AHR AGONISTS
20230127797 · 2023-04-27
Inventors
- Cheryl Ann CARSON (Indianapolis, CA)
- Christian Alexander Clarke (Fishers, IN)
- Douglas Linn GERNERT (Fishers, IN, US)
- Steven James GREEN (Indianapolis, IN, US)
- William Glen HOLLOWAY (Martinsville, IN, US)
Cpc classification
C07D401/12
CHEMISTRY; METALLURGY
C07D417/12
CHEMISTRY; METALLURGY
A61P43/00
HUMAN NECESSITIES
C07D211/88
CHEMISTRY; METALLURGY
C07D491/052
CHEMISTRY; METALLURGY
C07D403/12
CHEMISTRY; METALLURGY
C07D413/12
CHEMISTRY; METALLURGY
International classification
C07D401/12
CHEMISTRY; METALLURGY
C07D413/12
CHEMISTRY; METALLURGY
C07D417/12
CHEMISTRY; METALLURGY
Abstract
The present invention relates to certain substituted AHR agonist compounds, to pharmaceutical compositions comprising the compounds and to methods of using the compounds to treat immune-mediated diseases.
Claims
1. A compound of the formula: ##STR00143## wherein, R.sup.1 is selected from phenyl, C.sub.1-C.sub.6 alkyl, C.sub.3-C.sub.6 cycloalkyl, 5- to 6-membered heteroaryl optionally substituted by 1 to 2 R and 8- to 10-membered bicyclic heteroaryl optionally substituted with 1 to 2 R; R.sup.2 is selected from H and OH; X is selected from bond, —C(R.sup.3).sub.2- and —C(R.sup.3).sub.2C(R.sup.3).sub.2—; Y is selected from —C(R.sup.3).sub.2—, —O— and —N(RI)-; R.sup.3 is independently selected from H and C.sub.1-C.sub.3 alkyl; R.sup.i is selected from halogen, CH.sub.3, OCH.sub.3 and CF.sub.3; R.sup.j is C.sub.1-C.sub.3 alkyl, or a pharmaceutically acceptable salt thereof.
2. The compound according to claim 1, wherein R.sup.1 is selected from phenyl and 5- to 6-membered heteroaryl optionally substituted with 1 to 2 R.sup.i, or a pharmaceutically acceptable salt thereof.
3. The compound according to claim 1, wherein Y is —C(R.sup.3).sub.2—, or a pharmaceutically acceptable salt thereof.
4. The compound according to claim 1, wherein R.sup.3 is independently selected from H, CH.sub.3, CH.sub.2CH.sub.3 and CH(CH.sub.3).sub.2 or a pharmaceutically acceptable salt thereof.
5. The compound according to claim 1, wherein X is bond, or a pharmaceutically acceptable salt thereof.
6. The compound according to claim 1, which is selected from: ##STR00144## ##STR00145## or a pharmaceutically acceptable salt thereof.
7. The compound according to claim 1, which is selected from ##STR00146## or a pharmaceutically acceptable salt thereof.
8. The compound according to claim 1, which is selected from ##STR00147## or a pharmaceutically acceptable salt thereof.
9. The compound according to claim 1, which is selected from ##STR00148## or a pharmaceutically acceptable salt thereof
10. The compound according to claim 1, which is a free base.
11. A pharmaceutical composition, comprising a compound or a pharmaceutically acceptable salt thereof according to claim 1 with one or more pharmaceutically acceptable carriers, diluents, or excipients.
12. A method of treating an immune-mediated disease in a patient, comprising administering to a patient in need of such treatment an effective amount of a compound of claim 1.
13. A method of treating a disease or disorder selected from psoriasis, ulcerative colitis, Crohn's disease, graft-versus-host disease and multiple sclerosis in a patient, comprising administering to a patient in need of such treatment an effective amount of a compound of claim 1.
14. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, for use in therapy.
15. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, for use in the treatment of a disease or disorder selected from psoriasis, ulcerative colitis, Crohn's disease, graft-versus-host disease and multiple sclerosis.
Description
EXAMPLE 1
N-(5-Fluoropyrimidin-2-yl)−1,5-dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide
[0125] ##STR00033##
[0126] 1,5-Dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxylic acid (300 mg, 1.45 mmol), 5-fluoropyrimidin-2-amine (245 mg, 2.17 mmol), and pyridine (345 mg, 4.34 mmol) are dissolved in DCM (6 mL). Phosphoryl chloride (271 mg, 1.73 mmol) is added and stirred at 25° C. for 12 h. After this time, volatiles are evaporated under reduced pressure and the residue purified via silica gel chromatography (20 g, 0 100% EtOAc in petroleum ether) to give the product as a white solid (368.3 mg, 1.21 mmol, 84%). ES/MS (m/z): 303.1 (M+H). .sup.1H NMR (400.14 MHz, d6-DMSO): 12.97 (s, 1H), 8.79 (s, 2H), 8.34 (s, 1H), 3.57 (s, 3H), 3.24−3.12 (m, 3H), 2.40−2.33 (m, 1H), 1.69 1.65 (m, 1H), 1.23 (d, J=6.8 Hz, 3H).
[0127] The following compounds in Table 3 are prepared in a manner essentially analogous to the method of preparing N-(5-fluoropyrimidin-2-yl)−1,5-dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (Example 1) using the appropriate reagents and adjusting the reaction time to determine completion of the reaction.
TABLE-US-00003 TABLE 3 Examples 2 to 14 ES/MS Ex (m/z) No. Chemical Name Structure (M + H) 2 N-(5-Fluoro-2-pyridyl)- 1,5-dimethyl-2-oxo-6,7- dihydro-5H- cyclopenta[b]pyridine-3- carboxamide
Example 15
1, 5-Dimethyl-2-oxo-N-thiazol-2-yl-6, 7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide
[0128] ##STR00047##
[0129] 1,5-Dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxylic acid (500 mg, 2.4 mmol) and thiazol-2-amine (362 mg, 3.6 mmol) are dissolved in DCE (8 mL). 1-Propanephosphonic anhydride (1.7 mol/L in EtOAc) (3.07 g, 4.82 mmol) is added and the mixture heated to 80° C. for 16 h. After this time, the volatiles are evaporated under reduced pressure and residue stirred in McOH (5 mL) for 20 min. The formed solid is filtered to obtain the title product as an off-white solid (351.39 mg, 1.18 mmol, 49%). ES/MS (m/z): 290.3 (M+H). .sup.1H NMR (400.14 MHz, d6-DMSO): 13.49 (s, 1H), 8.38 (s, 1H), 7.52 (d, J=4.8 Hz, 1H), 7.28 (d, J=4.8 Hz, 1H), 3.585 (s, 3H), 3.30-2.93 (m, 3H), 2.37−2.33 (m, 1H), 1.69−1.61 (m, 1H), 1.22 (d, J=6.8 Hz, 3H).
[0130] The following compounds in Table 4 are prepared in a manner essentially analogous to the method of preparing 1,5-dimethyl-2-oxo-N-thiazol-2-yl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (Example 15) using the appropriate reagents and adjusting the reaction time to determine completion of the reaction.
TABLE-US-00004 TABLE 4 Examples 16 to 53 ES/MS Ex (m/z) No. Chemical Name Structure (M + H) 16 (5R)-N-Isothiazol-5-yl- 1,5-dimethyl-2-oxo- 6,7-dihydro-5H- cyclopenta[b]pyridine- 3-carboxamide
Example 54 & 55
N-(5-Fluorothiazol-2-yl)−1,5-dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer 1) and N-(5-Fluorothiazol-2-yl)−1,5-dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer 2)
[0131] ##STR00086##
[0132] N-(5-Fluorothiazol-2-yl)−1,5-dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (496 mg) is purified via chiral SFC chromatography (0.1% NI-140H - EtOH; Column: DAICEL CHIRALCEL.sup.® OJ (250 mm×30 mm, 10 μm); begin B: 35%; End B: 35%; Flow Rate: 80 mL/min.) to give isomer 1 as the first-off the column (200 mg) ES/MS (m/z): 308.3 (M+H). .sup.1H NMR (400.13 MHz, d6-DMSO): 13.44 (s, 1H), 8.34 (s, 1H), 7.36 (d, J=2.5 Hz, 1H), 3.58 (s, 3H), 3.25−3.19 (m, 3H), 2.41−2.32 (m, 1H), 1.69−1.63 (m, 1H), 1.24 (d, J=6.8 Hz, 3H), followed by isomer 2 (188 mg). ES/MS (m/z): 308.3 (M+H). .sup.1H NMR (400.13 MHz, d6-DMSO): 13.44 (s, 1H), 8.34 (s, 1H), 7.36 (d, J=2.5 Hz, 1H), 3.58 (s, 3H), 3.25−3.19 (m, 3H), 2.41 2.32 (m, 1H), 1.69−1.63 (m, 1H), 1.24 (d, J=6.8 Hz, 3H).
[0133] The following compounds in Table 5 are prepared in a manner essentially analogous to the method of preparing N-(5-fluorothiazol-2-yl)−1,5-dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer-1) and N-(5-fluorothiazol-2-yl)−1,5-dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer-2) (Example 54 & 55) using the appropriate reagents and adjusting the reaction time to determine completion of the reaction.
TABLE-US-00005 TABLE 5 Examples 56 to 71 and 112 to 115 ES/MS Ex (m/z) No. Chemical Name Structure (M + H) 56 N-(5-Fluoro-2- pyridyl)-1,5- dimethyl-2-oxo-6,7- dihydro-5H- cyclopenta[b] pyridine-3- carboxamide (isomer 1)
Example 72
[0134] ##STR00107##
[0135] (5 S)—N-(1 H-Indol-2-yl)−1, 5-dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide PGP-49,C.sub.1 (5 S)−1,5-Dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxylic acid (90 mg, 0.434 mmol), 2,3-dihydro-1H-indol-2-imine hydrochloride (120 mg, 0.676 mmol), and NMI (165 mg, 2 mmol) are dissolved in ACN (4 mL). TCFH (150 mg, 0.524 mmol) is added and stirred at 20° C. for 16 h. After this time volatiles are evaporated under reduced pressure and the residue purified via silica gel chromatography (20 g, 0 100% EtOAc in petroleum ether) to give the title product as a white solid (368.3 mg, 1.21 mmol, 84%). ES/MS (m/z): 322.0 (M+H). .sup.1H NMR (400.14 MHz, d6-DMSO): 12.70 (s, 1H), 11.40 (s, 1H), 8.36 (s, 1H), 7.41−7.39 (m, 1H), 7.32 (d, J=7.8 Hz, 1H), 7.01−6.93 (m, 2H), 6.48 (d, J=1.3 Hz, 1H), 3.59 (s, 3H), 3.26−3.25 (m, 1H), 3.13−3.08 (m, 2H), 2.44-2.42 (m, 1H), 1.70−1.64 (m, 1H), 1.25 (d, J=6.9 Hz, 3H).
[0136] The following compounds in Table 6 are prepared in a manner essentially analogous to the method of preparing (5 S)—N-(1H-indol-2-yl)−1,5-dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (Example 72) using the appropriate reagents and adjusting the reaction time to determine completion of the reaction.
TABLE-US-00006 TABLE 6 Examples 73 to 76 ES/MS Ex (m/z) No. Chemical Name Structure (M + H) 73 (5R)-N-(1H-indol-2- yl)-1,5-dimethyl-2- oxo-6,7-dihydro-5H- cyclopenta[b]pyridine- 3-carboxamide
Example 77 & 78
1,6-Dimethyl-2-oxo-N-phenyl-6, 7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer 1) and 1,6-Dimethyl-2-oxo-N-phenyl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer 2)0
[0137] ##STR00112##
[0138] Step A: 3-Methylcyclopentanone (4.4 mL, 38.9 mmol) is dissolved in a 2M solution of methylamine in THE (100 mL). Sodium sulfate (16.8 g, 116 mmol) is added and stirred for 18 h. The mixture is concentrated down to 5 mL total volume to give (Z)-N,3-dimethylcyclopentanimine residue. ES/MS (m/z): 111.1 (M+).
[0139] Step B: The residue from step A (4.3 g, 38.7 mmol) is dissolved in McOH (100 mL) and dimethyl methoxymethylenemalonate (13.8 g, 77.7 mmol) added and heated via a microwave to 140° C. for 3 h. The mixture is concentrated and purified via silica gel chromatography (0−10% McOH in DCM) to give methyl 1,6-dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxylate and methyl 1,5-dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxylate as a mixture of regio and stereoisomers (7.17 g, 32.4 mmol, 83%). ES/MS (m/z): 222.0 (M+). .sup.1H NMR (399.80 MHz, CDCl.sub.3): 1.17 (d, J=6.8 Hz, 3H), 1.24−1.21 (m, 3H), 2.42−2.34 (m, 2H), 2.55 (dd, J=6.5, 17.3 Hz, 1H), 2.70−2.61 (m, 2H), 2.99−2.87 (m, 3H), 3.15−3.07 (m, 2H), 3.49 (d, J=5.3 Hz, 6H), 3.86 (d, J=3.9 Hz, 6H), 8.03 (d, J=2.3 Hz, 2H).
[0140] Step C: The product from step B (7.17 g, 32.4 mmol) is dissolved in McOH (150 mL) and 5M aqueous sodium hydroxide solution is added and stirred for 18 h. The reaction is quenched with 5N aqueous HCl solution and extracted with a 3:1 mixture of DCM/isopropanol. The organic layers are combined and washed with brine, dried over anhydrous magnesium sulfate, filtered, and evaporated under vacuum to give 1,6-dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxylic acid and 1,5-dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxylic acid as a mixture of regio and stereoisomers (5.0 g, 24 mmol, 74%). ES/MS (m/z): 208.0 (M+).
[0141] Step D: The product from step C (300 mg, 1.45 mmol) is dissolved in DMF (10 mL). Aniline (0.3 mL, 3.3 mmol), DIEA (800 μL, 4.59 mmol), and T3P (50% by mass in dimethylformamide, 1.8 mL, 3.0 mmol) are added and heated to 100° C. in a microwave reactor for 1 h. The mixture is poured into a saturated solution of sodium bicarbonate (50 mL) and extracted with EtOAc (50 mL×3). The organic layers are combined and washed with brine, dried over anhydrous magnesium sulfate, filtered, evaporated under vacuum, and then purified via silica gel chromatography (0−100% EtOAc in hexanes) to give 1,6-dimethyl-2-oxo-N-phenyl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide and 1,5-dimethyl-2-oxo-N-phenyl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide as a mixture of regio and stereoisomers (94 mg, 0.33 mmol, 23%). ES/MS (m/z): 283.0 (M+). .sup.1H NMR (399.80 MHz, CDCl.sub.3): 1.21 (d, J=6.8 Hz, 4H), 1.29 (d, J=6.8 Hz, 2H), 2.52−2.43 (m, 2H), 2.60 (dd, J=6.5, 17.1 Hz, 1H), 2.75−2.66 (m, 1H), 3.01−2.90 (m, 1H), 3.18−3.03 (m, 1H), 3.22 (q, J=7.2 Hz, 2H), 3.60 (d, J=6.0 Hz, 6H), 7.10 (td, J=7.4, 1.1 Hz, 2H), 7.38−7.33 (m, 4H), 7.79−7.76 (m, 4H), 8.50 (d, J=10.3 Hz, 2H), 12.17 (d, J=4.9 Hz, 2H).
[0142] Step E: Examples 77 & 78: The product from step D (342 mg) is purified via chiral column chromatography (Chiralpak AD-H, 4.6×150 nm, 40% EtOH/CO2, 5 mL/min, 225 nm) to give isomer-1 as the first elution (86.6 mg), ES/MS (m/z): 283.0 (M+). .sup.1H NMR (399.80 MHz, d6-DMSO): 1.14 (d, J=6.7 Hz, 3H), 2.47−2.42 (m, 1H), 2.74−2.60 (m, 2H), 3.03 (dd, J=8.1, 15.0 Hz, 1H), 3.31−3.23 (m, 1H), 3.56 (s, 3H), 7.12-7.07 (m, 1H), 7.38−7.32 (m, 2H), 7.70 (dd, J=1.0, 8.6 Hz, 2H), 8.34 (s, 1H), 12.32 (s, 1H); followed by isomer-2 (82 mg). ES/MS (m/z): 283.0 (M+). .sup.1H NMR (399.80 MHz, d6-DMSO): 1.14 (d, J=6.7 Hz, 3H), 2.47−2.42 (m, 1H), 2.74−2.60 (m, 2H), 3.03 (dd, J=8.1, 15.0 Hz, 1H), 3.31−3.23 (m, 1H), 3.56 (s, 3H), 7.12−7.07 (m, 1H), 7.38−7.32 (m, 2H), 7.70 (dd, J=1.0, 8.6 Hz, 2H), 8.34 (s, 1H), 12.32 (s, 1H). (3.sup.d elution is example 37 and 4.sup.h elution is example 38).
[0143] The following compounds in Table 7 are prepared in a manner essentially analogous to the method of preparing 1,6-dimethyl-2-oxo-N-phenyl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer-1) and 1,6-dimethyl-2-oxo-N-phenyl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer-2) (Example 77 & 78) using the appropriate reagents and adjusting the reaction time to determine completion of the reaction.
TABLE-US-00007 TABLE 7 Examples 79 to 82 ES/MS Ex (m/z) No. Chemical Name Structure (M + H) 79 1,6-Dimethyl-2-oxo- N-pyrimidin-2-yl-6,7- dihydro-5H- cyclopenta[b]pyridine- 3-carboxamide
Examples 83 & 84 & 85 & 86
4-Hydroxy-1,6-dimethyl-2-oxo-N-pyrimidin-2-yl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer 1), 4-Hydroxy-1,6-dimethyl-2-oxo-N-pyrimidin-2-yl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer 2), 4-Hydroxy-1,5-dimethyl-2-oxo-N-pyrimidin-2-yl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer 1), and 4-Hydroxy-1, 5-dimethyl-2-oxo-N-pyrimidin-2-yl-6, 7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer 2)
[0144] ##STR00117##
[0145] A mixture of 4-hydroxy-1,5-dimethyl-2-oxo-6,7-dihydro-5H- PGP-51,C.sub.1 cyclopenta[b]pyridine-3-carboxylic acid and 4-hydroxy-1,6-dimethyl-2-oxo-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxylic acid (549 mg, 5.7 mmol) is dissolved in DCE (10 mL). T3P (1.7 mol/L in EtOAc) (4.89 g, 7.7 mmol) is added, and the mixture heated to 80° C. for 2 h. After this time, the volatiles are evaporated under reduced pressure and the residue purified via silica gel chromatography (20 g, and EtOAc in petroleum ether). The residue is further purified via chiral column (DAICEL CHIRALCEL OD, 250 mm×30 mm, 10 um; mobile phase: 0.1% NI-140H - EtOH; B %:40%−40%.). First-off the column is 4-hydroxy-1,6-dimethyl-2-oxo-N-pyrimidin-2-yl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer 1) as a white solid (150.66 mg, 0.5 mmol, 27%). ES/MS (m/z): 301.1 (M+H). .sup.1H NMR (400.15 MHz, d6-DMSO): 15.15 (s, 1H), 13.20 (s, 1H), 8.73 (d, J=4.9 Hz, 2H), 7.28 (t, J=4.9 Hz, 1H), 3.43 (s, 3H), 3.31−3.29 (m, 1H), 2.99−2.93 (m, 1H), 2.71−2.63 (m, 2H), 2.37−2.30 (m, 1H), 1.14 (d, J=6.8 Hz, 3H). Next off is 4-hydroxy-1,6-dimethyl-2-oxo-N-pyrimidin-2-yl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer 2) as a white solid (217 mg, 0.72 mmol, 39%). ES/MS (m/z): 301.3 (M+H). .sup.1H NMR (400.15 MHz, d6-DMSO): 15.15 (s, 1H), 13.20 (s, 1H), 8.73 (d, J=4.9 Hz, 2H), 7.28 (t, J=4.9 Hz, 1H), 3.43 (s, 3H), 3.31−3.29 (m, 1H), 2.99−2.93 (m, 1H), 2.71−2.63 (m, 2H), 2.37−2.30 (m, 1H), 1.14 (d, J=6.8 Hz, 3H). Third off the column is 4-hydroxy-1,5-dimethyl-2-oxo-N-pyrimidin-2-yl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer 1) as a white solid (78.5 mg, 0.26 mmol, 14%). ES/MS (m/z): 301.3 (M+H). .sup.1H NMR (400.15 MHz, d6-DMSO): 15.33−15.30 (m, 1H), 13.22 (s, 1H), 8.73 (d, J=4.8 Hz, 2H), 7.28 (t, J=4.9 Hz, 1H), 3.44 (s, 3H), 3.31-3.26 (m, 1H), 3.19−3.16 (m, 1H), 3.03−2.99 (m, 1H), 2.35−2.30 (m, 1H), 1.69−1.61 (m, 1H), 1.22 (d, J=6.9 Hz, 3H). Finally, 4-hydroxy-1,5-dimethyl-2-oxo-N-pyrimidin-2-yl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide (isomer 2) as a white solid (77 mg, 0.25 mmol, 14%). ES/MS (m/z): 301.1 (M+H). .sup.1H NMR (400.15 MHz, d6-DMSO): 15.33−15.30 (m, 1H), 13.22 (s, 1H), 8.73 (d, J=4.8 Hz, 2H), 7.28 (t, J=4.9 Hz, 1H), 3.44 (s, 3H), 3.31−3.26 (m, 1H), 3.19−3.16 (m, 1H), 3.03−2.99 (m, 1H), 2.35−2.30 (m, 1H), 1.69−1.61 (m, 1H), 1.22 (d, J=6.9 Hz, 3H).
[0146] The following compounds in Table 8 are prepared in a manner essentially analogous to the method of preparing 4-hydroxy-1,6-dimethyl-2-oxo-N-pyrimidin-2-yl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide and isomers, as well 4-hydroxy-1, 5-dimethyl-2-oxo-N-pyrimidin-2-yl-6,7-dihydro-5H-cyclopenta[b]pyridine-3-carboxamide isomers (Example 86 & 87 & 88 & 89) using the appropriate reagents and adjusting the reaction time to determine completion of the reaction.
TABLE-US-00008 TABLE 8 Examples 87 to 102 Ex ES/MS (m/z) No. Chemical Name Structure (M + H) 87 4-Hydroxy-1,6- dimethyl-2-oxo-N- (2-pyridyl)-6,7- dihydro-5H- cyclopenta[b] pyridine- 3-carboxamide (isomer 1)
Example 103
1-Methyl-2-oxo-N-pyrimidin-2-yl-5,6,7,8-tetrahydroquinoline-3-carboxamide
[0147] ##STR00134##
[0148] 1-Methyl-2-oxo-1,2,5,6,7,8-hexahydro-3-quinolinecarboxylic acid (250 mg, 1.14 mmol) is dissolved in DMF (2.2 mL). 2-Aminopyrimidine (100 mg, 1.02 mmol), DIPEA (463 μL, 2.6 mmol), and HATU (499 mg, 1.31 mmol) are added and stirred at RT for 15 h. The mixture is purified via reverse phase HPLC to give the product (33 mg, 0.16 mmol, 11%). ES/MS (m/z): 285.0 (M+H). .sup.1H NMR (400.13 MHz, d6-DMSO): 1.71-1.65 (m, 2H), 1.84−1.78 (m, 2H), 2.68−2.63 (m, 2H), 2.82 (t, J=6.3 Hz, 2H), 3.58 (s, 3H), 7.22 (t, J=4.8 Hz, 1H), 8.20 (s, 1H), 8.70 (d, J=4.8 Hz, 2H), 12.88 (s, 1H).
[0149] The following compounds in Table 9 are prepared in a manner essentially analogous to the method of preparing 1-methyl-2-oxo-N-pyrimidin-2-yl-5,6,7,8-tetrahydroquinoline-3-carboxamide (Example 103) using the appropriate reagents and adjusting the reaction time to determine completion of the reaction.
TABLE-US-00009 TABLE 9 Examples 104 to 109 ES/MS Ex (m/z) No. Chemical Name Structure (M + H) 104 1-Methyl-2-oxo-N- phenyl-5,6,7,8- tetrahydroquinoline-3- carboxamide
Example 110
4-Hydroxy-1,6,6-trimethyl-2-oxo-N-phenyl-7, 8-dihydro-5H-quinoline-3-carboxamide
[0150] ##STR00141##
Ethyl4-hydroxy-1,3,6,6-tetramethyl-2-oxo-7, 8-dihydro-5H-quinoline-3-carboxylate (181 mg, 0.62 mmol) is dissolved in toluene (3.5 mL). Aniline (0.1 mL, 1 mmol) is heated via microwave to 200° C. for 1 h. and concentrated under vacuum and purified via silica gel chromatography (0−50% EtOAc in hexanes) to give the title product (58.2 mg, 0.178 mmol, 29%). ES/MS (m/z): 327.2 (M+H). .SUP.1.H NMR (399.80 MHz, CDCl.SUB.3.): 1.03 (s, 6H), 1.65 (t, J=6.6 Hz, 2H), 2.37 (s, 2H), 2.68 (t, J=6.6 Hz, 2H), 3.54 (s, 3H), 7.17−7.13 (m, 1H), 7.39−7.35 (m, 3H), 7.70−7.68 (m, 2H), 12.60 (s, 1H)
EXAMPLE 111
[0151] 4-Hydroxy-N-isopentyl-1-methyl-2-oxo-5,6,7,8-tetrahydroquinoline-3-carboxamide
##STR00142##
[0152] 4-Hydroxy-N-isopentyl-2-oxo-5,6,7,8-tetrahydro- 1 H-quinoline-3-carboxamide (45.8 mg, 0.165 mmol) is dissolved in DMF (1 mL) and McOH (1 mL). Potassium carbonate (50 mg, 0.36 mmol) and iodomethane (23 μL, 0.34 mmol) are added and stirred for 24 h. The resulting mixture is purified via reverse phase HPLC to give the title product (37.3 mg, 0.128 mmol, 78%). ES/MS (m/z): 293.2 (M+H). .sup.1H NMR (400.13 MHz, d6-DMSO): 0.91 (d, J=6.6 Hz, 6H), 1.42 (q, J=7.1 Hz, 2H), 1.66−1.58 (m, 3H), 1.79−1.73 (m, 2H), 2.39 (t, J=6.1 Hz, 2H), 2.72−2.67 (m, 2H). .sup.1H NMR (400.13 MHz, d6-DMSO): 0.91 (d, J=6.6 Hz, 6H), 1.42 (q, J=7.1 Hz, 2H), 1.66−1.58 (m, 3H), 1.79-1.73 (m, 2H), 2.39 (t, J=6.1 Hz, 2H), 2.72−2.67 (m, 2H), 3.17 (d, J=5.3 Hz, 1H), 3.30 (d, J=5.3 Hz, 1H), 3.40 (s, 3H), 10.39 (t, J=5.5 Hz, 1H).
hAHR Nuclear Translocation Assay
[0153] Stable cell lines were established using Jump-In™ T-RExTM HEK293 Retargeting Kit (Life Technologies). hAhR cDNA was cloned into the pJTI R.sup.4 CMV-TO EGFP vector. The EGFP was cloned to the C-terminal of AHR to form AhR-EGFP chimera. The pJTI R4 CMV-TO AhR-EGFP vector was transfected using FuGENE® HD into Jump-InTm T-RExTM HEK293 cells. Transfected cells were selected using 2.5 mg/ml G418 for 10 to 14 days, then expanded, harvested, and suspended in freeze media (FBS with 8% DMSO) at 2×10.sup.7 cells/mL, and aliquots were stored in liquid nitrogen. One day before the assay date, cells were thawed and resuspended in DMEM with 5% FBS in the presence of 1 μg/mL Doxycycline and plated into ploy-L-Lysine coated CELLCARRIER-384 ULTRA Microplates (Perkin Elmer) at 12,000 to 15,000 cells per well and incubated at 37° C. and 5% CO2 overnight. On the assay date, compound was serially diluted (1:2) into 384-well nunc plates with DMSO using acoustic dispensing (ECHO). The dose response was a 20-point curve. Compound was resuspended in 40 μL of DMEM plus 0.1% BSA. The culture media was damped and 25 μL of DMEM plus 0.1% BSA was added, then 25 μL of compound in DMEM plus 0.1% BSA was added into cell plates. Cells were incubated with compounds at 37° C. and 5% CO2 for 45 min.
[0154] The final DMSO concentration was 0.2%. The media was damped after 45 minutes incubation. The cells were fixed with 40 μL of cold McOH (−20° C.) for 20 min. The McOH was damped and 50 μL of DPBS containing 1 μg/mL Hochst was added into the cell plates. The intensity of EGFP was quantitated by using OPERA PHENIX® or Operetta high content image system (Perkin Elmer) with 20x Water Objective and five field per well. The ratio of EGFP fluorescent intensity in nuclear over cytosol was analyzed using a 4-parameter nonlinear logistic equation to determine the potency of AhR agonists.
[0155] Table 10 shows the hAHR nuclear translocation assay EC5o values for the exemplified compounds.
TABLE-US-00010 TABLE 10 hAHR nuclear translocation assay EC.sub.50 values Example EC.sub.50 (nM) 1 9.78 2 9.78 3 0.340 4 1.82 5 16.9 6 4.57 7 8.26 8 1.42 9 191 10 13.7 11 64.7 12 7.90 13 4.67 14 26.8 15 1.93 16 1.35 17 1.34 18 1.32 19 1.04 20 5.23 21 4.72 22 4.10 23 11.5 24 2.45 25 1.91 26 2.87 27 6.41 28 3.68 29 2.23 30 1.62 31 2.40 32 0.845 33 2.64 34 7.46 35 4.42 36 2.78 37 1.97 38 1.01 39 0.378 40 2.37 41 2.65 42 9.41 43 0.973 44 2.38 45 16.2 46 31.5 47 0.803 48 11.3 49 2.03 50 8.27 51 4.35 52 6.40 53 0.607 54 0.566 55 0.691 56 2.44 57 1.83 58 3.59 59 3.59 60 396 61 42.0 62 1.84 63 2.55 64 1.54 65 1.22 66 0.525 67 0.500 68 0.137 69 0.161 70 0.961 71 0.189 72 1.79 73 0.275 74 19.0 75 17.7 76 1.12 77 3.49 78 0.297 79 20.3 80 10.9 81 0.574 82 25.1 83 233 84 8.67 85 4.1 86 7.79 87 27.3 88 0.897 89 2.33 90 2.89 91 4.86 92 1.44 93 1.06 94 0.533 95 1.21 96 0.600 97 0.709 98 0.342 99 2.12 100 2.17 101 6.01 102 2.59 103 27.0 104 0.502 105 0.640 106 25.9 107 108 108 2.50 109 1.04 110 2.27 111 10.3 112 0.865 113 11.2 114 2.44 115 2.96
[0156] The results of this assay demonstrate that the exemplified compounds are AhR agonists.