+).
(40) The free base (22 mg) was dissolved in 1 mL of acetonitrile. To the solution was added 0.2 mL of 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (8).
(41) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 7
Preparation of N-Methyl-N-{(1S)-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethyl}-2,2-diphenylacetamide, hydrochloride salt (9)
(42) ##STR00081##
(43) (1S)—N-Methyl-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethanamine (from example 5, step 3) (0.100 g, 0.22 mmol), 2,2-diphenylacetyl chloride (0.051 g, 0.22 mmol), and N,N-diisopropylethylamine (0.056 g, 0.44 mmol) were dissolved in 10 mL of dichloromethane. The mixture was stirred for 2 hours at room temperature and then 100 mL dichloromethane was added into the mixture. The resultant solution was washed with water and was dried over sodium sulfate. The solvent was removed and the residue was purified by flash chromatography. N-Methyl-N-{(1S)-2-[(3S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl]-1-phenylethyl}-2,2-diphenylacetamide was obtained (0.100 g, 70% yield). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.31 (m, 15H), 7.20 (d, 0.5H), 7.05 (d, 0.5H), 6.18 (m, 0.77H), 5.47 (s, 0.23H). 5.30 (s, 0.77H), 5.12 (m, 0.23H), 4.08 (m, 0.77H), 3.95 (m, 0.23H), 3.65 (m, 16H), 3.55 (m, 4H), 3.40 (s, 3H), 3.14 (m, 1H), 2.85 (m, 1H), 2.75 (m, 1H), 2.70 (s, 3H), 2.52 (m, 1H), 2.08 (m, 2H), 1.82 (m, 1H); MS (EI) for C.sub.38H.sub.52N.sub.2O.sub.7: 649 (MH.sup.+).
(44) The free base (95 mg) was dissolved in 1 mL of acetonitrile. To the solution was added 0.5 mL of 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (9).
(45) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 8
Preparation of 2-(3,4-Dichlorophenyl)-N-{(1S)-2-[ethyl(2-methoxyethyl)amino]-1-phenylethyl}-N-methylacetamide, hydrochloride salt (10)
(46) ##STR00082##
(47) 2-(3,4-Dichlorophenyl)-N-{(1S)-2-[ethyl(2-methoxyethyl)amino]-1-phenylethyl}-N-methylacetamide, hydrochloride salt (10) was prepared according to the following steps.
Step 1: Preparation of (1S)—N2-Ethyl-N2-(2-methoxyethyl)-N1-methyl-1-phenylethane-1,2-diamine
(48) ##STR00083##
(49) Boc-Phg-OH ((2S)-[(tert-Butoxycarbonyl)amino](phenyl)acetic acid) (0.5 g, 1.99 mmol), N-ethyl-2-methoxyethanamine (0.205 g, 1.99 mmol), and DPTS [1:1 salt of 4-(dimethylamino)pyridine and p-tolunesulfonic acid] (0.308 g, 1.0 mmol) were dissolved in 20 mL of dichloromethane. N,N′-Diisopropylcarbodiimide (0.376 g, 2.98 mmol) was added into the solution. The mixture was stirred for 1 hour. 150 mL of ethyl acetate was added into the reaction mixture. The resultant solution was washed with water (100 mL×3) and was dried over sodium sulfate. Evaporation of the solvent provided a residue which was dissolved in tetrahydrofuran. The tetrahydrofuran solution was added dropwise, over a period of 30 minutes, to a stirred suspension of lithium aluminum hydride (2.0 M, 3 mL, 6.0 mmol) at 0° C. The mixture was stirred for 16 hours at room temperature and then a 3N sodium carbonate solution was added carefully until effervescence ceased. The mixture was filtered through celite and the filtrate was concentrated. The residue was purified by flash chromatography. 1S)—N.sup.2-Ethyl-N.sup.2-(2-methoxyethyl)-N.sup.1-methyl-1-phenylethane-1,2-diamine. (0.261 g, 56% yield) was obtained. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.38 (m, 2H), 7.34 (m, 2H), 7.26 (m, 1H), 4.74 (br., 1H), 3.60 (m, 1H), 3.45 (m, 2H), 3.37 (s, 3H), 2.75 (m, 2H), 2.62 (m, 3H), 2.55 (m, 1H), 2.30 (s, 3H), 1.05 (m, 3H); MS (EI) for C.sub.14H.sub.24N.sub.2O: 237 (MH.sup.+).
Step 2: Preparation of 2-(3,4-Dichlorophenyl)-N-{(1S)-2-[ethyl(2-methoxyethyl)amino]-1-phenylethyl}-N-methylacetamide, hydrochloride salt (10)
(50) ##STR00084##
(51) (1S)—N.sup.2-Ethyl-N.sup.2-(2-methoxyethyl)-N.sup.1-methyl-1-phenylethane-1,2-diamine (Step 1) (0.080 g, 0.34 mmol), 2-(3,4-dichlorophenyl)acetic acid (0.069 g, 0.34 mmol), and N,N-diisopropylethylamine (0.040 g, 0.34 mmol) were dissolved in 10 mL of acetonitrile, and then O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU) (0.109 g, 0.34 mmol) was added into the solution. The reaction mixture was stirred for 2 hours and then dichloromethane (100 mL) was added. The resultant solution was washed with water (100 mL×2). The organic phase was dried over sodium sulfate and was concentrated. The residue was purified by flash chromatography. 2-(3,4-Dichlorophenyl)-N-{(1S)-2-[ethyl(2-methoxyethyl)amino]-1-phenylethyl}-N-methylacetamide was obtained (0.050 g, 35% yield). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.44-7.11 (m, 8H), 6.11 (m, 1H), 3.75 (m, 2H), 3.44 (m, 2H), 3.05 (m, 1H), 2.93 (m, 1H), 2.85 (m, 1H), 2.80 (m, 4H), 2.78 (m, 1H), 2.74 (m, 2H), 2.65 (m, 2H), 1.01 (m, 3H); MS (EI) for C.sub.22H.sub.28Cl.sub.2N.sub.2O.sub.2: 423 (MH.sup.+).
(52) The free base (20 mg) was dissolved in 1 mL of acetonitrile. To the solution was added 0.2 mL of 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (10).
(53) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 9
Preparation of N-[(1S)-2-[(3S)-3-Hydroxypyrrolidin-1-yl]-1-(3-{[(2-methoxyethoxy)acetyl]amino}phenyl)ethyl]-N-methyl-2,2-diphenylacetamide, hydrochloride salt (11)
(54) ##STR00085##
(55) N-[(1S)-2-[(3S)-3-Hydroxypyrrolidin-1-yl]-1-(3-{[(2-methoxyethoxy)acetyl]amino}phenyl)ethyl]-N-methyl-2,2-diphenylacetamide was prepared according to the following steps.
Step 1: Preparation of ((3S)-1-[(2S)-2-[(Diphenylacetyl)(methyl)amino]-2-(3-nitrophenyl)ethyl]pyrrolidin-3-yl diphenylacetate
(56) ##STR00086##
(57) (3S)-1-[(2S)-2-(Methylamino)-2-(3-nitrophenyl)ethyl]pyrrolidin-3-ol (0.400 g, 1.21 mmol (prepared according to Ghosh, A., Chemical Communications, no. 15, p. 1644 (2002)), 2,2-diphenylacetyl chloride (0.618 g, 2.41 mmol), and N,N-diisopropylethylamine (0.463 g, 3.62 mmol)) were dissolved in 40 mL of dichloromethane. The mixture was stirred for 2 hours at room temperature and then 100 mL dichloromethane was added into the mixture. The resultant solution was washed with water and was dried over sodium sulfate. The solvent was removed and the residue was purified by flash chromatography. ((3S)-1-[(2S)-2-[(Diphenylacetyl)(methyl)amino]-2-(3-nitrophenyl)ethyl]pyrrolidin-3-yl diphenylacetate (0.410 g, 52% yield) was obtained. MS (EI) for C.sub.41H.sub.39N.sub.3O.sub.5: 654 (MH.sup.+).
Step 2: Preparation of (3S)-1-{(2S)-2-(3-Aminophenyl)-2-[(diphenylacetyl)(methyl)amino]ethyl}pyrrolidin-3-yl diphenylacetate
(58) ##STR00087##
(59) ((3S)-1-[(2S)-2-[(Diphenylacetyl)(methyl)amino]-2-(3-nitrophenyl)ethyl]pyrrolidin-3-yl diphenylacetate (from step 1) (0.24 g, 0.37 mmol) was dissolved in 15 mL of tetrahydrofuran. The reduction reaction was performed by H-Cube (CatCart THS01131, 10% Pd/C, Flow rate 1 mL/Min). (3S)-1-{(2S)-2-(3-Aminophenyl)-2-[(diphenylacetyl)(methyl)amino]ethyl}pyrrolidin-3-yl diphenylacetate was obtained (0.2 g, 87% yield). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.40-7.20 (m, 20H), 7.10 (m, 1H), 6.65 (m, 1H), 6.60 (m, 2H), 6.08 (m, 1H), 5.30 (m, 1H), 5.25 (s, 1H), 5.08 (s, 1H), 3.62 (br., 2H), 3.10 (t, 1H), 3.00 (m, 2H), 2.82 (m, 1H), 2.68 (s, 3H), 2.65 (m, 1H), 2.35 (m, 1H), 2.25 (m, 1H), 1.85 (m, 1H); MS (EI) for C.sub.41H.sub.41N.sub.3O.sub.3: 624 (MH.sup.+).
Step 3: Preparation of N-[(1S)-2-[(3S)-3-Hydroxypyrrolidin-1-yl]-1-(3-{[(2-methoxyethoxy)acetyl]amino}phenyl)ethyl]-N-methyl-2,2-diphenylacetamide, hydrochloride salt (11)
(60) ##STR00088##
(61) (3S)-1-{(2S)-2-(3-Aminophenyl)-2-[(diphenylacetyl)(methyl)amino]ethyl}pyrrolidin-3-yl diphenylacetate (from step 2) (0.120 g, 0.192 mmol), 2-(2-methoxyethoxy)acetic acid (0.031 g, 0.231 mmol), and DPTS [1;1 salt of 4-(dimethylamino)pyridine and p-tolunesulfonic acid] (0.036 g, 0.115 mmol) were dissolved in 10 mL of dichloromethane. N,N-diisopropylcarbodiimide (0.073 g, 0.577 mmol) was added into the mixture. The reaction mixture was stirred for 2 hours. 100 mL of dichloromethane was added into the reaction mixture and the resultant solution was washed with water (100 mL×3). The solution was dried over sodium sulfate and was concentrated. The residue was dissolved in 6 mL of CH.sub.3CN/0.5N KOH (1:1). The mixture was stirred at 65° C. for 3 hours. The mixture was extracted with dichloromethane (50 mL×3). The dichloromethane solution was dried over sodium sulfate and was concentrated. The crude product was purified by flash chromatography. N-[(1S)-2-[(3S)-3-Hydroxypyrrolidin-1-yl]-1-(3-{[(2-methoxyethoxy)acetyl]amino}phenyl)ethyl]-N-methyl-2,2-diphenylacetamide (0.050 g, 49% yield) was obtained. .sup.1H NMR (500 MHz, MeOD): δ 7.64-7.50 (m, 2H), 7.40-7.20 (m, 1H), 7.10 (m, 1H), 6.10 (m, 0.8H), 5.52 (s, 0.2H), 5.45 (s, 0.8H), 5.24 (m, 0.2H), 4.35 (br., 0.8H), 4.22 (br., 0.2H), 4.24 (m, 2H), 3.78 (m, 2H), 3.65 (m, 2H), 3.45 (s, 3H), 3.34 (s, 1H), 3.24 (t, 1H), 3.10 (m, 1H), 2.92-2.80 (m, 2H), 2.28 (s, 3H), 2.60 (m, 1H), 2.45 (m, 1H), 2.14 (m, 1H), 1.74 (m, 1H); MS (EI) for C.sub.32H.sub.39N.sub.3O.sub.5: 546 (MH.sup.+).
(62) The free base (25 mg) was dissolved in 1 mL of acetonitrile. To the solution was added 0.2 mL of 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (11).
(63) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 10
Preparation of N-{3-[(1S)-1-[(Diphenylacetyl)(methyl)amino]-2-(pyrrolidin-1-yl)ethyl]phenyl}-2,5,8,11,14,17-hexaoxanonadecan-19-amide, hydrochloride salt (12)
(64) ##STR00089##
(65) N-{3-[(1S)-1-[(Diphenylacetyl)(methyl)amino]-2-(pyrrolidin-1-yl)ethyl]phenyl}-2,5,8,11,14,17-hexaoxanonadecan-19-amide may be prepared according to the following steps.
Step 1: Preparation of (S)—N-Methyl-N-(1-(3-nitrophenyl)-2-(pyrrolidin-1-yl)ethyl)-2,2-diphenylacetamide
(66) ##STR00090##
(67) (S)-1-[2-(Methylamino)-2-(3/4-nitrophenyl)ethyl]pyrrolidin (0.84 g, 3.37 mmol), (Portoghese, P., Journal of Medicinal Chemistry, (1994), vol. 37, pp. 4490-4498) was dissolved in anhydrous dichloromethane (14 mL). To the dark solution was added diisopropylethylamine (1.17 mL, 6.74 mmol) at 0° C. Diphenylacetyl chloride (0.95 g, 3.71 mmol) was dissolved in anhydrous dichloromethane (10 mL) and was added dropwise to the dark solution, maintaining the temperature below 5° C. The reaction mixture was allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the mixture was partitioned between dichloromethane (35 mL) and saturated sodium chloride (50 mL). The aqueous layer was extracted with dichloromethane (3×15 mL). The combined organic portion was washed with saturated sodium bicarbonate (2×25 mL) and saturated sodium chloride (25 mL). The combined organic portion was dried over anhydrous sodium sulfate (approximately 1.70 g) and concentrated in vacuo. The residue was purified on a column of silica gel using dichloromethane/methanol (9:1) as eluent to give (S)—N-methyl-N-(1-(3-nitrophenyl)-2-(pyrrolidin-1-yl)ethyl)-2,2-diphenylacetamide (0.62 g, 42% yield), as a light-yellow oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 8.13-7.23 (m, 14H), 6.17 (m, 1H), 5.25 (s, 1H), 3.09 (m, 1H), 2.86 (m, 1H), 2.75 (s, 3H), 2.72 (m, 2H), 2.52 (m, 2H), 1.77 (m, 4H); MS (EI) for C.sub.27H.sub.29N.sub.3O.sub.3: 444 (MH.sup.+).
(68) The compound was converted into the hydrochloride salt by dissolving the oil in acetonitrle and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt as a white powder.
Step 2: Preparation of (S)—N-Methyl-N-(1-(3-aminophenyl)-2-(pyrrolidin-1-yl)ethyl)-2,2-diphenylacetamide, dihydrochloride salt (51)
(69) ##STR00091##
(70) (S)—N-methyl-N-(1-(3-nitrophenyl)-2-(pyrrolidin-1-yl)ethyl)-2,2-diphenylacetamide (from step 1) (1.36 g, 3.07 mmol) was dissolved in ethyl acetate:ethanol (1:1, 30 mL each) and the reaction mixture was passed through a 10% Palladium/Carbon CatCart® cartridge (70 millimeters) in a continuous flow through the H-Cube™, in which hydrogen is generated in-situ from the electrolysis of water. The eluent was collected and was concentrated under reduced pressure to give a residue. The residue was purified on a column of silica gel using hexane/methanol (9:1) as eluent to give (S)—N-methyl-N-(1-(3-aminophenyl)-2-(pyrrolidin-1-yl)ethyl)-2,2-diphenylacetamide (0.47 g, 40% yield), as a light-yellow foam. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.32-7.04 (m, 14H), 6.71 (m, 1H), 6.57 (m, 2H), 6.10 (m, 1H), 5.26 (s, 1H), 3.61-3.40 (m, 2H), 2.95-2.30 (m, 5H), 1.70 (m, 3H), 1.69 (m, 2H); MS (EI) for C.sub.27H.sub.31N.sub.3O: 414 (MH.sup.+).
(71) The compound was converted into the dihydrochloride salt by dissolving the foam in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the dihydrochloride salt (51) as a white powder.
Step 3: N-{3-[(1S)-1-[(Diphenylacetyl)(methyl)amino]-2-(pyrrolidin-1-yl)ethyl]phenyl}-2,5,8,11,14,17-hexaoxanonadecan-19-amide, hydrochloride salt (12)
(72) ##STR00092##
(73) (S)—N-methyl-N-(1-(3-aminophenyl)-2-(pyrrolidin-1-yl)ethyl)-2,2-diphenylacetamide (from step 2) (75 mg, 0.18 mmol) was dissolved in anhydrous dichloromethane (1 mL) and to the light-yellow solution was added diisopropylethylamine (0.06 mL, 0.36 mmol), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (50 mg, 0.27 mmol) and 2,5,8,11,14,17-hexaoxanonadecan-19-oic acid (80 mg, 0.27 mmol). The yellow reaction mixture was stirred under nitrogen at room temperature. After approximately 17 hours at room temperature, the reaction mixture was diluted with dichloromethane (25 mL) and partitioned with water (30 mL). The aqueous layer was extracted with dichloromethane (2×15 mL). The combined organic portion was washed with 1.0 N hydrochloric acid (35 mL), water (35 mL), saturated sodium bicarbonate (35 mL), and saturated sodium chloride (35 mL). The combined organic portion was dried over anhydrous sodium sulfate (approximately 0.20 g) and concentrated in vacuo. The residue was purified on a column of silica gel using dichloromethane/methanol (9:1) as eluent to give N-{3-[(1S)-1-[(diphenylacetyl)(methyl)amino]-2-(pyrrolidin-1-yl)ethyl]phenyl}-2,5,8,11,14,17-hexaoxanonadecan-19-amide (35 mg, 27% yield), as a light-yellow oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 8.73 (bs, 1H), 7.53-7.06 (m, 14H), 6.15 (m, 1H), 5.25 (s, 1H), 4.11 (s, 2H), 3.76-3.54 (m, 21H), 3.38 (s, 3H), 2.75-2.60 (m, 5H), 2.50-2.42 (m, 3H), 1.75 (m, 4H); MS (EI) for C.sub.40H.sub.55N.sub.3O.sub.8: 706 (MH.sup.+).
(74) The compound was converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt (12) as a white powder.
(75) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 11
Preparation of N-Methyl-2-[4-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)phenyl]-N-[(1S)-1-phenyl-2-(pyrrolidin-1-yl)ethyl]acetamide, hydrochloride salt (13)
(76) ##STR00093##
(77) N-Methyl-2-[4-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)phenyl]-N-[(1S)-1-phenyl-2-(pyrrolidin-1-yl)ethyl]acetamide was prepared according to the following steps.
Step 1: Preparation of 2-(4-Hydroxyphenyl)-N-methyl-N-[(1S)-1-phenyl-2-(pyrrolidin-1-yl)ethyl acetamide, hydrochloride salt (52)
(78) ##STR00094##
(79) (1S)—N-Methyl-1-phenyl-2-pyrrolidin-1-yl)ethanamine (0.30 g, 1.46 mmol), (Singh, V. et. al., Journal of Organic Chemistry (1996), vol. 61, pp. 6108-6113) was dissolved in anhydrous acetonitrile (7 mL). The clear solution was stirred at 0° C., under nitrogen, followed by the addition of diiso-propylethylamine (0.57 mL, 3.23 mmol), 2-(4-hydroxyphenyl)acetic acid (0.25 g, 1.61 mmol) and O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.39 g, 1.61 mmol). The clear reaction mixture was allowed to equilibrate to room temperature and stirred under nitrogen. After approximately 17 hours at room temperature the mixture was concentrated in vacuo. The residue was dissolved in ethyl acetate (8 mL) and was washed with saturated sodium bicarbonate (2×15 mL) and saturated sodium chloride (15 mL). The combined organic portion was dried over anhydrous sodium sulfate (approximately 0.60 g) and concentrated in vacuo. The residue was purified on a column of silica gel using dichloromethane/methanol (9:1) as eluent to give 2-(4-hydroxyphenyl)-N-methyl-N-[(1S)-1-phenyl-2-(pyrroldin-1-yl)ethyl acetamide (0.18 g, 36% yield), as a light-yellow oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.37-7.02 (m, 9H), 6.67 (m, 2H), 6.15 (m, 1H), 3.76 (m, 2H), 2.77 (m, 6H), 2.52 (m, 2H), 1.75 (m, 4H); MS (EI) for C.sub.21H.sub.26N.sub.2O.sub.2: 339 (MH.sup.+).
(80) The compound was converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt (52) as a white powder.
Step 2: Preparation of N-Methyl-2-[4-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)phenyl]-N-[(1S)-1-phenyl-2-(pyrrolidin-1-yl)ethyl]acetamide, hydrochloride salt (13)
(81) ##STR00095##
(82) 2-(4-Hydroxyphenyl)-N-methyl-N-[(1S)-1-phenyl-2-(pyrroldin-1-yl)ethyl acetamide (from step 1) (0.10 g, 0.29 mmol) was dissolved in anhydrous acetone (10 mL) and to the clear solution was added potassium carbonate (0.10 g, 0.73 mmol), followed by 2,5,8,11,14-pentaoxahexadecan-16-yl methanesulfonate (0.10 g, 0.31 mmol). The light-yellow reaction mixture was stirred under nitrogen and heated to reflux. After approximately 17 hours the yellow reaction mixture was cooled to room temperature. The solvent was removed under reduced pressure and the residue was partitioned between water (10 mL) and dichloromethane (10 mL). The aqueous portion was extracted with dichloromethane (3×8 mL). The combined organic portion was washed with water (2×15 mL) and saturated sodium chloride (15 mL). The organic portion was dried over anhydrous sodium sulfate (approximately 0.25 g), filtered and concentrated under reduced pressure to give a yellow oil. The residue was purified on a column of silica gel using dichloromethane/methanol (9:1) as eluent to give N-methyl-2-[4-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)phenyl]-N-[(1S)-1-phenyl-2-(pyrrolidin-1-yl)ethyl]acetamide (89 mg, 53% yield), as a light-yellow oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.29-7.18 (m, 7H), 6.84 (m, 2H), 6.12 (m, 1H), 4.10 (m, 2H), 3.84 (m, 2H), 3.66 (m, 3H), 3.56-3.54 (m, 15H), 3.37 (s, 3H), 3.14-2.48 (m, 9H), 1.76 (m, 4H); MS (EI) for C.sub.32H.sub.48N.sub.2O.sub.7: 573 (MH.sup.+).
(83) The compound was converted into the hydrochloride salt (13) by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt as a white powder.
(84) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 12
Preparation of 2-(3,4-dichlorophenyl)-N-methyl-N-[(1S)-1-{3-[(methylsulfonyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]acetamide, hydrochloride salt (14)
(85) ##STR00096##
(86) 2-(3,4-Dichlorophenyl)-N-methyl-N-[(1S)-1-{3-[(methylsulfonyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]acetamide may be prepared according to the following steps.
Step 1: Preparation of 2-(3,4-dichlorophenyl)-N-methyl-N-[(1S)-1-(3-nitrophenyl)-2-(pyrrolidin-1-yl)ethyl]acetamide
(87) ##STR00097##
(88) (S)-1-[2-(Methylamino)-2-(3/4-nitrophenyl)ethyl]pyrroldin (0.66 g, 2.65 mmol), (Portoghese, P. Journal of Medicinal Chemistry, 1994, 37, 4490-4498) was dissolved in anhydrous acetonitrile (14 mL). To the dark solution was added diisopropylethylamine (1.03 mL, 5.82 mmol) at 0° C., followed by 3,4-dichlorophenylacetic acid (0.60 g, 2.91 mmol) and O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.96 g, 2.91 mmol). The reaction mixture was allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the reaction mixture was concentrated under reduced pressure. The residue was taken up in ethyl acetate (15 mL) and washed with saturated sodium bicarbonate (2×25 mL) and saturated sodium chloride (25 mL). The combined organic portion was dried over anhydrous sodium sulfate and concentrated under reduced pressure. Purification by chromatography gave 2-(3,4-dichlorophenyl)-N-methyl-N-[(1S)-1-(3-nitrophenyl)-2-(pyrrolidin-1-yl)ethyl acetamide (0.99 g, 86% yield), as a light-yellow oil. The compound was converted into the hydrochloride salt by dissolving the oil in acetonitrle and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt as a white powder. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 8.13 (m, 2H), 7.66 (m, 1H), 7.51 (m, 1H), 7.37 (m, 2H), 7.15 (m, 1H), 6.11 (m, 1H), 3.77 (m, 2H), 3.13 (m, 1H), 2.80 (m, 1H), 2.69 (s, 3H), 2.60 (m, 2H), 1.76 (m, 4H); MS (EI) for C.sub.21H.sub.23Cl.sub.2N.sub.3O.sub.3: 437 (MH.sup.+).
Step 2: Preparation of N-[(1S)-1-(3-aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-2-(3,4-dichlorophenyl)-N-methylacetamide
(89) ##STR00098##
(90) 2-(3,4-dichlorophenyl)-N-methyl-N-[(1S)-1-(3-nitrophenyl)-2-(pyrrolidin-1-yl)ethyl acetamide (0.80 g, 1.77 mmol), hydrazine hydrate (1.36 g, 21.34 mmol) and Raney nickel slurry (1.0 mL) in 95% ethanol (89 mL) was heated to 55° C. After approximately two hours the reaction was complete as indicated by TLC. The reaction mixture was filtered through Celite, and the Raney nickel was washed with hot methanol. The combined filtrates were concentrated under reduced pressure to give 0.60 g (83%) of N-[(1S)-1-(3-aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-2-(3,4-dichlorophenyl)-N-methylacetamide. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.38 (m, 2H), 7.17 (m, 1H), 7.10 (m, 1H), 6.63 (m, 3H), 6.01 (m, 1H), 3.66-3.79 (m, 4H), 3.14 (m, 1H), 2.46-2.72 (m, 5H), 2.47 (m, 3H), 1.74 (m, 4H); MS (EI) for C.sub.21H.sub.25Cl.sub.2N.sub.3O: 406 (MH.sup.+).
Step 3: 2-(3,4-Dichlorophenyl)-N-methyl-N-[(1S)-1-{3-[(methylsulfonyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]acetamide, hydrochloride salt (14)
(91) ##STR00099##
(92) N-[(1S)-1-(3-Aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-2-(3,4-dichlorophenyl)-N-methylacetamide (0.060 g, 0.15 mmol) was dissolved in dichloromethane (1.5 mL) and anhydrous pyridine (0.090 mL). To the cooled (0° C.) yellow solution there was added dropwise methanesulfonyl chloride (0.017 mL, 0.22 mmol). The yellow reaction mixture was allowed to stir at 0° C., with the color turning orange. The reaction mixture was allowed to equilibrate to room temperature. After approximately 17 hours at room temperature, the reaction mixture was diluted with dichloromethane (25 mL) and then added water (30 mL). The aqueous portion was extracted with dichloromethane (2×15 mL). The combined organic portions were washed with 1N hydrochloric acid, water, saturated sodium bicarbonate, water and brine (35 mL each). The organic portion was dried over anhydrous sodium sulfate, filtered, concentrated and purified by chromatography to give 0.045 g (63%) of 2-(3,4-dichlorophenyl)-N-methyl-N-[(1S)-1-{3-[(methylsulfonyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]acetamide as a yellow oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.07-7.34 (m, 7H), 6.06 (m, 1H), 3.79 (m, 1H), 3.70 (m, 1H), 3.23 (m, 1H), 2.88-3.10 (m, 6H), 2.76 (s, 3H), 2.58 (m, 2H), 1.79 (m, 5H); MS (EI) for C.sub.22H.sub.27Cl.sub.2N.sub.3O.sub.3S: 484 (MH.sup.+).
(93) The compound was converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt (14) as an off-white powder.
(94) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 13
Preparation of 2-(3,4-dichlorophenyl)-N-methyl-N-[(1S)-1-{3-[(2,5,8,11,14-pentaoxahexadecan-16-ylcarbamoyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]acetamide, hydrochloride salt (15)
(95) ##STR00100##
(96) To a solution of triphosgene (0.054 gm, 0.17 mmol) in anhydrous acetonitrile (3.5 mL), at −5° C., was added an acetonitrile (3.5 mL) solution of N-[(1S)-1-(3-aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-2-(3,4-dichlorophenyl)-N-methylacetamide (0.12 gm, 0.29 mmol) and triethylamine (0.055 mL, 0.38 mmol) over a period of ten minutes. The mixture was stirred for an additional 20 minutes at 0° C., and then a dichloroethane (2.8 mL) solution of 2,5,8,11,14-pentaoxahexadecan-16-amine (0.096 gm, 0.38 mmol) and triethylamine (0.055 mL, 0.38 mmol) was added, maintaining the temperature less than 5° C. The reaction mixture was allowed to equilibrate to room temperature overnight. After 20 hours the mixture was diluted with dichloromethane (40 mL) and transferred to a separatory funnel with water (35 mL). The aqueous portion was extracted with dichloromethane (3×25 mL). The combined organic portions were washed with water (2×40 mL) and saturated sodium chloride (50 mL). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to a residue. Purification by chromatography was carried out to give 0.074 gm (37%) of 2-(3,4-dichlorophenyl)-N-methyl-N-[(1S)-1-{3-[(2,5,8,11,14-pentaoxahexadecan-16-ylcarbamoyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]acetamide as a yellow oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 8.83 (m, 1H), 7.59-6.79 (m, 7H), 6.30 (m, 1H), 6.06 (m, 1H), 4.10 (m, 3H), 3.80 (m, 1H), 3.78-3.40 (m, 22H), 3.22 (m, 4H), 3.15 (m, 2H), 2.80 (m, 3H), 1.98 (m, 3H); MS (EI) for C.sub.33H.sub.48Cl.sub.2N.sub.4O.sub.7: 683 (MH.sup.+).
(97) The compound was converted into the hydrochloride salt (15) by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt as an off-white powder.
(98) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 14
Preparation of 2,2-Bis(4-chlorophenyl)-N-methyl-N-[(1S)-1-{3-[(2,5,8,11,14-pentaoxahexadecan-16-ylcarbamoyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]acetamide, hydrochloride salt. (16)
(99) ##STR00101##
(100) 2,2-Bis(4-chlorophenyl)-N-methyl-N-[(1S)-1-{3-[(2,5,8,11,14-pentaoxahexadecan-16-ylcarbamoyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]acetamide may be prepared according to the following steps.
Step 1: Preparation of 2,2-bis(4-chlorophenyl)-N-methyl-N-[(1S)-1-(3-nitrophenyl)-2-(pyrrolidin-1-yl)ethyl]acetamide
(101) ##STR00102##
(102) (S)-1-[2-(Methylamino)-2-(3/4-nitrophenyl)ethyl]pyrroldin (1.02 g, 4.09 mmol), (Portoghese, P. Journal of Medicinal Chemistry, 1994, 37, 4490-4498) was dissolved in anhydrous acetonitrile (20 mL). To the solution was added diisopropylethylamine (1.48 mL, 8.39 mmol) at 0° C. Diphenylacetyl chloride (1.03 g, 4.50 mmol) was taken up in anhydrous dichloromethane (10 mL) and added dropwise to the solution. The reaction mixture was allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the reaction mixture was concentrated under reduced pressure. The residue was partitioned between dichloromethane (15 mL) and water (15 mL). The aqueous layer was extracted with dichloromethane (3×15 mL). The combined organic portion was washed with saturated sodium bicarbonate (2×25 mL) and saturated sodium chloride (25 mL). The combined organic portion was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified on a column of silica gel using dichloromethane/methanol (9:1) as eluent to give 1.38 g (76%) of 2,2-bis(4-chloro-phenyl)-N-methyl-N-[(1S)-1-(3-nitrophenyl)-2-(pyrrolidin-1-yl)ethyl]acetamide as a yellow oil. MS (EI) for C.sub.27H.sub.27Cl.sub.2N.sub.3O.sub.3: 512 (MH.sup.+).
Step 2: Preparation of N-[(1S)-1-(3-aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-2,2-bis(4-chlorophenyl)-N-methylacetamide
(103) ##STR00103##
(104) 2,2-Bis(4-chlorophenyl)-N-methyl-N-[(1S)-1-(3-nitrophenyl)-2-(pyrrolidin-1-yl)ethyl]acetamide (0.60 g, 1.35 mmol) in ethyl acetate:ethanol (1:1; 13.5 mL each; 0.05 M) was pumped through the 10% Pd/C filled CatCart™ column in an H-Cube™ reactor operated at 20° C. and at a flow rate of 1.0 mL/min. The solvent was concentrated under reduced pressure to give 0.34 g (60%) of N-[(1S)-1-(3-aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-2,2-bis(4-chlorophenyl)-N-methylacetamide as a light-yellow oil. MS (EI) for C.sub.27H.sub.29Cl.sub.2N.sub.3O: 482 (MH.sup.+).
Step 3: Preparation of 2,2-bis(4-chlorophenyl)-N-methyl-N-[(1S)-1-{3-[(2,5,8,11,14-pentaoxahexadecan-16-ylcarbamoyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]acetamide, hydrochloride salt (16)
(105) ##STR00104##
(106) To a solution of triphosgene (0.054 gm, 0.17 mmol) in anhydrous acetonitrile (3.5 mL), at −5° C., was added an acetonitrile (3.5 mL) solution of N-[(1S)-1-(3-aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-2,2-bis(4-chlorophenyl)-N-methylacetamide (0.12 gm, 0.29 mmol) and triethylamine (0.055 mL, 0.38 mmol) over a period of ten minutes. The mixture was stirred for an additional 20 minutes at 0° C., and then a dichloroethane (2.8 mL) solution of 2,5,8,11,14-pentaoxahexa-decan-16-amine (0.096 gm, 0.38 mmol) and triethylamine (0.055 mL, 0.38 mmol) was added, maintaining the temperature less than 5° C. The reaction mixture was allowed to equilibrate to room temperature overnight. After 20 hours the mixture was diluted with dichloromethane (40 mL) and transferred to a separatory funnel with water (35 mL). The aqueous layer was extracted with dichloromethane (3×25 mL). The combined organic portion was washed with water (2×40 mL) and saturated sodium chloride (50 mL). The organic portion was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure and purified by chromatography to give 0.034 gm (43%) of 2,2-bis(4-chlorophenyl)-N-methyl-N-[(1S)-1-{3-[(2,5,8,11,14-pentaoxahexadecan-16-ylcarbamoyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]acetamide as a yellow oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.42 (m, 1H), 7.21-7.08 (m, 12H), 6.78 (m, 1H), 3.54-3.39 (m, 20H), 3.24-3.18 (m, 10H), 3.20 (m, 2H), 2.77 (m, 3H), 1.85 (m, 3H); MS (EI) for C.sub.39H.sub.52Cl.sub.2N.sub.4O.sub.7: 759 (MH.sup.+).
(107) The compound was converted into the hydrochloride salt (16) by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt as an off-white powder.
(108) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 15
Preparation of 2-(3,4-dichlorophenyl)-N-[(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-{3-[(methylsulfonyl)amino]phenyl}ethyl]-N-methylacetamide, hydrochloride salt. (17)
(109) ##STR00105##
(110) 2-(3,4-Dichlorophenyl)-N-[(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-1-{3-[(methylsulfonyl)amino]phenyl}ethyl]-N-methylacetamide may be prepared according to the following steps.
Step 1: Preparation of tert-butyl(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidine-1-carboxylate
(111) ##STR00106##
(112) (S)-Tert-butyl 3-hydroxypyrrolidine-1-carboxylate (1.0 g, 5.18 mmol) was taken up in tetrahydrofuran (30 mL) and then added to 60% sodium hydride (0.41 g, 10.3 mmol) and stirred for 30 minutes at room temperature. To the cloudy solution was added 1-bromo-2-(2-(2-methoxyethoxy)ethoxy)ethane (2.35 g, 10.3 mmol) in tetrahydrofuran (28 mL) and the temperature was stirred under nitrogen at room temperature. After approximately 17 hours at room temperature, the cloudy reaction mixture was concentrated under reduced pressure. The residue was diluted with methyl tert-butyl ether (100 mL) and transferred to a separatory funnel and then washed with saturated sodium chloride (2×150 mL), and dried over anhydrous sodium sulfate. The organic portion was filtered and concentrated under reduced pressure to give a light-yellow oil which was purified by chromatography to give 0.82 g (48%) of tert-butyl(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxyl}pyrrolidine-1-carboxylate as a light-yellow oil. .sup.1H NMR (500 MHz, DMSO-d6): δ 4.03 (m, 1H), 3.51-3.41 (m, 12H), 3.33-3.23 (m, 3H), 3.18 (m, 1H), 1.89 (m, 1H), 1.39 (s, 9H).
Step 2: Preparation of (3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidine
(113) ##STR00107##
(114) Anhydrous hydrochloric acid (4 M hydrochloric acid in dioxane; 3.3 mL) was added to (S)-tert-butyl 3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidine-1-carboxylate (0.80 g, 2.39 mmol) in dichloromethane (4 mL) at 0° C. The light-yellow reaction mixture was equilibrated to room temperature and stirred for two hours. The mixture was concentrated under reduced pressure, and the residual hydrochloric acid was removed by taking up in chloroform and evaporating the solvent. The light-brown residue was dried under high vacuum for three hours to give 0.64 g (quant.) of (3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidine as a clear oil. .sup.1H NMR (500 MHz, DMSO-d6): δ 9.10 (m, 1H), 4.21 (m, 1H), 3.54-3.41 (m, 10H), 3.34 (s, 3H), 3.23-3.17 (m, 8H), 2.02 (m, 1H), 1.92 (s, 1H); MS (EI) for C.sub.11H.sub.23NO.sub.4: 234 (MH.sup.+).
Step 3: Preparation of benzyl {(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)-ethoxy]ethoxy}-pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate
(115) ##STR00108##
(116) A solution of (2S)-{[(benzyloxy)carbonyl]amino}(phenyl)acetic acid (0.71 g, 2.48 mmol), (3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidine (0.67 g, 2.48 mmol) and diisopropylethyl amine (1.33 mL, 7.45 mmol) in anhydrous acetonitrile (16 mL) was stirred at room temperature for 15 minutes and then cooled to 0° C. O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.72 g, 2.98 mmol) was added and the light-brown reaction mixture was stirred at 0° C. for one hour, and then equilibrated to room temperature. After approximately 17 hours at room temperature the orange mixture was concentrated under reduced pressure. The residue was purified by chromatography to give 0.93 g (75%) of benzyl {(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)-ethoxy]ethoxy}-pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate as a clear oil. MS (EI) for C.sub.27H.sub.36N.sub.2O.sub.7: 501 (MH.sup.+).
Step 4: Preparation of (1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-N-methyl-1-phenylethanamine
(117) ##STR00109##
(118) Benzyl {(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)-ethoxy]ethoxy}-pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate (0.77 g, 1.53 mmol) was dissolved in anhydrous tetrahydrofuran (15 mL) and cooled to 0° C. To this solution was added lithium aluminum hydride (2 M in tetrahydrofuran, 1.53 mL, 3.08 mmol) carefully. The cloudy reaction mixture was then heated under reflux (oil bath at 70° C.) for two hours. The cloudy reaction mixture was allowed to equilibrate to room temperature. After approximately 18 hours at room temperature the cloudy mixture was cooled with an ice bath. The excess lithium aluminum hydride was quenched with two drops of ethyl acetate. The reaction mixture was treated with water (52 μL), followed by 4N NaOH (52 μL). After five minutes water (154 μL) was added and stirred for 15 minutes. A white precipitate was filtered off and the filtrate was dried over anhydrous sodium sulfate, and then filtered and concentrated under reduced pressure. The residue was purified by chromatography to give 0.34 g (60%) of benzyl {(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)-ethoxy]ethoxy}-pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate as a light-yellow semi-solid. MS (EI) for C.sub.20H.sub.34N.sub.2O.sub.4: 367 (MH.sup.+).
Step 5: Preparation of (1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-N-methyl-1-(3-nitrophenyl)ethanamine
(119) ##STR00110##
(120) To an ice-cold solution of benzyl {(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)-ethoxy]ethoxy}-pyrrolidin-1-yl]-2-oxo-1-phenylethyl}carbamate (0.60 gm, 2.94 mmol) in concentrated sulfuric acid (11 mL) was added 70% nitric acid (0.28 mL, 4.41 mmol) with vigorous stirring. The stirring with ice-cooling continued for 45 minutes and the reaction mixture was made basic by careful addition of 4N sodium hydroxide and water with stirring and ice-cooling. The reaction mixture was partitioned between ethyl acetate (45 mL) and water (60 mL). The aqueous layer was extracted with ethyl acetate (3×20 mL). The organic portion was washed with water and saturated sodium chloride (60 mL each) and dried over anhydrous sodium sulfate to give 0.25 g (67%) of (1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-N-methyl-1-(3-nitrophenyl)ethanamine as a light yellow oil. MS (EI) for C.sub.20H.sub.33N.sub.3O.sub.6: 412 (MH.sup.+).
Step 6: Preparation of 2-(3,4-dichlorophenyl)-N-[(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-(3-nitrophenyl)ethyl]-N-methylacetamide
(121) ##STR00111##
(122) To a stirring solution of (1S)-2-[(3.5)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-N-methyl-1-(3-nitrophenyl)ethanamine (0.66 gm, 2.65 mmol) in dry acetonitrile (13.2 mL; 0.20M) at 0° C. under nitrogen was added diisopropylethylamine (1.03 mL, 5.82 mmol), 3,4-dichlorophenylacetic acid (0.60 gm, 2.91 mmol) and O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.96 gm, 2.91 mmol). The dark reaction mixture was allowed to equilibrate to room temperature, and stirred under nitrogen. After approximately 17 hours at room temperature, the dark mixture was concentrated under reduced pressure. The dark oil was taken up in ethyl acetate and washed with saturated sodium bicarbonate and saturated sodium chloride. The organic portion was filtered and concentrated under reduced pressure. The residue was purified by chromatography to give 0.15 g (42%) of 2-(3,4-dichlorophenyl)-N-[(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-1-(3-nitrophenyl)ethyl]-N-methylacetamide as a light-yellow oil. MS (EI) for C.sub.28H.sub.37Cl.sub.2N.sub.3O.sub.7: 598 (MH.sup.+).
Step 7: Preparation of 2-(3,4-dichlorophenyl)-N-[(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-(3-aminophenyl)ethyl]-N-methylacetamide
(123) ##STR00112##
(124) To a three-neck, 100 mL round bottom flask equipped with a condenser, thermometer, and magnetic stirrer was added (3,4-dichlorophenyl)-N-[(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-(3-nitrophenyl)ethyl]-N-methylacetamide compound (0.15 g, 0.25 mmol) in 9 mL absolute ethanol. To the light-yellow solution was added hydrazine hydrate (0.19 g, 3.05 mmol) and Raney nickel slurry (13 drops) and heated to 55° C. (oil bath temp at 64° C.). The light-yellow reaction turned clear and after approximately two hours at 55° C. the reaction was complete as indicated by LC-MS. The reaction mixture was filtered through Celite, and the Raney nickel was washed with hot methanol. The combined filtrates were concentrated under reduced pressure to give a light-yellow oil. Purification by chromatography gave 0.12 g (82%) of 2-(3,4-dichlorophenyl)-N-[(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-(3-aminophenyl)ethyl]-N-methylacetamide as a clear oil. MS (EI) for C.sub.28H.sub.39Cl.sub.2N.sub.3O.sub.5: 568 (MH.sup.+).
Step 8: Preparation of 2-(3,4-dichlorophenyl)-N-[(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-{3-[(methylsulfonyl)amino]phenyl}ethyl]-N-methylacetamide, hydrochloride salt (17)
(125) ##STR00113##
(126) 2-(3,4-Dichlorophenyl)-N-[(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-1-(3-aminophenyl)ethyl]-N-methylacetamide (0.11 g, 0.20 mmol) was dissolved in dichloromethane (4 mL) and anhydrous pyridine (0.12 mL, 1.55 mmol). To the cooled (0° C.) clear solution there was added dropwise methanesulfonyl chloride (0.024 mL, 0.31 mmol), maintaining the temperature less than 10° C. The yellow reaction mixture was allowed to stir at 0° C., with the color turning orange. The reaction mixture was allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the reaction mixture was diluted with dichloromethane (10 mL) and partitioned with water (15 mL). The organic portion was extracted with dichloromethane (3×5 mL). The organic portion was washed with saturated sodium chloride. The organic portion was filtered and concentrated under reduced pressure. Purification by chromatography gave 0.085 g (63%) of product as a clear oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.32-6.98 (m, 7H), 5.95 (m, 1H), 3.96 (m, 1H), 3.63-3.45 (m, 16H), 3.30 (s, 3H), 2.97 (m, 1H), 2.93 (s, 3H), 2.75-2.40 (m, 6H), 1.97 (m, 1H), 1.71 (m, 2H); MS (EI) for C.sub.29H.sub.41Cl.sub.2N.sub.3O.sub.7S: 646 (MH.sup.+).
(127) The compound was converted into the hydrochloride salt (17) by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt as an off-white powder.
(128) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 16
Preparation of 2-(3,4-dichlorophenyl)-N-[(1S)-1-(3-{[(2-methoxyethyl)sulfonyl]amino}phenyl)-2-(pyrrolidin-1-yl)ethyl]-N-methylacetamide, hydrochloride salt. (18)
(129) ##STR00114##
(130) N-[(1S)-1-(3-Aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-2-(3,4-dichloro-phenyl)-N-methylacetamide (0.10 g, 0.24 mmol) was dissolved in dichloromethane (2.5 mL) and anhydrous pyridine (0.15 mL). To the cooled (0° C.) yellow solution there was added dropwise 2-methoxyethanesulfonyl chloride (0.050 g, 0.31 mmol). The yellow reaction mixture was allowed to stir at 0° C., with the color turning orange. The reaction mixture was allowed to equilibrate to room temperature. The orange mixture was diluted with dichloromethane (15 mL) and partitioned with water (15 mL). The aqueous layer was extracted with dichloromethane (3×5 mL) and washed with saturated sodium chloride (2×15 mL). The organic portion was dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gave 0.072 g (55%) of 2-(3,4-dichlorophenyl)-N-[(1S)-1-(3-{[(2-methoxyethyl)sulfonyl]amino}phenyl)-2-(pyrrolidin-1-yl)ethyl]-N-methylacetamide as a light-yellow oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.37-7.14 (m, 7H), 6.05 (m, 1H), 3.81-3.71 (m, 4H), 3.41 (s, 3H), 3.21 (m, 2H), 3.14 (m, 1H), 2.80 (m, 3H), 2.69 (s, 3H), 2.49 (m, 3H), 1.75 (m, 4H); MS (EI) for C.sub.29H.sub.41Cl.sub.2N.sub.3O.sub.7S: 646 (MH.sup.+).
(131) The compound was converted into the hydrochloride salt (18) by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt as an off-white powder.
(132) The example described above may be modified to introduce oligomers of various lengths (at the methoxyethyl substituent) as disclosed herein.
Example 17
Preparation of 2-(3,4-dichlorophenyl)-N-methyl-N-{(1S)-2-(pyrrolidin-1-yl)-1-[3-({[2-(2,2,2-trifluoroethoxy)ethyl]sulfonyl}amino)phenyl]ethyl}acetamide, hydrochloride salt. (19)
(133) ##STR00115##
(134) N-[(1S)-1-(3-Aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-2-(3,4-dichloro-phenyl)-N-methylacetamide (0.048 g, 0.12 mmol) was dissolved in dichloromethane (2 mL) and anhydrous pyridine (0.072 mL, 0.88 mmol). To the cooled (0° C.) clear solution there was added dropwise 2-(2,2,2-trifluoroethoxy)ethanesulfonyl chloride (0.070 g, 0.29 mmol), maintaining the temperature less than 10° C. The yellow reaction mixture was allowed to stir at 0° C., with the color turning orange. The reaction mixture was allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the orange mixture was diluted with dichloromethane (15 mL) and partitioned with water (15 mL). The aqueous layer was extracted with dichloromethane (3×5 mL) and washed with saturated sodium chloride (2×15 mL). The organic portion was dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gave 0.039 g (55%) of 2-(3,4-dichlorophenyl)-N-methyl-N-{(1S)-2-(pyrrolidin-1-yl)-1-[3-({[2-(2,2,2-trifluoroethoxy)ethyl]sulfonyl}amino)phenyl]ethyl}acetamide as a clear oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.29-7.07 (m, 7H), 5.96 (m, 1H), 3.98 (m, 2H), 3.82 (m, 2H), 3.69 (m, 2H), 3.21 (m, 2H), 3.10 (m, 1H), 2.63 (m, 5H), 2.42 (m, 4H), 1.68 (m, 4H); MS (EI) for C.sub.25H.sub.30Cl.sub.2F.sub.3N.sub.3O.sub.4S: 596 (MH.sup.+).
(135) The compound was converted into the hydrochloride salt (19) by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt as an off-white powder.
(136) The example described above may be modified to introduce oligomers of various lengths (at the trifluoroethoxyethyl substituent) as disclosed herein.
Example 18
Preparation of 2-(3,4-dichlorophenyl)-N-[(1S)-1-[3-({[2-(2-methoxyethoxy)ethyl]sulfonyl}amino)phenyl]-2-(pyrrolidin-1-yl)ethyl]-N-methylacetamide, hydrochloride salt. (20)
(137) ##STR00116##
(138) N-[(1S)-1-(3-Aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-2-(3,4-dichloro-phenyl)-N-methylacetamide (0.052 g, 0.12 mmol) was dissolved in dichloromethane (2.5 mL) and anhydrous pyridine (0.078 mL, 0.96 mmol). To the cooled (0° C.) clear solution there was added dropwise 2-(2-methoxyethoxy)ethanesulfonyl chloride (0.082 g, 0.38 mmol), maintaining the temperature less than 10° C. The yellow reaction mixture was allowed to stir at 0° C., with the color turning orange. The reaction mixture was allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the orange mixture was diluted with dichloromethane (15 mL) and partitioned with water (15 mL). The aqueous layer was extracted with dichloromethane (3×5 mL) and washed with saturated sodium chloride (2×15 mL). The organic portion was dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gave 31 mg (42%) of 2-(3,4-dichlorophenyl)-N-[(1S)-1-[3-({[2-(2-methoxyethoxy)ethyl]sulfonyl}amino)phenyl]-2-(pyrrolidin-1-yl)ethyl]-N-methylacetamide as a clear oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.31-7.07 (m, 7H), 6.01 (m, 1H), 3.87 (m, 2H), 3.70-3.58 (m, 6H), 3.37 (s, 3H), 3.20-3.05 (m, 2H), 2.68 (m, 4H), 2.46 (m, 2H), 1.68 (m, 4H), 1.58 (m, 4H); MS (EI) for C.sub.26H.sub.35Cl.sub.2N.sub.3O.sub.5S: 572 (MH.sup.+).
(139) The compound was converted into the hydrochloride salt (20) by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt as an off-white powder.
(140) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 19
Preparation of N-[(1S)-1-{3-[({2-[2-(2-butoxyethoxy)ethoxy]ethyl}sulfonyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]-2-(3,4-dichlorophenyl)-N-methylacetamide, hydrochloride salt. (21)
(141) ##STR00117##
(142) N-[(1S)-1-(3-Aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-2-(3,4-dichloro-phenyl)-N-methylacetamide (0.048 g, 0.11 mmol) was dissolved in dichloromethane (2 mL) and anhydrous pyridine (0.072 mL, 0.88 mmol). To the cooled (0° C.) clear solution there was added dropwise 2-[2-(2-butoxyethoxy)ethoxy]ethanesulfonyl chloride (0.042 g, 0.17 mmol), maintaining the temperature less than 10° C. The yellow reaction mixture was allowed to stir at 0° C., with the color turning orange. The reaction mixture was allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the orange mixture was diluted with dichloromethane (15 mL) and partitioned with water (15 mL). The aqueous layer was extracted with dichloromethane (3×5 mL) and washed with saturated sodium chloride (2×15 mL). The organic portion was dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gave 0.026 mg (54%) of N-[(1S)-1-{3-[({2-[2-(2-butoxyethoxy)ethoxy]ethyl}sulfonyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]-2-(3,4-dichlorophenyl)-N-methylacetamide as a clear oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.32-7.07 (m, 7H), 6.00 (m, 1H), 3.87 (m, 2H), 3.84-3.62 (m, 10H), 3.50 (m, 2H), 3.28 (m, 2H), 3.27 (m, 2H), 2.92 (m, 2H), 2.76 (m, 4H), 2.43 (m, 2H), 1.67 (m, 4H), 1.33 (m, 2H), 1.18 (m, 2H), 0.73 (m, 3H); MS (EI) for C.sub.31H.sub.45Cl.sub.2N.sub.3O.sub.6S: 658 (MH.sup.+).
(143) The compound was converted into the hydrochloride salt (21) by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt as an off-white powder.
(144) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 20
Preparation of 2-(3,4-dichlorophenyl)-N-[(1S)-1-(3-{[(4-hydroxy-1,1-dioxidotetrahydrothiophen-3-yl)sulfonyl]amino}phenyl)-2-(pyrrolidin-1-yl)ethyl]-N-methylacetamide, hydrochloride salt. (20)
(145) ##STR00118##
(146) N-[(1S)-1-(3-Aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-2-(3,4-dichloro-phenyl)-N-methylacetamide (0.038 g, 0.0.094 mmol) was dissolved in dichloromethane (1.8 mL) and anhydrous pyridine (0.057 mL, 0.70 mmol). To the cooled (0° C.) clear solution there was added dropwise 4-hydroxytetrahydrothiophene-3-sulfonyl chloride 1,1-dioxide (0.069 g, 0.28 mmol), maintaining the temperature less than 10° C. The yellow reaction mixture was allowed to stir at 0° C., with the color turning orange. The reaction mixture was allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the orange mixture was diluted with dichloromethane (15 mL) and partitioned with water (15 mL). The aqueous layer was extracted with dichloromethane (3×5 mL) and washed with saturated sodium chloride (2×15 mL). The organic portion was dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gave 26 mg (46%) of 2-(3,4-dichlorophenyl)-N-[(1S)-1-(3-{[(4-hydroxy-1,1-dioxidotetrahydrothiophen-3-yl)sulfonyl]amino}phenyl)-2-(pyrrolidin-1-yl)ethyl]-N-methylacetamide as a clear oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.32-6.95 (m, 7H), 5.89 (m, 1H), 4.82 (m, 2H), 3.90-3.41 (m, 4H), 3.40-2.85 (m, 6H), 2.81-2.49 (m, 6H), 1.75 (m, 4H), 1.15 (m, 1H); MS (EI) for C.sub.25H.sub.31Cl.sub.2N.sub.3O.sub.6S.sub.2: 604 (MH.sup.+).
(147) The compound was converted into the hydrochloride salt (20) by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt as an off-white powder.
Example 21
Preparation of 2-(3,4-difluorophenyl)-N-methyl-N-[(1S)-1-phenyl-2-(pyrrolidin-1-yl)ethyl]acetamide, hydrochloride salt. (23)
(148) ##STR00119##
(149) (1S)—N-Methyl-1-phenyl-2-pyrrolidin-1-yl)ethanamine (0.060 g, 0.29 mmol), (Singh, V. et. al., Journal of Organic Chemistry (1996), vol. 61, pp. 6108-6113) was dissolved in anhydrous acetonitrile (1.5 mL). The clear solution was stirred at 0° C., under nitrogen, followed by the addition of diisopropylethylamine (0.11 mL, 0.64 mmol), (3,4-difluorophenyl)acetic acid (0.057 g, 0.32 mmol) and O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.078 g, 0.32 mmol). The clear reaction mixture was allowed to equilibrate to room temperature and stirred under nitrogen. After approximately 17 hours at room temperature the mixture was concentrated in vacuo. The residue was dissolved in ethyl acetate (2 mL) and was washed with saturated sodium bicarbonate (2×10 mL) and saturated sodium chloride (10 mL). The combined organic portion was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified on a column of silica gel using dichloromethane/methanol (9:1) as eluent to give 45 mg (43%) of 2-(3,4-difluorophenyl)-N-methyl-N-[(1S)-1-phenyl-2-(pyrrolidin-1-yl)ethyl]acetamide as a light-yellow oil. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.36-7.03 (m, 8H), 6.14 (m, 1H), 3.81 (m, 2H), 3.78 (m, 1H), 3.21 (m, 1H), 2.82-2.72 (m, 3H), 2.50 (m, 2H), 1.77 (m, 3H), 1.60 (m, 3H); MS (EI) for C.sub.21H.sub.24F.sub.2N.sub.2O: 359 (MH.sup.+).
(150) The compound was converted into the hydrochloride salt (23) by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt as a white powder.
Example 22
Preparation of ({(3S)-1-[(2S)-2-{[(3,4-dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]pyrrolidin-3-yl}oxy)acetic acid, hydrochloride salt. (24)
(151) ##STR00120##
(152) ({(3S)-1-[(2S)-2-{[(3,4-Dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]pyrrolidin-3-yl}oxy)acetic acid may be prepared according to the following steps.
Step 1: Preparation of (2-hydroxy-1-phenyl-ethyl)-methyl-carbamic acid tert-butyl ester
(153) ##STR00121##
(154) To a solution of lithium aluminum hydride (1.3 g) in tetrahydrofuran at 0° C. was added a solution of tert-butoxycarbonylamino-phenyl-acetic acid (2 g, 7.96 mmol) in tetrahydrofuran (40 mL) drop wise. The mixture was stirred at room temperature for 15 minutes, before heating in a 78° C. oil bath overnight. The mixture was then cooled in ice water bath, 6 mL 15% sodium hydroxide aqueous solution was added slowly. Tetrahydrofuran (25 mL) was added to dilute the mixture. After stirring at room temperature for thirty minutes, the mixture was filtered and washed with ethyl acetate. The filtrate was concentrated and taken into a mixture of dioxane/water (20 mL). 1 N sodium hydroxide was added (10 mL). Di-tert-butyl dicarbonate (1.7 g) was added. The mixture was stirred at room temperature for four hours. It was then concentrated and re-dissolved in ethyl acetate. After washing with water and brine, it was dried over sodium sulfate (anhydrous), filtered and concentrated to afford (2-hydroxy-1-phenyl-ethyl)-methyl-carbamic acid tert-butyl ester (1.4 g). The crude material was stored in freezer and used without further purification. LC-MS (ESI, MH.sup.+): 252.1.
Step 2: Preparation of {1-[2-(tert-butoxycarbonyl-methyl-amino)-2-phenyl-ethyl]-pyrrolidin-3-yloxy}-acetic acid ethyl ester
(155) ##STR00122##
(156) (2-Hydroxy-1-phenyl-ethyl)-methyl-carbamic acid tert-butyl ester (700 mg, 2.8 mmol) was dissolved in dichloromethane (8 mL) and cooled in ice water bath. Dess Martin reagent (Oakwood, 621 mg, 2.8 mmol) was added in small portions. The mixture was stirred at room temperature for ninety minutes. LC/MS analysis showed >90% conversion. Sodium bicarbonate (aq. Sat. 10 mL) was then added, followed by aqueous sodium thiosulfate solution (3 mL) and dichloromethane (15 mL). The mixture was stirred at room temperature for 15 minutes. Layers were separated and the dichloromethane layer was dried over anhydrous sodium sulfate, filtered and concentrated to give the crude aldehyde (700 mg). LC-MS (ESI, MH.sup.+): 250.2. This material was used immediately in the next step. Freshly prepared crude aldehyde (700 mg, 2.81 mmol) was dissolved in dichloromethane (25 mL) and stirred under nitrogen atmosphere. The reaction was cooled in ice bath and ethyl-(S)-pyrrolidine-3-yloxy-acetate hydrochloride salt (875 mg, 3.55 mmol (the preparation of which is described in Example 28) in dichloromethane (5 mL) was added, followed by diisopropylethylamine (0.62 mL) and sodium triacetoxyborohydride (1.1 g, 5.2 mmol). The mixture was stirred overnight at room temperature. Dichloromethane (60 mL) was added and the solution was washed with 10% sodium bicarbonate, brine, dried over sodium sulfate and concentrated to give 1.2 g of crude {1-[2-(tert-Butoxycarbonyl-methyl-amino)-2-phenyl-ethyl]-pyrrolidin-3-yloxy}-acetic acid ethyl ester. LC-MS (ESI, MH.sup.+): 407.5.
Step 3/4: Preparation of crude [1-(2-{[2-(3,4-dichloro-phenyl)-acetyl]-methyl-amino}-2-phenyl-ethyl)-pyrrolidin-3-yloxy]-acetic acid ethyl ester
(157) ##STR00123##
(158) Crude {1-[2-(tert-Butoxycarbonyl-methyl-amino)-2-phenyl-ethyl]-pyrrolidin-3-yloxy}-acetic acid ethyl ester (1.2 g) was dissolved in dichloromethane/trifluoroacetic acid (5 mL each). The mixture was stirred at room temperature for one hour. LCMS show the reaction was completed. The mixture was then concentrated. The crude amine was dissolved in dimethylformamide (10 mL) under nitrogen atmosphere. 3,4-Dichlorophenyl acetic acid (590 mg, 2.88 mmol) was added, followed by diisopropylethylamine (2.8 mL, 16.0 mmol). The reaction mixture was cooled in an ice bath. HATU (1.0 g, 2.88 mmol) was added in one portion. The reaction was continued at room temperature for two hours. LCMS showed major product (M+H: 493.3). Ice water (20.0 mL) was added and the reaction mixture was extracted with ether (30.0 mL×2). The combined organic layer was washed with brine, dried over sodium sulfate and concentrated to give 1.0 g crude [1-(2-{[2-(3,4-Dichloro-phenyl)-acetyl]-methyl-amino}-2-phenyl-ethyl)-pyrrolidin-3-yloxy]-acetic acid ethyl ester.
Step 5: Preparation of [1-(2-{[2-(3,4-dichloro-phenyl)-acetyl]-methyl-amino}-2-phenyl-ethyl)-pyrrolidin-3-yloxy]-acetic acid as hydrochloride salt
(159) ##STR00124##
(160) Crude [1-(2-{[2-(3,4-Dichloro-phenyl)-acetyl]-methyl-amino}-2-phenyl-ethyl)-pyrrolidin-3-yloxy]-acetic acid ethyl ester (1 g, 2.03 mmol) was dissolved in 20 mL tetrahydrofuran. 3.5 mL of 2N lithium hydroxide was added. The mixture was stirred at room temperature for one hour. Aqueous hydrochloric acid (5 mL, 2%) was then added to adjust the pH to 3. Ethyl acetate (30 mL) was added and the mixture was washed with brine, dried over sodium sulfate and concentrated. The crude residue was purified by reverse phase high pressure liquid chromatography. The crude residue was purified by reverse phase high pressure liquid chromatography. The residue was diluted with 0.1 N hydrochloride and lyophilized to give 200 mg [1-(2-{[2-(3,4-Dichloro-phenyl)-acetyl]-methyl-amino}-2-phenyl-ethyl)-pyrrolidin-3-yloxy]-acetic acid as hydrochloride salt. .sup.1H NMR (500 MHz, DMSO-d6): δ 10.85 (bs, 1H), 7.62-7.26 (m, 8H), 6.17 (m, 1H), 4.35 (m, 2H), 4.15-4.01 (m, 3H), 3.89-3.25 (m, 5H), 3.17 (m, 2H), 2.78 (m, 2H), 2.40-1.95 (2H); MS (EI) for C.sub.23H.sub.26Cl.sub.2N.sub.2O.sub.4: 465.4 (MH.sup.+).
Example 23
2-(3,4-Dichlorophenyl)-N-{(1S)-2-[(3S)-3-(2-hydroxyethoxy)pyrrolidin-1-yl]-1-phenylethyl}-N-methylacetamide, hydrochloride salt. (25)
(161) ##STR00125##
(162) Using a synthetic approach similar to the one employed in Example 22, 2-(3,4-dichlorophenyl)-N-{(1S)-2-[(3S)-3-(2-hydroxyethoxy)pyrrolidin-1-yl]-1-phenylethyl}-N-methylacetamide, hydrochloride salt (25) was prepared. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.50-7.17 (m, 8H), 6.45 (m, 1H), 4.14-3.98 (m, 5H), 3.75-3.52 (m, 4H), 3.34-3.15 (m, 4H), 2.87 (m, 3H), 2.40 (m, 1H), 2.25 (m, 2H); MS (EI) for C.sub.23H.sub.28Cl.sub.2N.sub.2O.sub.3: 451.3 (MH.sup.+).
(163) The example described above may be modified to introduce oligomers of various lengths (at the 2-hydroxyethoxy substituent) as disclosed herein.
Example 24
2-(3,4-Dichlorophenyl)-N-[(1S)-2-{(3S)-3-[2-(2-hydroxyethoxy)ethoxy]pyrrolidin-1-yl}-1-phenylethyl]-N-methylacetamide, hydrochloride salt. (26)
(164) ##STR00126##
(165) Using a synthetic approach similar to the one employed in Example 22, 2-(3,4-dichlorophenyl)-N-[(1S)-2-{(3S)-3-[2-(2-hydroxyethoxy)ethoxy]pyrrolidin-1-yl}-1-phenylethyl]-N-methylacetamide, hydrochloride salt (26) was prepared. .sup.1H NMR (500 MHz, DMSO-d6): δ 7.57-7.24 (m, 8H), 6.13 (m, 1H), 4.30 (m, 2H), 4.12 (m, 2H), 3.90-3.35 (m, 12H), 2.77 (m, 3H), 2.31 (m, 2H), 2.06 (m, 2H); MS (EI) for C.sub.25H.sub.32Cl.sub.2N.sub.2O.sub.4: 495.4 (MH.sup.+).
(166) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 25
2-(3,4-Dichlorophenyl)-N-[(1S)-2-[(3S)-3-{2-[2-(2-hydroxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-phenylethyl]-N-methylacetamide, hydrochloride salt. (27)
(167) ##STR00127##
(168) Using a synthetic approach similar to the one employed in Example 22, 2-(3,4-dichlorophenyl)-N-{(1S)-2-[(3S)-3-{2-[2-(2-hydroxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-1-phenylethyl}-N-methylacetamide, hydrochloride salt (27) was prepared. .sup.1H NMR (500 MHz, DMSO-d6): δ 7.60-7.23 (m, 8H), 6.14 (m, 1H), 4.33-4.01 (m, 4H), 3.83 (m, 1H), 3.51-3.36 (m, 15H), 2.77 (m, 3H), 2.34 (m, 1H), 2.16 (m, 2H), 2.10 (m, 1H), 1.95 (m, 1H); MS (EI) for C.sub.27H.sub.36Cl.sub.2N.sub.2O.sub.5: 538.2 (MH.sup.+).
(169) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 26
[2-({(3S)-1-[(2S)-2-{[(3,4-Dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]pyrrolidin-3-yl}oxy)ethoxy]acetic acid, hydrochloride salt. (28)
(170) ##STR00128##
(171) Using a synthetic approach similar to the one employed in Example 22, [2-({(3S)-1-[(2S)-2-{[(3,4-dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]pyrrolidin-3-yl}oxy)ethoxy]acetic acid, hydrochloride salt (28) was prepared. .sup.1H NMR (500 MHz, DMSO-d6): δ 7.61-7.22 (m, 8H), 6.15 (m, 1H), 4.35-4.23 (m, 2H), 4.10-3.84 (m, 2H), 3.65-3.27 (m, 10H), 2.77 (m, 3H), 2.35 (m, 1H), 2.20-1.90 (m, 3H); MS (EI) for C.sub.25H.sub.30Cl.sub.2N.sub.2O.sub.5: 509.5 (MH.sup.+).
(172) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 27
{2-[2-({(3S)-1-[(2S)-2-{[(3,4-Dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]pyrrolidin-3-yl}oxy)ethoxy]ethoxy}acetic acid, hydrochloride salt. (29)
(173) ##STR00129##
(174) Using a synthetic approach similar to the one employed in Example 22, {2-[2-({(3S)-1-[(2S)-2-{[(3,4-dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]pyrrolidin-3-yl}oxy)ethoxy]ethoxy}acetic acid, hydrochloride salt (29) was prepared. .sup.1H NMR (500 MHz, DMSO-d6): δ 7.62-7.23 (m, 8H), 6.15 (m, 1H), 4.33-4.25 (m, 2H), 4.13-3.85 (m, 2H), 3.98-3.29 (m, 15H), 2.75 (m, 3H), 2.37 (m, 1H), 2.15 (m, 1H), 1.97 (m, 1H); MS (EI) for C.sub.27H.sub.34Cl.sub.2N.sub.2O.sub.6: 553.3 (MH.sup.+).
(175) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 28
Preparation of Pyrrolidine-Based Compounds Potentially Useful in Synthetic Approaches
Preparation of (S)-3-methoxycarbonylmethoxy-pyrrolidine-1-carboxylic acid benzyl ester
(176) ##STR00130##
(S)-3-Methoxycarbonylmethoxy-pyrrolidine-1-carboxylic acid benzyl ester
(177) To a stirred solution of cbz-(S)-pyrrolidin-3-ol (5.4 g, 24 mmol, 1 eq) in anhydrous tetrahydrofuran (100 mL) at 0° C., sodium hydride (60% in oil, 2.4 g, 60 mmol, 2.5 eq) was added and the solution was stirred at room temperature for thirty minutes. Methyl bromoacetate (7.3 g, 4.4 mL, 48 mmol, 2 eq) was added to the solution and then further stirred at room temperature for three hours, by which time LC-MS analysis revealed the formation of a new product. Saturated aqueous ammonium chloride solution (10 mL) was added to the mixture and the mixture was extracted with ether (3×30 mL). The combined organic layers were washed with saturated sodium bicarbonate solution (30 mL), brine (30 mL), dried over magnesium sulfate and concentrated in vacuo. The residue was purified by column chromatography on silica gel using a gradient of solvents, hexane: ethyl acetate from 5-50% ethyl acetate to give the desired product (6.6 g, 94.3%) as light brown oil. LC-MS (ESI, MH.sup.+): 294.1. .sup.1H NMR (250 MHz, CDCl.sub.3) δ 7.6-7.1 (5H, m), 5.15 (2H, s), 4.25-4.05 (3H, m), 3.85-3.4 (7H, m), 2.2-1.7 (4H, m).
Preparation of (S)-3-(2-methoxycarbonylmethoxy-ethoxy)-pyrrolidine-1-carboxylic acid benzyl ester
(178) ##STR00131##
(S)-3-(2-Methoxycarbonylmethoxy-ethoxy)-pyrrolidine-1-carboxylic acid benzyl ester
(179) To a stirred solution of (S)-3-(2-hydroxy-ethoxy)-pyrrolidine-1-carbooxylic acid benzyl ester (9 g, 34 mmol, 1 eq) in anhydrous tetrahydrofuran (100 mL) at 0° C., sodium hydride (60% in oil, 2.0 g, 51 mmol, 1.5 eq) was added and the solution was stirred at room temperature for 30 minutes. Methyl bromoacetate (6.3 g, 3.8 mL, 41 mmol, 1.2 eq) was added to the solution and then further stirred at room temperature for three hours, by which time LC-MS analysis indicated that no starting material left. Saturated aqueous ammonium chloride solution (60 mL) was added to the mixture and the mixture was extracted with ether (3×60 mL). The combined organic layers were washed with saturated sodium bicarbonate solution (50 mL), brine (50 mL), dried over magnesium sulfate and concentrated in vacuo. The residue was purified by column chromatography on silica gel using a gradient of solvents, hexane: ethyl acetate from 10-60% ethyl acetate to give(S)-3-(2-Methoxycarbonylmethoxy-ethoxy)-pyrrolidine-1-carboxylic acid benzyl ester as light brown oil (9 g, 78.3%). LC-MS: 338.2 (M.sup.++1), 675.3 (2 M.sup.++1), 692.5 (2M.sup.++hydrogen O).
Preparation of (S)-3-[2-(2-ethoxycarbonylmethoxy-ethoxy)-ethoxy]-pyrrolidine-1-carboxylic acid benzyl ester
(180) ##STR00132##
(S)-3-[2-(2-Ethoxycarbonylmethoxy-ethoxy)-ethoxy]-pyrrolidine-1-carboxylic acid benzyl ester
(181) To a stirred solution of (S)-3-[2-(2-hydroxy-ethoxy)-ethoxy]-pyrrolidine-1-carboxylic acid benzyl ester (2.4 g, 7.7 mmol, 1 eq) in anhydrous tetrahydrofuran (30 mL) at 0° C., sodium hydride (60% in oil, 468 mg, 11.7 mmol, 1.5 eq) was added and the solution was stirred at room temperature for thirty minutes. Ethyl bromoacetate (1.54 g, 1.02 mL, 9.2 mmol, 1.2 eq) was added to the solution and then further stirred at room temperature for three hours, by which time LC-MS analysis indicated that no starting material left. Saturated aqueous ammonium chloride solution (60 mL) was added to the mixture and the mixture was extracted with ether (3×60 mL). The combined organic layers were washed with saturated sodium bicarbonate solution (50 mL), brine (50 mL), dried over magnesium sulfate and concentrated in vacuo. The residue was purified by column chromatography on silica gel using a gradient of solvents, hexane: ethyl acetate from 10-60% ethyl acetate to give the desired product (S)-3-[2-(2-Ethoxycarbonylmethoxy-ethoxy)-ethoxy]-pyrrolidine-1-carboxylic acid benzyl ester (2.4 g, 80%) as light brown oil. LC-MS: 396.3 (M.sup.++1), 413.4 (M.sup.++hydrogenO).
Preparation of (S)-3-{2-[2-(2-benzyloxy-ethoxy)-ethoxy]-ethoxyl}-pyrrolidine-1-carboxylic acid benzyl ester
(182) ##STR00133##
(S)-3-{2-[2-(2-Benzyloxy-ethoxy)-ethoxy]-ethoxyl}-pyrrolidine-1-carboxylic acid benzyl ester
(183) To a solution of (S)-3-hydroxy-pyrrolidine-1-carboxylic acid benzyl ester (3.5 g, 15 mmol, 1.2 eq) in 60 mL of anhydrous tetrahydrofuran, NaH (60% in oil, 800 mg, 20 mmol) was added and the solution was stirred at room temperature for thirty minutes. Toluene-4-sulfonic acid 2-[2-benzyolxy-ethoxy)-ethoxy]ethyl ester (5 g, 13 mmol 1 eq) was then added. The mixture was heated at 50° C. overnight. LC-MS analysis revealed the formation of a new product. Saturated aqueous ammonium chloride solution (10 mL) was added to the mixture and the aqueous layer was extracted with ether (4×15 mL). The combined organic layers were washed with saturated sodium bicarbonate solution (4×10 mL), brine (4×10 mL), dried over magnesium sulfate and concentrated in vacuo. The residue was purified by column chromatography on silica gel using a gradient of hexane/ethyl acetate (10-60% ethyl acetate) to give (S)-3-{2-[2-(2-Benzyloxy-ethoxy)-ethoxy]-ethoxyl}-pyrrolidine-1-carboxylic acid benzyl ester (4 g, 69.6%) as light brown oil.
Preparation of (S)-3-[2-(2-hydroxy-ethoxy)-ethoxy]-pyrrolidine-1-carboxylic acid benzyl ester
(184) ##STR00134##
S)-3-[2-(2-Hydroxy-ethoxy)-ethoxy]-pyrrolidine-1-carboxylic acid benzyl ester
(185) To a stirred solution of (S)-3-(2-Methoxycarbonylmethoxy-ethoxy)-pyrrolidine-1-carboxylic acid benzyl ester (3.1 g, 9.2 mmol, 1 eq) in anhydrous tetrahydrofuran (100 mL) at 0° C., lithium aluminum hydride (240 mg, 11 mmol, 1.2 eq) was added and allowed to warm to room temperature and stirred overnight. The reaction was quenched by the portion wise addition of sodium sulfate decahydrate (20 g) and stirred for several hours. The heterogeneous mixture was filtered and washing the solid with tetrahydrofuran and the filtrate was concentrated under reduced pressure to give compound (S)-3-[2-(2-hydroxy-ethoxy)-ethoxy]-pyrrolidine-1-carboxylic acid benzyl ester (2.4 g, 88.9%) as oil which was used subsequently in next step. LC-MS: 310.3 (M.sup.++1), 619.6 (2 M+1), 636.7 (2M.sup.++hydrogenO).
Preparation of 2-{2-[2-((S)-pyrrolidin-3-yloxy)-ethoxy]-ethoxyl}-ethanol
(186) ##STR00135##
2-{2-[2-((S)-Pyrrolidin-3-yloxy)-ethoxy]-ethoxyl}-ethanol
(187) (S)-3-{2-[2-(2-Benzyloxy-ethoxy)-ethoxy]-ethoxy}-pyrrolidine-1-carboxylic acid benzyl ester (2.4 g) was dissolved in 40 mL of ethanol in a hydrogenation flask, 0.8 g of Pd/C (10%) was added. The mixture was hydrogenated on a Parr shaker (50 psi) for 24 hours. The mixture was purged with nitrogen and filtered through a cclitc pad washed with more ethanol. The filtrate was concentrated and concentrated. The crude product was purified by reverse phase prep HPLC to give pure product 650 mg (48%). LC-MS: 220.0 (M.sup.++1), 439.5 (2M.sup.++1).
Example 29
Preparation of N-[(1S)-1-{3-[({2-[2-(2-Butoxyethoxy)ethoxy]ethyl}sulfonyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]-N-methyl-2-(pyridin-2-yl)acetamide, dihydrochloride salt. (30)
(188) ##STR00136##
(189) N-[(1S)-1-{3-[({2-[2-(2-Butoxyethoxy)ethoxy]ethyl}sulfonyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]-N-methyl-2-(pyridin-2-yl)acetamide may be prepared according to the following steps.
Step 1: Preparation of N-Methyl-N-[(1S)-1-(3-nitrophenyl)-2-(pyrrolidin-1-yl)ethyl]-2-(pyridin-2-yl)acetamide
(190) ##STR00137##
(191) (S)-1-[2-(Methylamino)-2-(3/4-nitrophenyl)ethyl]pyrroldin (0.25 g, 1.00 mmol), (Portoghese, P. Journal of Medicinal Chemistry, 1994, 37, 4490-4498) was dissolved in anhydrous acetonitrile (5 mL). To the yellow solution was added diisopropylethylamine (0.55 mL, 3.11 mmol) at 0° C., followed by pyridin-2-ylacetic acid (0.19 g, 1.10 mmol) and O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.26 g, 1.10 mmol). The reaction mixture was allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the reaction mixture was concentrated under reduced pressure. The residue was taken up in ethyl acetate (15 mL) and washed with saturated sodium bicarbonate (2×25 mL) and saturated sodium chloride (25 mL). The combined organic portion was dried over anhydrous sodium sulfate (approximately 1.70 g) and concentrated in vacuo. The residue was purified on a column of silica gel using dichloromethane/methanol (9:1) as eluent to give 0.18 g (49%) of N-methyl-N-[(1S)-1-(3-nitrophenyl)-2-(pyrrolidin-1-yl)ethyl]-2-(pyridin-2-yl)acetamide as a clear oil. MS (EI) for C.sub.20H.sub.24N.sub.4O.sub.3: 369 (MH.sup.+).
Step 2: Preparation of N-[(1S)-1-(3-aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-N-methyl-2-(pyridin-2-yl)acetamide
(192) ##STR00138##
(193) A mixture of N-methyl-N-[(1S)-1-(3-nitrophenyl)-2-(pyrrolidin-1-yl)ethyl]-2-(pyridin-2-yl)acetamide (0.13 g, 0.35 mmol), hydrazine hydrate (0.27 g, 4.23 mmol) and Raney Ni slurry (0.57 mL) in 95% ethanol (17.5 mL) was heated to 55° C. After approximately two hours at 55° C. the reaction was complete as indicated by TLC. The reaction mixture was filtered through Celite, and the Raney Ni was washed with hot methanol. The filtrate was concentrated under reduced pressure to give a brown residue. Purification by chromatography gave 0.11 g (92%) of N-[(1S)-1-(3-aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-N-methyl-2-(pyridin-2-yl)acetamide as a light-yellow oil. MS (EI) for C.sub.20H.sub.26N.sub.4O: 339 (MH.sup.+).
Step 3: Preparation of N-[(1S)-1-{3-[({2-[2-(2-butoxyethoxy)ethoxy]ethyl}sulfonyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]-N-methyl-2-(pyridin-2-yl)acetamide, dihydrochloride salt (30)
(194) ##STR00139##
(195) N-[(1S)-1-(3-Aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-N-methyl-2-(pyridin-2-yl)acetamide (0.11 g, 0.32 mmol) was dissolved in dichloromethane (6.5 mL) and anhydrous pyridine (0.19 mL, 2.43 mmol). To the cooled (0° C.) clear solution there was added dropwise 2-[2-(2-butoxyethoxy)ethoxy]ethanesulfonyl chloride (0.24 g, 0.81 mmol), maintaining the temperature less than 10° C. The yellow reaction mixture was allowed to stir at 0° C., with the color turning orange. After approximately 17 hours at room temperature the orange mixture was diluted with dichloromethane (15 mL) and partitioned with water (15 mL). The aqueous layer was extracted with dichloromethane (3×5 mL) and washed with saturated sodium chloride (2×15 mL). The organic portion was dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gave 0.11 g (57%) of N-[(1S)-1-{3-[({2-[2-(2-butoxyethoxy)ethoxy]ethyl}sulfonyl)amino]phenyl}-2-(pyrrolidin-1-yl)ethyl]-N-methyl-2-(pyridin-2-yl)acetamide as a clear oil.
(196) The compound was converted into the dihydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution was lyophilized to give the hydrochloride salt as an off-white powder.
(197) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 30
Preparation of N-[(1S)-2-Amino-1-phenylethyl]-2-(3,4-dichlorophenyl)-N-methylacetamide, hydrochloride salt. (31)
(198) ##STR00140##
(199) N-[(1S)-2-Amino-1-phenylethyl]-2-(3,4-dichlorophenyl)-N-methylacetamide may be prepared according to the following steps.
Step 1: Preparation of benzyl[(1S)-2-amino-2-oxo-1-phenylethyl]carbamate
(200) ##STR00141##
(201) A solution of (2S)-{[(benzyloxy)carbonyl]amino}(phenyl)acetic acid (2 g, 7.01 mmol), N-methylmorpholine (780 mg, 7.71 mmol) in tetrahydrofuran (32 mL) was cooled to −10° C. Ethyl chloroformate (837 mg, 7.71 mmol) was added and the solution was stirred for one hour at room temperature. The reaction mixture was cooled to −10° C. and aqueous ammonium hydroxide (28%) (524 μl, 7.71 mmol) was added. The mixture was stirred for three hours at −10° C. The solvent was removed under reduced pressure and the residue was dissolved in ethyl acetate. The organic portion, after being washed with saturated sodium bicarbonate solution, water and saturated sodium chloride, was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was crystallized from ethyl acetate-hexane to give 1.7 g (85%) of benzyl[(1S)-2-amino-2-oxo-1-phenylethyl]carbamate as a white solid.
Step 2: Preparation of (1S)—N1-methyl-1-phenylethane-1,2-diamine
(202) ##STR00142##
(203) Benzyl[(1S)-2-amino-2-oxo-1-phenylethyl]carbamate (1.7 g, 5.97 mmol) was dissolved in tetrahydrofuran (34 mL). The solution was cooled to 0° C. and lithium aluminum hydride (1.13 g, 29.89 mmol) was added. The mixture was stirred for 15 minutes at 0° C., followed by 16 hours stirring at 65° C. The reaction mixture was cooled to 0° C. and 3N aqueous sodium carbonate (100 mL) was added carefully until effervescence ceased. The precipitated solid was filtered and washed with ethyl acetate (100 mL). The filtrate was concentrated and the residue was dissolved in ethyl acetate (150 mL). The product was extracted into 1N aqueous hydrochloric acid (2×10 mL) and washed with tert-butyl methyl ether (3×15 mL). The pH of the aqueous portion was adjusted to 9, and the product was extracted into ethyl acetate. The organic portion was washed with saturated sodium chloride, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by chromatography to give 265 mg (30%) of (1S)—N.sup.1-methyl-1-phenylethane-1,2-diamine.
Step 3: Preparation of tert-butyl[(2S)-2-(methylamino)-2-phenylethyl]carbamate
(204) ##STR00143##
(205) To a solution of (1S)—N.sup.1-methyl-1-phenylethane-1,2-diamine (258 mg, 1.717 mmol) in dichloromethane (4 mL) at room temperature, was added a solution of di-tert-butyl dicarbonate (337 mg, 1.54 mmol) in dichloromethane (1 mL). The reaction mixture was stirred for two hours and concentrated under reduced pressure. The residue was purified by chromatography to give 260 mg (61%) of tert-butyl[(2S)-2-(methylamino)-2-phenylethyl]carbamate.
Step 4: Preparation of tert-butyl[(2S)-2-{[(3,4-dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]carbamate
(206) ##STR00144##
(207) To a solution of tert-butyl[(2S)-2-(methylamino)-2-phenylethyl]carbamate (254 mg, 1.01 mmol) in acetonitrile (4 mL) was added 3,4-dichlorophenyl acetic acid (229 mg, 1.11 mmol), 1-hydroxybenzotriazole hydrate (164 mg, 1.21 mmol), diisopropylethyl amine (265 μl, 1.52 mmol) and N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (292 mg, 1.522 mmol) at room temperature. The mixture was stirred for two hours and the solvent was removed under reduced pressure. The residue was dissolved in dichloromethane (15 mL), and washed with 10% aqueous sodium carbonate (4×15 mL), 10% aqueous ammonium chloride (4×15 mL), and saturated sodium chloride (15 mL). The organic portion was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by chromatography to give 280 mg (63%) of tert-butyl[(2S)-2-{[(3,4-dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]carbamate.
Step 5: Preparation of N-[(1S)-2-amino-1-phenylethyl]-2-(3,4-dichlorophenyl)-N-methylacetamide, hydrochloride salt (31)
(208) ##STR00145##
(209) To a solution of tert-butyl[(2S)-2-{[(3,4-dichlorophenyl)acetyl](methyl)amino}-2-phenylethyl]carbamate (256 mg, 0.585 mmol) in isopropyl alcohol (2 mL) was added 4M hydrochloric acid in isopropyl alcohol (2 mL). The reaction mixture was stirred for two hours at room temperature and was concentrated under reduced pressure to give 195 mg (99%) of N-[(1S)-2-amino-1-phenylethyl]-2-(3,4-dichlorophenyl)-N-methylacetamide hydrochloride as a white solid. .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 7.90 (bs, 3H), 7.2-7.6 (m, 8H), 5.95 (dd, 1H), 3.8 (quartet, 2H), 3.55 (dd, 2H), 2.75 (s, 3H); MS (EI): 337.09 (free base).
Example 31
Preparation of 2-(3,4-Dichlorophenyl)-N-{(1S)-2-[(3R,4R)-3,4-dihydroxypyrrolidin-1-yl]-1-phenylethyl}-N-methylacetamide, hydrochloride salt (32)
(210) ##STR00146##
(211) 2-(3,4-Dichlorophenyl)-N-{(1S)-2-[(3R,4R)-3,4-dihydroxypyrrolidin-1-yl]-1-phenylethyl}-N-methylacetamide hydrochloride salt was prepared according to the following steps.
Step 1: Preparation of benzyl {(1S)-2-[(3R,4R)-3,4-dihydroxypyrrolidin-1-yl]-1-phenylethyl}carbamate
(212) ##STR00147##
(213) (S)-2-(((Benzyloxy)carbonyl)amino)-2-phenylacetic acid (250 mg, 0.87 mmol), (3R,4R)-pyrrolidine-3,4-diol (90 mg, 0.87 mmol), and N,N-diisopropylethylamine (0.30 mL, 1.75 mmol) were dissolved in 18 mL of tetrahydrofuran. The mixture was stirred for ten minutes at room temperature and then O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU) (448 mg, 1.4 mmol) was added into the solution. The reaction mixture was stirred for three hours and then concentrated under reduced pressure. The resulting residue was dissolved in methylene chloride (50 mL) and washed with brine (25 mL×2). The organic phase was separated and dried over anhydrous sodium sulfate and concentrated. The obtained residue was then purified by flash chromatography to yield benzyl {(1S)-2-[(3R,4R)-3,4-dihydroxypyrrolidin-1-yl]-1-phenylethyl}carbamate (250 mg, 0.68 mmol, yield 77%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.36 (m, 2H), 7.34 (m, 8H), 6.24-6.22 (d, 1H), 5.39 (m, 1H), 5.10 (d, 1H), 5.03 (d, 1H), 4.20 (m, 1H), 4.10 (m, 1H), 3.89 (m, 1H), 3.67 (m, 3H), 3.11 (m. 2H); MS (EI) for C.sub.20H.sub.24N.sub.2O.sub.4: 371 (MH.sup.+).
Preparation of (3R,4R)-1-[(2S)-2-(Methylamino)-2-phenylethyl]pyrrolidine-3,4-diol
(214) ##STR00148##
(215) Step 2: Benzyl {(1S)-2-[(3R,4R)-3,4-dihydroxypyrrolidin-1-yl]-1-phenylethyl}carbamate (250 mg, 0.68 mmol) was dissolved in tetrahydrofuran (5 mL). The mixture was cooled to 0° C. Lithium aluminum hydride solution in tetrahydrofuran (1M, 2.7 mL, 2.7 mmol) was added to the above mixture and the solution was stirred for 15 minutes at 0° C. and then heated to 65° C. and maintained at that temperature for 14 hours. Sodium sulfate decahydrate was added cautiously until effervescence ceased. The solid was removed by filtration and was washed with methylene chloride (10 mL). The solution was dried over anhydrous sodium sulfate and concentrated under vacuum to obtain the crude product which was used for next step without further purification.
Preparation of 2-(3,4-Dichlorophenyl)-N-{(1S)-2-[(3R,4R)-3,4-dihydroxypyrrolidin-1-yl]-1-phenylethyl}-N-methylacetamide hydrochloride (32)
(216) ##STR00149##
(217) (3R,4R)-1-[(2S)-2-(Methylamino)-2-phenylethyl]pyrrolidine-3,4-diol (150 mg, 0.64 mmol), 2-(3,4-dichlorophenyl)acetic acid (131 mg, 0.64 mmol), and N,N-diisopropylethylamine (0.23 ml, 1.28 mmol) were dissolved in methylene chloride (5 mL). The resulting mixture was stirred for 15 minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU) (227 mg, 0.89 mmol) was added into the solution. The reaction mixture was stirred for 18 hours at room temperature. The solution was added in methylene chloride (50 mL) and washed with water and dried over sodium sulfate. Evaporation of solvent and purification of the resulting residue by flash chromatography gave product as free base, which was then dissolved in 1M hydrochloride in acetonitrile to afford product as hydrochloride salt 2-(3,4-Dichlorophenyl)-N-{(1S)-2-[(3R,4R)-3,4-dihydroxypyrrolidin-1-yl]-1-phenylethyl}-N-methylacetamide hydrochloride (32) (40 mg, 0.094 mmol, 14% yield for two steps). .sup.1H NMR (500 MHz, MeOD): δ 7.43 (m, 2H), 7.40 (m, 3H), 7.30 (m, 2H), 7.25 (m, 1H), 6.31 (m, 1H), 4.28 (d, 2H), 4.17 (m, 1H), 3.90-3.74 (m, 4H), 3.74 (m, 1H), 3.61 (m, 1H), 3.48 (d, 1H), 2.81 (s, 3H); MS (EI) for C.sub.21H.sub.24Cl.sub.2N.sub.2O.sub.3: 423 (MH.sup.+).
Example 32
Preparation of 2-(3,4-Dichlorophenyl)-N-{(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-[4-(trifluoromethyl)phenyl]ethyl}-N-methylacetamide, hydrochloride salt (33)
(218) ##STR00150##
(219) 2-(3,4-Dichlorophenyl)-N-{(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-[4-(trifluoromethyl)phenyl]ethyl}-N-methylacetamide hydrochloride was prepared according to the following steps.
Step 1: Preparation of (2S)-[(tert-butoxycarbonyl)amino][4-(trifluoromethyl)phenyl]acetic acid
(220) ##STR00151##
(221) 4-(Trifluoromethyl)-L-phenylglycine (250 mg, 1.14 mmol) and sodium bicarbonate (168 mg, 2 mmol) were added in a mixture of 10 mL of dioxane and 10 mL water. Boc anhydride (300 mg, 1.37 mg) was added into the solution. The mixture was stirred at room temperature for 18 hours. 2N HCl solution was added to adjust to pH 5. The resulting solution was added to methylene chloride (50 mL) and washed with brine (25 mL×2). The organic phase was separated and dried over anhydrous sodium sulfate and concentrated. The obtained residue was then purified by flash chromatography to yield (2S)-[(tert-butoxycarbonyl)amino][4-(trifluoromethyl)phenyl]acetic acid (340 mg, 1.07 mmol, 93%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 8.25 (s, 1H), 7.64 (d, 2H), 7.58 (d, 2H), 5.22 (d, 1H), 3.73 (s, 1H), 1.24 (s, 9H).
Step 2: Preparation of tert-butyl {(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-2-oxo-1-[4-(trifluoromethyl)phenyl]ethyl}carbamate
(222) ##STR00152##
(223) (2S)-[(tert-Butoxycarbonyl)amino][4-(trifluoromethyl)phenyl]acetic acid (293 mg, 1.02 mmol), (S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidine (210 mg, 0.90 mmol), and N,N-diisopropylethylamine (0.31 mL, 1.80 mmol) were dissolved in 18 mL of tetrahydrofuran. The mixture was stirred for 10 minutes at room temperature and then O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU) (376 mg, 1.17 mmol) was added into the solution and the solution was stirred for three hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was dissolved in methylene chloride (50 mL) and washed with brine (25 mL×2). The solution was dried over anhydrous sodium sulfate and concentrated. The obtained residue was then purified by flash chromatography to yield tert-butyl {(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-2-oxo-1-[4-(trifluoromethyl)phenyl]ethyl}carbamate (380 mg, 0.71 mmol, yield 79%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.62 (m, 2H), 7.55 (m, 2H), 6.12 (dd, 1H), 5.40 (t, 1H), 4.07 (m, 1H), 3.77 (m, 1H), 3.70-3.55 (m, 12H), 3.44 (m, 2H), 3.38 (s, 3H), 2.17-1.76 (m, 2H), 1.41 (d, 9H); MS (EI) for C.sub.25H.sub.37F.sub.3N.sub.2O.sub.7: 535 (MH.sup.+).
Step 3: Preparation of 2-(3,4-dichlorophenyl)-N-{(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-[4-(trifluoromethyl)phenyl]ethyl}-N-methylacetamide hydrochloride salt (33)
(224) ##STR00153##
(225) A solution of tert-butyl {(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-2-oxo-1-[4-(trifluoromethyl)phenyl]ethyl}carbamate (380 mg, 0.71 mmol) in tetrahydrofuran (10 mL) was added dropwise to a stirred 1.0 M solution of lithium aluminum hydride (2.84 mL, 2.84 mmol) at room temperature. The mixture was stirred for thirty minutes at room temperature and then was heated to 65° C. for 14 hours. After cooling down to room temperature, sodium sulfate decahydrate was added cautiously until effervescence ceased. The solid was removed by filtration and was washed with methylene chloride (20 mL). The solution was dried over anhydrous sodium sulfate and concentrated under vacuum to obtain the crude product, which was used for next step without further purification. The above obtained crude, 2-(3,4-dichlorophenyl)acetic acid (0.127 g, 0.62 mmol), and N,N-diisopropylethylamine (0.22 g, 1.24 mmol) were dissolved in 3 mL of acetonitrile. The mixture was stirred for 10 minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU) (0.222 g, 0.87 mmol) was added into the solution. The reaction mixture was stirred for four hours at room temperature and was concentrated. The residue was dissolved in methylene chloride (20 mL), was washed with brine (20 mL). The organic phase was separated and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded 2-(3,4-Dichlorophenyl)-N-{(1S)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-1-[4-(trifluoromethyl)phenyl]ethyl}-N-methylacetamide (0.080 g, 0.129 mmol, 20% yield). The free base as dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford product as the hydrochloride salt (33). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.60 (d, 2H), 7.43 (d, 2H), 7.42 (d, 1H), 7.29 (s, 1H), 7.18 (d, 1H), 6.12 (m, 1H), 4.06 (m, 1H), 3.81 (m, 1H), 3.72-3.53 (m, 12H), 3.39 (s, 3H), 3.15 (m, 1H), 3.03 (m, 1H), 2.87 (m, 1H), 2.78 (m, 1H), 2.74 (s, 1H), 2.54 (m, 1H), 2.08 (m, 2H), 1.86 (m, 1H); MS (EI) for C.sub.29H.sub.37Cl.sub.2F.sub.3N.sub.2O.sub.5: 621 (MH.sup.+).
(226) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 33
Preparation of (S)—N-(1-(4-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)phenyl)-2-(pyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide, hydrochloride salt (34)
(227) Using the procedure outlined in the schematic below, the named compound was prepared.
(228) ##STR00154##
Step 1: Preparation of (S)-2-((tert-butoxycarbonyl)amino)-2-(4-hydroxyphenyl)acetic acid
(229) ##STR00155##
(230) 4-Hydroxy-L-phenylglycine (4.00 g, 23.93 mmol) and sodium bicarbonate (6.04 g, 71.80 mmol) was dissolved in water/tetrahydrofuran (70 mL/70 mL). The solution was cooled in an ice-bath and then di-tert-butyl dicarbonate (7.83 g, 35.90 mmol) was added into the solution. After stirring at room temperature overnight the mixture was washed with ether (50 mL). The aqueous portion was acidified to pH 4 with 2N HCl and extracted with dichloromethane (3×100 mL). The organic portion was washed with saturated sodium chloride (60 mL) and dried over anhydrous sodium sulfate. The organic portion was filtered, concentrated, and dried under high vacuum to give 6.35 g product as a white solid (yield: 99%). .sup.1H NMR (500 MHz, CD.sub.3OD): δ 7.22 (d, 2H), 6.77 (d, 2H), 5.07 (br, 1H), 1.45 (s, 9H); MS (EI) for C.sub.13H.sub.17NO.sub.5: 266 (MH.sup.+).
Step 2: Preparation of (S)-tert-butyl(1-(4-hydroxyphenyl)-2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate
(231) ##STR00156##
(232) (S)-2-((Tert-butoxycarbonyl)amino)-2-(4-hydroxyphenyl)acetic acid (0.50 g, 1.87 mmol) and N,N-diisopropylethylamine (0.72 g, 5.61 mmol) were dissolved in 20 mL of dichloromethane. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.72 g, 2.25 mmol) and pyrrolidine (0.16 g, 2.25 mmol) in 10 mL of dichloromethane were added into the solution. The reaction mixture was stirred for 16 hours. Dichloromethane (100 mL) was added into the reaction mixture, and the resulted solution was washed with saturated sodium chloride solution (50 mL×2). The solution was dried over sodium sulfate and concentrated. The crude product was purified by flash chromatography and 0.36 g of product was obtained (yield: 60%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.19 (d, 2H), 6.82 (d, 2H), 5.92 (d, 1H), 5.28 (m, 1H), 3.50 (m, 2H), 3.40 (m, 1H), 3.06 (m, 1H), 1.80 (m, 4H), 1.36 (s. 9H); MS (EI) for C.sub.17H.sub.24N.sub.2O.sub.4: 321 (MH.sup.+).
Step 3: Preparation of (S)-tert-butyl(1-(4-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)phenyl)-2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate
(233) ##STR00157##
(234) A solution of (S)-tert-butyl(1-(4-hydroxyphenyl)-2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate (0.30 g, 0.94 mmol) and 16-bromo-2,5,8,11,14-pentaoxahexadecane (0.59 g, 1.87 mmol) in acetone (20 mL) was added sodium carbonate (0.39 g, 2.81 mmol). The mixture was stirred at 70° C. and reaction was completed after six hours. The reaction mixture was cooled to room temperature and 150 mL of dichloromethane was added. The solution was washed with water (150 mL×2). Organic phase was dried over sodium sulfate and was then concentrated under reduced pressure. The residue was purified by column chromatography to provide product (0.51 g, yield: 98%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.32 (d, 2H), 6.88 (d, 2H), 6.02 (d, 1H), 5.31 (m, 1H), 4.12 (m, 2H), 3.87 (m, 2H), 3.74 (m, 2H), 3.72 (m, 10H), 3.55 (m, 4H), 3.52 (m, 2H), 3.44 (m, 1H), 3.40 (s, 3H), 3.05 (m, 1H), 1.80 (m, 4H)′ 1.41 (s, 9H); MS (EI) for C.sub.28H.sub.46N.sub.2O.sub.9: 555 (MH.sup.+).
Step 4: Preparation of (S)-1-(4-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)phenyl)-N-methyl-2-(pyrrolidin-1-yl)ethanamine
(235) ##STR00158##
(236) (S)-tert-butyl(1-(4-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)phenyl)-2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate (0.50 g, 0.90 mmol) in tetrahydrofuran (10 mL) was added dropwise to a stirred 2.0 M solution of lithium aluminum hydride (3 mL, 6.0 mmol) at room temperature. The mixture was stirred for thirty minutes at room temperature and then was heated to 65° C. for four hours. A 3N solution of sodium carbonate was added cautiously until effervescence ceased. The solid was filtered out and washed with dichloromethane (100 mL). The filtrate was concentrated and the residue was dissolved in 150 mL of dichloromethane. The resulted solution was washed with saturated sodium chloride solution (100 mL) and dried over sodium sulfate. The product was obtained after removing solvent (0.33 g, yield 81%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.27 (d, 2H), 6.89 (d, 2H), 4.13 (m, 2H), 3.85 (m, 2H), 3.75 (m, 2H), 3.68 (m, 10H), 3.60 (m, 1H), 3.55 (m, 4H), 3.40 (s, 3H), 2.62 (m, 2H), 2.46 (m, 2H), 2.30 (s, 3H), 2.28 (m, 1H), 2.10 (m, 2H)′ 1.80 (m, 4H); MS (EI) for C.sub.24H.sub.42N.sub.2O.sub.6: 455 (MH.sup.+).
Step 5: Preparation of (S)—N-(1-(4-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)phenyl)-2-(pyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide, hydrochloride salt (34)
(237) ##STR00159##
(238) (S)-1-(4-(2,5,8,11,14-Pentaoxahexadecan-16-yloxy)phenyl)-N-methyl-2-(pyrrolidin-1-yl)ethanamine (0.33 g, 0.73 mmol), 2-(3,4-dichlorophenyl)acetic acid (0.15 g, 0.73 mmol), and N,N-diisopropylethylamine (0.19 g, 1.47 mmol) were dissolved in 5 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.28 g, 0.88 mmol) was added into the solution. The reaction mixture was stirred for four hours at room temperature. Dichloromethane (150 mL) was added into the solution and the solution was washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded the target compound (0.22 g, yield: 47%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.35 (d, 2H), 7.18 (d, 2H), 7.10 (m, 1H), 6.85 (d, 2H), 6.04 (m, 0.82H), 4.98 (m, 0.18H), 4.10 (m, 2H), 3.84 (m, 2H), 3.70 (m, 3H), 3.66 (m, 14H), 3.52 (m, 2H), 3.35 (s, 3H), 3.20 (m, 1H), 2.75 (m, 2H), 2.65 (s, 3H), 2.50 (br., 2H), 1.75 (m, 4H); MS (EI) for C.sub.32H.sub.46Cl.sub.2N.sub.2O.sub.7: 641 (MH.sup.+).
(239) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 34
Preparation of (S)—N-(1-(4-(2,5,8,11,14-pentaoxahexadecan-16-ylamino)phenyl)-2-(pyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide, dihydrochloride salt (35)
(240) Using the procedure outlined in the schematic below, the named compound was prepared.
(241) ##STR00160##
Step 1: Preparation of (S)-4-(1-((tert-butoxycarbonyl)amino)-2-oxo-2-(pyrrolidin-1-yl)ethyl)phenyl trifluoromethanesulfonate
(242) ##STR00161##
(243) (S)-Tert-butyl(1-(4-hydroxyphenyl)-2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate (0.40 g, 1.25 mmol) and caesium carbonate (0.61 g, 1.87 mmol) were dissolved in tetrahydrofuran. Then, 1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide (0.67 g, 1.87 mmol) was added to the mixture and the reaction was stirred under nitrogen at 70° C. for five hours. The tetrahydrofuran was removed by rotavap. To the residue was added dichloromethane (100 mL), followed by 100 mL of water. The organic phase was washed with brine and dried over sodium sulfate. The solvent was removed and the crude product was purified by flash chromatography yielded the target compound (0.45 g, yield: 79%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.49 (d, 2H), 7.23 (d, 2H), 6.13 (d, 1H), 5.40 (d, 1H), 3.56 (m, 2H), 3.42 (m, 1H), 3.04 (m, 1H), 1.80 (m, 4H), 1.39 (s, 9H); MS (EI) for C.sub.18H.sub.23F.sub.3N.sub.2O.sub.6S: 453 (MH.sup.+).
Step 2: Preparation of (S)-tert-butyl(1-(4-(2,5,8,11,14-pentaoxahexadecan-16-ylamino)phenyl)-2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate
(244) ##STR00162##
(245) (S)-4-(1-((Tert-butoxycarbonyl)amino)-2-oxo-2-(pyrrolidin-1-yl)ethyl)phenyl trifluoromethanesulfonate (0.20 g, 0.44 mmol) and 2,5,8,11,14-pentaoxahexadecan-16-amine (0.36 g, 1.44 mmol) were dissolved in anhydrous toluene (10 mL). Caesium carbonate (0.47 g, 1.44 mmol), di-tert-butyl(2′,4′,6′-triisopropyl-3,6-dimethoxy-[1,1′-biphenyl]-2-yl)phosphine (0.048 g, 0.096 mmol), and diacetoxypalladium (0.011 g, 0.049 mmol) were added. The starting material was purged with nitrogen. The mixture was slight yellow after a few minutes. The mixture was stirred at 90° C. for five hours. Water (10 mL) and 2 mL of saturated sodium chloride solution were added into the reaction mixture. The mixture was extracted with dichloromethane (3×40 mL). The combined organic solution was dried over sodium sulfate and then concentrated. The residue was purified with flash column chromatography (0.13 g, yield 49%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.13 (d, 2H), 6.53 (d, 2H), 5.90 (d, 1H), 5.20 (m, 1H), 3.60 (m, 15H), 3.52 (m, 5H), 3.45 (m, 1H), 3.32 (m, 3H), 3.24 (m, 2H), 3.05 (m, 1H), 1.75 (m, 5H), 1.35 (s, 9H); MS (EI) for C.sub.28H.sub.47N.sub.3O.sub.8: 554 (MH.sup.+).
Step 3: Preparation of (S)—N-(4-(1-(methylamino)-2-(pyrrolidin-1-yl)ethyl)phenyl)-2,5,8,11,14-pentaoxahexadecan-16-amine
(246) ##STR00163##
(247) (S)-Tert-butyl(1-(4-(2,5,8,11,14-pentaoxahexadecan-16-ylamino)phenyl)-2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate (0.15 g, 0.26 mmol) in tetrahydrofuran (3 mL) was added dropwise to a stirred 2.0 M solution of lithium aluminum hydride (1 mL, 2.0 mmol). The mixture was stirred for thirty minutes at room temperature and then was heated to 65° C. for four hours. A 3N solution of sodium carbonate was added cautiously until effervescence ceased. The solid was filtered out and washed with dichloromethane (100 mL). The filtrate was concentrated and the residue was dissolved in 150 mL of dichloromethane. The resulting solution was dried over sodium sulfate. The product was obtained after removing solvent (0.10 g, yield 84%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.27 (m, 2H), 6.58 (m, 2H), 3.65 (m, 16H), 3.51 (m, 2H), 3.50 (m, 1H), 3.35 (s, 3H), 3.30 (m, 2H), 3.12 (m, 2H), 2.85 (m, 1H), 2.60 (m, 2H), 2.40 (m, 2H), 2.25 (m, 3H), 1.75 (m, 4H); MS (EI) for C.sub.24H.sub.43N.sub.3O.sub.5: 454 (MH.sup.+).
Step 4: Preparation of (S)—N-(1-(4-(2,5,8,11,14-pentaoxahexadecan-16-ylamino)phenyl)-2-(pyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide, dihydrochloride salt (35)
(248) ##STR00164##
(249) (S)—N-(4-(1-(Methylamino)-2-(pyrrolidin-1-yl)ethyl)phenyl)-2,5,8,11,14-pentaoxahexadecan-16-amine (0.10 g, 0.22 mmol), 2-(3,4-dichlorophenyl)acetic acid (0.041 g, 0.20 mmol), and N,N-diisopropylethylamine (0.056 g, 0.44 mmol) were dissolved in 3 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.085 g, 0.27 mmol) was added into the solution. The reaction mixture was stirred for four hours at 0° C. Dichloromethane (100 mL) was added into the solution and the solution was washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded the target compound (0.040 g, yield: 28%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford product as the dihydrochloride salt (35). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.35 (d, 2H), 7.18 (d, 2H), 7.10 (m, 1H), 6.58 (d, 2H), 6.05 (m, 0.8H), 4.96 (m, 0.2H), 4.25 (br. 1H). 3.82 (m, 1H), 3.66 (m, 17H), 3.55 (m, 2H), 3.40 (s, 3H), 3.30 (m, 2H), 2.95 (m, 1H), 2.78 (m, 2H), 2.70 (m, 3H), 2.50 (m, 2H), 1.80 (m, 4H); MS (EI) for C.sub.32H.sub.47Cl.sub.2N.sub.3O.sub.6: 640 (MH.sup.+).
(250) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 35
Preparation of (S)-2-(3,4-dichlorophenyl)-N-(1-(4-(2-hydroxyethoxy)phenyl)-2-(pyrrolidin-1-yl)ethyl)-N-methylacetamide, hydrochloride salt (36)
(251) Using the procedure outlined in the schematic below, the named compound was prepared.
(252) ##STR00165##
Step 1: Preparation of (S)-tert-butyl(1-(4-(2-hydroxyethoxy)phenyl)-2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate
(253) ##STR00166##
(254) A solution of (S)-tert-butyl(1-(4-hydroxyphenyl)-2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate (0.30 g, 0.94 mmol) and 2-bromoethanol (0.37 g, 2.81 mmol) in acetone (15 mL) was added potassium carbonate (0.39 g, 2.81 mmol). The mixture was stirred at 70° C. for six hours. 2-Bromoethanol (0.5 g) and 0.4 g of potassium carbonate were added. The mixture was stirred overnight at 70° C. After this period, the reaction mixture was cooled to room temperature and 150 mL of dichloromethane was added. The solid was filtered out and solution was washed with brine (150 mL×2). The organic phase was dried over sodium sulfate and was then concentrated under reduced pressure. The residue was purified by column chromatography to provide product (0.27 g, yield: 80%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.30 (d, 2H), 6.85 (d, 2H), 6.05 (d, 1H), 5.30 (m, 1H), 4.05 (m, 2H), 3.95 (m, 2H), 3.52 (m, 2H), 3.40 (m, 1H), 3.06 (m, 1H), 1.80 (m, 5H), 1.35 (s, 9H); MS (EI) for C.sub.19H.sub.28N.sub.2O.sub.5: 365 (MH.sup.+).
Step 2: Preparation of (S)-2-(4-(1-(methylamino)-2-(pyrrolidin-1-yl)ethyl)phenoxy)ethanol
(255) ##STR00167##
(256) (S)-Tert-butyl(1-(4-(2,5,8,11,14-pentaoxahexadecan-16-ylamino)phenyl)-2-oxo-2-(pyrrolidin-1-yl)ethyl)carbamate (0.24 g, 0.66 mmol) was dissolved in tetrahydrofuran (5 mL). A 2.0 M solution of lithium aluminum hydride (2 mL, 4.0 mmol) was added into the solution at room temperature. The mixture was stirred for thirty minutes at room temperature and then was heated to 65° C. for four hours. A 3N solution of sodium carbonate was added cautiously until effervescence ceased. The solid was filtered out and washed with dichloromethane (100 mL). The filtrate was concentrated and the residue was dissolved in 150 mL of dichloromethane. The resulted solution was washed with sodium bicarbonate solution and dried over sodium sulfate. The product was obtained after removing solvent (0.66 g, yield 38%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.24 (d, 2H), 6.86 (d, 2H), 4.05 (m, 2H), 3.92 (m, 2H), 3.50 (m, 3H), 2.86 (t, 1H), 2.60 (m, 2H), 2.45 (m, 2H), 2.30 (s, 3H), 1.75 (m, 4H); MS (EI) for C.sub.15H.sub.24N.sub.2O.sub.2: 265 (MH.sup.+).
Step 3: Preparation of: (S)-2-(3,4-dichlorophenyl)-N-(1-(4-(2-hydroxyethoxy)phenyl)-2-(pyrrolidin-1-yl)ethyl)-N-methylacetamide, hydrochloride salt (36)
(257) ##STR00168##
(258) (S)-2-(4-(1-(Methylamino)-2-(pyrrolidin-1-yl)ethyl)phenoxy)ethanol (0.066 g, 0.25 mmol), 3,4-dichlorophenylacetic acid (0.049 g, 0.237 mmol), and N,N-diisopropylethylamine (0.064 g, 0.50 mmol) were dissolved in 3 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.096 g, 0.30 mmol) was added into the solution. The reaction mixture was stirred for four hours at 0° C. Dichloromethane (100 mL) was added into the solution and the solution was washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded the target compound (0.05 g, yield: 44%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (36). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.37 (m, 2H), 7.25 (m, 2H), 7.15 (m, 1H), 6.90 (m, 2H), 6.09 (m, 0.8H), 5.02 (m, 0.2H), 4.09 (m, 2H), 3.97 (m, 2H), 3.28 (m, 1H), 3.70 (m, 1H), 3.20 (br., 1H), 2.75 (m, 5H), 2.52 (m, 3H), 1.80 (m, 4H); MS (EI) for C.sub.23H.sub.28Cl.sub.2N.sub.2O.sub.3: 451 (MH.sup.+).
Example 36
Preparation of 2-(3,4-Dichlorophenyl)-N-{(1S)-1-[4-(2-hydroxyethoxy)phenyl]-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]ethyl}-N-methylacetamide, hydrochloride (37)
(259) Using the procedure outlined in the schematic below, the named compound was prepared.
(260) ##STR00169##
Step 1: Preparation of tert-butyl((S)-1-(4-hydroxyphenyl)-2-((S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl)-2-oxoethyl)carbamate
(261) ##STR00170##
(262) (S)-3-(2-(2-(2-Methoxyethoxy)ethoxy)ethoxy)pyrrolidine HCl salt (0.505 g, 1.87 mmol) and N,N-diisopropylethylamine (0.84 g, 6.55 mmol) were dissolved in 20 mL of dichloromethane. (S)-2-((Tert-butoxycarbonyl)amino)-2-(4-hydroxyphenyl)acetic acid (0.50 g, 1.87 mmol) was added into the solution and the mixture was cooled in an ice-bath under stirring. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.72 g, 2.25 mmol) was added into the solution. The reaction mixture was stirred for four hours at room temperature. Dichloromethane (150 mL) was added into the mixture and resultant solution was washed with water (2×100 mL) and dried over sodium sulfate. The crude product was purified by flash chromatography and 0.72 g product was obtained (yield: 80%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.45 (m, 1H), 7.28 (m, 2H), 6.81 (m, 2H), 5.98 (m, 1H), 5.25 (m, 1H), 4.00 (m, 1H), 4.00 (m, 1H), 3.65 (m, 6H), 3.52 (m, 3H), 3.42 (s, 3H), 3.35 (m, 3H), 3.20 (m, 2H), 2.90 (m, 1H), 2.00 (m, 1H), 1.90 (m, 1H), 1.42 (s, 9H); MS (EI) for C.sub.24H.sub.38N.sub.2O.sub.8: 483 (MH.sup.+).
Step 2: Preparation of tert-butyl((S)-1-(4-(2-((tert-butyldimethylsilyl)oxy)ethoxy)phenyl)-2-((S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl)-2-oxoethyl)carbamate
(263) ##STR00171##
(264) A solution of tert-butyl((S)-1-(4-hydroxyphenyl)-2-((S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl)-2-oxoethyl)carbamate (0.54 g, 1.12 mmol) and (2-bromoethoxy)(tert-butyl)dimethylsilane (1.34 g, 5.60 mmol) in acetone (20 mL) was added potassium carbonate (0.77 g, 5.60 mmol). The mixture was stirred at 70° C. for twenty hours. After this period, the reaction mixture was cooled to room temperature and 150 mL of dichloromethane was added. The solution was washed with water (150 mL×2). The organic phase was dried over sodium sulfate and was then concentrated under reduced pressure. The residue was purified by column chromatography (0.62 g, yield: 86%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.31 (m, 2H), 6.88 (d, 2H), 6.00 (dd, 1H), 5.30 (m, 1H), 4.08 (m, 3H), 3.96 (m, 2H), 3.85 (m, 2H), 3.62 (m, 10H), 3.45 (m, 2H), 3.40 (s, 3H), 3.25 (m, 1H), 2.10 (m, 1H), 1.95 (m, 1H), 1.80 (m, 1H), 1.40 (s, 9H), 0.90 (s, 9H), 0.10 (m, 6H); MS (EI) for C.sub.32H.sub.56N.sub.2O.sub.9Si; 641 (MH.sup.+).
Step 3: Preparation of 2-(4-((S)-2-((S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl)-1-(methylamino)ethyl)phenoxy)ethanol
(265) ##STR00172##
(266) Tert-butyl((S)-1-(4-(2-((tert-butyldimethylsilyl)oxy)ethoxy)phenyl)-2-((S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl)-2-oxoethyl)carbamate (0.52 g, 0.80 mmol) was dissolved in tetrahydrofuran (6 mL). A 2.0 M solution of lithium aluminum hydride (2.8 mL, 5.6 mmol) was added into the solution at room temperature. The mixture was stirred at 65° C. for four hours. A 3N solution of sodium carbonate was added cautiously until effervescence ceased. Dichloromethane (50 mL) was added into the mixture. The solid was filtered out and washed with dichloromethane (100 mL). The filtrate was washed with saturated sodium chloride solution and dried over sodium sulfate. The product was obtained after removing solvent (0.23 g, yield 60%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.25 (m, 2H), 6.88 (m, 2H), 4.06 (m, 3H), 3.95 (m, 2H), 3.65 (m, 9H), 3.55 (m, 5H), 3.40 (s, 3H), 3.30 (br., 2H), 2.82 (m, 3H), 2.55 (m, 1H), 2.44 (m, 1H), 2.30 (m, 1H), 2.06 (m, 1H), 1.80 (m, 1H); MS (EI) for C.sub.22H.sub.38N.sub.2O.sub.6; 427 (MH.sup.+).
Step 4: Preparation of 2-(3,4-Dichlorophenyl)-N-{(1S)-1-[4-(2-hydroxyethoxy)phenyl]-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]ethyl}-N-methylacetamide, hydrochloride salt (37)
(267) ##STR00173##
(268) 2-(4-((S)-2-((S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidin-1-yl)-1-(methylamino)ethyl)phenoxy)ethanol (0.21 g of 80%, 0.39 mmol), 3,4-dichlorophenylacetic acid (0.081 g, 0.39 mmol), and N,N-diisopropylethylamine (0.10 g, 0.79 mmol) were dissolved in 5 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.15 g, 0.47 mmol) was added into the solution. The reaction mixture was stirred for one hour at 0° C. and then overnight. Dichloromethane (150 mL) was added into the solution and the solution was washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded product (0.20 g, yield: 83%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (37). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.36 (m, 2H), 7.18 (m, 2H), 7.05 (m, 1H), 6.82 (m, 2H), 6.00 (m, 0.83H), 4.96 (m, 0.17H), 4.02 (m, 3H), 3.90 (m, 1H), 3.76 (m, 1H), 3.65 (m, 9H), 3.55 (m, 4H), 3.45 (s, 3H), 3.14 (m, 1H), 3.00 (m, 1H), 2.80 (m, 3H), 2.65 (s, 3H), 2.62 (m, 1H), 2.50 (m, 2H), 2.00 (m, 1H), 1.80 (m, 1H); MS (EI) for C.sub.30H.sub.42Cl.sub.2N.sub.2O.sub.7: 613 (MH.sup.+).
(269) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 37
Preparation of 2-(3,4-dichlorophenyl)-N—((S)-1-(4-(2-hydroxyethoxy)phenyl)-2-((S)-3-hydroxypyrrolidin-1-yl)ethyl)-N-methylacetamide, hydrochloride salt (38)
(270) Using the procedure outlined in the schematic below, the named compound was prepared.
(271) ##STR00174## ##STR00175##
Step 1: Preparation of (2S)-2-((tert-butoxycarbonyl)amino)-2-(4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)acetic acid
(272) ##STR00176##
(273) (S)-2-((Tert-butoxycarbonyl)amino)-2-(4-hydroxyphenyl)acetic acid (2.67 g, 10.0 mmol) was dissolved in 70 mL of dimethylformamide and the solution was cooled in an ice bath. Sodium hydride (0.88 g, 60% in mineral oil, 22.0 mmol) was added in portions. The mixture was stirred for thirty minutes before 2-(2-bromoethoxy)tetrahydro-2H-pyran (2.30 g, 11.0 mmol) in 30 mL of dimethylformamide was added portionally. The reaction mixture was stirred at room temperature for 17 hours. Sodium hydride (0.16 g) was added and reaction was continued for one hour and then diluted with ice/water. The mixture was extracted with ethyl acetate (100 mL×2). The aqueous layer was cooled in an ice bath and acidified using 1.5 M aqueous potassium hydrogen sulfate to pH 2-3. The resulting mixture was extracted with ethyl acetate (100 mL×2). The organic phase was washed with water, brine, and dried over sodium sulfate. The product (3.3 g) was obtained (yield: 84%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.32 (m, 2H), 6.92 (m, 2H), 5.55 (m, 0.5H), 5.28 (m, 0.5H), 4.74 (m, 1H), 4.15 (m, 2H), 4.05 (m, 1H), 3.90 (m, 1H), 3.85 (m, 1H), 3.55 (m, 1H), 1.85 (m, 1H), 1.75 (m, 1H), 1.60 (m, 5H), 1.45 (s, 5H), 1.28 (s, 4H); MS (EI) for C.sub.20H.sub.29NO.sub.7; 394 (MH.sup.+).
Step 2: Preparation of tert-butyl((1S)-2-((S)-3-hydroxypyrrolidin-1-yl)-2-oxo-1-(4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)ethyl)carbamate
(274) ##STR00177##
(275) (2S)-2-((tert-butoxycarbonyl)amino)-2-(4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)acetic acid (3.20 g, 8.09 mmol), (s)-pyrrolidin-3-ol (0.78 g, 8.90 mmol), and N,N-diisopropylethylamine (2.07 g, 16.18 mmol) were dissolved in 18 mL of acetonitrile. The mixture was stirred for 10 min. at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (3.12 g, 9.71 mmol) was added into the solution. The reaction mixture was stirred for two hours under an ice bath (15% starting materials remaining) and the reaction was continued at room temperature for four hours (no change). O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (120 mg) was added and the reaction was completed after one hour. Dichloromethane (200 mL) was added into the reaction mixture, and the resulted solution was washed with water (200 mL×3). The solution was dried over sodium sulfate and concentrated. The product was obtained after dried under vacuum (3.40 g, yield: 90%). .sup.1H NMR (500 MHz, CDCl3): δ 7.30 (m, 2H), 6.90 (m, 2H), 5.96 (dd, 1H), 5.30 (dd, 1H), 4.70 (m, 1H), 4.45 (m, 1H), 4.15 (m, 1H), 4.05 (m, 1H), 3.90 (m, 1H), 3.80 (m, 1H), 3.70 (m, 2H), 3.55 (m, 2H), 3.35 (m, 0.5H), 3.10 (m, 0.5H), 1.96 (m, 1H), 1.85 (m, 2H), 1.75 (m, 1H), 1.60 (m, 2H), 1.55 (m, 3H), 1.40 (s, 9H); MS (EI) for C.sub.24H.sub.36N.sub.2O.sub.7; 465 (MH.sup.+).
Step 3: Preparation of (3S)-1-((2S)-2-(methylamino)-2-(4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)ethyl)pyrrolidin-3-ol
(276) ##STR00178##
(277) Tert-butyl((1S)-2-((S)-3-hydroxypyrrolidin-1-yl)-2-oxo-1-(4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)ethyl)carbamate (1.33 g, 2.85 mmol) was dissolved in tetrahydrofuran (20 mL). A 2.0 M solution of lithium aluminum hydride (8.0 mL, 16 mmol) was added into the solution at room temperature. The mixture was stirred at 65° C. for four hours. A 3N solution of sodium carbonate was added cautiously until effervescence ceased. Ethyl acetate (100 mL) was added into the mixture. The solid was filtered out and washed with ethyl acetate (100 mL). The filtrate was washed with saturated sodium chloride solution and dried over sodium sulfate. The product was obtained after removing solvent (0.84 g, yield: 71%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.28 (m, 2H), 6.92 (m, 2H), 4.74 (m, 1H), 4.34 (m, 1H), 4.16 (m, 2H), 4.06 (m, 1H), 3.92 (m, 1H), 3.84 (m, 1H), 3.55 (m, 2H), 3.02 (m, 1H), 2.85 (m, 1H), 2.65 (m, 2H), 2.30 (m, 3H), 2.20 (m, 2H), 1.80 (m, 4H), 1.60 (m, 5H); MS (EI) for C.sub.20H.sub.32N.sub.2O.sub.4; 365 (MH.sup.+).
Step 4: Preparation of 2-(3,4-dichlorophenyl)-N—((S)-1-(4-(2-hydroxyethoxy)phenyl)-2-((S)-3-hydroxypyrrolidin-1-yl)ethyl)-N-methylacetamide, hydrochloride salt (38)
(278) ##STR00179##
(279) (3S)-1-((2S)-2-(methylamino)-2-(4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)ethyl)pyrrolidin-3-ol (0.092 g of 80%, 0.20 mmol), 3,4-dichlorophenylacetic acid (0.041 g, 0.20 mmol), and N,N-diisopropylethylamine (0.052 g, 0.40 mmol) were dissolved in 3 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.078 g, 0.24 mmol) was added into the solution. The reaction mixture was stirred for three hours under an ice-bath. Dichloromethane (150 mL) was added into the solution and the resultant solution was washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded an oil (0.090 g, 0.16 mmol), which was dissolved in 10 mL of CH.sub.3OH. p-Toluenesulfonic acid (0.056 g, 0.32 mmol) was added into the solution. The mixture was stirred for sixty minutes at room temperature. Dichloromethane (100 mL) was added into the solution and the solution was washed with sodium carbonate (10%), water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded product as an oil (0.046 g, yield: 60%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (38). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.40 (m, 2H), 7.22, 7.05 (dd, 2H), 7.15 (m, 1H), 6.90 (m, 2H), 6.05 (m, 0.84H), 5.00 (m, 0.16H), 4.31 (m, 1H), 4.10 (m, 2H), 3.96 (m, 2H), 3.72 (m, 2H), 3.10 (m, 2H), 2.75 (m, 2H), 2.70 (s, 3H), 2.55 (m, 1H), 2.35 (m, 2H), 2.20 (m, 2H), 1.72 (m, 1H); MS (EI) for C.sub.23H.sub.28Cl.sub.2N.sub.2O.sub.4: 467 (MH.sup.+).
(280) The example described above may be modified to introduce various substituted phenyl moieties (such as difluorophenyl, 4-trifluorophethyl) and heterocycles (such as pyridine, thoazole, benzofuran).
(281) The example described above may be modified to introduce oligomers of various lengths (at the 2-hydroxyethoxy substituent) as disclosed herein.
Example 38
Preparation of (S)—N-(2-(4-(2,5,8,11,14-pentaoxahexadecan-16-yl)piperazin-1-yl)-1-phenylethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide, dihydrochloride salt (39)
(282) Using the procedure outlined in the schematic below, the named compound was prepared.
(283) ##STR00180##
Step 1: Preparation of (S)-tert-butyl 4-(2-(((benzyloxy)carbonyl)amino)-2-phenylacetyl)piperazine-1-carboxylate
(284) ##STR00181##
(285) (S)-2-((Tert-butoxycarbonyl)amino)-2-phenylacetic acid (0.70 g, 2.45 mmol), 4-dimethylaminopyridine/p-toluenesulfonic acid (1:1 salt) (0.38 g, 1.23 mmol) and N,N′-diisopropylcarbodiimide (0.62 g, 4.91 mmol) were dissolved in 15 mL of dichloromethane. To the solution was added 1-Boc-piperazine (0.46 g, 2.45 mmol). The mixture was stirred for four hours at room temperature. Dichloromethane (200 mL) was added into the reaction mixture, and the resulted solution was washed with water (200 mL×3). The solution was dried over sodium sulfate and concentrated. The purification of the residue by flash chromatography yielded the target compound (0.9 g, yield: 81%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.38 (m, 10H), 6.38 (m, 1H), 5.60 (d, 1H), 5.65 (m, 1H), 5.03 (m, 1H), 3.70 (m, 1H), 3.50 (m, 2H), 3.30 (m, 4H), 2.72 (m, 1H), 1.42 (s. 9H); MS (EI) for C.sub.25H.sub.31N.sub.3O.sub.5: 398 [(M-Bu)H.sup.+].
Step 2: Preparation of (S)-benzyl(2-(4-(2,5,8,11,14-pentaoxahexadecan-16-yl)piperazin-1-yl)-2-oxo-1-phenylethyl)carbamate
(286) ##STR00182##
(287) (S)-Tert-butyl 4-(2-(((benzyloxy)carbonyl)amino)-2-phenylacetyl)piperazine-1-carboxylate (0.90 g, 1.98 mmol) was dissolved in 9 mL of dichloromethane, and then 3 mL of trifloroacetic acid was added. The mixture was stirred for 1.5 hours and then was adjusted to pH 10 with saturated sodium bicarbonate. The resulting solution was extracted with dichloromethane (200 mL×3). The organic solution was washed with brine, dried over sodium sulfate, and concentrated (0.66 g, yield: 94%). MS (EI) for C.sub.20H.sub.23N.sub.3O.sub.3: 354 (MH.sup.+).
(288) A solution of above product [(S)-benzyl(2-oxo-1-phenyl-2-(piperazin-1-yl)ethyl)carbamate] (0.19 g, 0.54 mmol) and 16-bromo-2,5,8,11,14-pentaoxahexadecane (0.51 g, 1.62 mmol) in acetonitrile (10 mL) was added potassium carbonate (0.22 g, 1.61 mmol). The mixture was stirred at 90° C. for four hours. The solution was washed with water (150 mL×2). The organic phase was dried over sodium sulfate and was then concentrated under reduced pressure. The residue was purified by column chromatography to provide product (0.20 g, yield: 63%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.32 (m, 10H), 6.42 (m, 1H), 5.58 (m, 1H), 5.10 (m, 1H), 5.00 (m, 1H), 3.60 (m, 19H), 3.40 (m, 1H), 3.35 (s, 3H), 3.25 (m, 1H), 2.50 (m, 4H), 2.35 (m, 2H), 1.80 (m. 1H); MS (EI) for C.sub.31H.sub.45N.sub.3O.sub.8: 588 (MH.sup.+).
Step 3: Preparation of (S)-2-(4-(2,5,8,11,14-pentaoxahexadecan-16-yl)piperazin-1-yl)-N-methyl-1-phenylethanamine
(289) ##STR00183##
(290) (S)-Benzyl(2-(4-(2,5,8,11,14-pentaoxahexadecan-16-yl)piperazin-1-yl)-2-oxo-1-phenylethyl)carbamate (0.20 g, 0.33 mmol) in tetrahydrofuran (3 mL) was added dropwise to a stirred 2.0 M solution of lithium aluminum hydride (1.5 mL, 3.0 mmol) at room temperature. The mixture was stirred at 65° C. for four hours. A 3N solution of sodium carbonate was added cautiously until effervescence ceased. The solid was filtered out and washed with dichloromethane (100 mL). The filtrate was washed with water and then dried over sodium sulfate. The product was obtained after removing solvent (0.14 g, yield: 93%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.30 (m, 4H), 7.20 (m, 1H), 4.62 (s, 1H), 3.60 (m, 17H), 3.52 (m, 2H), 3.35 (s, 3H), 2.50 (m, 7H), 2.40 (m, 2H), 2.30 (m, 1H), 2.24 (s, 3H), 1.62 (m. 1H); MS (EI) for C.sub.24H.sub.43N.sub.3O.sub.5: 454 (MH.sup.+).
Step 4: Preparation of (S)—N-(2-(4-(2,5,8,11,14-pentaoxahexadecan-16-yl)piperazin-1-yl)-1-phenylethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide, dihydrochloride salt (39)
(291) ##STR00184##
(292) (S)-2-(4-(2,5,8,11,14-Pentaoxahexadecan-16-yl)piperazin-1-yl)-N-methyl-1-phenylethanamine (0.16 g, 0.35 mmol), 3,4-dichlorophenylacetic acid (0.080 g, 0.39 mmol), and N,N-diisopropylethylamine (0.090 g, 0.70 mmol) were dissolved in 3 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.14 g, 0.42 mmol) was added into the solution. The reaction mixture was stirred overnight. Dichloromethane (100 mL) was added into the solution and washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded the target compound (0.12 g, yield: 93%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford the product as a dihydrochloride salt (39). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.33 (m, 2H), 7.33, 7.10 (m, 5H), 7.18 (m, 1H), 6.13 (m, 0.82H), 5.00 (m, 0.18H), 3.75 (m, 1H), 3.65 (m, 18H), 3.52 (m, 2H), 3.35 (s, 3H), 2.90 (m, 1H), 2.80 (m, 2H), 2.70 (s, 3H), 2.60 (m, 2H), 2.43 (br., 6H); MS (EI) for C.sub.32H.sub.47Cl.sub.2N.sub.2O.sub.6: 640 (MH.sup.+).
(293) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 39
Preparation of (S)-2-(3,4-dichlorophenyl)-N-(2-(3,3-difluoropyrrolidin-1-yl)-1-phenylethyl)-N-methylacetamide, hydrochloride salt (40)
(294) Using the procedure outlined in the schematic below, the named compound was prepared.
(295) ##STR00185##
Step 1: Preparation of (S)-benzyl(2-(3,3-difluoropyrrolidin-1-yl)-2-oxo-1-phenylethyl)carbamate
(296) ##STR00186##
(297) (S)-2-(((Benzyloxy)carbonyl)amino)-2-phenylacetic acid (0.48 g, 1.69 mmol) and 4-dimethylaminopyridine/p-toluenesulfonic acid (1:1 salt) (0.26 g, 0.85 mmol) were dissolved in 20 mL dichloromethane. N,N′-diisopropylcarbodiimide (0.43 g, 3.38 mmol) was added into the solution. Finally 3,3-difluoropyrrolidine HCl (0.25 g, 1.69 mmol) and N,N-diisopropylethylamine (0.432 g, 3.38 mmol) in 5 mL of dichloromethane were added. The mixture was stirred for two hours at room temperature. Dichloromethane (200 mL) was added into the reaction mixture, and the resulted solution was washed with water (200 mL×3). The solution was dried over sodium sulfate and concentrated. The purification twice of the residue by flash chromatography yielded the target compound (0.22 g, yield: 35%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.36 (m, 10H), 6.45 (m, 1H), 5.50 (m, 0.5H), 5.35 (m, 0.5H), 5.10 (m, 2H), 3.90 (m, 1H), 3.76 (m, 2H), 3.62 (m, 0.5H), 3.30 (m, 0.5H), 2.25 (m. 2H); MS (EI) for C.sub.20H.sub.20F.sub.2N.sub.2O.sub.3: 375 (MH.sup.+).
Step 2: Preparation of (S)-2-(3,3-difluoropyrrolidin-1-yl)-N-methyl-1-phenylethanamine
(298) ##STR00187##
(299) (S)-Benzyl(2-(3,3-difluoropyrrolidin-1-yl)-2-oxo-1-phenylethyl)carbamate (0.22 g, 0.59 mmol) in tetrahydrofuran (3 mL) was added dropwise to a stirred 2.0 M solution of lithium aluminum hydride (2 mL, 4.0 mmol) at room temperature. The mixture was stirred for thirty minutes at room temperature and then was heated to 65° C. for four hours. A 3N solution of sodium carbonate was added cautiously until effervescence ceased. The solid was filtered out and washed with dichloromethane (100 mL). The filtrate was concentrated and the residue was dissolved in 150 mL of dichloromethane. The resulted solution was washed with saturated sodium chloride solution (100 mL) and dried over sodium sulfate. The product was obtained after removing solvent (0.12 g, yield 85%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.35 (m, 5H), 3.54 (m, 1H), 3.10 (m, 2H), 2.82 (m, 3H), 2.75 (m. 1H), 2.40 (m. 1H), 2.30 (m, 1H), 2.28 (s, 3H); MS (EI) for C.sub.13H.sub.18F.sub.2N.sub.2: 241 (MH.sup.+).
Step 3: Preparation of (S)-2-(3,4-dichlorophenyl)-N-(2-(3,3-difluoropyrrolidin-1-yl)-1-phenylethyl)-N-methylacetamide, hydrochloride salt (40)
(300) ##STR00188##
(301) (S)-2-(3,3-Difluoropyrrolidin-1-yl)-N-methyl-1-phenylethanamine (0.059 g, 0.25 mmol), 3,4-dichlorophenylacetic acid (0.055 g, 0.27 mmol), and N,N-diisopropylethylamine (0.063 g, 0.491 mmol) were dissolved in 3 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.095 g, 0.30 mmol) was added into the solution. The reaction mixture was stirred overnight. Dichloromethane (100 mL) was added into the solution and the solution was washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded the target compound (0.040 g, yield: 38%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (40). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.40 (m, 7H), 7.15 (m, 1H), 6.10 (m, 0.85H), 5.30 (m, 0.15H), 3.75 (m, 2H), 3.18 (m, 2H), 3.02 (m, 1H), 2.90 (m, 1H), 2.75 (m. 2H), 2.70 (s. 3H), 2.28 (m, 2H); MS (EI) for C.sub.21H.sub.22Cl.sub.2F.sub.2N.sub.2O: 427 (MH.sup.+).
Example 40
Preparation of N—((S)-2-((S)-3-(2,5,8,11,14-pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl)-1-phenylethyl)-N-methyl-2-(2-nitrophenyl)acetamide, hydrochloride salt (41)
(302) Using the procedure outlined in the schematic below, the named compound was prepared.
(303) ##STR00189##
(304) (S)-2-((S)-3-(2,5,8,11,14-Pentaoxahexadecan-16-yloxy)pyrrolidin-1-yl)-N-methyl-1-phenylethanamine (0.10 g, 0.22 mmol), 2-(3,4-dichlorophenyl)acetic acid (0.040 g, 0.22 mmol), and N,N-diisopropylethylamine (0.056 g, 0.44 mmol) were dissolved in 3 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.085 g, 0.26 mmol) was added into the solution. The reaction mixture was stirred for four hours at room temperature and concentrated. The residue was dissolved in dichloromethane (100 mL) and the solution was washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded the target compound (0.09 g, yield:66%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (41). .sup.1H NMR (500 MHz, CDCl3): δ 8.10 (d, 1H), 7.60 (m, 1H), 7.45 (m, 2H), 7.35 (m, 5H), 6.05 (m, 0.82H), 5.18 (m, 0.18H), 4.10 (m, 3H), 3.60 (m, 20H), 3.38 (s, 3H), 3.16 (m, 1H), 3.05 (m, 1H), 2.85 (s, 3H), 2.80 (m, 2H), 2.55 (m, 2H), 2.10 (m, 1H), 1.82 (m, 1H); MS (EI) for C.sub.32H.sub.47N.sub.3O.sub.9: 618 (MH.sup.+).
(305) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 41
Preparation of N—((S)-2-((S)-3-hydroxypyrrolidin-1-yl)-1-phenylethyl)-N-methyl-2-(2-nitrophenyl)acetamide, hydrochloride salt (42)
(306) Using the procedure outlined in the schematic below, the named compound was prepared.
(307) ##STR00190##
(308) (S)-1-((S)-2-(Methylamino)-2-phenylethyl)pyrrolidin-3-ol (0.15 g, 0.68 mmol), 2-(2-nitrophenyl)acetic acid (0.12 g, 0.68 mmol), and N,N-diisopropylethylamine (0.17 g, 1.36 mmol) were dissolved in 5 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.26 g, 0.82 mmol) was added into the solution. The reaction mixture was stirred for four hours at room temperature and concentrated. The residue was dissolved in dichloromethane (100 mL) and the solution was washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded the target compound (0.13 g, yield:50%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (42). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 8.10 (m, 1H), 7.60 (m, 1H), 7.35 (m, 7H), 6.05 (m, 0.9H), 5.20 (m, 0.1H), 4.38 (d, 0.5H), 4.30 (m, 1H), 4.20 (d, 0.5H), 4.05 (d, 0.5H), 3.88 (d, 0.5H), 3.20 (m, 3H), 3.00 (m, 1H), 2.85 (s, 3H), 2.75 (m, 1H), 2.55 (m, 2H), 2.20 (m, 1H), 1.82 (m, 1H); MS (EI) for C.sub.21H.sub.25N.sub.3O.sub.4: 384 (MH.sup.+).
Example 42
Preparation of 2-(2-aminophenyl)-N—((S)-2-((S)-3-hydroxypyrrolidin-1-yl)-1-phenylethyl)-N-methylacetamide, dihydrochloride salt (43)
(309) Using the procedure outlined in the schematic below, the named compound was prepared.
(310) ##STR00191##
(311) N—((S)-2-((S)-3-Hydroxypyrrolidin-1-yl)-1-phenylethyl)-N-methyl-2-(2-nitrophenyl)acetamide was dissolved in 10 mL of tetrahydrofuran. The reduction reaction was performed by H-Cube (CatCart THS01131, 10% Pd/C, Flow rate 1 mL/Min). The product was purified by biotage and 0.02 g of product was obtained (yield: 27%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford the product as a dihydrochloride salt (43). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.30 (m, 5H), 7.10 (m, 2H), 6.72 (m, 2H), 6.10 (m, 0.8H), 5.30 (s, 0.2H). 4.25 (m, 1H), 3.70 (m, 1H), 3.65 (m, 1H), 3.20 (m, 1H), 3.04 (m, 1H), 2.90 (m, 1H), 2.85 (s, 3H), 2.75 (m, 2H), 2.50 (m, 2H), 2.15 (m, 1H), 1.65 (m, 1H); MS (EI) for C.sub.21H.sub.27N.sub.3O.sub.2: 354 (MH.sup.+).
Example 43
Preparation of 2-(2,4-Difluorophenyl)-N-methyl-N-[(1S)-1-phenyl-2-(pyrrolidin-1-yl)ethyl]acetamide, hydrochloride salt (44)
(312) Using the procedure outlined in the schematic below, the named compound was prepared.
(313) ##STR00192##
(314) (S)—N-Methyl-1-phenyl-2-(pyrrolidin-1-yl)ethanamine (0.062 g, 0.30 mmol), 2-(2,4-difluorophenyl)acetic acid (0.052 g, 0.30 mmol), and N,N-diisopropylethylamine (0.078 g, 0.61 mmol) were dissolved in 2 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.12 g, 0.36 mmol) was added into the solution. The reaction mixture was stirred for two hours at 0° C. Dichloromethane (100 mL) was added into the solution and the solution was washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded product (0.078 g, yield 72%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (44). .sup.1H NMR (HCl salt, 500 MHz, CDCl.sub.3): δ 7.5 (m, 1H), 7.40 (m, 3H), 7.24 (m, 2H), 6.82 (m, 2H), 6.36 (d, 1H), 4.10 (m, 3H), 4.00 (t, 1H), 3.80 (d, 1H), 3.25 (m, 1H), 2.98 (s, 3H), 2.90 (m, 2H), 2.38 (m, 1H), 2.28 (m, 1H), 2.05 (m, 2H); MS (EI) for C.sub.21H.sub.24F.sub.2N.sub.2O: 359 (MH.sup.+).
Example 44
Preparation of N-Methyl-N-[(1S)-1-phenyl-2-(pyrrolidin-1-yl)ethyl]-2-(pyridine-2-yl)acetamide, dihydrochloride salt (45)
(315) Using the procedure outlined in the schematic below, the named compound was prepared.
(316) ##STR00193##
(317) (S)—N-Methyl-1-phenyl-2-(pyrrolidin-1-yl)ethanamine (0.064 g, 0.31 mmol), 2-(pyridin-2-yl)acetic acid hydrochloride (0.054 g, 0.31 mmol), and N,N-diisopropylethylamine (0.080 g, 0.63 mmol) were dissolved in 2 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.12 g, 0.38 mmol) was added into the solution. The reaction mixture was stirred for two hours at 0° C. Dichloromethane (100 mL) was added into the solution and the solution was washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded product (0.063 g, yield 62%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford product as the dihydrochloride salt (45). .sup.1H NMR (HCl salt, 500 MHz, CDCl.sub.3): δ 8.62 (m, 1H), 8.50 (m, 1H), 8.35 (m, 1H), 7.80 (m, 1H), 7.45 (m, 2H), 7.39 (m, 1H), 7.30 (m, 2H), 6.35 (m, 1H), 4.95 (d, 1H), 4.74 (d, 1H), 4.10 (m, 2H), 3.96 (m, 1H), 3.40 (m, 1H), 3.00 (m, 2H), 2.94 (s, 3H), 2.35 (m, 1H), 2.25 (m, 1H), 2.05 (m, 2H); MS (EI) for C.sub.20H.sub.25N.sub.3O: 324 (MH.sup.+).
Example 45
Preparation of 2-(6-Chloropyridin-3-yl)-N-methyl-N-[(1S)-1-phenyl-2-(pyrrolidin-1-yl)ethyl]acetamide, dihydrochloride salt (46)
(318) Using the procedure outlined in the schematic below, the named compound was prepared.
(319) ##STR00194##
(320) (S)—N-Methyl-1-phenyl-2-(pyrrolidin-1-yl)ethanamine (0.050 g, 0.25 mmol), 2-(6-chloropyridin-3-yl)acetic acid (0.042 mg, 0.25 mmol), and N,N-diisopropylethylamine (0.063 g, 0.49 mmol) were dissolved in 2 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.094 g, 0.29 mmol) was added into the solution. The reaction mixture was stirred for two hours at 0° C. Dichloromethane (100 mL) was added into the solution and the solution was washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded product (0.075 g, yield 86%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford the product as a dihydrochloride salt (46). .sup.1H NMR (HCl salt, 500 MHz, CDCl.sub.3): δ 8.72 (d, 1H), 8.60 (s, 1H), 7.64 (d, 1H), 7.40 (m, 3H), 7.24 (m, 2H), 6.35 (d, 1H), 4.66 (d, 1H), 4.10 (m, 3H), 3.70 (d, 1H), 3.35 (m, 1H), 3.05 (m, 2H), 2.94 (s, 3H), 2.38 (m, 1H), 2.25 (m, 1H), 2.10 (m, 2H); MS (EI) for C.sub.20H.sub.24ClN.sub.3O: 358 (MH.sup.+).
Example 46
Preparation of N—((S)-1-(3-aminophenyl)-2-((S)-3-hydroxypyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide, dihydrochloride salt (47)
(321) Using the procedure outlined in the schematic below, the named compound was prepared.
(322) ##STR00195##
Step 1: Preparation of 2-(3,4-dichlorophenyl)-N—((S)-2-((S)-3-hydroxypyrrolidin-1-yl)-1-(3-nitrophenyl)ethyl)-N-methylacetamide
(323) ##STR00196##
(324) (S)-1-((S)-2-(Methylamino)-2-(3-nitrophenyl)ethyl)pyrrolidin-3-ol (0.47 g, 1.77 mmol), 3,4-dichlorophenylacetic acid (0.31 g, 1.51 mmol), and N,N-diisopropylethylamine (0.45 g, 3.54 mmol) were dissolved in 3 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.68 g, 2.13 mmol) was added into the solution. The reaction mixture was stirred for two hours at 0° C. Dichloromethane (100 mL) was added into the solution and the solution was washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded product (0.48 g, yield: 60%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 8.10 (m, 2H), 7.62 (m, 1H), 7.50 (m, 1H), 7.35 (m, 2H), 7.12 (m, 1H), 6.06 (m, 0.9H), 5.10 (m, 0.1H), 4.30 (m, 1H), 3.64 (m, 2H), 3.15 (m, 1H), 2.90 (m, 2H), 2.80 (m, 2H), 2.75 (s, 3H), 2.60 (m, 1H), 2.40 (m, 1H), 2.10 (m, 1H), 1.70 (m, 1H); MS (EI) for C.sub.21H.sub.23Cl.sub.2N.sub.3O.sub.4: 452 (MIA
Step 2: Preparation of N—((S)-1-(3-aminophenyl)-2-((S)-3-hydroxypyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide, dihydrochloride salt (47)
(325) ##STR00197##
(326) To a three-neck, 100 mL round bottom flask equipped with a condenser, thermometer and magnetic stirrer was added 2-(3,4-dichlorophenyl)-N—((S)-2-((S)-3-hydroxypyrrolidin-1-yl)-1-(3-nitrophenyl)ethyl)-N-methylacetamide (0.42 g, 0.93 mmol) in 20 mL absolute ethanol. To the light-yellow solution was added hydrazine hydrate (0.72 g, 11.7 mmol) and Raney Ni slurry (40 drops) and heated to 55° C. The light-yellow reaction turned clear and after approximately two hours at 55° C. The reaction mixture was filtered through Celite, and the Raney Ni was washed with hot methanol. The combined filtrate was concentrated under reduced pressure to give the aniline product as oil (0.26 g, yield: 66%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford the product as a dihydrochloride salt (47).sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.40 (m, 2H), 7.14 (m, 2H), 6.15 (m, 1H), 6.10 (m, 2H), 6.00 (m, 0.86H), 4.92 (m, 0.14H), 4.30 (m, 1H), 3.70 (m, 4H), 3.15 (m, 2H), 2.20 (m, 5H), 2.32 (m, 1H), 2.15 (m, 1H), 1.74 (m, 1H); MS (EI) for C.sub.21H.sub.25Cl.sub.2N.sub.3O.sub.2: 422 (MH.sup.+).
Example 47
Preparation of N-(3-{(1S)-1-{[(3,4-Dichlorophenyl)acetyl](methyl)amino}-2-[(3S)-3-hydroxypyrrolidin-1-yl]ethyl}phenyl)-N2-2,5,8,11,14,17,20,23-octaoxatetracosan-1-oylvalinamide, hydrochloride salt (48)
(327) Using the procedure outlined in the schematic below, the named compound was prepared.
(328) ##STR00198##
Step 1: Preparation of (3S)-1-((2S)-2-(3-(2-((tert-butoxycarbonyl)amino)-3-methylbutanamido)phenyl)-2-(2-(3,4-dichlorophenyl)-N-methylacetamido)ethyl)pyrrolidin-3-yl 2-((tert-butoxycarbonyl)amino)-3-methylbutanoate
(329) ##STR00199##
(330) N—((S)-1-(3-Aminophenyl)-2-((S)-3-hydroxypyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide (0.15 g, 0.36 mmol), 2-((tert-butoxycarbonyl)amino)-3-methylbutanoic acid (0.17 g, 0.78 mmol), and 4-dimethylaminopyridine/p-toluenesulfonic acid (1:1 salt) (0.11 g, 0.36 mmol) were dissolved in 10 mL of dichloromethane. N,N′-diisopropylcarbodiimide (0.22 g, 1.77 mmol) was added into the mixture. The reaction mixture was stirred for three hours. Dichloromethane (100 mL) was added into the reaction mixture and the resulted solution was washed with water (100 mL×3). The solution was dried over sodium sulfate and concentrated. Product was obtained after purified by flash chromatography.
(331) The above product [(3S)-1-((2S)-2-(3-(2-((tert-butoxycarbonyl)amino)-3-methylbutanamido)phenyl)-2-(2-(3,4-dichlorophenyl)-N-methylacetamido)ethyl)pyrrolidin-3-yl 2-((tert-butoxycarbonyl)amino)-3-methylbutanoate] was dissolved in 3 mL of acetonitrile, and then 3 mL of 0.5 N potassium hydroxide was added into the mixture. The reaction mixture was stirred for two hours at 65° C. Dichloromethane (100 mL) was added into the reaction mixture and the resulted solution was washed with water (100 mL×3). The solution was dried over sodium sulfate and concentrated. The crude product was purified by flash chromatography and 0.94 g (yield: 43%) product was obtained as white solid. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 8.92 (m, 1H), 7.50 (m, 1H), 7.37 (m, 3H), 7.15 (m, 2H), 6.95 (m, 1H), 6.00 (m, 0.84H), 5.45 (m, 1H), 4.98 (m, 0.16H), 4.30 (m, 1H), 4.10 (m, 1H), 3.70 (m, 2H), 3.00 (m, 3H), 2.75 (m, 2H), 2.68 (s, 3H), 2.55 (m, 1H), 2.35 (m, 1H), 2.10 (m, 2H), 1.70 (m, 1H), 1.42 (s, 9H), 1.00 (m, 6H); MS (EI) for C.sub.31H.sub.42Cl.sub.2N.sub.4O.sub.5: 621 (MH.sup.+).
Step 2: Preparation of N-(3-{(1S)-1-{[(3,4-Dichlorophenyl)acetyl](methyl)amino}-2-[(3S)-3-hydroxypyrrolidin-1-yl]ethyl}phenyl)-N2-2,5,8,11,14,17,20,23-octaoxatetracosan-1-oylvalinamide, hydrochloride salt (48)
(332) ##STR00200##
(333) The product of step 1 [tert-butyl(1-((3-((S)-1-(2-(3,4-dichlorophenyl)-N-methylacetamido)-2-((S)-3-hydroxypyrrolidin-1-yl)ethyl)phenyl)amino)-3-methyl-1-oxobutan-2-yl)carbamate] (0.094 g, 0.21 mmol) was dissolved in 2 mL of dioxane, and then 2 mL of 4 N HCl in dioxane was added into the mixture. The reaction mixture was stirred for two hours. The solvent was removed and 0.079 g product was obtained as white solid, which was dissolved in 10 mL of dichloromethane. To the solution was added 2,5-dioxopyrrolidin-1-yl 2,5,8,11,14,17,20-heptaoxadocosan-22-yl carbonate (0.11 g, 1.5 mmol) and N,N-diisopropylethylamine (0.19 g, 1.5 mmol). The resulted solution was stirred at room temperature for three hours. A colorless oil was obtained after removing solvent. The crude product was purified by flash chromatography and 0.080 g of product was obtained after the purification (yield 60%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (48). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 9.00 (br., 1H), 7.55 (m, 1H), 7.40 (m, 1H), 7.35 (m, 2H), 7.22 (m, 1H), 7.15 (m, 1H), 6.90 (m, 1H), 6.12 (m, 1H), 5.30 (m, 1H), 4.40 (m, 1H), 4.20 (m, 3H), 3.88 (m, 1H), 3.60 (m, 28H), 3.46 (s, 3H), 3.10 (m, 2H), 2.75 (s, 3H), 2.20 (m, 3H), 1.90 (m, 1H), 1.60 (m, 1H), 1.00 (m, 6H); MS (EI) for C.sub.42H.sub.64Cl.sub.2N.sub.4O.sub.12: 887 (MH.sup.+).
(334) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 48
Preparation of 2,5,8,11,14,17,20-Heptaoxadocosan-22-yl {1-[(3-{(1S)-1-{[(3,4dichlorophenyl)acetyl](methyl)amino}-2-[(3S)-3-hydroxypyrrolidin-1yl]ethyl}phenyl)carbamoyl]cyclopropyl}carbamate, hydrochloride salt (49)
(335) Using the procedure outlined in the schematic below, the named compound was prepared.
(336) ##STR00201##
Step 1: Preparation of tert-butyl(1-((3-((S)-1-(2-(3,4-dichlorophenyl)-N-methylacetamido)-2-((S)-3-hydroxypyrrolidin-1-yl)ethyl)phenyl)carbamoyl)cyclopropyl)carbamate
(337) ##STR00202##
(338) N—((S)-1-(3-Aminophenyl)-2-((S)-3-hydroxypyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide (0.10 g, 0.24 mmol), 1-((tert-butoxycarbonyl)amino)cyclopropanecarboxylic acid (0.11 g, 0.52 mmol), and 4-dimethylaminopyridine/p-toluenesulfonic acid (1:1 salt) (0.073 g, 0.24 mmol) were dissolved in 20 mL of dichloromethane. N,N′-Diisopropylcarbodiimide (0.15 g, 1.18 mmol) was added into the mixture. The reaction mixture was stirred for three hours. Dichloromethane (100 mL) was added into the reaction mixture and the resulted solution was washed with water (100 mL×3). The solution was dried over sodium sulfate and concentrated. Crude product was dissolved in 2 mL of acetonitrile and 2 mL of 0.5 N potassium hydroxide was added. The mixture was stirred at 65° C. for two hours. Dichloromethane (100 mL) was added into the reaction mixture and the resulted solution was washed with water (100 mL×3). The solution was dried over sodium sulfate and concentrated. The product was obtained after purified by flash chromatography (0.046 g, yield: 32%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 8.55 (br., 1H), 7.56 (m, 1H), 7.42 (m, 2H), 7.30 (m, 2H), 7.20 (m, 1H), 7.02 (m, 1H), 6.05 (m, 0.83H), 5.40 (m, 1H), 5.02 (m, 0.17H), 4.40 (m, 1H), 3.72 (m, 2H), 3.15 (m, 1H), 3.06 (m, 1H), 2.75 (m, 4H), 2.68 (m, 1H), 2.35 (m, 3H), 2.15 (m, 1H), 1.72 (m, 1H), 1.65 (m, 2H), 1.50 (s, 9H), 1.20 (m, 2H); MS (EI) for C.sub.30H.sub.38Cl.sub.2N.sub.4O.sub.5: 605 (MH.sup.+).
Step 2: Preparation of 2,5,8,11,14,17,20-Heptaoxadocosan-22-yl{1-[(3-{(1S)-1-{[(3,4dichlorophenyl)acetyl](methyl)amino}-2-[(3S)-3-hydroxypyrrolidin-1yl]ethyl}phenyl)carbamoyl]cyclopropyl}carbamate, hydrochloride salt
(339) ##STR00203##
(340) Tert-butyl(1-((3-((S)-1-(2-(3,4-dichlorophenyl)-N-methylacetamido)-2-((S)-3-hydroxypyrrolidin-1-yl)ethyl)phenyl)carbamoyl)cyclopropyl)carbamate (0.045 g, 0.074 mmol) was dissolved in 2 mL of dioxane and 2 mL 4 N HCl in dioxane was added. The mixture was stirred at 20° C. for two hours. The solvent was removed and the residue (35 mg, 0.069 mmol) was dissolved in 10 mL of dichloromethane. To the solution was added 2,5-dioxopyrrolidin-1-yl 2,5,8,11,14,17,20-heptaoxadocosan-22-yl carbonate (0.040 g, 0.083 mmol) and N,N-diisopropylethylamine (0.12 mL, 0.7 mmol). The resulted solution was stirred at room temperature for three hours. Dichloromethane (100 mL) was added into the reaction mixture. The resulted solution was washed with water (100 mL), 5% sodium bicarbonate (100 mL), and then water (100 mL) again. The organic phase was dried over sodium sulfate. A colorless oil was obtained after removing the solvent. The crude product was purified by biotage and 0.015 g of product was obtained after the purification (yield: 25%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford product as hydrochloride salt (49). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 8.84 (br., 1H), 7.65 (s, 1H), 7.55 (br., 1H), 7.50 (m, 2H), 7.30 (m, 1H), 7.20 (m, 1H), 7.00 (m, 1H), 6.15 (br., 1H), 4.30 (br., 1H), 3.60 (m, 29H), 3.58 (m, 3H), 3.40 (s, 3H), 2.80 (br., 4H), 2.28 (br., 3H), 1.70 (m, 2H), 1.18 (m, 4H); MS (EI) for C.sub.41H.sub.60Cl.sub.2N.sub.4O.sub.12: 871 (MH.sup.+).
(341) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 49
Preparation of 2-(3,4-dichlorophenyl)-N-[(1S)-2-[(3S)-3-hydroxypyrrolidin-1-yl]-1-(3-{[(2-methoxyethoxy)acetyl]amino}phenyl)ethyl]-N-methylacetamidee, hydrochloride salt (50)
(342) Using the procedure outlined in the schematic below, the named compound was prepared.
(343) ##STR00204##
Step 1: Preparation of (S)-1-((S)-2-(2-(3,4-dichlorophenyl)-N-methylacetamido)-2-(3-nitrophenyl)ethyl)pyrrolidin-3-yl 2-(3,4-dichlorophenyl)acetate
(344) ##STR00205##
(345) (S)-1-((S)-2-(methylamino)-2-(3-nitrophenyl)ethyl)pyrrolidin-3-ol (0.30 g, 1.13 mmol), 2-(3,4-dichlorophenyl)acetic acid (0.46 g, 2.26 mmol), and 4-dimethylaminopyridine/p-toluenesulfonic acid (1:1 salt) (0.35 g, 1.13 mmol) were dissolved in 20 mL of dichloromethane. N,N′-Diisopropylcarbodiimide (0.71 g, 5.65 mmol) was added into the mixture. The reaction mixture was stirred for three hours. Dichloromethane (100 mL) was added into the reaction mixture and the resulted solution was washed with water (100 mL×3). The solution was dried over sodium sulfate and concentrated. Crude product was purified by flash chromatography. 0.40 g of product (yield: 55%) was obtained. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 8.15 (m, 2H), 7.65 (m, 1H), 7.54 (m, 1H), 7.40 (m, 4H), 7.13 (m, 2H), 6.10 (m, 1H), 5.20 (m, 1H), 3.75 (m, 2H), 3.55 (m, 1H), 3.12 (m, 1H), 2.95 (m, 2H), 2.85 (m, 2H), 2.75 (s, 3H), 2.66 (m, 1H), 2.45 (m, 1H), 2.21 (m, 1H), 1.72 (m, 1H); MS (EI) for C.sub.29H.sub.27Cl.sub.4N.sub.3O.sub.5: 638 (MH.sup.+).
Step 2: Preparation of (S)-1-((S)-2-(3-aminophenyl)-2-(2-(3,4-dichlorophenyl)-N-methylacetamido)ethyl)pyrrolidin-3-yl 2-(3,4-dichlorophenyl)acetate
(346) ##STR00206##
(347) (S)-1-((S)-2-(N-methyl-2,2-diphenylacetamido)-2-(3-nitrophenyl)ethyl)pyrrolidin-3-yl 2,2-diphenylacetate (0.40 g, 0.63 mmol) was dissolved in 15 mL of tetrahydrofuran. The reduction reaction was performed by H-Cube. CatCart THS01131, 10% Pd/C, Flow rate 1 mL/Min. The product was purified by flash chromatography (0.32 g, yield: 84%). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.40 (m, 4H), 7.15 (m, 3H), 6.65 (m, 3H), 6.00 (m, 0.82H), 5.20 (m, 1H), 4.95 (m, 0.18H), 3.72 (m, 4H), 3.12 (m, 1H), 2.95 (m, 2H), 2.72 (s, 3H), 2.58 (m, 2H), 2.40 (m, 1H), 2.22 (m, 1H), 1.80 (m, 1H); MS (EI) for C.sub.29H.sub.29Cl.sub.4N.sub.3O.sub.3: 608 (MH.sup.+).
Step 3: Preparation of 2-(3,4-dichlorophenyl)-N—((S)-2-((S)-3-hydroxypyrrolidin-1-yl)-1-(3-(2-(2-methoxyethoxy)acetamido)phenyl)ethyl)-N-methylacetamide, hydrochloride salt (50)
(348) ##STR00207##
(349) (S)-1-((S)-2-(3-Aminophenyl)-2-(2-(3,4-dichlorophenyl)-N-methylacetamido)ethyl)pyrrolidin-3-yl 2-(3,4-dichlorophenyl)acetate (0.12 g, 0.20 mmol), 2-(2-methoxyethoxy)acetic acid (0.032 g, 0.24 mmol), and 4-dimethylaminopyridine/p-toluenesulfonic acid (1:1 salt) (0.037 g, 0.12 mmol) were dissolved in 10 mL of dichloromethane. N,N-diisopropylcarbodiimide (0.1 mL) was added into the mixture. The reaction mixture was stirred for two hours. Dichloromethane (100 mL) was added into the reaction mixture and the resulted solution was washed with water (100 mL×3). The solution was dried over sodium sulfate and concentrated. Crude product was dissolved in 6 mL of acetonitrile/0.5 N potassium hydroxide (1:1). The mixture was stirred at 65° C. for three hours and then extracted with dichloromethane (50 mL×3). The organic solution was dried over sodium sulfate and concentrated. The crude product was purified by flash chromatography and 0.020 g (54%) product was obtained. The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford the product as a hydrochloride salt (50). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.60 (m, 2H), 7.45 (m, 2H), 7.32 (m, 1H), 7.23 (m, 1H), 7.10 (m, 1H), 6.04 (m, 0.87H), 5.20 (m, 0.13H), 4.35 (m, 1H), 4.13 (s, 2H), 3.80 (m, 4H), 3.65 (m, 2H), 3.45 (s, 3H), 3.35 (m 2H), 3.10 (m, 1H), 2.95 (m, 1H), 2.72 (m, 4H), 2.68 (m, 1H), 2.55 (m, 1H), 2.12 (m, 1H), 1.75 (m, 1H); MS (EI) for C.sub.26H.sub.33Cl.sub.2N.sub.3O.sub.5: 538 (MH.sup.+).
(350) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 50
Preparation of 2-methoxy-N-methyl-2-phenyl-N—((S)-1-phenyl-2-(pyrrolidin-1-yl)ethyl) acetamide, hydrochloride salt (53)
(351) ##STR00208##
(352) (S)—N-Methyl-1-phenyl-2-(pyrrolidin-1-yl)ethanamine (0.050 g, 0.245 mmol), 2-methoxy-2-phenylacetic acid (0.045 g, 0.270 mmol), and N,N-diisopropylethylamine (0.05 mL) were dissolved in 3 mL of acetonitrile. The mixture was stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.078 g, 0.245 mmol) was added into the solution. The reaction mixture was stirred for 16 hours and then concentrated. The residue was dissolved in dichloromethane (50 mL) and washed with water (50 mL) and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded the target compound in base form (0.030 g, yield: 35%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford the product as the hydrochloride salt (53). .sup.1H NMR (500 MHz, HCl salt in MeOD): δ 7.40 (m, 5H), 7.28 (m, 3H), 7.05 (m, 2H), 6.33 (m, 1H), 5.24 (s, 1H), 4.15 (m, 1H), 4.00 (br., 1H), 3.78 (m, 2H), 3.40 (s, 3H), 3.30 (br., 2H), 2.55 (s, 3H), 2.20 (m, 4H); MS (EI) for C.sub.22H.sub.28N.sub.2O.sub.2: 353.2 (MH.sup.+).
Example 51
Preparation of N—((S)-2-((S)-3-hydroxypyrrolidin-1-yl)-1-phenylethyl)-N-methyl-2-(4-(trifluoromethyl)phenyl)acetamide, hydrochloride salt (54)
(353) ##STR00209##
(354) (S)-1-((S)-2-(Methylamino)-2-phenylethyl)pyrrolidin-3-ol dihydrochloride (0.050 g, 0.170 mmol), 2-(4-(trifluoromethyl)phenyl)acetic acid (0.032 g, 0.155 mmol), and N,N-diisopropylethylamine (0.2 mL) were dissolved in 3 mL of acetonitrile. The mixture was stirred for 10 min. at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.060 g, 0.190 mmol) was added into the solution. The reaction mixture was stirred for 4 hours and then was concentrated. The residue was dissolved in dichloromethane (50 mL) and washed with saturated sodium bicarbonate (30 mL), brine (30 mL) and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yielded the target compound in base form (0.040 g, yield: 63%). The free base was dissolved in acetonitrile. To the solution was added 1N hydrochloride. The mixture was lyophilized to afford the product as the hydrochlorise salt (54). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.60 (m, 2H), 7.45 (m, 2H), 7.32 (m, 4.6H), 7.12 (m, 0.4H), 6.14 (m, 0.84H), 5.05 (m, 0.16H), 4.30 (m, 1H), 3.85 (m, 2H), 3.20 (m, 1H), 3.12 (m, 1H), 2.78 (m, 2H), 2.74 (s, 3H), 2.67 (m, 1H), 2.32 (m, 2H), 2.25 (m, 1H), 1.72 (m, 1H); MS (EI) for C.sub.22H.sub.25F.sub.3N.sub.2O.sub.2: 407.2 (MH.sup.+).
Example 52
Preparation of 2-(3,4-dichlorophenyl)-N—((S)-2-((S)-3-((2-(2-(2-methoxyethoxy)ethoxy)ethyl)amino)pyrrolidin-1-yl)-1-phenylethyl)-N-methylacetamide dihydrochloride salt (55)
(355) ##STR00210##
(356) 2-(3,4-Dichlorophenyl)-N—((S)-2-((S)-3-((2-(2-(2-methoxyethoxy)ethoxy)ethyl)amino)pyrrolidin-1-yl)-1-phenylethyl)-N-methylacetamide dihydrochloride salt (55) was synthesized according to the following steps.
Step 1: Preparation of (S)-2-(((benzyloxy)carbonyl)amino)-2-phenylacetic acid
(357) ##STR00211##
(358) To a solution (S)-2-amino-2-phenylacetic acid (20 g, 0.132 mol) in water (500 mL) was added sodium carbonate (27.97 g, 0.264 mol) and sodium bicarbonate (11.1 g, 0.132 mol) at ambient temperature. The mixture was stirred to get a clear solution. Acetone (40 mL) was added and the resulting slightly turbid solution was cooled (in an ice water bath) to 15-20° C. Cbz-Cl (28.15 g, 0.165 mol) was added slowly, with stirring, and the reaction mixture allowed to warm to ambient temperature. After stirring for an additional three hours, the mixture was extracted with methyl tertbutyl ether (100 mL). The pH of aqueous layer was adjusted to 2 using aqueous HCl. The resulting oil was extracted into ethyl acetate (100 mL×2). The combined organic layer was washed with water and concentrated under vacuum to get (S)-2-(((benzyloxy)carbonyl)amino)-2-phenylacetic acid (30.2 g, 80% yield) as a white solid.
Step 2: Preparation of benzyl((S)-2-((R)-3-hydroxypyrrolidin-1-yl)-2-oxo-1-phenylethyl)carbamate
(359) ##STR00212##
(360) (S)-2-(((Benzyloxy)carbonyl)amino)-2-phenylacetic acid (10.36 g, 36.3 mmol), (R)-pyrrolidin-3-ol (3.48 g, 39.93 mmol) and diisopropyl ethyl amine (14.0 g, 108.9 mmol) were dissolved in acetonitrile (80 mL). The mixture was stirred for fifteen minutes at 22-25° C. and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU) (14.0 g, 43.56 mmol) was added into the solution. The reaction mixture was stirred for one hour at 0° C. and then at four hours at ambient temperature. At the end of four hours, the reaction mixture was concentrated under reduced pressure. The residue was dissolved in dichloromethane (50 mL) and washed with brine (25 mL×2). The dichloromethane solution was dried over anhydrous sodium sulfate and concentrated. The obtained residue, when purified by column chromatography yielded ((S)-2-((R)-3-hydroxypyrrolidin-1-yl)-2-oxo-1-phenylethyl)carbamate (7.5 g, 58% yield).
Step 3: Preparation of (R)-1-((S)-2-(methylamino)-2-phenylethyl)pyrrolidin-3-ol
(361) ##STR00213##
(362) ((S)-2-((R)-3-Hydroxypyrrolidin-1-yl)-2-oxo-1-phenylethyl)carbamate (7.0 g, 19.75 mmol) was dissolved in tetrahydrofuran (55 mL). The mixture was cooled to 0° C. and LAH (3.74 g, 98.75 mmol) was added. The mixture was stirred for fifteen minutes at 0° C., followed by sixteen hours of stirring at 65° C. The reaction mass was cooled to 0° C. and 3N aq. sodium carbonate solution (150 mL) was added (cautiously) until effervescence ceased. The precipitated solid was filtered and washed with ethyl acetate (100 mL). In the filtrate, the organic layer was separated, and from the organic layer the product was extracted into 1N aq. HCl (2×25 mL). The acidic aqueous layer was washed with methyl tertbutyl ether (3×15 mL). The pH of the aqueous layer was adjusted to 9, and the product was extracted into ethyl acetate. The organic layer was washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum to obtain the crude compound. The crude compound, upon purification using column chromatography, yielded (R)-1-((S)-2-(methylamino)-2-phenylethyl)pyrrolidin-3-ol (2.5 g, 58% yield).
Step 4: Preparation of 2-(3,4-dichlorophenyl)-N—((S)-2-((R)-3-hydroxypyrrolidin-1-yl)-1-phenylethyl)-N-methylacetamide
(363) ##STR00214##
(364) To a solution of (R)-1-((S)-2-(methylamino)-2-phenylethyl)pyrrolidin-3-ol (2.5 mg, 11.35 mmol) in acetonitrile (40 mL) were added 3,4-dichlorophenyl acetic acid (2.56 g, 12.48 mmol), HOBt.H.sub.2O (1.84 g, 13.62 mmol), diisopropyl ethyl amine (2.2 g, 17.02 mmol) and EDC.HCl (3.26 g, 17.02 mmol) at ambient temperature. The mixture was stirred for two hours and acetonitrile was distilled off. The crude compound, dissolved in dichloromethane (25 mL), was washed with 10% sodium carbonate (4×25 mL), 10% ammonium chloride (4×25 mL), and brine (25 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The resultant gummy mass, upon purification by column chromatography, yielded 2-(3,4-dichlorophenyl)-N—((S)-2-((R)-3-hydroxypyrrolidin-1-yl)-1-phenylethyl)-N-methylacetamide (2.9 g, 63% yield).
Step 5: Preparation of (R)-1-((S)-2-(2-(3,4-dichlorophenyl)-N-methylacetamido)-2-phenylethyl)pyrrolidin-3-yl methanesulfonate
(365) ##STR00215##
(366) To a solution of 2-(3,4-dichlorophenyl)-N—((S)-2-((R)-3-hydroxypyrrolidin-1-yl)-1-phenylethyl)-N-methylacetamide (0.5 g, 1.23 mmol) in dichloromethane (5 mL) were added triethyl amine (1.48 mmol) and MsCl (1.35 mmol) at ambient temperature. The mixture was stirred for two hours and quenched with water (10 mL). The organic layer was separated and washed with 5% aq. ammonium chloride (5 mL×2) followed by brine (5 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum to give (R)-1-((S)-2-(2-(3,4-dichlorophenyl)-N-methylacetamido)-2-phenylethyl)pyrrolidin-3-yl methanesulfonate (0.53 g, 90% yield) as light yellow color gum.
Step 6: Preparation of N—((S)-2-((S)-3-azidopyrrolidin-1-yl)-1-phenylethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide
(367) ##STR00216##
(368) To a solution of (R)-1-((S)-2-(2-(3,4-dichlorophenyl)-N-methylacetamido)-2-phenylethyl)pyrrolidin-3-yl methanesulfonate (0.5 g, 1.03 mmol) in DMF (5 mL) was added sodium azide (0.1 g, 1.54 mmol). The mixture was heated to 60° C. for three hours. After cooling down to ambient temperature, the mixture was concentrated under reduced pressure. To the residue, water (10 mL) was added and the product was extracted into dichloromethane (10 mL×2). The combined organic layer was washed with brine (20 mL) and concentrated under vacuum to afford N—((S)-2-((S)-3-azidopyrrolidin-1-yl)-1-phenylethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide (0.39 g, 88% yield) as brown color gum.
Preparation of N—((S)-2-((S)-3-aminopyrrolidin-1-yl)-1-phenylethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide
(369) ##STR00217##
(370) A solution of N—((S)-2-((S)-3-azidopyrrolidin-1-yl)-1-phenylethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide (0.35 g, 0.81 mmol) and water (87 μL, 4.86 mmol) in tetrahydrofuran (3.5 mL) was cooled to 0° C. (in an ice-bath). PPh.sub.3 (0.425 g, 1.62 mmol) was added into the mixture, as a solid, in small portions. After the addition, the mixture was slowly warmed to ambient temperature. It was then heated to 50° C. for five hours. At the end of five hours of stirring, the mixture was concentrated under vacuum and to it water (7 mL) and dichloromethane (10 mL) were charged. The layers were separated. The pH of the aqueous layer was adjusted with 1N HCl to 2, and then washed with dichloromethane (10 mL×2). The aqueous phase was then basified, with 6N NaOH, to pH 10. The basic aqueous layer was extracted with dichloromethane (10 mL×3), and the organic layers were combined. The combined organic layer was washed with brine (10 mL), dried over anhydrous sodium sulfate and concentrated under vacuum to give N—((S)-2-((S)-3-aminopyrrolidin-1-yl)-1-phenylethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide (0.23 g, 70% yield) as light yellow color gum.
Step 8: Preparation of 2-(3,4-dichlorophenyl)-N—((S)-2-((S)-3-((2-(2-(2-methoxyethoxy)ethoxy)ethyl)amino)pyrrolidin-1-yl)-1-phenylethyl)-N-methylacetamide dihydrochloride salt (55)
(371) ##STR00218##
(372) To a solution of N—((S)-2-((S)-3-aminopyrrolidin-1-yl)-1-phenylethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide (150 mg, 0.37 mmol) in acetonitrile (3 mL) was added mPEG.sub.3-NH.sub.2 (66 mg, 0.407 mmol) and potassium carbonate (153 mg, 1.11 mmol). The mixture was heated to 50° C. for 18 hours. After 18 hours, the mixture was concentrated under vacuum and the resulting crude was dissolved in ethyl acetate (10 mL). The organic layer, after washes with aq. ammonium chloride (10 mL), water (10 mL) and brine (10 mL), was dried over anhydrous sodium sulfate and concentrated under vacuum to give the crude compound. The crude compound post purification by column chromatography afforded 2-(3,4-dichlorophenyl)-N—((S)-2-((S)-3-((2-(2-(2-methoxyethoxy)ethoxy)ethyl)amino)pyrrolidin-1-yl)-1-phenylethyl)-N-methylacetamide as the free base (0.081 g, 40% yield). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.61 (m, 1H), 7.40-7.27 (m, 7H), 5.22 (m, 1H), 4.20 (t, 2H), 3.70-3.45 (m, 10H), 3.32 (s, 3H), 3.24 (m, 2H), 3.04 (m, 1H), 2.79-2.72 (m, 2H), 2.62 (s, 3H), 2.46-2.30 (m, 4H), 1.54-1.79 (m, 2H); MS (EI) for C.sub.28H.sub.39Cl.sub.2N.sub.3O.sub.4: 553 (MH.sup.+). The free base was dissolved in 4M hydrochloride in 2-propanol. The mixture was concentrated to afford product as dihydrochloride salt (55).
(373) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 53
Preparation of 2-(2-(2-methoxyethoxy)ethoxy)ethyl((S)-1-((S)-2-(2-(3,4-dichlorophenyl)-N-methylacetamido)-2-phenylethyl)pyrrolidin-3-yl)carbamate hydrochloride salt (56)
(374) ##STR00219##
(375) 2-(2-(2-methoxyethoxy)ethoxy)ethyl((S)-1-((S)-2-(2-(3,4-dichlorophenyl)-N-methylacetamido)-2-phenylethyl)pyrrolidin-3-yl)carbamate hydrochloride salt (56) was synthesized as described below.
Step 1: Preparation of 2-(2-(2-methoxyethoxy)ethoxy)ethyl((S)-1-((S)-2-(2-(3,4-dichlorophenyl)-N-methylacetamido)-2-phenylethyl)pyrrolidin-3-yl)carbamate hydrochloride salt (56)
(376) ##STR00220##
(377) To a solution of N—((S)-2-((S)-3-aminopyrrolidin-1-yl)-1-phenylethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide (prepared in accordance with Step 7 of Example 52) (200 mg, 0.492 mmol) in dichloromethane (3 mL) was added 2,5-dioxopyrrolidin-1-yl(2-(2-(2-methoxyethoxy)ethoxy)ethyl) carbonate (165 mg, 0.54 mmol) and triethyl amine (103 μL, 0.74 mmol) at ambient temperature. The mixture was stirred for 16 hours at the same temperature. After 16 hours, the mixture was washed with water (10 mL×2) and brine (10 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum to yield 2-(2-(2-methoxyethoxy)ethoxy)ethyl((S)-1-43)-2-(2-(3,4-dichlorophenyl)-N-methylacetamido)-2-phenylethyl)pyrrolidin-3-yl)carbamate free base (225 mg, 77% yield). .sup.1H NMR (500 MHz, CDCl.sub.3): δ 8.03 (br, s, 1H), 7.6-7.27 (m, 8H), 5.22 (m, 1H), 4.20 (t, 2H), 3.80-3.45 (m, 11H), 3.37 (m, 2H), 3.30 (s, 3H), 3.04 (m, 1H), 2.79 (m, 1H), 2.72 (s, 3H), 2.60-2.3 (m, 4H), 1.94-1.70 (m, 2H); MS (EI) for C.sub.29H.sub.39Cl.sub.2N.sub.3O.sub.6: 597 (MH.sup.+). The free base was dissolved in 4M hydrochloride in 2-propanol. The mixture was concentrated to afford the product as the hydrochloride salt (56).
(378) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 54
Preparation of (S)—N-(1-(3-((N-(2,5,8,11,14,17,20-heptaoxadocosan-22-yl)sulfamoyl)amino)phenyl)-2-(pyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide hydrochloride salt (57)
(379) ##STR00221##
(380) (S)—N-(1-(3-((N-(2,5,8,11,14,17,20-heptaoxadocosan-22-yl)sulfamoyl)amino)phenyl)-2-(pyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide hydrochloride salt (57) was synthesized as given below.
Step 1: Preparation of 2,5,8,11,14,17,20-heptaoxadocosan-22-ylsulfamoyl chloride
(381) ##STR00222##
(382) To a solution of mPEG.sub.7-NH.sub.2 (2.5 g, 7.36 mmol) in dichloromethane (25 mL) was added triethyl amine (2.15 μL, 15.5 mmol). The mixture was cooled to −78° C. A solution of sulfuryl chloride (1.2 mL, 14.72 mmol) in dichloromethane (12 mL) was slowly added to the above reaction mixture, making sure temperature of reaction mixture did not increase above −50° C. After the addition, the reaction mixture was slowly warmed to ambient temperature and thereafter stirred for additional two hours. The reaction mixture was then quenched into ice-cold water and stirred for 15 minutes. The organic layer was separated, dried over anhydrous sodium sulfate, and concentrated under vacuum to give 2,5,8,11,14,17,20-heptaoxadocosan-22-ylsulfamoyl chloride (2.5 g, 77% yield) which was used directly for the next step.
Step 2: Preparation of (S)—N-(1-(3-((N-(2,5,8,11,14,17,20-heptaoxadocosan-22-yl)sulfamoyl)amino)phenyl)-2-(pyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide hydrochloride salt (57)
(383) ##STR00223##
(384) To A solution of (S)—N-(1-(3-aminophenyl)-2-(pyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide (100 mg, 0.246 mmol), triethyl amine (106 μL, 0.738 mmol) in dichloromethane (2.5 mL) was cooled to 0° C. A mixture of 2,5,8,11,14,17,20-heptaoxadocosan-22-ylsulfamoyl chloride (0.215 g, 0.492 mmol) in dichloromethane (1 mL) was slowly added to the above mixture. After the addition, the reaction mixture was slowly warmed to ambient temperature and stirred for 18 hours. The reaction mixture was washed with water (10 mL), 5% aqueous ammonium chloride (10 mL) and brine (1 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum to give a crude product. The crude product was purified by column chromatography to yield (S)—N-(1-(3-((N-(2,5,8,11,14,17,20-heptaoxadocosan-22-yl)sulfamoyl)amino)phenyl)-2-(pyrrolidin-1-yl)ethyl)-2-(3,4-dichloropheny))-N-methylacetamide (85 mg, 43% yield) a light yellow gum, as free base. .sup.1H NMR (500 MHz, CDCl.sub.3): δ 7.74 (br, s 1H), 7.61 (m, 1H), 7.25-7.15 (m, 3H), 7.10-6.95 (m, 3H), 5.11 (m, 1H), 4.0 (br, s, 1H), 3.96 (t, 2H), 3.83-3.45 (m, 24H), 3.31 (s, 3H), 3.37 (s, 2H), 3.04 (m, 1H), 2.96 (t, 2H), 2.79 (m, 1H), 2.72 (s, 3H), 2.55-2.41 (m, 2H), 2.33-2.22 (m, 2H); MS (EI) for C.sub.36H.sub.56Cl.sub.2N.sub.4O.sub.10S: 808 (MH.sup.+). The free base was dissolved in 4M hydrochloride in 2-propanol. The mixture was concentrated to afford product as hydrochloride salt (57).
(385) The example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Example 55
Radioligand Competition Binding and cAMP Accumulation Assays
(386) Radio Competition Binding Assay.
(387) The binding affinities of certain kappa agonist compounds (compounds of the present invention were tested as HCl salts) were evaluated using radioligand binding assays in membranes prepared from CHO-K1 cells expressing recombinant human kappa (KOR) or mu (MOR) opioid receptors.
(388) Competition binding experiments were conducted by incubating membrane protein to equilibrium in triplicate in the presence of a fixed concentration of radioligand and increasing concentrations of test compound for evaluation of binding to KOR or single concentration (10 μM) of test compound for evaluation of binding to MOR in 101 μL final volume. The radioligands used were specific for each receptor type, and the assay conditions are described in Table 1. Following incubations, the membranes were rapidly filtered through GF/B filter plate (presoaked with 0.5% polyethyleneimine), washed five times with cold 50 mM Tris-HCl, pH 7.5, and the bound radioactivity was then measured by liquid scintillation counting. Non-specific binding was measured in the presence of excess ligand; this value was subtracted from the total binding to yield the specific binding at each test concentration. Assay conditions are reported in Table 1 below.
(389) TABLE-US-00001 TABLE 1 Non- Receptor Membrane specific Receptor Source Protein Radioligand K.sub.d binding Methods Kappa Human 2.5 μg/well [.sup.3H] 0.3 nM U-50488 Reaction in 50 mM Tris-HCl Opioid recombinant Diprenorphine (10 μM) (pH 7.5), 5 mM MgCl.sub.2, 0.05% in CHO-K1 (1 nM) BSA at room temperature for 1 h cells with shaking Mu Opioid Human 5 μg/well [.sup.3H] Naloxone — Naloxone Reaction in 50 mM Tris-HCl recombinant (4 nM) (10 μM) (pH 7.5). 5 mM MgCl.sub.2 at room in CHO-K1 temperature for 1 h with shaking cells
(390) For KOR binding, IC.sub.50 (concentration of test compound required to inhibit 50% of specific binding) values were obtained from non-linear regression analysis of dose-response curves, using GraphPad's Prism 5.01 software, and were calculated for those compounds that showed >50% inhibition of specific binding at the highest concentration tested. K.sub.i (affinity of test compound) was obtained using the Cheng Prusoff correction using experimental K.sub.d (affinity of radioligand) values that were previously determined under these assay conditions. For MOR binding, compounds were tested at one concentration, 10 μM, to evaluate its ability to inhibit specific radioligand binding. The values are expressed as percent inhibition of specific binding and greater than 50% inhibition of binding was considered to be significant. Data are expressed as means of one experiment in triplicate determination and reported in Table 2.
(391) cAMP Accumulation Assay.
(392) Inhibition of cAMP accumulation by select compounds was measured in forskolin-stimulated CHO-K1 cells stably expressing KOR. CHO-K1 cells stably expressing KOR were harvested using Invitrogen Cell Dissociation Buffer, and then centrifuged at 1200 rpm for five minutes. The supernatant was aspirated and cells were resuspended in assay buffer to a density of 4×10.sup.5 cells/mL. Cells (25 μL) were added into a white half-area 96 well plate. Fourteen point serial dilutions of test compounds were done in assay buffer (PBS with 0.5 mM IBMX). One of ketazocine, ICI-199441, or asimadoline was used as a positive control for each assay. Compound (12.5 μL) was added to the cells in duplicate for each test concentration. The cells were then stimulated with 12.5 forskolin at a final concentration 20 μM. Cells were incubated for 45 minutes in a 37° C., 5% CO.sub.2 water jacketed incubator. CisBio HTRF cAMP assay reagent was used for cAMP quantitation. Two hours after substrate addition, signal at 665/615 nm was measured using the Perkin Elmer Victor X4 HTRF reader. Data analysis was carried out by use of GraphPad Prism, sigmoidal dose-response (variable slope) curve fitting. Certain compounds of the present invention (as HCl salts) were tested as described above. Data is reported in Table 2 below.
(393) TABLE-US-00002 TABLE 2 Compound Kappa receptor Kappa receptor Number binding Mu receptor % inhibition Mu receptor activity (not example IC50 at single concentration binding EC50 cAMP number) (nM) Ki (nM) % inhibition @ 10 μM IC50 (nM) (nM) ICI-199,441 0.35 0.08 96.3 136.6 0.05 Asimadoline 11.5 2.6 46.3 11560 0.41 U50, 488H 30.23 6.98 58.6 5829.00 1.24 1 7.5 1.7 79.1 1.2 10 82.7 19.1 82.8 3.1 3 0.13 0.03 99.8 10.6 0.03 4 8.9 2.1 82 20410 0.42 7 298.1 68.79 68.6 15620 9 5 1194 275.5 35.6 75.7 6 1105 255 20.6 31240 56 9 8974 2071 36.9 178 12 849.3 196 46.5 53340 36.12 8 1480 341.6 47.5 74.71 13 1320 304.7 69.9 11 58.12 13.41 45.2 5.02 14 0.34 0.08 99.3 0.05 15 2.33 0.54 84.1 0.42 16 62.75 14.48 27.1 15.1 17 1568 361.9 81.9 67.32 18 0.57 0.13 97.8 0.09 19 0.69 0.16 99.1 0.08 20 0.6 0.14 97.1 0.15 21 0.18 0.04 96.5 0.2 22 0.32 0.07 90.2 0.03 23 2.28 0.3 75.0 0.09 24 3592 468.5 −7.9 158300 264.60 25 31.84 4.15 57.5 4512 4.07 26 1132 147.6 35.1 38680 14.35 27 106.1 13.84 50.4 9215 4.38 28 2100 273.9 4.9 73360 147.70 29 580 110.9 30.5 25940 41.22 30 463.6 60.47 85.5 38.9 31 59.82 7.8 95.6 177.8 16.4 32 1.08 0.25 78.0 0.18 33 1237 285.4 84.1 34 106.6 24.64 17.6 13 35 24.68 5.7 30.6 2.59 36 0.43 0.1 64.9 0.029 37 98.14 22.64 57.8 4.15 38 0.49 0.06 87.2 0.03 39 18490 4268 9.6 1955 40 34.59 7.98 55.6 2.34 41 5195 1199 20.5 42 2.9 0.67 67.4 0.47 43 10.53 2.43 47.3 1.12 44 4.86 1.12 67.7 0.93 45 48.55 11.2 50.4 30.1 46 3.95 0.91 73.7 0.92 47 0.23 0.05 101 0.02 48 3.48 0.8 73.2 0.43 49 1.61 0.37 78.1 0.11 50 0.52 0.12 101 0.06 51 62.17 14.35 54.5 5.18 52 66.42 15.33 64.8 13.1 53 12.22 2.82 88 0.52 54 0.16 0.04 101.4 0.009
(394) Asimadoline, ICI-199,441 and U50,488 were assayed as known kappa opioid agonists. As is evident from Table 2, several of the tested compounds have affinity for the kappa opioid receptor. Further, for the compounds tested at the mu opioid receptor (where an IC50 was generated), there is a selectivity for the kappa opioid receptor over the mu receptor.
(395) The data in Table 2 further indicates that the tested compounds were effective in reducing cAMP in cells following KOR binding, indicating that the compounds function as agonists at the kappa opioid receptor.
Example 56
Assessment of Hepatic Clearance in Humans and Rats Using Hepatocytes
(396) All test compounds were obtained from Nektar Therapeutics (San Francisco, Calif., USA). Human and rat cryopreserved hepatocytes were purchased from Xenotech LLC (Lenexa, Kans., USA). Doxepin, acetonitrile, and formic acid were purchased from Sigma-Aldrich (St. Louis, Mo., USA).
(397) Test compounds were incubated with species-specific, thawed hepatocytes for up to 0, 30, 60, 90, and 120 minutes at 37° C., 5% CO.sub.2, and 75% relative humidity. Incubations were carried out at a final hepatocyte concentration of 0.5 million viable cells/mL and a final test article concentration of 500 nM. Final DMSO concentration was 0.01% (v/v). At each sampling time, 100 μL incubations were terminated by addition of 100 μL of ice-cold acetonitrile containing 200 nM doxepin as an internal standard and 0.1% (v/v) formic acid. Samples were immediately placed in an ice bath until centrifugation at 4,000 g for 30 min at 4° C. Sample supernatant was collected and stored at −70° C. prior to LC/MS/MS analysis. The slope of the linear regression from natural log percentage remaining of test compounds versus incubation time relationships was used in calculation of in vitro intrinsic clearance. In vivo hepatic clearance (Table 3) of test compounds was predicted from in vitro intrinsic clearance using the well-stirred model.
(398) The data in Table 3 show that hepatic clearance of all the compounds in humans is predicted to be moderate to high and to be similar to that in rats except for the test compounds 34, 35, and 36. For the test compounds 34, 35, and 36, higher hepatic clearance is predicted in humans than in rats.
(399) TABLE-US-00003 TABLE 3 Compound In vitro Hepatocyte Clearance Number Rat Human (not example CL % hep Blood CL % hep number) (mL/min/kg) flow (mL/min/kg) Blood flow ICI-199,441 44.0 73.0 19.7 85.0 Asimadoline 51.0 85.0 18.2 79.0 1 50.1 83.0 19.0 82.0 10 49.72 83 20.62 89 3 51.0 85.0 20.7 89.0 4 49 82 19.4 84 5 49.1 82 20.44 88 6 48.82 81 20.05 86 19 26.8 45 11.7 50 34 15.4 26 16.4 71 35 26.5 44 15 65 36 14.3 24 16.5 71
(400) Based on the data reported in Table 4, there is comparable in-vitro clearance in both rat and human liver hepatocytes across all the compounds relative to known kappa agonists (ICI-199,441 and Asimadoline), where higher clearance is predicted in the rat relative to human.
Prophetic Example 1
Preparation of 2-(3,4-dichlorophenyl)-N-{(1S)-1-(3-{[(2-hydroxypropyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methylacetamide, hydrochloride salt
(401) ##STR00224##
(402) 2-(3,4-Dichlorophenyl)-N-{(1S)-1-(3-{[(2-hydroxypropyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methylacetamide may be prepared according to the following steps.
Step 1: Preparation of N-{(1S)-1-(3-{[(2-{[tert-butyl(dimethyl)silyl]oxyl}propyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide
(403) ##STR00225##
(404) N-[(1S)-1-(3-Aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-2-(3,4-dichloro-phenyl)-N-methylacetamide is dissolved in dichloromethane and anhydrous pyridine (7.5 equivalents). To the ice-cold solution is added 2-{[tert-butyl(dimethyl)silyl]oxy}propane-1-sulfonyl chloride (2.0 equivalents), dropwise, maintaining the temperature less than 10° C. The yellow reaction mixture is allowed to stir at 0° C., with the color turning orange. The reaction mixture is allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the orange mixture is diluted with dichloromethane and partitioned with water. The aqueous portion is extracted with dichloromethane and washed with saturated sodium chloride. The organic portion is dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gives N-{(1S)-1-(3-{[(2-{[tert-butyl(dimethyl)silyl]oxy}propyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide.
Step 2: Preparation of 2-(3,4-dichlorophenyl)-N-{(1S)-1-(3-{[(2-hydroxypropyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methylacetamide, hydrochloride salt
(405) ##STR00226##
(406) To a solution of N-{(1S)-1-(3-{[(2-{[tert-butyl(dimethyl)silyl]oxy}propyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide in tetrahydrofuran is added tetrabutylammonium fluoride (1.2 equivalents), and the resulting solution is stirred at room temperature for one hour. The solution is diluted with water, and the mixture is extracted three times with dichloromethane. The combined organic portions are dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by chromatography to give 2-(3,4-dichlorophenyl)-N-{(1S)-1-(3-{[(2-hydroxypropyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methylacetamide. The compound is converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution is lyophilized to give the hydrochloride salt.
Prophetic Example 2
Preparation of N-[(1S)-1-(3-{[(2-hydroxypropyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl]-N-methyl-2,2-diphenylacetamide, hydrochloride salt
(407) ##STR00227##
(408) N-{(1S)-1-(3-{[(2-Hydroxypropyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide may be prepared according to the following steps.
Step 1: Preparation of N-{(1S)-1-(3-{[(2-{[tert-butyl(dimethyl)silyl]oxy}propyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide
(409) ##STR00228##
(410) N-[(1S)-1-(3-Aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-N-methyl-2,2-diphenylacetamide is dissolved in dichloromethane and anhydrous pyridine (7.5 equivalents). To the ice-cold solution is added 2-{[tert-butyl(dimethyl)silyl]oxy}propane-1-sulfonyl chloride (2.0 equivalents), dropwise, maintaining the temperature less than 10° C. The yellow reaction mixture is allowed to stir at 0° C., with the color turning orange. The reaction mixture is allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the orange mixture is diluted with dichloromethane and partitioned with water. The aqueous layer is extracted with dichloromethane and washed with saturated sodium chloride. The organic portion is dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gives N-{(1S)-1-(3-{[(2-{[tert-butyl(dimethyl)silyl]oxy}propyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide.
Step 2: Preparation of N-{(1S)-1-(3-{[(2-hydroxypropyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide
(411) ##STR00229##
(412) To a solution of N-{(1S)-1-(3-{[(2-{[tert-butyl(dimethyl)silyl]oxy}propyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide in tetrahydrofuran is added tetrabutylammonium fluoride (1.2 equivalents), and the resulting solution is stirred at room temperature for one hour. The solution is diluted with water, and the mixture is extracted three times with dichloromethane. The combined organic portions are dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue is purified by chromatography to give N-{(1S)-1-(3-{[(2-hydroxypropyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide. The compound is converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution is lyophilized to give the hydrochloride salt.
Prophetic Example 3
Preparation of 2-(3,4-dichlorophenyl)-N-{(1S)-2-[(3R)-3-hydroxypyrrolidin-1-yl]-1-[3-({[2-(2,2,2-trifluoroethoxy)ethyl]sulfonyl}amino)phenyl]ethyl}-N-methylacetamide, hydrochloride salt
(413) ##STR00230##
(414) N-{(1S)-1-(3-Aminophenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide is dissolved in dichloromethane and anhydrous pyridine (7.5 equivalents). To the ice-cold solution is added 2-(2,2,2-trifluoroethoxy)ethanesulfonyl chloride (2.0 equivalents), dropwise, maintaining the temperature less than 10° C. The yellow reaction mixture is allowed to stir at 0° C., with the color turning orange. The reaction mixture is allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the orange mixture is diluted with dichloromethane and partitioned with water. The aqueous portion is extracted with dichloromethane and washed with saturated sodium chloride. The organic portion is dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gives 2-(3,4-dichlorophenyl)-N-{(1S)-2-[(3R)-3-hydroxypyrrolidin-1-yl]-1-[3-({[2-(2,2,2-trifluoroethoxy)ethyl]sulfonyl}amino)phenyl]ethyl}-N-methylacetamide.
(415) The compound is converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution is lyophilized to give the hydrochloride salt.
Prophetic Example 4
Preparation of N-{(1S)-2-[(3R)-3-hydroxypyrrolidin-1-yl]-1-[3-({[2-(2,2,2-trifluoroethoxy)ethyl]sulfonyl}amino)phenyl]ethyl}-N-methyl-2,2-diphenylacetamide, hydrochloride salt
(416) ##STR00231##
(417) N-[(1S)-1-(3-Aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-N-methyl-2,2-diphenylacetamide is dissolved in dichloromethane and anhydrous pyridine (7.5 equivalents). To the ice-cold solution is added 2-(2,2,2-trifluoroethoxy)ethanesulfonyl chloride (2.0 equivalents), dropwise, maintaining the temperature less than 10° C. The yellow reaction mixture is allowed to stir at 0° C., with the color turning orange. The reaction mixture is allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the orange mixture is diluted with dichloromethane and partitioned with water. The aqueous layer is extracted with dichloromethane and washed with saturated sodium chloride. The organic portion is dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gives N-{(1S)-2-[(3R)-3-hydroxypyrrolidin-1-yl]-1-[3-({[2-(2,2,2-trifluoroethoxy)ethyl]sulfonyl}amino)phenyl]ethyl}-N-methyl-2,2-diphenylacetamide.
(418) The compound is converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution is lyophilized to give the hydrochloride salt
Prophetic Example 5
Preparation of N-{(1S)-1-(3-{[(4-cyanophenyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide, hydrochloride salt
(419) ##STR00232##
(420) N-{(1S)-1-(3-Aminophenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide is dissolved in dichloromethane and anhydrous pyridine (7.5 equivalents). To the ice-cold solution is added 4-cyanobenzenesulfonyl chloride (2.0 equivalents) dropwise, maintaining the temperature less than 10° C. The yellow reaction mixture is allowed to stir at 0° C., with the color turning orange. The reaction mixture is allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the orange mixture is diluted with dichloromethane and partitioned with water. The aqueous portion is extracted with dichloromethane and washed with saturated sodium chloride. The organic portion is dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gives N-{(1S)-1-(3-{[(4-cyanophenyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide.
(421) The compound is converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution is lyophilized to give the hydrochloride salt.
Prophetic Example 6
Preparation of N-{(1S)-1-(3-{[(4-cyanophenyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide, hydrochloride salt
(422) ##STR00233##
(423) N-[(1S)-1-(3-Aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-N-methyl-2,2-diphenylacetamide is dissolved in dichloromethane and anhydrous pyridine (7.5 equivalents). To the ice-cold solution is added 4-cyanobenzenesulfonyl chloride (2.0 equivalents) dropwise, maintaining the temperature less than 10° C. The yellow reaction mixture is allowed to stir at 0° C., with the color turning orange. The reaction mixture is allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the orange mixture is diluted with dichloromethane and partitioned with water. The aqueous portion is extracted with dichloromethane and washed with saturated sodium chloride. The organic portion is dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gives N-{(1S)-1-(3-{[(4-cyanophenyl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide.
(424) The compound is converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution is lyophilized to give the hydrochloride salt.
Prophetic Example 7
Preparation of N-{(1S)-1-(3-{[(6-cyanopyridin-3-yl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide, hydrochloride salt
(425) ##STR00234##
(426) N-{(1S)-1-(3-Aminophenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide is dissolved in dichloromethane and anhydrous pyridine (7.5 equivalents). To the ice-cold solution is added 6-cyanopyridine-3-sulfonyl chloride (2.0 equivalents) dropwise, maintaining the temperature less than 10° C. The yellow reaction mixture is allowed to stir at 0° C., with the color turning orange. The reaction mixture is allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the orange mixture is diluted with dichloromethane and partitioned with water. The aqueous portion is extracted with dichloromethane and washed with saturated sodium chloride. The organic portion is dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gives N-{(1S)-1-(3-{[(6-cyanopyridin-3-yl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide.
(427) The compound is converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution is lyophilized to give the hydrochloride salt.
Prophetic Example 8
Preparation of N-{(1S)-1-(3-{[(6-cyanopyridin-3-yl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide, hydrochloride salt
(428) ##STR00235##
(429) N-[(1S)-1-(3-Aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-N-methyl-2,2-diphenylacefamide is dissolved in dichloromethane and anhydrous pyridine (7.5 equivalents). To the ice-cold solution is added 6-cyanopyridine-3-sulfonyl chloride (2.0 equivalents) dropwise, maintaining the temperature less than 10° C. The yellow reaction mixture is allowed to stir at 0° C., with the color turning orange. The reaction mixture is allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the orange mixture is diluted with dichloromethane and partitioned with water. The aqueous portion is extracted with dichloromethane and washed with saturated sodium chloride. The organic portion is dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gives N-{(1S)-1-(3-{[(6-cyanopyridin-3-yl)sulfonyl]amino}phenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide.
(430) The compound is converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution is lyophilized to give the hydrochloride salt.
Prophetic Example 9
Preparation of N-{(1S)-1-[3-({[4-cyano-3-(trifluoromethyl)phenyl]sulfonyl}amino)phenyl]-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide, hydrochloride salt
(431) ##STR00236##
(432) N-{(1S)-1-(3-Aminophenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide is dissolved in dichloromethane and anhydrous pyridine (7.5 equivalents). To the ice-cold solution is added 4-Cyano-3-(trifluoromethyl)benzenesulfonyl chloride (2.0 equivalents) dropwise, maintaining the temperature less than 10° C. The yellow reaction mixture is allowed to stir at 0° C., with the color turning orange. The reaction mixture is allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the orange mixture is diluted with dichloromethane and partitioned with water. The aqueous portion is extracted with dichloromethane and washed with saturated sodium chloride. The organic portion is dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gives N-{(1S)-1-[3-({[4-cyano-3-(trifluoromethyl)phenyl]sulfonyl}amino)phenyl]-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide.
(433) The compound is converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution is lyophilized to give the hydrochloride salt.
Prophetic Example 10
Preparation of N-{(1S)-1-[3-({[4-cyano-3-(trifluoromethyl)phenyl]sulfonyl}amino)phenyl]-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide, hydrochloride salt
(434) ##STR00237##
(435) N-[(1S)-1-(3-Aminophenyl)-2-(pyrrolidin-1-yl)ethyl]-N-methyl-2,2-diphenylacetamide is dissolved in dichloromethane and anhydrous pyridine (7.5 equivalents). To the ice-cold solution is added 4-Cyano-3-(trifluoromethyl)benzenesulfonyl chloride (2.0 equivalents) dropwise, maintaining the temperature less than 10° C. The yellow reaction mixture is allowed to stir at 0° C., with the color turning orange. The reaction mixture is allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the orange mixture is diluted with dichloromethane and partitioned with water. The aqueous portion is extracted with dichloromethane and washed with saturated sodium chloride. The organic portion is dried over anhydrous sodium sulfate, filtered and concentrated to an orange residue. Purification by chromatography gives N-{(1S)-1-[3-({[4-cyano-3-(trifluoromethyl)phenyl]sulfonyl}amino)phenyl]-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide.
(436) The compound is converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution is lyophilized to give the hydrochloride salt.
Prophetic Example 11
Preparation of 2-(3,4-dichlorophenyl)-N-{(1S)-1-{3-[(2,5,8,11,14,17,20-heptaoxadocosan-22-ylcarbamoyl)amino]phenyl}-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methylacetamide, hydrochloride salt
(437) ##STR00238##
(438) To a solution of triphosgene (0.50 equivalents) in anhydrous acetonitrile, at −5° C., is added an acetonitrile solution of N-{(1S)-1-(3-aminophenyl)-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide (1.0 equivalent) and triethylamine (1.30 equivalents) over a period of 10 minutes. The mixture is stirred for an additional 20 minutes at 0° C., and then a dichloroethane solution of 2,5,8,11,14,17,20-heptaoxadocosan-22-amine and triethylamine (1.30 equivalents) is added, maintaining the temperature less than 5° C. The reaction mixture is allowed to equilibrate to room temperature overnight. After approximately 17 hours the mixture is diluted with dichloromethane (40 mL) and partitioned with water. The aqueous portion is extracted with dichloromethane three times. The combined organic portions are washed with water and saturated sodium chloride. The organic portion is dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. Purification by chromatography gives 2-(3,4-dichlorophenyl)-N-{(1S)-1-{3-[(2,5,8,11,14,17,20-heptaoxadocosan-22-ylcarbamoyl)amino]phenyl}-2-[(3R)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methylacetamide.
(439) The compound is converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution is lyophilized to give the hydrochloride salt.
(440) The prophetic example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Prophetic Example 12
Preparation of N-{1-[4-(2-hydroxyethyl)phenyl]-2-[(3S)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide, hydrochloride salt
(441) ##STR00239##
(442) N-{1-[4-(2-hydroxyethyl)phenyl]-2-[(3S)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide may be prepared according to the following steps.
Step 1: Preparation of N-(2-[(3S)-3-Hydroxypyrrolidin-1-yl]-1-{4-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]phenyl}ethyl)-N-methyl-2,2-diphenylacetamide
(443) ##STR00240##
(444) (3S)-1-[(2S)-2-(Methylamino)-2-{4-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]phenyl}ethyl]pyrrolidin-3-ol is dissolved in anhydrous dichloromethane. To the solution is added diisopropylethylamine (2.0 equivalents) at 0° C. Diphenylacetyl chloride (1.10 equivalents) is dissolved in anhydrous dichloromethane and is added dropwise to the above solution, maintaining the temperature below 5° C. The reaction mixture is allowed to equilibrate to room temperature. After approximately 17 hours at room temperature the mixture is partitioned between dichloromethane and water. The aqueous layer is extracted with dichloromethane three times. The combined organic portion is washed with saturated sodium bicarbonate, and saturated sodium chloride. The organic portion is dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue is purified by chromatography to give N-(2-[(3S)-3-hydroxypyrrolidin-1-yl]-1-{4-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]phenyl}ethyl)-N-methyl-2,2-diphenylacetamide.
Step 2: Preparation of N-{1-[4-(2-hydroxyethyl)phenyl]-2-[(3S)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide, hydrochloride salt
(445) ##STR00241##
(446) N-(2-[(3S)-3-Hydroxypyrrolidin-1-yl]-1-{4-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]phenyl}ethyl)-N-methyl-2,2-diphenylacetamide and 4-methylbenzenesulfonic acid (2.0 equivalents) are dissolved in methanol. The mixture is stirred for one hour at room temperature. After one hour the reaction mixture is diluted with dichloromethane and partitioned with water. The aqueous layer is extracted with dichloromethane three times. The combined organic portions are washed with aqueous sodium carbonate, water and saturated sodium chloride. The organic portion is dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue is purified by chromatography to give N-{1-[4-(2-hydroxyethyl)phenyl]-2-[(3S)-3-hydroxypyrrolidin-1-yl]ethyl}-N-methyl-2,2-diphenylacetamide.
(447) The compound is converted into the hydrochloride salt by dissolving the oil in acetonitrile and adding 1N hydrochloric acid. The solution is lyophilized to give the hydrochloride salt.
(448) The prophetic example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Prophetic Example 13
Preparation of 2-(3,4-Dichlorophenyl)-N-{(1S)-1-[4-(2-hydroxyethoxy)phenyl]-2-[(3S)-3-(2-hydroxyethoxy)pyrrolidin-1-yl]ethyl}-N-methylacetamide
(449) Using the procedure outlined in the schematic below, the named compound can be prepared.
(450) ##STR00242##
Step 1: Preparation of tert-butyl((1S)-2-((S)-3-(2-hydroxyethoxy)pyrrolidin-1-yl)-2-oxo-1-(4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)ethyl)carbamate
(451) ##STR00243##
(452) (S)-2-(Pyrrolidin-3-yloxy)ethanol (0.26 g, 2.00 mmol) and N,N-diisopropylethylamine (0.51 g, 4.00 mmol) are dissolved in 20 mL of dichloromethane. (2S)-2-((Tert-butoxycarbonyl)amino)-2-(4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)acetic acid (0.79 g, 2.00 mmol) is added into the solution and the mixture is cooled in an ice-bath under stirring. Finally, O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.77 g, 2.40 mmol) is added into the solution. The reaction mixture is stirred for four hours at 0° C. Dichloromethane (150 mL) is added into the mixture and resultant solution is washed with water (2×100 mL) and dried over sodium sulfate. The residue is purified by column chromatography.
Step 2: Preparation of 2-(((3S)-1-((2S)-2-(methylamino)-2-(4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)ethyl)pyrrolidin-3-yl)oxy)ethanol
(453) ##STR00244##
(454) Tert-butyl((1S)-2-((S)-3-(2-hydroxyethoxy)pyrrolidin-1-yl)-2-oxo-1-(4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)ethyl)carbamate (0.51 g, 1.00 mmol) is dissolved in tetrahydrofuran (6 mL). A 2.0 M solution of lithium aluminum hydride (2.8 mL, 5.6 mmol) is added into the solution at room temperature. The mixture is stirred at 65° C. for four hours. A 3N solution of sodium carbonate is added cautiously until effervescence ceased. Dichloromethane (50 mL) is added into the mixture. The solid is filtered out and washed with dichloromethane (100 mL). The filtrate is washed with saturated sodium chloride solution and dried over sodium sulfate. The product is obtained after removing solvent.
Step 3: Preparation 2-(3,4-dichlorophenyl)-N—((S)-1-(4-(2-hydroxyethoxy)phenyl)-2-((S)-3-(2-hydroxyethoxy)pyrrolidin-1-yl)ethyl)-N-methylacetamide
(455) ##STR00245##
(456) 2-(((3S)-1-((2S)-2-(Methylamino)-2-(4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)ethyl)pyrrolidin-3-yl)oxy)ethanol (0.082 g, 0.20 mmol), 3,4-dichlorophenylacetic acid (0.041 g, 0.20 mmol), and N,N-diisopropylethylamine (0.051 g, 0.40 mmol) are dissolved in 5 mL of acetonitrile. The mixture is stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.077 g, 0.24 mmol) is added into the solution. The reaction mixture is stirred for three hours under an ice-bath. Dichloromethane (150 mL) is added into the solution and the resultant solution is washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yield an oil, which is dissolved in 10 mL of methanol. p-Toluenesulfonic acid (0.056 g, 0.32 mmol) is added into the solution. The mixture is stirred for 60 minutes at room temperature. Dichloromethane (100 mL) is added into the solution and the solution is washed with sodium carbonate (10%), water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yield product.
(457) The third step of reaction described above may be modified to introduce various substituted phenyl (such as difluorophenyl, 4-trifluorophethyl) and heterocycles (such as pyridine, thoazole, benzofuran).
(458) The prophetic example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Prophetic Example 14
Preparation of (S)—N-(1-(3-(N-(2,5,8,11,14,17,20-heptaoxadocosan-22-yl)sulfamoyl)phenyl)-2-(pyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide hydrochloride
(459) ##STR00246##
(460) (S)—N-(1-(3-(N-(2,5,8,11,14,17,20-Heptaoxadocosan-22-yl)sulfamoyl)phenyl)-2-(pyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide hydrochloride can be synthesized as per following procedures.
Step 1: Preparation of (S)-benzyl(2-oxo-1-phenyl-2-(pyrrolidin-1-yl)ethyl)carbamate
(461) ##STR00247##
(462) To a stirred and cooled solution (0° C.) of (S)-2-(((Benzyloxy)carbonyl)amino)-2-phenylacetic acid (3 g, 10.51 mmol), pyrrolidine (0.822 g, 11.57 mmol) and diisopropyl ethyl amine (2.75 mL, 15.76 mmol) in acetonitrile (45 mL) is charged O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (TBTU) (5.06 g, 15.76 mmol). The solution is stirred for one hour at 0° C. and then for four hours at ambient temperature. A work up step comprising concentration of the reaction mixture, will give a crude product, which is dissolved in dichloromethane (50 mL). Following washes with brine (25 mL×2) and drying over anhydrous sodium sulfate, the dichloromethane layer is concentrated. The residue obtained is purified by column chromatography to afford (S)-benzyl(2-oxo-1-phenyl-2-(pyrrolidin-1-yl)ethyl)carbamate.
Step 2: Preparation of (S)—N-methyl-1-phenyl-2-(pyrrolidin-1-yl)ethanamine
(463) ##STR00248##
(464) (S)-benzyl(2-oxo-1-phenyl-2-(pyrrolidin-1-yl)ethyl)carbamate (2.5 g, 7.4 mmol) is dissolved in tetrahydrofuran (25 mL) and the solution is cooled to 0° C. To the cooled solution, LAH (1.40 g, 37 mmol) is charged. After stirring the reaction for 15 minutes at 0° C., it is warmed to 65° C. and stirred for 16 hours. After cooling the reaction mass to 0° C., 3N aq. sodium carbonate solution (75 mL) is charged to it, until effervescence ceases. Precipitated solid is filtered and the organic layer is separated (in the filtrate). The product is extracted into 1N aq. HCl (25 mL×2), form the organic phase. The combine aqueous acidic layer is washed with methyl tertbutyl ether (15 mL×3). The pH of the aqueous layer is adjusted to 9, and the product is extracted into ethyl acetate (20 mL×2). The organic layer is washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum to afford the crude compound. The crude is purified using column chromatography to afford (S)—N-methyl-1-phenyl-2-(pyrrolidin-1-yl)ethanamine.
Step 3: Preparation of (S)-(9H-fluoren-9-yl)methyl methyl(1-phenyl-2-(pyrrolidin-1-yl)ethyl)carbamate
(465) ##STR00249##
(466) (S)—N-Methyl-1-phenyl-2-(pyrrolidin-1-yl)ethanamine (2.2 g, 10.76 mmol), diisopropyl ethyl amine (1.87 mL, 10.76 mmol) are dissolved in acetonitrile (20 mL), and the solution is cooled to 0° C. FMOC-Cl (2.78 g, 10.76 mmol) is charged to the cooled solution. After the addition, the mixture is warmed to ambient temperature, and the reaction is stirred for two hours. At the end of two hours, the reaction mixture is concentrated under reduced pressure and the crude mass is dissolved in ethyl acetate (20 mL). The ethyl acetate solution, after washes with water (10 mL×2) and brine (10 mL), is dried over anhydrous sodium sulfate. Concentration of the organic layer under vacuum would affords the crude compound. The crude is purified by column chromatography to afford (S)-(9H-fluoren-9-yl)methyl methyl(1-phenyl-2-(pyrrolidin-1-yl)ethyl)carbamate.
Step 4: Preparation of (S)-(9H-fluoren-9-yl)methyl(1-(3-(chlorosulfonyl)phenyl)-2-(pyrrolidin-1-yl)ethyl)(methyl)carbamate
(467) ##STR00250##
(468) A solution of (S)-(9H-fluoren-9-yl)methyl methyl(1-phenyl-2-(pyrrolidin-1-yl)ethyl)carbamate (2.0 g, 4.67 mmol) in dichloromethane (20 mL) is cooled to −20° C. The cooled solution is charged with a solution of chlorosulfonic acid (0.655 g, 5.62 mmol) in dichloromethane (2 mL) at −20° C. (over 15 minutes). The contents are slowly warmed to ambient temperature and stirred for 16 hours at that temperature. The reaction mixture is quenched into a stirring mixture of dichloromethane (20 mL), crushed ice and water. The organic layer is separated, washed with brine, dried over anhydrous sodium sulfate and concentrated under vacuum to afford crude (S)-(9H-fluoren-9-yl)methyl(1-(3-(chlorosulfonyl)phenyl)-2-(pyrrolidin-1-yl)ethyl)(methyl)carbamate.
Step 5: Preparation of (S)-(9H-fluoren-9-yl)methyl(1-(3-(N-(2,5,8,11,14,17,20-heptaoxadocosan-22-yl)sulfamoyl)phenyl)-2-(pyrrolidin-1-yl)ethyl)(methyl)carbamate
(469) ##STR00251##
(470) A mixture of (S)-(9H-fluoren-9-yl)methyl(1-(3-(chlorosulfonyl)phenyl)-2-(pyrrolidin-1-yl)ethyl)(methyl)carbamate (1.0 g, 1.90 mmol), TEA (0.32 mL, 2.28 mmol) in DCM (10 mL) is reacted with mPEG.sub.7-NH.sub.2 (0.65 g, 1.90 mmol) at ambient temperature. After five hours of stirring, the reaction mixture is washed with H.sub.2O (10 mL×2) and brine (10 mL). The organic layer, post drying over anhydrous sodium sulfate, is concentrated under vacuum to afford the crude product. The crude product, upon purification using column chromatography affords (S)-(9H-fluoren-9-yl)methyl(1-(3-(N-(2,5,8,11,14,17,20-heptaoxadocosan-22-yl)sulfamoyl)phenyl)-2-(pyrrolidin-1-yl)ethyl)(methyl)carbamate.
Step 6: Preparation of (S)—N-(2,5,8,11,14,17,20-heptaoxadocosan-22-yl)-3-(1-(methylamino)-2-(pyrrolidin-1-yl)ethyl)benzenesulfonamide
(471) ##STR00252##
(472) To a solution of (5)-(9H-fluoren-9-yl)methyl(1-(3-(N-(2,5,8,11,14,17,20-heptaoxadocosan-22-yl)sulfamoyl)phenyl)-2-(pyrrolidin-1-yl)ethyl)(methyl)carbamate (1.00 g, 1.20 mmol) in acetonitrile, TEA (0.34 mL, 2.4 mmol) is charged at ambient temperature. The mixture would is stirred for three hours. After three hours, the reaction mixture is concentrated under vacuum. The crude product, upon purification by column chromatography, efforts (S)—N-(2,5,8,11,14,17,20-heptaoxadocosan-22-yl)-3-(1-(methylamino)-2-(pyrrolidin-1-yl)ethyl)benzenesulfonamide.
Step 7: Preparation of (S)—N-(1-(3-(N-(2,5,8,11,14,17,20-heptaoxadocosan-22-yl)sulfamoyl)phenyl)-2-(pyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide hydrochloride
(473) ##STR00253##
(474) To a solution of (S)—N-(2,5,8,11,14,17,20-heptaoxadocosan-22-yl)-3-(1-(methylamino)-2-(pyrrolidin-1-yl)ethyl)benzenesulfonamide (200 mg, 0.33 mmol) in acetonitrile (5 mL), 3,4-dichlorophenyl acetic acid (75 mg, 0.363 mmol), HOBt.H.sub.2O (53 mg, 0.396 mmol), DIPEA (115 μL, 0.66 mmol) and EDC.HCl (76 mg, 0.396 mmol), are charged at ambient temperature. The reaction mixture, after stirring for two hours, is concentrated under vacuum. The residue is dissolved in DCM (10 mL), and is washed with 10% sodium carbonate (10 mL×2), 10% NH.sub.4Cl (10 mL×2), and brine (10 mL), respectively. The organic layer, after drying over anhydrous sodium sulfate, is concentrated under vacuum. The resultant gummy mass upon purification by column chromatography affords (S)—N-(1-(3-(N-(2,5,8,11,14,17,20-heptaoxadocosan-22-yl)sulfamoyl)phenyl)-2-(pyrrolidin-1-yl)ethyl)-2-(3,4-dichlorophenyl)-N-methylacetamide free base. The free base is dissolved in 4M hydrochloride in 2-propanol and the mixture would be concentrated to afford product as hydrochloride salt.
(475) The prophetic example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Prophetic Example 15
Preparation of N-{(1S)-2-(Azetidin-1-yl)-1-[4-(2-hydroxyethoxy)phenyl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide
(476) Using the procedure outlined in the schematic below, the named compound can be prepared.
(477) ##STR00254##
Step 1: Preparation of tert-Butyl[(1S)-2-(azetidin-1-yl)-2-oxo-1-{4-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]phenyl}ethyl]carbamate
(478) ##STR00255##
(479) Azetidine (0.057 g, 1.00 mmol) and N,N-diisopropylethylamine (0.256 g, 2.00 mmol) are dissolved in 20 mL of dichloromethane. (2S)-2-((tert-butoxycarbonyl)amino)-2-(4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)acetic acid (0.395 g, 1.00 mmol) is added into the solution and the mixture is cooled in an ice-bath under stirring. Finally, 0-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborale (0.385 g, 1.20 mmol) is added into the solution. The reaction mixture is stirred for four hours at 0° C. Dichloromethane (150 mL) is added into the mixture and resultant solution is washed with water (2×100 mL) and dried over sodium sulfate. The residue is purified by column chromatography.
Step 2: Preparation of (1S)-2-(azetidin-1-yl)-N-methyl-1-(4-(2-((tetrahydro-2H-pyran-2 yl)oxy)ethoxy)phenyl)ethanamine
(480) ##STR00256##
(481) Tert-butyl al S)-2-(azetidin-1-yl)-2-oxo-1-(4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)ethyl)carbamate (0.20 g, 0.46 mmol) is dissolved in tetrahydrofuran (3 mL). A 2.0 M solution of lithium aluminum hydride (“LAM”) (1.5 mL, 3.0 mmol) is added into the solution at room temperature. The mixture is stirred at 65° C. for four hours. A 3N solution of sodium carbonate is added cautiously until effervescence ceases. Dichloromethane (50 mL) is added into the mixture. The solid is filtered out and washed with dichloromethane (100 mL). The filtrate is washed with saturated sodium chloride solution and dried over sodium sulfate. The product is obtained after removing solvent.
Step 3: Preparation of N-{(1S)-2-(Azetidin-1-yl)-1-[4-(2-hydroxyethoxy)phenyl]ethyl}-2-(3,4-dichlorophenyl)-N-methylacetamide
(482) ##STR00257##
(483) (1S)-2-(Azetidin-1-yl)-N-methyl-1-(4-(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)phenyl)ethanamine (0.20 g, 0.60 mmol), 3,4-dichlorophenylacetic acid (0.120 g, 0.60 mmol), and N,N-diisopropylethylamine (0.15 g, 1.20 mmol) are dissolved in 5 mL of acetonitrile. The mixture is stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (0.23 g, 0.72 mmol) is added into the solution. The reaction mixture is stirred for three hours under an ice-bath. Dichloromethane (150 mL) is added into the solution and the resultant solution is washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yield an oil, which is dissolved in 10 mL of methanol. P-Toluenesulfonic acid (0.20 g, 1.2 mmol) is added into the solution. The mixture is stirred for sixty minutes at room temperature. Dichloromethane (100 mL) is added into the solution and the solution is washed with sodium carbonate (10%), water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yield product.
(484) The example described above may be modified to introduce various substituted azetidines (such as azetidin-3-ol, azetidin-3-amine, and azetidin-3-one) and heterocycles to replace dichlorophenyl (such as pyridine, thoazole, benzofuran).
(485) The prophetic example described above may be modified to introduce oligomers of various lengths as disclosed herein.
Prophetic Example 16
Preparation of 2-(3,4-dichlorophenyl)-N-methyl-N-{(1S)-1-phenyl-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]ethyl}acetamide
(486) ##STR00258##
Step 1: Preparation of 2-(2-(2-(trifluoromethoxy)ethoxy)ethoxy)ethyl methanesulfonate
(487) ##STR00259##
Step 1a: Preparation of O-(2-(2-(2-(benzyloxy)ethoxy)ethoxy)ethyl)S-methyl carbonodithioate
(488) ##STR00260##
(489) 2-(2-(2-(Benzyloxy)ethoxy)ethoxy)ethanol (T. Shiyama et al., Bioorg. Med. Chem. (2004), #12, 2831) (13.0 g, 54.1 mmol) is dissolved in 250 mL of tetrahydrofuran and sodium hydride (2.6 g of 60% in mineral oil, 65 mmol) is added into the solution. The mixture is stirred for ten minutes at room temperature and then cooled to 0° C. Carbon disulfide (4.25 mL, 70.33 mmol) is then added under stirring. After stirring for one hour, methyl iodide (4.38 mL, 70.33 mmol) is added drop wise at 0° C. The mixture is stirred for 18 hours at ambient temperature, and concentrated. The residue is dissolved in ethyl acetate (200 mL), the ethyl acetate solution is washed with water (2×300 mL), brine (300 mL) and dried over anhydrous sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yields O-(2-(2-(2-(benzyloxy)ethoxy)ethoxy)ethyl)S-methyl carbonodithioate.
Step 1b: Preparation of 2-(2-(2-(trifluoromethoxy)ethoxy)ethoxy)ethanol
(490) ##STR00261##
(491) To a cooled (to −78° C.) suspension of 1,3-dibromo-5,5-dimethylhydantoin (38.63 g, 136.35 mmol) in dichloromethane (300 mL), HF-pyridine (45.5 mL, 1600 mmol) and O-(2-(2-(2-(benzyloxy)ethoxy)ethoxy)ethyl)S-methyl carbonodithioate (15.0 g, 45.45 mmol) is added into the reaction mixture at −78° C. After the addition, the mixture is stirred at −78° C. for one hour, warmed to 0° C., and stirred for two hours. Dichloromethane (150 mL) is added into the reaction mixture. The solution is washed with water (100 mL×3), saturated sodium bisulfate and dried over anhydrous sodium sulfate. The crude product is then dissolved in methanol and hydrogenated at ambient temperature with addition of 10% Pd/C (50% wet) (1.5 g) under a hydrogen atmosphere. The reaction mixture is filtered and concentrated under vacuum to give 2-(2-(2-(trifluoromethoxy)ethoxy)ethoxy)ethanol.
Step 1c: Preparation of 2-(2-(2-(trifluoromethoxy)ethoxy)ethoxy)ethyl methanesulfonate
(492) ##STR00262##
(493) To a mixture of 2-(2-(2-(trifluoromethoxy)ethoxy)ethoxy)ethanol (8.0 g, 36.7 mmol) and TEA (6.64 mL, 47.7 mmol) in dichloromethane (150 mL), is added methane sulfonyl chloride (3.12 mL, 40.37 mmol) at 0° C. The reaction mixture is stirred at ambient temperature for one hour. Dichloromethane (150 mL) is added into the reaction mixture, is washed with water (200 mL×3) and is dried over anhydrous sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography yields 24242-(trifluoromethoxy)ethoxy)ethoxy)ethyl methanesulfonate.
Step 2: Preparation of (3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxy}ethoxy)pyrrolidine
(494) ##STR00263##
(495) (S)-tert-Butyl-3-hydroxypyrrolidine-1-carboxylate is dissolved in tetrahydrofuran and the solution is cooled to 0° C. Sodium hydride (60% in mineral oil, 1.2 equivalents) is added and the reaction mixture is stirred for thirty minutes. 2-(2-(2-(Trifluoromethoxy)ethoxy)ethoxy)ethyl methanesulfonate (1.5 equivalents), dissolved in tetrahydrofuran, is added to the above mixture while maintaining the temperature at 0° C. The reaction mixture is stirred for an additional one hour at 0° C., and then allowed to equilibrate to room temperature. After overnight stirring at room temperature, the reaction mixture is concentrated under reduced pressure. The residue is dissolved in dichloromethane and the resulting solution is washed with water. The organic portion is dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue is dissolved in dichloromethane/trifluoroacetic acid (2:1) and stirred for four hours at room temperature. The solution is concentrated under reduced pressure. The residue is dissolved in water and the pH of the solution is adjusted to 9 with sodium carbonate. The solution is saturated with sodium chloride and extracted with dichloromethane. The organic portion is dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue is purified by chromatography to give (3S)-3-(2-{2-[2(trifluoromethoxy)ethoxy]ethoxyl}ethoxy)pyrrolidine.
Step 3: Preparation of benzyl {(1S)-2-oxo-1-phenyl-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxyl}ethoxy)pyrrolidin-1-yl]ethyl}carbamate
(496) ##STR00264##
(497) (3S)-3-(2-{2-[2(Trifluoromethoxy)ethoxy]ethoxy}ethoxy)pyrrolidine (1.0 equivalent), (S)-2-(((benzyloxy)carbonyl)amino)-2-phenylacetic acid (1.0 equivalent) and diisopropylethylamine (3.0 equivalents) are dissolved in acetonitrile. The solution is stirred for 15 minutes at 22-25° C. and is then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (1.2 equivalents) is added to the solution. The reaction mixture is stirred for one hour at 0° C. and then for four hours at room temperature. The reaction mixture is concentrated under reduced pressure. The resulting residue is dissolved in dichloromethane and washed with brine. The solution is dried over anhydrous sodium sulfate and concentrated. The residue is then purified by chromatography to give benzyl {(1S)-2-oxo-1-phenyl-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxyl}ethoxy)pyrrolidin-1-yl]ethyl}carbamate.
Step 4: Preparation of (1S)—N-methyl-1-phenyl-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxyl}ethoxy)pyrrolidin-1-yl]ethanamine
(498) ##STR00265##
(499) Benzyl {(1S)-2-oxo-1-phenyl-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]ethyl}carbamate is dissolved in tetrahydrofuran. The mixture is cooled to 0° C. Lithium aluminum hydride (2.0 M in tetrahydrofuran, 5.0 equivalents) is added to the above mixture and the mixture is stirred for 15 minutes at 0° C. and then heated to 65° C. and maintained at that temperature for four hours. A 3N aqueous sodium carbonate solution is added cautiously until effervescence ceases. The solid is removed by filtration and is washed with dichloromethane. The filtrate is concentrated under reduced pressure and the residue is dissolved in dichloromethane. The solution is washed with brine, and dried over anhydrous sodium sulfate. The organic portion is filtered and concentrated under reduced pressure. The residue is purified by chromatography to give (1S)—N-methyl-1-phenyl-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]ethanamine.
Step 5: Preparation of 2-(3,4-dichlorophenyl)-N-methyl-N-{(1S)-1-phenyl-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]ethyl}acetamide
(500) ##STR00266##
(501) (1S)—N-methyl-1-phenyl-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxyl}ethoxy)pyrrolidin-1-yl]ethanamine (1.0 equivalent), 2-(3,4-dichlorophenyl)acetic acid (1.0 equivalent), and N,N-diisopropylethylamine (2.0 equivalents) are dissolved in acetonitrile. The resulting solution is stirred for 15 minutes at 22-25° C. and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (1.2 equivalents) is added to the solution. The reaction mixture is stirred for four hours at 22-25° C. and is then concentrated under reduced pressure. The residue is dissolved in dichloromethane and is washed with water. The organic portion is dried over anhydrous sodium sulfate. The organic portion is filtered and concentrated under reduced pressure. The residue is purified by chromatography to give 2-(3,4-dichlorophenyl)-N-methyl-N-{(1S)-1-phenyl-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]ethyl}acetamide.
Prophetic Example 17
Preparation of 2-(3,4-Dichlorophenyl)-N-{(1S)-1-[4-(2-hydroxyethoxy)phenyl]-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]ethyl}-N-methylacetamide
(502) ##STR00267##
Step 1: Preparation of tert-butyl {(1S)-1-(4-hydroxyphenyl)-2-oxo-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]ethyl}carbamate
(503) ##STR00268##
(504) (3S)-3-(2-{2-[2-(Trifluoromethoxy)ethoxy]ethoxyl}ethoxy)pyrrolidine (1.0 equivalent) and N,N-diisopropylethylamine (2.0 equivalents) are dissolved in dichloromethane. (S)-2-((Tert-butoxycarbonyl)amino)-2-(4-hydroxyphenyl)acetic acid (1.0 equivalent) is added to the solution and the mixture is cooled in an ice-bath under stirring. 0-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (1.2 equivalents) is added and the reaction mixture is stirred for four hours at room temperature. Dichloromethane is added and the resultant mixture is washed with water. The organic portion is dried over anhydrous sodium sulfate, and is then filtered and concentrated under reduced pressure. The residue is purified by chromatography to give tert-butyl {(1S)-1-(4-hydroxyphenyl)-2-oxo-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxyl}ethoxy)pyrrolidin-1-yl]ethyl}carbamate.
Step 2: Preparation of tert-butyl {(1S)-1-[4-(2-{[tert-butyl(dimethyl)silyl]oxy}ethoxy)phenyl]-2-oxo-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxyl}ethoxy)pyrrolidin-1-yl]ethyl}carbamate
(505) ##STR00269##
(506) To a solution of tert-butyl {(1S)-1-(4-hydroxyphenyl)-2-oxo-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]ethyl}carbamate (1.0 equivalent) and (2-bromoethoxy)(tert-butyl)dimethylsilane (5.0 equivalents) in acetone is added potassium carbonate (5.0 equivalents). The mixture is stirred at 70° C. overnight. After this period, the reaction mixture is cooled to room temperature and dichloromethane is added and the resultant mixture is washed with water. The organic portion is dried over anhydrous sodium sulfate, and is then filtered and concentrated under reduced pressure. The residue is purified by chromatography to give tert-butyl {(1S)-1-[4-(2-{[tert-butyl(dimethyl)silyl]oxy}ethoxy)phenyl]-2-oxo-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxyl}ethoxy)pyrrolidin-1-yl]ethyl}carbamate.
Step 3: Preparation of 2-(4-{(1S)-1-(Methylamino)-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]ethyl}phenoxy)ethanol
(507) ##STR00270##
(508) tert-Butyl{(1S)-1-[4-(2-{[tert-butyl(dimethyl)silyl]oxy}ethoxy)phenyl]-2-oxo-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]ethyl}carbamate (1.0 equivalent) is dissolved in tetrahydrofuran. A 2.0 M solution of lithium aluminum hydride (7.0 equivalents) is added to the solution at room temperature. The mixture is stirred at 65° C. for four hours. A 3N solution of sodium carbonate is added cautiously until effervescence ceases. Dichloromethane is added to the mixture. The solid is filtered and washed with dichloromethane. The filtrate is washed with saturated sodium chloride solution and is dried over sodium sulfate. The organic portion is filtered and concentrated under reduced pressure to give 2-(4-{(1S)-1-(methylamino)-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxyl}ethoxy)pyrrolidin-1-yl]ethyl}phenoxy)ethanol.
Step 4: Preparation of 2-(3,4-dichlorophenyl)-N-{(1S)-1-[4-(2-hydroxyethoxy)phenyl]-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxyl}ethoxy)pyrrolidin-1-yl]ethyl}-N-methylacetamide
(509) ##STR00271##
(510) 2-(4-{(1S)-1-(methylamino)-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]ethyl}phenoxy)ethanol (1.0 equivalent), 3,4-dichlorophenylacetic acid (1.0 equivalent), and N,N-diisopropylethylamine (2.0 equivalents) are dissolved in acetonitrile. The solution is stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (1.2 equivalents) is added to the solution. The reaction mixture is stirred for one hour at 0° C. and then overnight. Dichloromethane is added and the mixture is washed with water and dried over sodium sulfate. The organic portion is dried over anhydrous sodium sulfate, and is then filtered and concentrated under reduced pressure. The residue is purified by chromatography to give 2-(3,4-dichlorophenyl)-N-{(1S)-1-[4-(2-hydroxyethoxy)phenyl]-2-[(3S)-3-(2-{2-[2-(trifluoromethoxy)ethoxy]ethoxyl}ethoxy)pyrrolidin-1-yl]ethyl}-N-methylacetamide.
Prophetic Example 18
Preparation of 2-(3,4-Dichlorophenyl)-N-(2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-{4-[(2-methoxyethyl)amino]phenyl}ethyl)-N-methylacetamide
(511) ##STR00272##
(512) Using the procedure outlined in the schematic below, the named compound can be prepared.
(513) ##STR00273##
Step 1: Preparation of tert-Butyl{1-(4-hydroxyphenyl)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-2-oxoethyl}carbamate
(514) ##STR00274##
(515) [(tert-Butoxycarbonyl)amino](4-hydroxyphenyl)acetic acid (1.0 equivalent), (S)-3-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)pyrrolidine (1.0 equivalent) and DIPEA (3.0 equivalents) are dissolved in acetonitrile. The above mixture is stirred for 15 minutes at 22-25° C. and then is cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (1.2 equivalents) is added into the solution. The reaction mixture is stirred for one hour at 0° C. and then at four hours at room temperature. The reaction mixture is concentrated under reduced pressure. The resulting residue is dissolved in DCM and washed with brine. The solution is dried over anhydrous sodium sulfate and is concentrated. The obtained residue is then purified by column chromatography to give tert-butyl {1-(4-hydroxyphenyl)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-2-oxoethyl}carbamate.
Step 2: Preparation of 4-{1-[(tert-butoxycarbonyl)amino]-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-2-oxoethyl}phenyl trifluoromethanesulfonate
(516) ##STR00275##
(517) (tert-Butyl{1-(4-hydroxyphenyl)-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-2-oxoethyl}carbamate (1.0 equivalent) and cesium carbonate (1.5 equivalents) are dissolved in tetrahydrofuran. Then, 1,1,1-trifluoro-N-phenyl-N-((trifluoromethyl)sulfonyl)methanesulfonamide (1.5 equivalents) is added to the mixture and the reaction is stirred under nitrogen at 70° C. for five hours. The tetrahydrofuran is removed under reduced pressure. To the residue is added dichloromethane, followed by water. The organic portion is washed with brine and is dried over sodium sulfate. The solvent is removed and the crude product is purified by chromatography to give 4-{1-[(tert-Butoxycarbonyl)amino]-2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-2-oxoethyl}phenyl trifluoromethanesulfonate.
Step 3: Preparation of tert-butyl(2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-1-{4-[(2-methoxyethyl)amino]phenyl}-2-oxoethyl)carbamate
(518) ##STR00276##
(519) 4-{1-[(tert-Butoxycarbonyl)amino]-2-[(3S)-3-{2-[2-(2methoxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-2-oxoethyl}phenyl trifluoromethanesulfonate (1.0 equivalent) and 2-methoxyethanamine (3.0 equivalents) are dissolved in anhydrous toluene. Cesium carbonate (3.0 equivalents), di-tert-butyl(2′,4′,6′-triisopropyl-3,6-dimethoxy-[1,1′-biphenyl]-2-yl)phosphine (0.2 equivalents), and diacetoxypalladium (0.1 equivalents) are added. The reaction mixture is purged with nitrogen. The mixture is stirred at 90° C. for five hours. Water and saturated sodium chloride solution are added into the reaction mixture. The mixture is extracted with dichloromethane. The organic portion is dried over anhydrous sodium sulfate and is then concentrated under reduced pressure. The residue is purified by chromatography to give tert-butyl(2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-1-{4-[(2-methoxyethyl)amino]phenyl}-2-oxoethyl)carbamate.
Step 4: Preparation of 4-{2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-(methylamino)ethyl}-N-(2-methoxyethyl)aniline
(520) ##STR00277##
(521) tert-Butyl(2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-{4-[(2-methoxyethyl)amino]phenyl}-2-oxoethyl)carbamate (1.0 equivalent) in tetrahydrofuran is added dropwise to a stirred 2.0 M solution of lithium aluminum hydride (7.0 equivalents). The mixture is stirred for thirty minutes at room temperature and then is heated to 65° C. for four hours. A 3N solution of sodium carbonate is added cautiously until effervescence ceases. The solid is filtered out and washed with dichloromethane. The filtrate is concentrated and the residue is dissolved in dichloromethane. The resulting solution is dried over sodium sulfate. The organic portion is filtered and concentrated under reduced pressure. The residue is purified by chromatography to give 4-{2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-1-(methylamino)ethyl}-N-(2-methoxyethyl)aniline.
Step 5: Preparation of 2-(3,4-Dichlorophenyl)-N-(2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-{4-[(2-methoxyethyl)amino]phenyl}ethyl)-N-methylacetamide
(522) ##STR00278##
(523) 4-{2-[(3S)-3-{2-[2-(2-Methoxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-1-(methylamino)ethyl}-N-(2-methoxyethyl)aniline (1.1 equivalents), 2-(3,4-dichlorophenyl)acetic acid (1.0 equivalent), and N,N-diisopropylethylamine (2.2 equivalents) are dissolved in acetonitrile. The mixture is stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (1.3 equivalents) is added into the solution. The reaction mixture is stirred for four hours at 0° C. Dichloromethane is added into the solution and the solution is washed with water and dried over sodium sulfate. Evaporation of the solvent and purification of the residue by flash chromatography gives 2-(3,4-Dichlorophenyl)-N-(2-[(3S)-3-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-{4-[(2-methoxyethyl)amino]phenyl}ethyl)-N-methylacetamide.
Prophetic Example 19
Preparation of 2-(3,4-dichlorophenyl)-N-{(1S)-2-[(3S)-3-{2-[2-(2-hydroxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-[4-(2-hydroxyethoxy)phenyl]ethyl}-N-methylacetamide
(524) ##STR00279##
(525) Using the procedure outlined in the schematic below, the named compound can be prepared.
(526) ##STR00280##
Step 1: Preparation of (2S)-{[(Benzyloxy)carbonyl]aminol 14-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]phenyl}acetic acid
(527) ##STR00281##
(528) (2S)-{[(Benzyloxy)carbonyl]amino}(4-hydroxyphenyl)acetic acid is dissolved in dimethylformamide and the solution is cooled in an ice bath. Sodium hydride (60% in mineral oil, 2.2 equivalents) is added in portions. The mixture is stirred for thirty minutes at 0° C. and then 2-(2-bromoethoxy)tetrahydro-2H-pyran (1.1 equivalents) in dimethylformamide is added in portions. The reaction mixture is stirred at room temperature overnight. The reaction mixture is diluted with ice/water. The mixture is extracted with ethyl acetate. The aqueous layer is cooled in an ice bath and acidified using 1.5 M aqueous potassium hydrogen sulfate to pH 2-3. The resulting mixture is extracted with ethyl acetate. The organic portion is washed with water, brine, and dried over sodium sulfate. The organic portion is filtered and concentrated under reduced pressure to give (2S)-{[(Benzyloxy)carbonyl]amino}{4-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]phenyl}acetic acid.
Step 2: Preparation of (3S)-3-(2-{2-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]ethoxyl}ethoxy)pyrrolidin
(529) ##STR00282##
(530) Benzyl(3S)-3-hydroxypyrrolidine-1-carboxylate (6.0 g, 32.04 mmol) is dissolved in tetrahydrofuran. The solution is cooled to 0° C. and sodium hydride (60% in mineral oil, 1.2 equivalents) is added in portions. The solution is stirred at 0° C. for 30 minutes. 2-{2-[2-(Tetrahydro-2H-pyran-2-yloxy)ethoxy]ethoxyl}ethyl methanesulfonate (Lu, J. Journal of Drug Targeting, 2010, vol. 18, #7, 520-535) (1.5 equivalents) in tetrahydrofuran is added to the solution while maintaining the temperature at 0° C. The reaction mixture is stirred for additional one hour at 0° C. and is allowed to warm to 22-25° C. The reaction is then heated to 60° C. and is stirred for six hours. The reaction mixture is concentrated under reduced pressure. The residue is dissolved in dichloromethane and the resulting solution is washed with water. The organic portion is dried over anhydrous sodium sulfate and concentrated under reduced pressure. Benzyl(3S)-3-(2-{2-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]ethoxy}ethoxy)pyrrolidine-1-carboxylate is dissolved in ethanol in a hydrogenation flask, and charged with Pd/C (10%). The mixture is hydrogenated on a Parr shaker (50 psi) for 24 hours. The mixture is purged with nitrogen and filtered through a celite pad. The celite pad is further washed with more ethanol. The filtrate is concentrated under reduced pressure to give (3S)-3-(2-{2-[2-(Tetrahydro-2H-pyran-2-yloxy)ethoxy]ethoxy}ethoxy)pyrrolidine.
Step 3: Preparation of benzyl(2-oxo-2-[(3R)-3-(2-{2-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]ethoxyl}ethoxy)pyrrolidin-1-yl]-1-{4-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]phenyl}ethyl)carbamate
(531) ##STR00283##
(532) (2S)-{[(Benzyloxy)carbonyl]amino}{4-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]phenyl}acetic acid (1.0 equivalent), (3S)-3-(2-{2-[2-(Tetrahydro-2H-pyran-2-yloxy)ethoxy]ethoxy}ethoxy)pyrrolidine (1.1 equivalents), and N,N-diisopropylethylamine (2.0 equivalents) are dissolved in acetonitrile. The mixture is stirred for ten minutes at room temperature and is then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (1.2 equivalents) is added into the solution. The reaction mixture is stirred for two hours under an ice bath and the reaction is continued at room temperature for four hours. Dichloromethane is added into the reaction mixture, and the resulting solution is washed with water. The solution is dried over sodium sulfate and concentrated. The residue is purified by chromatography to give benzyl(2-oxo-2-[(3R)-3-(2-{2-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]ethoxyl}ethoxy)pyrrolidin-1-yl]-1-{4-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]phenyl}ethyl)carbamate.
Step 4: Preparation of N-methyl-2-[(3R)-3-(2-{2-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]-1-{4-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]phenyl}ethanamine
(533) ##STR00284##
(534) tert-Butyl(2-oxo-2-[(3R)-3-(2-{2-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]-1-{4-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]phenyl}ethyl)carbamate (1.0 equivalent) is dissolved in tetrahydrofuran. A 2.0 M solution of lithium aluminum hydride (6.0 equivalents) is added into the solution at room temperature. The mixture is stirred at 65° C. for four hours. A 3N solution of sodium carbonate is added cautiously until effervescence ceases. Ethyl acetate is added into the mixture. The solid is filtered out and washed with ethyl acetate. The filtrate is washed with saturated sodium chloride solution and is dried over sodium sulfate. The organic portion is filtered and concentrated under reduced pressure. The residue is purified by chromatography to give N-Methyl-2-[(3R)-3-(2-{2-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]-1-{4-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]phenyl}ethanamine.
Step 5: Preparation of 2-(3,4-dichlorophenyl)-N-{(1S)-2-[(3S)-3-{2-[2-(2-hydroxyethoxy)ethoxy]ethoxy}pyrrolidin-1-yl]-1-[4-(2-hydroxyethoxy)phenyl]ethyl}-N-methylacetamide
(535) ##STR00285##
(536) N-Methyl-2-[(3R)-3-(2-{2-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]ethoxy}ethoxy)pyrrolidin-1-yl]-1-{4-[2-(tetrahydro-2H-pyran-2-yloxy)ethoxy]phenyl}ethanamine (1.0 equivalent), 3,4-dichlorophenylacetic acid (1.0 equivalent), and N,N-diisopropylethylamine (2.0 equivalents) are dissolved in acetonitrile. The mixture is stirred for ten minutes at room temperature and then cooled to 0° C. O-(Benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate (1.2 equivalents) is added into the solution. The reaction mixture is stirred for three hours under an ice-bath. Dichloromethane is added into the solution and the resultant solution is washed with water and dried over anhydrous sodium sulfate. The organic portion is filtered and is concentrated under reduced pressure. The residue is dissolved in methanol. p-Toluenesulfonic acid (2.0 equivalents) is added into the solution. The mixture is stirred for one hour at room temperature. Dichloromethane is added into the solution and the solution is washed with sodium carbonate (10%), water and is dried over anhydrous sodium sulfate. The organic portion is filtered and concentrated under reduced pressure. The residue is purified by chromatography to give 2-(3,4-dichlorophenyl)-N-{(1S)-2-[(3S)-3-{2-[2-(2-hydroxyethoxy)ethoxy]ethoxyl}pyrrolidin-1-yl]-1-[4-(2-hydroxyethoxy)phenyl]ethyl}-N-methylacetamide.
Prophetic Example 20
In Situ Rat Brain Perfusion
(537) The in situ perfusion experiment measures the relative permeability of compounds across a model of the blood-brain barrier. In situ perfusion of opioids into rat brain is performed as described in Summerfield et al., J Pharmacol Exp Ther 322: 205-213 (2007).
(538) Adult male Sprague Dawley rats are used for the study. Rats are anaesthetized and the left common carotid artery is surgically cannulated for perfusion. Test compounds are perfused at concentrations of 5-50 μM in a Krebs Ringer perfusion buffer (pH 7.4). Atenolol and antipyrine are included as low and moderate permeability markers, respectively. At the end of a 30 second perfusion, the brains are removed, the left brain hemisphere is excised and homogenized. Test compounds are extracted and analyzed using LC-MS/MS. The brain permeability of the test compounds is calculated as follows:
P=Kin/S,
where P is the permeability in cm/s, Kin is the unidirectional transfer constant (ml/min/gram), and S is the luminal area of the brain vascular space.
(539) The relative permeability as determined in the in situ brain perfusion experiment provides information regarding the rates at which compounds enter the central nervous system from the periphery. It is used to characterize and compare the degree to which compounds of the present invention penetrate the BBB as compared to known compounds or analogs of the tested compounds.
Prophetic Example 21
Acetic Acid Writhing
(540) An analgesic assay may be used to determine whether a given compound can reduce and/or prevent visceral pain in mice. The assay utilizes CD-1 male mice (5-8 mice per group), each mouse being approximately 0.015-0.030 kg on the study day. Mice are treated according to standard protocols.
(541) Mice are given a single “pretreatment” dose of a compound of the present invention, a known compound, such as morphine (which is a known analgesic which has been shown to reduce writhing behavior in this model), or control solution (IV, SC, IP or orally) fifteen to thirty minutes prior to the administration of the acetic acid solution. The animal is given an IP injection of an irritant (acetic acid) that induces “writhing” which may include: contractions of the abdomen, twisting and turning of the trunk, arching of the back and the extension of the hindlimbs. Mice are given a single IP injection (0.1 mL/10 g bodyweight) of a 0.5% acetic acid solution. After the injection the animals are returned to their observation enclosure and their behavior is observed. Contractions are counted between 0 and 20 minutes after the injection. The animals are used once. Each test article may be dose at multiple doses to determine a dose response curve.