Phenyl-3,4-dihydroisoquinolin-2(1h)-yl-ethan-1-one derivatives as dopamine d1receptor positive allosteric modulators

Abstract

The invention provides certain (phenyl)-3,4-dihydroisoquinolin-2(1H)-yl)ethan-1-one related compounds of formula I as D1 positive allosteric modulators (PAMs), and pharmaceutical compositions thereof. The invention further provides methods of using a compound of formula I, to treat certain symptoms of dopaminergic CNS disorders including Parkinson's disease, Schizophrenia, ADHD or Alzheimer's disease.

##STR00001##

Claims

1. A compound of the formula: ##STR00061## wherein: R.sup.1 is ##STR00062## R.sup.2 is —F or —Cl; R.sup.3 is —F or —Cl; and R.sup.4 is —H or —F; provided that when R.sup.1 is ##STR00063## then R4 is —F.

2. The compound according to claim 1 of formula: wherein: ##STR00064## R.sup.1 is ##STR00065## R.sup.2 is —F or —Cl; R.sup.3 is —F or —Cl; and R.sup.4 is —H or —F; provided that when R.sup.1 is ##STR00066## then R4 is —F.

3. The compound according to claim 1 which is: ##STR00067##

4. A pharmaceutical composition comprising a compound according to claim 1, and a pharmaceutically acceptable carrier, diluent or excipient.

5. A pharmaceutical composition according to claim 4 comprising: ##STR00068## and a pharmaceutically acceptable carrier, diluent or excipient.

6-11. (canceled)

12. A method of treating a dopaminergic CNS disorder comprising administering to a patient in need thereof an effective amount of a compound which is 2-(2-chloro-6-fluoro-phenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-(3-hydroxy-3-methyl-butyl)-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethanone.

13-14. (canceled)

15. The method of claim 12, wherein the dopaminergic CNS disorder is Parkinson's disease.

16. The method of claim 12, wherein the dopaminergic CNS disorder is Alzheimer's disease.

17. A method of treating a dopaminergic CNS disorder comprising administering to a patient in need thereof a compound of claim 1 in simultaneous, separate, or sequential combination with a dopamine precursor.

18. The method of claim 17, wherein the dopaminergic CNS disorder is Parkinson's disease.

19. The method of claim 17, wherein the dopaminergic CNS disorder is Alzheimer's disease.

20. A method of treating a dopaminergic CNS disorder comprising administering to a patient in need thereof a compound of claim 1 in simultaneous, separate, or sequential combination with a dopamine agonist.

21. The method of claim 20, wherein the dopaminergic CNS disorder is Parkinson's disease.

22. The method of claim 20, wherein the dopaminergic CNS disorder is Alzheimer's disease.

Description

EXAMPLES 1 AND 2

2-(2,6-Dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 1

and

2-(2,6-Dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 2

[0167] ##STR00055##

[0168] A 1M solution of tetrabutylammonium formate in THF (0.8 mL) is added to a solution of 1-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-[(trans-2-(2-hydroxy-2-methyl-propyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]-2-(2,6-dichlorophenyl)ethanone (170 mg, 0.3 mmol) in THE (2.5 mL) and the resulting mixture is stirred for 30 min. The reaction mixture is concentrated under reduced pressure. The resulting residue is purified by flash chromatography on silica gel, eluting with a gradient of 0-65% EtOAc in hexanes with further purification by flash chromatography on silica gel, eluting with a gradient of 0-50% EtOAc in DCM, to give the title compound, 2-(2,6-Dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 2 (20 mg, 14% yield) after solvent evaporation of the desired chromatographic fractions. The mixed fractions from the additional flash chromatography are further purified by reversed phase chromatography on 18C silica gel, using a gradient of 5-95% water containing ammonium bicarbonate in acetonitrile) to give the title compound, 2-(2,6-Dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 1 (26 mg, 18% yield) after solvent evaporation of the desired chromatographic fractions. ESMS (m/z) for each: 462 (M+1). .sup.1H nmr (400 MHz, dmso-d.sub.6) Isomer 1: δ 0.66-1.01 (m, 4H), 1.08-1.27 (m, 6H), 1.40-1.89 (m, 1H), 2.00-2.08 (m, 1H), 2.66-3.04 (m, 2H), 3.20-3.32 (m, 2H), 3.63-3.75 (m, 1H), 4.09-4.31 (m, 3H), 4.38-4.50 (m, 1H), 4.98-5.25 (m, 2H), 6.88-7.16 (m, 3H), 7.33 (t, 1H), 7.48 (d, 2H). .sup.1H nmr (400 MHz, dmso-d.sub.6) Isomer 2: δ 0.66-1.01 (m, 4H), 1.06-1.31 (m, 5H), 1.40-1.89 (m, 2H), 1.99-2.05 (m, 1H), 2.67-3.00 (m, 2H), 3.24-3.32 (m, 2H), 3.66-3.77 (m, 1H), 4.09-4.01 (m, 3H), 4.40-4.49 (m, 1H), 4.98-5.25 (m, 2H), 6.92-7.15 (m, 3H), 7.34 (t, 1H), 7.48 (d, 2H).

EXAMPLES 3 AND 4

2-(2,6-Dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[cis-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 1

and

2-(2,6-Dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[cis-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 2

[0169] ##STR00056##

[0170] Using essentially the method described in Examples 1 and 2, using 1-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-[cis-2-(2-hydroxy-2-methyl-propyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]-2-(2,6-dichlorophenyl)ethenone Isomer 1 (100 mg, 173 mmol) gives the title compound 2-(2,6-Dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[cis-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 1 (61 mg, 76% yield). ESMS (m/z): 462 (M+1). .sup.1H nmr (400 MHz, dmso-d.sub.6) Isomer 1: δ 0.55 (s, 1H), 0.65 (s, 2H), 0.94-1.00 (m, 2H), 1.07-1.22 (m, 7H), 1.40-1.49 (m, 2H), 1.98-2.07 (m, 1H), 2.60-3.15 (m, 2H), 3.20-3.29 (m, 1H), 3.53-3.65 (m, 1H), 4.09-4.34 (m, 2H), 4.37-4.48 (m, 1H), 4.98-5.34 (m, 2H), 7.00-7.22 (m, 3H), 7.30-7.37 (m, 1H), 7.45-7.51 (m, 2H).

[0171] Using essentially the method described in Examples 1 and 2, using 1-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-[cis-2-(2-hydroxy-2-methyl-propyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]-2-(2,6-dichlorophenyl)ethenone Isomer 2 (60 mg, 104 mmol), gives the title compound 2-(2,6-Dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[cis-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 2, (42 mg, 87% yield). ESMS for each (m/z): 462 (M+1). .sup.1H nmr (400 MHz, dmso-d.sub.6) Isomer 1: δ 0.81 (s, 3H), 0.83-0.94 (m, 4H), 1.14-1.26 (m, 5H), 1.28-1.37 (m, 1H), 1.50 (d, 1H), 2.09-2.18 (m, 1H), 2.66-3.05 (m, 2H), 3.27-3.32 (m, 1H), 3.56-3.67 (m, 1H), 4.08-4.32 (m, 2H), 4.39-4.48 (m, 1H), 4.96-5.26 (m, 2H), 7.00-7.16 (m, 3H), 7.33 (t, 1H), 7.48 (d, 2H).

EXAMPLES 5 AND 6

2-(2,6-Dichlorophenyl)-1-[(1S,3R)-5-[4,4-difluoro-3-hydroxy-3-methyl-butyl]-3-(hydroxymethyl)-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 1

and

2-(2,6-Dichlorophenyl)-1-[(1S,3R)-5-[4,4-difluoro-3-hydroxy-3-methyl-butyl]-3-(hydroxymethyl)-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 2

[0172] ##STR00057##

[0173] A mixture of 2-(2,6-dichlorophenyl)-1-[(1S,3R)-5-[(3R)-4,4-difluoro-3-hydroxy-3-methyl-butyl]-3-(hydroxymethyl)-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethanone and 2-(2,6-dichlorophenyl)-1-[(1S,3R)-5-[(3S)-4,4-difluoro-3-hydroxy-3-methyl-butyl]-3-(hydroxymethyl)-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethanone (493 mg, 0.75 mmol) is dissolved in THE (12 mL). A 1M solution of tetrabutylammonium fluoride in THE (3 mL, 3 mmol) is added and the resulting mixture is stirred at RT for 1 h. The mixture is diluted with EtOAc and washed with saturated aqueous NaCl. The organic layer is separated, dried over Na.sub.2SO.sub.4, filtered, and the filtrate is concentrated under reduced pressure. The resulting residue is purified by flash chromatography on silica gel, using a gradient of 0-75% EtOAc in hexanes, to give a mixture of the title as a white foam (342 mg, 97% yield) after evaporation of the desired chromatographic fractions. ESMS (m/z): 470 (M+1).

[0174] The two diastereomers are further purified and separated by chiral SFC (CHIRALCEL® OD-H column, 21×250 mm), eluting with methanol:CO.sub.2 (15:85) at a flow rate of 80 mL/min and temperature of 40° C., to give Isomer 1 (156 mg, 46% yield) and Isomer 2 (141 mg, 41% yield) after solvent evaporation of the desired chromatographic fractions. [0175] Isomer 1: ESMS (m/z): 470 (M+H). Analytical SFC t.sub.R: 1.618 min (CHIRALCEL® OD-H column, 4×150 mm, 15% methanol/CO.sub.2, 5 mL/min, 225 nm). [0176] Isomer 2: ESMS (m/z): 470 (M+H). Analytical SFC t.sub.R: 2.213 min (CHIRALCEL® OD-H column, 4×150 mm, 15% methanol/CO.sub.2, 5 mL/min, 225 nm).

EXAMPLE 7

2-(2-chloro-6-fluoro-phenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-(3-hydroxy-3-methyl-butyl)-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethanone

[0177] ##STR00058##

[0178] 1-((1S,3R)-3-(((tert-Butyldiphenylsilyl)oxy)methyl)-6-fluoro-5-(3-hydroxy-3-methylbutyl)-1-methyl-3,4-dihydroisoquinolin-2(1H)-yl)-2-(2-chloro-6-fluorophenyl)ethan-1-one (5.3 g, 7.7 mmol) is dissolved in THF (129 mL) and a 1M solution of tetrabutylammonium fluoride in THF (23 mL, 23 mmol) is added. The mixture is stirred at RT for 1 h. The mixture is diluted with EtOAc, washed with saturated aqueous NaCl, and the layers are separated. The organic extract is dried over Na.sub.2SO.sub.4, filtered, and the filtrate is concentrated under reduced pressure. The resulting residue is purified by flash chromatography on silica gel, eluting with a gradient of 50-100% EtOAc in hexanes, to obtain the title compound as a white solid (2.75 g, 79% yield) after solvent evaporation of the desired chromatographic fractions. ESMS (m/z): 452 (M+1).

EXAMPLES 8 AND 9

2-(2-chloro-6-fluoro-phenyl)-1-[(1R,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 1

and

2-(2-chloro-6-fluoro-phenyl)-1-[(1R,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 2

[0179] ##STR00059##

[0180] A mixture of 1-[(1R,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]-2-(2-chloro-6-fluoro-phenyl)ethanone Isomer 1 and 1-[(1R,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]-2-(2-chloro-6-fluoro-phenyl)ethanone Isomer 2 (1.1 g, 1.5 mmol) is dissolved in THE (10 mL) and the mixture is cooled to 0° C. A 1M solution of tetrabutylammonium fluoride in THF (3 mL, 3 mmol) is added dropwise and the reaction mixture is stirred while warming to RT overnight. The reaction mixture is diluted with EtOAc and washed sequentially with water and saturated aqueous NaCl. The resulting layers are separated, and the organic layer is dried over MgSO.sub.4, filtered, and the filtrate is concentrated under reduced pressure. The resulting residue is purified by flash chromatography on silica gel, eluting with a gradient of 0-100% methyl tert-butyl ether in hexanes, to obtain a mixture of the title compounds as amber oil (490 mg, 69% yield) after solvent evaporation of the desired chromatographic fractions. ESMS (m/z): 464 (M+1).

[0181] The two diastereomers are further purified and separated by chiral SFC (PHENOMENEX® LUX® Cellulose-2 column, 21×250 mm, 20% isopropanol/CO.sub.2, 80 mL/min, 40° C.) to obtain the 2-(2-chloro-6-fluoro-phenyl)-1-[(1R,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 1 (136 mg, 28% yield; analytical HPLC t.sub.R: 2.769 min) and 2-(2-chloro-6-fluoro-phenyl)-1-[(1R,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 2 (121 mg, 25% yield; analytical HPLC t.sub.R: 3.368 min) as white solids after solvent evaporation of the desired chromatographic fractions.

[0182] Isomer 1: ESMS (m/z): 464 (M+H). .sup.1H NMR (400 MHz, CDCl.sub.3): δ 0.94-1.17 (m, 2H), 1.17-1.25 (m, 3H), 1.25-1.44 (m, 4H), 1.60 (s, 3H), 1.80-1.90 (m, 1H), 2.07-2.37 (m, 1H), 2.91-3.23 (m, 3H), 3.47-3.57 (m, 1H), 3.92 (t, J=16.1 Hz, 1H), 4.05 (s, 2H), 4.42-4.66 (m, 1H), 5.04 (q, J=7.0 Hz, 0.5H), 5.21 (q, J=7.0 Hz, 0.5H). 6.83-7.09 (m, 3H), 7.19-7.26 (m, 2H). Analytical SCF t.sub.R 2.769 min (PHENOMENEX® LUX® Cellulose-2 column, 4×150 mm, 20% isopropanol/CO.sub.2, 5 mL/min, 225 nm).

[0183] Isomer 2: ESMS (m/z): 464 (M+H). .sup.1H NMR (400 MHz, CDCl.sub.3): δ 0.71-0.81 (m, 1H), 1.22 (s, 3H), 1.24 (s, 3H), 1.33-1.44 (m, 4H), 1.48-1.59 (m, 1H), 1.59-1.69 (m, 2H), 1.69-1.81 (m, 1H), 2.87-3.08 (m, 1H), 3.08-3.31 (m, 1H), 3.47-3.57 (m, 1H), 3.70-3.95 (m, 1H), 3.97-4.11 (m, 2H), 4.43-4.60 (m, 1H), 5.01-5.26 (m, 1H), 6.84-7.08 (m, 3H), 7.20-7.26 (m, 2H). Analytical SFC t.sub.R 3.368 min (PHENOMENEX® LUX® Cellulose-2 column, 4×150 mm, 20% isopropanol/CO.sub.2, 5 mL/min, 225 nm).

EXAMPLES 10 AND 11

2-(2,6-dichlorophenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 1

and

2-(2,6-dichlorophenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 2

[0184] ##STR00060##

[0185] A mixture of 1-[(1S,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]-2-(2,6-dichlorophenyl)ethanone Isomer 1 and 1-[(1S,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]-2-(2,6-dichlorophenyl)ethanone Isomer 2 (333 mg, 0.5 mmol) is dissolved in THE (4.6 mL) and the mixture is cooled to 0° C. A 1M solution of tetrabutylammonium fluoride solution in THE (0.50 mL, 0.50 mmol) is added dropwise and the resulting reaction mixture is warmed to RT with stirring overnight. The reaction mixture is diluted with EtOAc and washed sequentially with water and saturated aqueous NaCl. The layers are separated, and the organic phase is dried over MgSO.sub.4, filtered, and the filtrated is concentrated under reduced pressure. The resulting residue is purified by flash chromatography on silica gel, eluting with a gradient of 0-100% methyl tert-butyl ether in hexanes, to obtain a mixture of the title compounds as amber oil (211 mg, 72% yield) after solvent evaporation of the desired chromatographic fractions. ESMS (m/z): 480 (M+1).

[0186] The two diastereomers are further purified and separated by chiral SFC (PHENOMENEX® LUX® Cellulose-2 column, 21×250 mm, 20% IPA/CO.sub.2, 80 mL/min, 40° C.) to obtain the title compounds 2-(2,6-dichlorophenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 1 (41 mg, 21% yield; analytical HPLC t.sub.R: 3.095 min) and 2-(2,6-dichlorophenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, Isomer 2 (47 mg, 24% yield; analytical HPLC t.sub.R: 3.793 min) as white solids after solvent evaporation of the desired chromatographic fractions.

[0187] Isomer 1: ESMS (m/z): 480 (M+H). .sup.1H NMR (400 MHz, CDCl.sub.3): δ 0.92-1.07 (m, 2H), 1.20-1.27 (m, 5H), 1.60-1.66 (d, J=6.6 Hz, 1H), 1.80-1.90 (m, 1H). 2.93-3.23 (m, 2H), 3.47-3.60 (m, 1H), 3.81-4.00 (m, 1H), 4.23 (s, 2H), 4.44-4.70 (m, 1H), 5.06 (q, J=6.7 Hz, 0.5H), 5.23 (q, J=6.7 Hz, 0.5H), 6.84-7.04 (m, 2H), 7.16-7.23 (m, 1H), 7.36 (d, J=8.1 Hz, 2H). Analytical SFC t.sub.R 3.095 min (PHENOMENEX® LUX® Cellulose-2 column, 4.6×150 mm, 20% IPA/CO.sub.2, 5 mL/min, 225 nm).

[0188] Isomer 2: ESMS (m/z): 480 (M+H). .sup.1H NMR (400 MHz, CDCl.sub.3): δ 0.70-0.83 (m, 1H), 1.09-1.18 (m, 1H), 1.23 (d, J=6.1 Hz, 1H), 1.32 (s, 3H), 1.35-1.46 (m, 4H), 1.64 (d, J=6.7 Hz, 2H), 1.69-1.82 (m, 1H), 2.90-3.10 (m, 1H), 3.16-3.37 (m, 1H), 3.50-3.61 (m, 1H), 3.70-4.00 (m, 1H), 4.23 (s, 2H), 4.48-4.63 (m, 1H), 5.09 (q, J=6.7 Hz, 0.6H), 5.24 (q, J=6.7 Hz, 0.4H). 6.85-7.05 (m, 2H), 7.15-7.23 (m, 1H), 7.36 (d, J=8.0 Hz, 2H). Analytical SFC t.sub.R 3.793 min (PHENOMENEX® LUX® Cellulose-2 column, 4.6×150 mm, 20% IPA/CO.sub.2, 5 mL/min, 225 nm).

[0189] Human D1 Receptor PAM Assay

[0190] The PAM activity of the compounds of the present invention may be measured essentially as described in Svensson et al., An Allosteric Potentiator of the Dopamine D1 Receptor Increases Locomotor Activity in Human D1 Knock-in Mices without Casusing Stereotypy or Tachyphylaxis. J. Pharmacol. Exp. Ther. (2017) 360:117-128.

[0191] More specifically, HEK293 cells that stably express the human D1 receptor (Accession number NM_000794) are generated by gene transduction using the pBABE-bleo retroviral vector and selected with Zeocin™ (InvivoGen). At approximately 80% confluency, the cells are harvested using TrypLE™ Express (Gibco), suspended in FBS plus 8% DMSO, and stored in liquid nitrogen. On the day of the assay, cells are thawed and resuspended in STIM buffer (Hanks Balanced Salt Solution supplemented with 0.1% BSA, 20 mM HEPES, 500 μM IBMX, and 100 μM ascorbic acid).

[0192] Test compound is serially diluted (1:2) with DMSO into assay plates (ProxiPlate-384 Plus, PerkinElmer) using acoustic dispensing (Labcyte) to provide 20 concentrations for full response curves. Test compound (80 nL) is added to 5 μL STIM buffer containing 2000 cells, and 5 μL of a 2×concentration dopamine solution in STIM buffer that will generate an EC20 level response (24 nM in stock solution, or 12 nM final) and a final DMSO concentration in the well of 0.8%. Plates are incubated at room temperature for a total reaction time of 60 min.

[0193] cAMP production is quantified using HTRF® detection (Cisbio) according to the manufacturer's instructions. Generally, lysis buffer containing anti-cAMP cryptate (5 μL) and D2-conjugate (from HTRF® kit)(5 μL) is added to the wells, plates are incubated for an additional 60-90 min, and the time-resolved fluorescence is detected using an EnVision™ plate reader (PerkinElmer). Fluorescence data is converted to cAMP concentrations using a cAMP standard curve and analyzing using a 4-parameter non-linear logistic equation (Genedata Screener, version 13.0.5-standard). For potentiator mode concentration-response curves, results are expressed as percent of the window between a response at EC.sub.20 concentration of dopamine alone (normalized to 0%) and the maximum response to dopamine (defined by response to 5 μM dopamine, final concentration, normalized as 100%). Absolute EC.sub.50 values are calculated based on the maximum and minimum responses of the control agonist (dopamine). The % Potentiation (% Top) is determined from the fitted top of the concentration response curve. The absolute EC.sub.50 and % Top for certain Example compounds are showed in the following Table 1:

TABLE-US-00001 TABLE 1 Abs EC.sub.50 (nM) % Top Compound (SEM, N) (SE, N) 2-(2,6-Dichlorophenyl)-1-[(1S,3R)-3- 8.28 (0.669, n = 3) 96.4 (5.10, n = 3) (hydroxymethyl)-5-[trans-2-(1- hydroxy-1-methyl-ethyl)cyclopropyl]- 1-methyl-3,4-dihydro-1H-isoquinolin- 2-yl]ethenone, Isomer 2 2-(2,6-Dichlorophenyl)-1-[(1S,3R)-5- 7.96 (1.66, n = 8) 101 (2.79, n = 8) [4,4-difluoro-3-hydroxy-3-methyl- butyl]-3-(hydroxymethyl)-1-methyl- 3,4-dihydro-1H-isoquinolin-2- yl]ethenone, Isomer 1 2-(2-chloro-6-fluoro-phenyl)-1- 13.2 (3.03, n = 3) 82.0 (6.80, n = 3) [(1R,3R)-6-fluoro-3-(hydroxymethyl)- 5-[trans-2-(1-hydroxy-1-methyl- ethyl)cyclopropyl]- 1-methyl-3,4-dihydro-1H-isoquinolin- 2-yl]ethenone, Isomer 2 2-(2,6-dichlorophenyl)-1-[(1S,3R)-6- 5.19 (0.976, n = 3) 92.7 (4.57, n = 3) fluoro-3-(hydroxymethyl)-5-[trans-(1- hydroxy-1-methyl-ethyl)cyclopropyl]- 1-methyl-3,4-dihydro-1H-isoquinolin- 2-yl]ethenone, Isomer 2 2-(2-chloro-6-fluoro-phenyl)-1- 10.2 (1.40, n = 15) 90.9 (2.69, n = 15) [(1S,3R)-6-fluoro-3-(hydroxymethyl)- 5-(3-hydroxy-3-methyl-butyl)-1- methyl-3,4-dihydro-1H-isoquinolin-2- yl]ethanone (Example 7) 2-(2,6-Dichlorophenyl)-1-[(1S,3R)- 12.1 (1.76, n = 3) 94.2 (5.95, n = 3) 3-(hydroxymethyl)-5-[trans-2-(1- hydroxy-1-methyl- ethyl)cyclopropyl]-1-methyl-3,4- dihydro-1H-isoquinolin-2- yl]ethenone, Isomer 1 2-(2,6-Dichlorophenyl)-1-[(1S,3R)- 11.2 (2.27, n = 3) 91.9 (3.95, n = 3) 3-(hydroxymethyl)-5-[cis-2-(1- hydroxy-1-methyl- ethyl)cyclopropyl]-1-methyl-3,4- dihydro-1H-isoquinolin-2- yl]ethenone, Isomer 1 2-(2,6-Dichlorophenyl)-1-[(1S,3R)- 86.7 (29.4, n = 3) 93.2 (7.36, n = 3) 3-(hydroxymethyl)-5-[cis-2-(1- hydroxy-1-methyl- ethyl)cyclopropyl]-1-methyl-3,4- dihydro-1H-isoquinolin-2- yl]ethenone, Isomer 2 2-(2,6-Dichlorophenyl)-1-[(1S,3R)- 12.7 (1.83, n = 7) 97.0 (3.44, n = 7) 5-[4,4-difluoro-3-hydroxy-3-methyl- butyl]-3-(hydroxymethyl)-1-methyl- 3,4-dihydro-1H-isoquinolin-2- yl]ethenone, Isomer 2 2-(2-chloro-6-fluoro-phenyl)-1- 33.6 (7.85, n = 3) 78.9 (1.43, n = 3) [(1R,3R)-6-fluoro-3- (hydroxymethyl)-5-[trans-2-(1- hydroxy-1-methyl- ethyl)cyclopropyl]-1-methyl-3,4- dihydro-1H-isoquinolin-2- yl]ethenone, Isomer 1 2-(2,6-dichlorophenyl)-1-[(1S,3R)- 12.3 (2.21, n = 3) 86.9 (4.11, n = 3) 6-fluoro-3-(hydroxymethyl)-5- [trans-(1-hydroxy-1-methyl- ethyl)cyclopropyl]-1-methyl-3,4- dihydro-1H-isoquinolin-2- yl]ethenone, Isomer 1

[0194] The absolute EC.sub.50 values provided for the above Example compounds in Table 1 illustrate the potentiation of human D1 receptor signaling in response to dopamine, and illustrate the activity of the compounds of Claim 1 as positive allosteric modulators of the human dopamine D1 receptor. Example compounds 1-6, and 8-11 of the present invention represent chiral compounds, and as described in the Examples herein, have been made and tested as individual stereoisomers. See Examples 1-6, 8-11, and Table 1 above. The combined data for individual stereoisomers (Abs EC.sub.50 ranging from 5.6 nM to 86.7 nM for Examples 1-6, 8-11) demonstrate that each individual stereoisomer represents a D1 positive allosteric modulator embodiment of the present invention. The characterization and determination of the absolute stereochemistry of the individual stereoisomers of the Examples provided herein is within the skill of the art for the ordinary artisan, and methods for such determinations are well known in medicinal chemistry literature (See e.g Chiral Analysis (Second Edition) Advances in Spectroscopy, Chromatography and Emerging Methods, 2018). For example, absolute configurations are commonly determined by NMR on the basis of the use of CDAs: diastereomeric derivatives involving covalent binding between the chiral auxiliary and the enantiomeric substrates adopt a preferred conformation which can be predicted on the basis of the differential shielding that is caused by an aromatic ring incorporated into the chiral discriminating reagent.

Generation of Human D1 Receptor Knock-In Mouse

[0195] A transgenic mouse in which the murine dopamine 1 (D1) receptor is replaced by its human counterpart may be generated by standard techniques (see generally Svensson et al., J. Pharmacol. Exp. Ther. (2017) 360:117-128). For example, mouse genomic fragments are subcloned from the RP23 bacterial artificial chromosome library and recloned into a PGK-neo targeting vector. The mouse open reading frame is replaced with the human D1 receptor open reading frame in exon 2. A neo selection marker upstream of exon 2 is flanked by frt sites for later removal. The flanking of exon 2 by loxP selection sites allows for the option to generate D1 knock-out mice by crossing with mice expressing the cre nuclease gene.

[0196] The C57BL/6 N embryonic stem cell line B6-3 is grown on a mitotically inactivated feeder layer of mouse embryonic fibroblasts in high-glucose DMEM with 20% FBS and 2×10.sup.6 unit/l leukemia inhibitory factor. Ten million embryonic stem cells plus 30 micrograms of linearized vector DNA are electroporated and subjected to G418 selection (200 μg/ml). Clones are isolated and analyzed by Southern blotting.

[0197] A clone containing the expected size insert is inserted into blastocysts and the resulting mice are genotyped by PCR. A male chimera is crossed with a female containing theflp nuclease gene to eliminate the selection marker. Progeny containing the human D1 receptor without the selection marker are identified by PCR. A male heterozygote is mated with female C57BL/6 mice. Male and female progeny containing the human D1 receptor are mated and homozygotes are identified by PCR. Behavior and reproduction of the homozygotes is found to be normal, and the colony is maintained in the homozygote state for succeeding generations.

Basal (Habituated) Locomotor Activity

[0198] The in vivo efficacy of the present compounds may be demonstrated to act through the D1 receptor using mouse locomotor activity. Locomotor activity is measured using an automated system to track movement in mice. Monitoring of mouse locomotor activity behaviors take place in transparent plastic shoebox cages having dimensions of 45×25×20 cm, with a 1 cm depth of wood chips for absorbent bedding, and covered with a ventilated filtered plastic cage top. Cages were placed in a rectangular frame containing a grid of 12 photocell beams in an 8×4 configuration (Kinder Scientific, Poway, CA) that is positioned 2.5 centimeters from the floor of the cage for the detection of body movements (ambulations) and recorded by computer.

[0199] Male human D1 receptor knock-in mice are placed in chambers and allowed to habituate to the chambers for 60 min. During the habituation period, the mice show decreasing locomotion over time, as expected. Following administration of a compound of the invention, animal movement is found to increase in a dose-dependent fashion.

[0200] The mice are randomly assigned to treatment groups. In the dose response study, each mouse is placed individually into one of the locomotor activity boxes for a 60 min. habituation period. The mice are then dosed orally using test compound in a 20% hydroxypropyl-betacyclodextrin vehicle and using a 10 mL/kg dose volume. After dosing, the mice are placed back into the LMA boxes and the total number of ambulations is recorded per 10 min interval for each mouse over a 60 min measurement period. Statistical analysis is carried out using one-way ANOVA followed by post-hoc analysis using Dunnett's Comparison test.

[0201] The compound of Example 7 is assayed essentially as described above and found to increase basal movement in a dose dependent manner (Table 2 below).

TABLE-US-00002 TABLE 2 Basal Locomotor Activity Example 7 (Total Ambulations for 60 min) (dose, mg/kg, PO) Means (SEM, % SE), N = 8/group 0.0 421 (138, 33%) (Vehicle - 20% hydroxypropyl- beta-cyclodextrin) 3.0 538 (101, 19%) 6.0 1111 (410, 37%) 10 1471*** (149, 10%) 30 3937**** (393, 10%) 60 4613**** (502, 11%) Statistical analysis is done on Total Ambulation data after Log10 Transformation. One-way ANOVA: ****p < 0.0001, (Dunnett's Multiple Comparison Test: compared to Vehicle Control on log10 transformed data: ***p < 0.001, ****p < 0.0001)

[0202] The Basal Locomotor Activity data for Example 7 shown in Table 2 illustrates that compounds of the invention, and Example 7 in particular, are effective in locomotor activation of animals that are habituated to the environment. This activity is believed to be the result of central activation of D1 receptors via allosteric potentiation (See e.g. Svensson et al., J. Pharmacol. Exp. Ther. (2017) 360:117-128). The data provided in Table 2 for Examples 7 illustrate the pharmacologically advantageous in vivo efficacy of the compounds of the invention for the potentiation of endogenous dopamine mediated responses. The data provided in Table 2 for Examples 7, further illustrates the pharmacologically advantageous oral bioavailabilitiy of Examples 7 and the compounds of formula I.

Plasma and Brain Levels

[0203] Example 7, 2-(2-chloro-6-fluoro-phenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-(3-hydroxy-3-methyl-butyl)-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, was orally dosed to male mouse from 3 mg/kg to 60 mg/kg in fed condition, and the plasma and brain concentration was determined 1 hr post-dose. The fraction unbound of the compound was determined in vitro as described previously (Zamek-Gliszczynski M J, et al., Validation of 96-well equilibrium dialysis with non-radiolabeled drug for definitive measurement of protein binding and application to clinical development of highly-bound drugs, J. Pharm. Sci. (2011) 100: 2498-2507). The ratio (Kpuu) of unbound brain concentration (Cu, brain) vs. unbound plasma concertation (Cu, plasma) was determined as described previously (Raub T J, et al., Brain Exposure of Two Selective Dual CDK4 and CDK6 Inhibitors and the Antitumor Activity of CDK4 and CDK6 Inhibition in Combination with Temozolomide in an Intracranial Glioblastoma Xenograft. Drug Metab. Dispos. (2015) 43:1360-71). The data presented below in Table 3 for Example 7 are averages from 3 animals at each dose. “Con.” refers to concentration.

TABLE-US-00003 TABLE 3 Plasma Brain Cu, Cu, Dose Time con. con. fu, fu, plasma brain mg/kg hr nM nM plasma brain nM nM Kpuu 3 1 353 18 0.0141 0.0249 4.97 0.448 0.126 6 1 682 45.1 0.0141 0.0249 9.62 1.12 0.143 10 1 1210 84.9 0.0141 0.0249 17.1 2.11 0.122 30 1 4980 557 0.0141 0.0249 70.1 13.9 0.194 60 1 9080 1290 0.0141 0.0249 128 32.1 0.248

[0204] Compounds of the invention, for instance Example 7, 2-(2-chloro-6-fluoro-phenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-(3-hydroxy-3-methyl-butyl)-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, show an advantageous combination of pharmacological properties, such as potentiation of human D1 receptor signaling in response to dopamine, high oral in vivo availability, central nervous system availability, and in vivo efficacy in locomotor activation of animals that are habituated to the environment. For instance Example 7 demonstrates potentiation of human D1 receptor signaling in response to dopamine (10.2±1.40 nM (n=15)), and significant in vivo efficacy when orally administered at 10, 30, and 60 mg/kg PO, in locomotor activation of human D1 receptor knock-in mice that are habituated to the environment, illustrating the favorable oral bioavailability of this compound. Further, Example 7 is generally well tolerated when administered in vivo to normal rats over a broad dose range, and shows an advantageous lack of toxicity in this in vivo experiment. Thus, Example 7 demonstrates an advantageous combination of favorable pharmacological properties supporting possible use as an orally administered therapeutic agent for dopamine D1 receptor potentiaion, and treatment for Parkinson's disease, Schizophrenia, ADHD, and/or Alzheimer's disease.

Determination of Fraction of Compound Cleared Through CYP3A4 (fm.sub.CYP3A4) Metabolism

[0205] The fraction of overall clearance of a drug via oxidative metabolism by the cytochrome P450 (CYP) is an important indication for potential victim drug-drug interaction mediated adverse effects. (see generally Ogu, Chris C., and Maxa, Jan L., Drug interactions due to cytochrome P450, BUMC Proceedings 2000; 13:421-423.) The greater fraction of overall clearance for a given drug that goes though a CYP oxidation pathway, particularly exclusively through a single CYP, as for example CYP3A4, the greater the potential that the drug may be found susceptible to undesired victim drug-drug interactions when used in therapy. To determine the fm.sub.CYP3A4 (the fraction of metabolism via CYP3A4), the fraction of P450-mediated oxidation, (fm.sub.CYP) is first determined in human hepatocytes. The relative contribution of CYP3A4 mediated oxidation (fm.sub.CYP3A4_RCP) over other P450 s is then determined using a recombinant CYP phenotyping assay (RCP). The fm.sub.CYP3A4 is calculated using the following equation:

fm.sub.CYP3A4=fm.sub.CYP×fm.sub.CYP3A4_RCP

Determination of Fraction of Cytochrome P450 (CYP)-Mediated Oxidative Metabolism (fm.sub.CYP) in Human Hepatocytes

[0206] The relative contribution of P450 s in the overall metabolism of a compound was determined from the clearance of the drug in an assay using human hepatocytes in the presence and absence of the pan P450 inhibitor 1-aminobenzotriazole (ABT). The intrinsic clearance was determined in cryopreserved human hepatocytes essentially as described in McGinnity D F, et al. (2004) Evaluation of fresh and cryopreserved hepatocytes as in vitro drug metabolism tools for the prediction of metabolic clearance. Drug Metab Dispos 32:1247-1253. Generally, the assay incubations contain 0.3 μM test compound and 10.sup.6 cells/mL hepatocytes, with or without 0.5 hr. pre-incubation with ABT (1 mM). Parent compound loss is measured by LC/MS after 15, 30, 60 and 90 min. incubation. The intrinsic clearance (μL/min/10.sup.6 cells) is calculated using the following equation for both ±ABT. The fraction of metabolism by CYP is the percentage of inhibition by ABT.

CL.sub.int=−k.sub.dep×incubation volume/10.sup.6 cells,

in which the k.sub.dep, the substrate depletion constant (min.sup.−1), is the slope determined using linear regression from the log transformed % remaining on the y-axis vs time on the x-axis (min.sup.−1).

Determination of Fraction of CYP3A4-Mediated Metabolism Using a Recombinant CYP Phenotyping Assay (Fm.SUB.CYP3A4_RCP.)

[0207] The relative fraction of a compound's metabolism contributed by a given CYP, as for example CYP3A4, of the total P450 metabolism of the compound may be determined as follows:

[0208] A panel of 9 human recombinant CYPs (rCYPs) provided as Supersomes (BD Gentest, Woburn, MA), including rCYPs 1A2, 2B6, 2C8, 2C9, 2C19, 2D6, 2J2, 3A4, and 3A5, are evaluated for compound metabolism (see generally Cannady E A, et al. (2015) Evacetrapib: in vitro and clinical disposition, metabolism, excretion, and assessment of drug interaction potential with strong CYP3A and CYP2C8 inhibitors. Pharmacol Res Perspect 3:e00179). Incubations and calculations are performed essentially as described in Wickremsinhe E R, et al. (2014), Disposition and metabolism of LY2603618, a Chk-1 inhibitor following intravenous administration in patients with advanced and/or metastatic solid tumors. Xenobiotica 44:827-841, except that incubations are carried out for 2 hr., given the low turnover rate of the test compounds. Intrinsic clearance (CL.sub.int) in rCYPs is scaled to human liver microsme (HLM-scaled CL.sub.int) according to equation below:

HLM-scaled CL.sub.int=−k.sub.dep×(incubation volume/pmol rCYP)×RAF

in which k.sub.dep is the substrate depletion rate constant (min.sup.−1), pmol rCYP is the lot-specific amount of rCYP in the incubation, and relative activity factor (RAF) is a relative activity factor (pmol/mg) appropriate for the rCYPs/HLMs pair. The rate constant is determined in-house by experiment, the slope determined using linear regression from the log transformed % remaining on the y-axis vs time on the x-axis (min.sup.−1). pmol rCYP and RAF are vender supplied constenats for the supersome.

[0209] The fraction of CYP3A4-mediated metabolism (fm.sub.CYP3A4_RCP) is determined by dividing the HLM-scaled CL.sub.int of CYP3A4 by the sum of the HLM-scaled CL.sub.1n.sub.t for each P450 subtype assayed (Cannady et al., 2015).

Hepatocyte Metabolite Profile

[0210] The hepatocyte metabolic profile is determined to confirm the formation of metabolites via P450-mediated oxidative and Phase II enzyme (e.g., UDP-glucuronosyltransferases and sulfotransferases) mediated non-oxidative pathways. It is determined in human hepatocytes essentially described in Zhou X, et al. (2016), Difference in the Pharmacokinetics and Hepatic Metabolism of Antidiabetic Drugs in Zucker Diabetic Fatty and Sprague-Dawley Rats. Drug Metab Dispos 44:1184-1192). In short, incubations are performed in a CO.sub.2 incubator at 37° C. using a 24-well plate containing 250,000 cells/well. A stock solution of test compound is added to medium to give a final incubation concentration of 2 μM. The incubations including the media and cells are quenched with an equal volume of acetonitrile after 4 hr. Samples are processed and analyzed using LC/MS essentially as presented in Zhou et al. 2016.

Calculation of Fraction of Compound Cleared Through CYP3A4 Metabolism (Fm.SUB.CYP3A4.)

[0211] The renal and biliary excretion of test compounds is tested in bile-duct cannulated dogs essentially according to Burkey J L, et al. (2002) Disposition of LY333531, a selective protein kinase C beta inhibitor, in the Fischer 344 rat and beagle dog. Xenobiotica 32:1045-1052. The compound of Example 7 is tested essentially as described and its renal and biliary excretion is found to be negligible. The fraction of compound cleared through CYP3A4 metabolism is calculated using the equation below:

fm.sub.CYP3A4=fm.sub.CYP×fm.sub.3A4_RCP

[0212] where fm.sub.CYP is the fraction of compound metabolized via CYP measured in hepatocyte CL.sub.int±ABT assay

[0213] fm.sub.3A4_RCP is the fraction of compound metabolized via CYP3A4 in recombinant CYP phenotyping assay

Determination of Fraction Escaping First-Pass Gut Metabolism (F.SUB.G.)

[0214] The estimation of fraction metabolized in the gut may be determined using the following equation found in Yang J, et al. (2007) Prediction of intestinal first-pass drug metabolism. Curr Drug Metab 8:676-684:

F.sub.G=Q.sub.gut/(Q.sub.gut+fu.sub.gut×Cl.sub.int CYP3A,gut)

Q.sub.gut=Q.sub.villi×Cl.sub.perm/(Q.sub.villi+Cl.sub.perm)

Cl.sub.perm═P.sub.app×Intestinal surface area

Cl.sub.int,CYP3A,gut=CL.sub.int,CYP3A,liver/fu.sub.mic×microsomal protein in gut (mg)×0.4 (in house calculation)

[0215] Where Q.sub.gut is a hybrid flow term dependent upon the villous blood flow and permeability of the compound

[0216] fu.sub.gut is the fraction of drug unbound in the enterocyte

[0217] Q.sub.villi is villous blood flow

[0218] Cl.sub.perm is the clearance term defining permeability through the enterocyte

[0219] P.sub.app is the passive permeability measured in MDCKII cell line in house

[0220] fu.sub.mic is the fraction of drug unbound in human liver microsomes

[0221] Human intestine parameters are from Yang et al., 2007, and Gertz M, et al. (2010) Prediction of human intestinal first-pass metabolism of 25 CYP3A substrates from in vitro clearance and permeability data. Drug Metab Dispos 38:1147-1158.

Mechanistic Static Model for Estimation of Victim DDI by CYP3A4 Inhibition

[0222] The potential change in drug AUC exposure magnitude of test compounds in the presence of itraconazole may be determined by employing a mechanistic static model essentially as described in Han B, et al. (2013) Prediction of CYP3A Mediated Drug-Drug Interactions: Estimation of Gut Wall and Hepatic Contributions. ASCPT Annual Meeting, Indianapolis, IN:

AUC.sub.PO,inh/AUC.sub.PO=[1/(A×fm+(1−fm))]×[1/(X×(1−F.sub.G)+F.sub.G)]

A=1/(1+[I].sub.h/Ki)

X=1/(1+[I].sub.gut/Ki)

[0223] Where fm is fraction metabolized by hepatic CYP3A4

[0224] F.sub.G is fraction that escapes intestinal metabolism in enterocytes

[0225] Ki is the dissociation constant of itraconazole from the enzyme

[0226] [I].sub.h is the concentration of itraconazole in the liver

[0227] [I].sub.gut is the concentration of itraconazole in gut

[0228] The parameters of itraconazole as CYP3A4 inhibitor (A=0.10 and X=0.10) are from Olkkola K T, et al. (1994) Midazolam should be avoided in patients receiving the systemic antimycotics ketoconazole or itraconazole. Clin Pharmacol Ther 55:481-485.

[0229] The compound of Examples 7, representative of compounds of intervention is tested essentially as described above and is found to be primarily metabolized through non-oxidative processes in the hepatocytes as opposed to through oxidative metabolism by P450 (fractions metabolized by CYPs being 32.3%). The compound was not examined in RCP assay, but the worst-case senarior of 100% metabolized by CYP3A4 was assumed for predicting AUC ratio in the presence of intraconazole. The predicted AUC ratio was less than 2. Thus, the compounds of the invention are believed to present minimal victim drug-drug interaction risk via any CYP, including CYP3A4 (See Table 4.).

TABLE-US-00004 TABLE 4 fm.sub.CYP3A4, F.sub.G and predicted AUC ratio for the compounds of Ex. 7 Compound Ex. 7 fm.sub.CYP 0.322 fm.sub.3A4.sub.—.sub.RCP 1 fm.sub.CYP3A4 0.322 CL.sub.int, CYP3A, liver μl/min/mg 25.5 fu.sub.mic 0.392 P.sub.app 10.sup.−6 cm/s 37.9 F.sub.G 0.78 Predicted AUC ratio 1.76 (with itraconazole)