Kynurenine-3-monooxygenase inhibitors, pharmaceutical compositions, and methods of use thereof
09822058 · 2017-11-21
Assignee
Inventors
- Leticia M. Toledo-Sherman (Santa Monica, CA, US)
- Celia Dominguez (Los Angeles, CA)
- Michael Prime (Abingdon, GB)
- William Leonard Mitchell (Market Rasen, GB)
- Peter Johnson (Abingdon, GB)
- Naomi Went (Didcot, GB)
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International classification
C07D203/10
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C07C69/757
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C07D203/08
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C07C229/48
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C07C323/62
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C07D413/10
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C07D405/10
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C07D307/54
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C07D305/08
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C07D305/06
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C07D295/155
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C07D207/337
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C07D271/07
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C07D207/08
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C07D263/58
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C07D317/60
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C07D403/10
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Abstract
Certain compounds, or pharmaceutically acceptable salts or prodrugs thereof, are provided herein. Also provided are pharmaceutical compositions comprising at least one compound, or pharmaceutically acceptable salt or prodrug thereof, described herein and one or more pharmaceutically acceptable vehicle. Methods of treating patients suffering from certain diseases and disorders responsive to the inhibition of KMO activity are described, which comprise administering to such patients an amount of at least one compound, or pharmaceutically acceptable salt or prodrug thereof, described herein effective to reduce signs or symptoms of the disease or disorder are disclosed. These diseases include neurodegenerative disorders such as Huntington's disease. Also described are methods of treatment include administering at least one compound, or pharmaceutically acceptable salt or prodrug thereof, described herein as a single active agent or administering at least one compound, or pharmaceutically acceptable salt or prodrug thereof, described herein in combination with one or more other therapeutic agents. Also provided are methods for screening compounds capable of inhibiting KMO activity.
Claims
1. A compound of Formula I: ##STR00172## or a pharmaceutically acceptable salt or prodrug thereof, wherein: R.sub.1 is ##STR00173## R.sub.11 is chloro and R.sub.12 is Z—R.sub.20, wherein: Z is —O—, —S—, —S(O)—, —S(O).sub.2—, or —CR.sub.21R.sub.22—, R.sub.21 and R.sub.22 are independently selected from hydrogen, lower alkyl, hydroxyl, and lower alkoxy, or R.sub.21 and R.sub.22, taken together with the carbon to which they are attached, form an optionally substituted 3- to 7-membered cycloalkyl or heterocycloalkyl, and R.sub.20 is optionally substituted C.sub.1-C.sub.6 alkyl, optionally substituted cycloalkyl or optionally substituted heterocycloalkyl; R.sub.13 is hydrogen or halo; X is —CR.sub.2R.sub.3—; R.sub.2 and R.sub.3 are independently selected from hydrogen, optionally substituted amino, hydroxyl, lower alkoxy, and optionally substituted lower alkyl, or R.sub.2 and R.sub.3, taken together with the carbon to which they are attached, form an optionally substituted 5- to 7-membered cycloalkyl or optionally substituted 5- to 7-membered heterocycloalkyl; L is —C(O)O—; R.sub.5 is hydrogen; and R.sub.6 and R.sub.7 are independently selected from hydrogen, halo, optionally substituted amino, hydroxyl, lower alkoxy, and optionally substituted lower alkyl.
2. The compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.2 is hydrogen, hydroxyl, lower alkoxy, amino optionally substituted with one or more alkyl groups, or lower alkyl optionally substituted with one or more groups independently selected from halo, hydroxyl, lower alkoxy, and amino optionally substituted with one or more alkyl groups.
3. The compound of claim 2, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.2 is hydrogen, amino, methylamino, dimethylamino, hydroxyl, methyl, methoxy, difluoromethyl, trifluoromethyl, hydroxymethyl, methoxymethyl, aminomethyl, (methylamino)methyl, or (dimethylamino)methyl.
4. The compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.3 is hydrogen or lower alkyl.
5. The compound of claim 4, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.3 is hydrogen or methyl.
6. The compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.2 and R.sub.3 are hydrogen.
7. The compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.2 is methyl and R.sub.3 is hydrogen.
8. The compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.2 and R.sub.3 are methyl.
9. The compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.2 and R.sub.3, taken together with the carbon to which they are attached form an optionally substituted 5- or 6-membered cycloalkyl or optionally substituted 5- or 6-membered heterocycloalkyl.
10. The compound of claim 9, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.2 and R.sub.3, taken together with the carbon to which they are attached, form an optionally substituted cyclopentyl or optionally substituted pyrrolidin-3-yl.
11. The compound of claim 10, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.2 and R.sub.3, taken together with the carbon to which they are attached, form a cyclopentyl or pyrrolidin-3-yl.
12. The compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof, wherein Z is —O—.
13. The compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof, wherein Z is —S—.
14. The compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof, wherein Z is —S(O).sub.2—.
15. The compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof, wherein Z is —CR.sub.21R.sub.22—.
16. The compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.20 is methyl, difluoromethyl, trifluoromethyl, ethyl, 2,2,2-trifluoro-1-methyl-ethyl, (2R)-butan-2-yl, (2S)-butan-2-yl, cyclopropyl, cyclobutyl, cyclopentyl, isopropyl, or oxetan-3-yl.
17. The compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.1 is 3-chloro-4-(cyclopropanesulfinyl)phenyl, 3-chloro-4-(cyclopropanesulfonyl)phenyl, 3-chloro-4-cyclopropoxy-phenyl, 3-chloro-4-(cyclopropylsulfanyl)phenyl, 3-chloro-4-isopropoxy-phenyl, 3-chloro-4-(oxetan-3-yloxy)phenyl, 3-chloro-4-(trifluoromethoxy)phenyl, 3-chloro-4-cyclopropoxyphenyl, 4-(aziridin-1-ylmethyl)-3-chlorophenyl, 4-[(2R)butan-2-yloxy]-3-chlorophenyl, or 4-[(2S)-butan-2-yloxy]-3-chlorophenyl.
18. The compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.7 is hydrogen, aminomethyl, methoxymethyl, methyl, 1-aminoethyl, 1-methoxy-ethyl, methoxy, or halo.
19. The compound of claim 18, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.7 is methyl.
20. The compound of claim 18, or a pharmaceutically acceptable salt or prodrug thereof, wherein R.sub.7 is hydrogen.
21. A compound of formula: ##STR00174## ##STR00175## or a pharmaceutically acceptable salt or prodrug thereof.
22. A pharmaceutical composition comprising at least one compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof, and at least one pharmaceutically acceptable excipient.
23. A method of treating a condition or disorder mediated by Kynurenine 3-mono-oxygenase activity in a subject in need of such a treatment which method comprises administering to the subject a therapeutically effective amount of at least one compound of claim 1, or a pharmaceutically acceptable salt or prodrug thereof.
24. The method of claim 23 wherein said condition or disorder involves a neurodegenerative pathology.
Description
EXAMPLES
(1) The compounds, pharmaceutically acceptable salts and prodrugs thereof, described herein, compositions, and methods described herein are further illustrated by the following non-limiting examples.
(2) As used herein, the following abbreviations have the following meanings. If an abbreviation is not defined, it has its generally accepted meaning. CDI=carbonyldiimidazole DCM=dichloromethane DME=dimethyl ether DMEM=Dulbecco's modified Eagle's medium DMF=N,N-dimethylformamide DMSO=dimethylsulfoxide EDC.HCl=1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride EtOH=ethanol Et.sub.2O=diethylether EtOAc=ethyl acetate g=gram hr=hour hrs=hours HOBt=1-Hydroxybenzotriazol LiHMDS=lithium hexamethyl-disilazide LC/MS=liquid chomatography/mass spectrometry mg=milligram min=minutes mL=milliliter mmol=millimoles mM=millimolar ng=nanogram nm=nanometer nM=nanomolar PBS=phosphate buffered saline rt=room temperature TBME=t-butyl methyl ether THF=tetrahydrofuran TMOF=trimethylorthoformate μL=microliter μM=micromolar 1 g/1 ml=1 vol
EXPERIMENTAL
(3) Commercially available reagents and solvents (HPLC grade) were used without further purification.
(4) Thin-layer chromatography (TLC) analysis was performed with Kieselgel 60 F.sub.254 (Merck) plates and visualized using UV light. Microwave reactions were carried out using CEM focussed microwaves.
(5) .sup.1H NMR spectra were recorded on a Bruker DRX 500 MHz spectrometer or Bruker DPX 250 MHz spectrometer in deuterated solvents.
(6) Analytical HPLC-MS was performed on Agilent HP1100 and Shimadzu 2010, systems using reverse phase Atlantis dC18 columns (5 μm, 2.1×50 mm), gradient 5-100% B ( A=water/0.1% formic acid, B=acetonitrile/0.1% formic acid) over 2 or 3.5 minutes, injection volume 3 μl, flow=1.0 ml/min.
(7) UV spectra were recorded at 215 nm using a Waters 2996 photo diode array or on the Shimadzu 2010 system. UV spectra were recorded at 215 nm using a Waters 2487 dual wavelength UV detector or the Shimadzu 2010 system. Mass spectra were obtained over the range m/z 150 to 850 at a sampling rate of 2 scans per second using Waters ZMD and over m/z 100 to 1000 at a sampling rate of 2 Hz using Electrospray ionisation, by a Shimadzu 2010 LC-MS system, or analytical HPLC-MS was performed on Agilent HP1100 and Shimadzu 2010, systems using reverse phase Water Atlantis dC18 columns (3 μm, 2.1×100 mm), gradient 5-100% B (A=water/0.1% formic acid, B=acetonitrile/0.1% formic acid) over 7 min, injection volume 3 μl, flow=0.6 ml/min. UV spectra were recorded at 215 nm using a Waters 2996 photo diode array or on the Shimadzu 2010 system.
Example 1
(8) (+)-(1S,2S)-Cyclopropane-1,2-dicarboxylic acid monomethyl ester was prepared as described in EP1475385, 2004
(9) ##STR00005##
Step 1, Method 1: (1S,2S)-Methyl 2-(carbonochloridoyl)cyclopropane-1-carboxylate
(10) DMF (0.16 mL, 2.08 mmol) was added dropwise to a stirred solution of (+)-(1S,2S)-cyclopropane-1,2-dicarboxylic acid monomethyl ester (3.0 g, 20.8 mmol) in DCM (80 mL) at room temperature under a nitrogen atmosphere. Oxalyl chloride (5.45 mL, 62.4 mmol) was added dropwise to the reaction mixture over 30 minutes and the reaction was stirred at room temperature for 1 hour. After this time, the reaction mixture was concentrated and the resulting residue was co-evaporated with DCM (3×20 mL) to give the title compound (3.42 g, 98% yield) as a yellow oil which was used directly without further purification.
Step 2, Method 1: (1S,2S)-Methyl 2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(11) See, e.g., Naoaki et al, Bioorganic and Medicinal Chemistry Letters, 2005, 121-124.
(12) A mixture of (3,4-dichlorophenyl)boronic acid (4.82 g, 25.24 mmol) and potassium phosphate hydrate (3:1:1) (7.27 g, 31.56 mmol) in Toluene (40 mL) was degassed with nitrogen for 15 minutes. Dichloropalladium; bis(triphenylphosphane) (0.29 g, 0.42 mmol) was then added at room temperature followed by a mixture of methyl (1S,2S)-2-(carbonochloridoyl)cyclopropane-1-carboxylate (3.42 g, 21.04 mmol) in Toluene (20 mL). The reaction mixture was then warmed to 80° C. and stirred at this temperature under nitrogen for 5 hours. After this time the reaction mixture was cooled to room temperature and filtered. The solid was washed with DCM (3×50 mL) and the filtrate concentrated under reduced pressure. The resulting residue was then purified by flash column chromatography (elution; 100% Heptane to 8:2 Heptane:EtOAC) to give the title compound as a brown oil/solid which was further purified by flash column chromatography (elution; 100% Heptane to 9:1 Heptane:EtOAC) to give the title compound (2.16 g, 35% yield) as a pale yellow solid. Tr=2.22 min m/z (ES.sup.+) (M+H.sup.+) 273.
Intermediate 2, Step 2, Methyl (1S,2S)-2-[(3,5-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(13) Tr=4.88 min (7 minute method) m/z (ES.sup.+) (M+H.sup.+) 273.
Intermediate 3, Step 2, Methyl (1S,2S)-2-[(3-chloro-4-fluorophenyl)carbonyl]cyclopropane-1-carboxylate
(14) Tr=2.10 min (3.5 minute method) m/z (ES.sup.+) (M+H.sup.+) 257.
Intermediate 4, Step 2, Methyl (1S,2S)-2-{[3-chloro-4-(trifluoromethyl)phenyl]carbonyl}cyclopropane-1-carboxylate
(15) Tr=2.26 min (3.5 minute method) m/z (ES.sup.−) (M−H) 305.
Intermediate 5, Step 2, Methyl (1S,2S)-2-{[3-chloro-4-(propan-2-yloxy)phenyl]carbonyl}cyclopropane-1-carboxylate
(16) Tr=2.25 min (3.5 minute method) m/z (ES+) (M+H+) 297.
Step 3, Method 1: (1S,2S)-2-[(3-Chloro-4-fluorophenyl)carbonyl]cyclopropane-1-carboxylic acid
(17) NaOH (2M solution, 0.75 mL, 1.5 mmol) was added in one portion to a stirred solution of (1S,2S)-methyl 2-[(3-chloro-4-fluorophenyl)carbonyl]cyclopropane-1-carboxylate (0.08 g, 0.3 mmol) in THF (3 mL) and the mixture was stirred at room temperature for 68 hours. After this time, the reaction mixture was concentrated and the resulting residue was partitioned between water (5 mL) and diethyl ether (5 mL). The aqueous layer was removed, acidified to pH 1 with 2M HCl and extracted with ethyl acetate (3×5 mL). The combined organic extracts were combined, dried (MgSO.sub.4), filtered and concentrated. The resulting residue was purified by prep HPLC to give the title compound (0.02 g, 20% yield) as a white solid.
(18) δ.sub.H (500 MHz, DMSO) 8.21-8.31 (1 H, m) 8.01-8.16 (1 H, m) 7.60 (1 H, dd) 3.23-3.28 (1 H, m) 2.04-2.20 (1 H, m) 1.41-1.56 (2 H, m). Tr=3.67 min m/z (ES.sup.+) (M+H.sup.+) 241, 243.
(19) The following compounds were prepared substantially as described above.
(1S,2S)-2-[(3,5-Dichlorophenyl)carbonyl]cyclopropane-1-carboxylic acid
(20) Tr=4.13 min m/z (ES.sup.+) (M+H.sup.+) 260.
(1S,2S)-2-[(3-Chloro-4-fluorophenyl)carbonyl]cyclopropane-1-carboxylic acid
(21) δ.sub.H (500 MHz, DMSO) 8.21-8.31 (1 H, m) 8.01-8.16 (1 H, m) 7.60 (1 H, dd) 3.23-3.28 (1 H, m) 2.04-2.20 (1 H, m) 1.41-1.56 (2 H, m). Tr=3.67 min m/z (ES.sup.+) (M+H.sup.+) 241, 243.
(1S,2S)-2-{[3-Chloro-4-(trifluoromethyl)phenyl]carbonyl}cyclopropane-1-carboxylic acid
(22) δ.sub.H (500 MHz, CDCl.sub.3) 8.12 (1 H, s) 7.99 (1 H, d) 7.85 (1 H, d) 3.19 (1 H, ddd) 2.47 (1 H, ddd) 1.74 (2 H, ddd). Tr=4.11 min m/z (ES.sup.+) (M−H.sup.+) 291.
(1S,2S)-2-[(3-Chloro-4-methylphenyl)carbonyl]cyclopropane-1-carboxylic acid
(23) δ.sub.H (500 MHz, CDCl.sub.3) 7.89 (1 H, d) 7.79 (1 H, dd) 7.47 (1 H, d) 3.20 (1 H, ddd) 2.46 (3 H, s) 2.37-2.43 (1 H, m) 1.68 (2 H, dddd). Tr=3.89 min m/z (ES.sup.+) (M−H.sup.+) 239.
(1S,2S)-2-{[3-Chloro-4-(propan-2-yloxy)phenyl]carbonyl}cyclopropane-1-carboxylic acid
(24) δ.sub.H (500 MHz, CDCl.sub.3) 8.07 (1 H, d) 7.92 (1 H, dd) 6.99 (1 H, d) 4.71 (1 H, dt) 3.16 (1 H, ddd) 2.32-2.44 (1 H, m) 1.59-1.74 (2 H, m) 1.44 (6 H, d). Tr=4.03 min m/z (ES.sup.+) (M+H.sup.+) 283, 285.
Example 2
(25) ##STR00006##
Step 1,4-Bromo-2-chloro-1-cyclopropoxybenzene
(26) Bromocyclopropane (14.6 g, 24.0 mmol) was added in one portion to a stirred solution of 4-bromo-2-chlorophenol (5.0 g, 24.0 mmol) and caesium dicarbonate (19.6 g, 60.0 mmol) in dimethyl acetamide (80 mL). The mixture was heated to 150° C. and stirred at this temperature for 16 hours. After this time, a further portion of bromocyclopropane (14.6 g, 24.0 mmol) was added and the mixture stirred at 150° C. for a further 24 hours. The reaction was then cooled to room temperature, poured onto ice-water (200 mL) and extracted with TBME (3×200 mL). The organic layers were combined and washed sequentially with water (2×100 mL) and brine (50 mL). The organic layer was removed, dried (MgSO.sub.4), filtered and concentrated. The resulting residue was purified by flash column chromatography (elution: heptane, then 33% heptane, 67% DCM) to give the title compound (5.0 g, 84% yield) as a colourless oil. Tr=2.41 min m/z (ES.sup.+) (M+H.sup.+) 248.
Step 2, 2-(3-Chloro-4-cyclopropoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane
(27) See, e.g., Pushpinder et al, Journal of Medicinal Chemistry, 2006, 35-38.
(28) A mixture of 4-bromo-2-chloro-1-cyclopropoxybenzene (6.77 g, 0.027 mol), bis -(pinacolato)diboron (8.33 g, 0.033 mol) and potassium acetate (8.05 g, 0.082 mol) in DMSO (70 mL) was degassed with nitrogen at room temperature for 20 minutes. After this time Pd(Cl).sub.2(dppf).sub.2 (1.12 g, 0.001 mol) was added in one portion and the reaction mixture was heated at 80° C. and stirred at this temperature under a nitrogen atmosphere for 6 hours. The reaction was then cooled to room temperature, and partitioned between ethyl acetate (100 mL) and water (50 mL). The organic layer was separated, washed sequentially with water (2×100 ml) and brine (50 mL). The organic layer was removed, dried (MgSO.sub.4), filtered and concentrated. The resulting residue was purified by flash column chromatography (elution: 12% DCM, 88% heptane, then 100% DCM) to give the title compound (3.69 g, 46% yield) as a yellow gum. Tr=2.57 min m/z (ES.sup.+) (M+H.sup.+) No ionisation.
Step 3, Methyl (1S,2S)-2-[(3-chloro-4-cyclopropoxyphenyl)carbonyl]cyclopropane-1-carboxylate
(29) See, e.g., Shionogi Co. Ltd, 2007, EP1839655.
(30) A mixture of 2-(3-chloro-4-cyclopropoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.2 g, 1.23 mmol), methyl (1S,2S)-2-(carbonochloridoyl)cyclopropane-1-carboxylate (0.31 g, 1.48 mmol), potassium phosphate hydrate (0.39 g, 1.85 mmol) and dichloropalladium; bis(triphenylphosphane) (0.02 g, 0.025 mmol) in Toluene (6 mL) was degassed with nitrogen for 15 minutes. The reaction mixture was then heated to 80° C. and stirred at this temperature under nitrogen for 2 hours. After this time the reaction mixture was cooled to room temperature and concentrated. The resulting residue was suspended in water (20 mL) and extracted with ethyl acetate (3×25 mL). The combined organic extracts were dried (Na.sub.2SO.sub.4), filtered and concentrated. The resulting residue was then purified by using a Biotage Isolera (50 g silica gel cartridge, elution; 2% EtOAc 1CV; 2% to 20% EtOAc over 10 CV; 20% EtOAc 5 CV) to give the title compound (0.12 g, 32% yield) as a white crystalline solid. Tr=2.15 min m/z (ES.sup.+) (M+H.sup.+) 295, 297.
Step 4, (1S,2S)-2-[(3-Chloro-4-cyclopropoxyphenyl)carbonyl]cyclopropane-1-carboxylic acid
(31) NaOH (2M solution, 0.97 mL, 1.9 mmol) was added in one portion to a stirred solution of methyl (1S,2S)-2-[(3-chloro-4-cyclopropoxyphenyl)carbonyl]cyclopropane-1-carboxylate (0.15 g, 0.39 mmol) in THF (4 mL) and the mixture was stirred at room temperature for 4 hours. After this time, the reaction mixture was concentrated and the resulting residue was partitioned between water (20 mL) and diethyl ether (20 mL). The aqueous layer was removed, acidified to pH 1 with 2M HCl and extracted with ethyl acetate (3×20 mL). The combined organic extracts were combined, dried (MgSO.sub.4), filtered and concentrated. The resulting residue was purified by prep HPLC to give the title compound (0.06 g, 52% yield) as a white solid.
(1S,2S)-2-[(3-Chloro-4-cyclopropoxyphenyl)carbonyl]cyclopropane-1-carboxylic acid
(32) δ.sub.H (500 MHz, DMSO) 11.88-13.36 (m, 1 H) 8.02-8.20 (m, 2 H) 7.48-7.64 (m, 1 H) 4.03-4.18 (m, 1 H) 3.15-3.25 (m, 1 H) 2.01-2.14 (m, 1 H) 1.29-1.56 (m, 2 H) 0.66-0.96 (m, 4 H). Tr=4.00 min m/z (ES.sup.+) (M+H.sup.+) 281, 283.
Example 3
(33) ##STR00007##
Step 1, Sodium (1S,2S)-2-{[3-chloro-4-(propan-2-yloxy)phenyl]carbonyl}cyclopropane-1-carboxylate
(34) (1S,2S)-2-{[3-Chloro-4-(propan-2-yloxy)phenyl]carbonyl}cyclopropane-1-carboxylic acid (0.14 g, 0.51 mmol) was stirred in NaOH (2M solution, 0.23 mL, 0.46 mmol) for 1 hour. After this time, diethyl ether was added (2 mL), the organic layer was removed and discarded. The aqueous layer was concentrated to give the title compound (0.097 g, 62% yield) as an off-white solid.
Sodium (1S,2S)-2-{[3-chloro-4-(propan-2-yloxy)phenyl]carbonyl}cyclopropane-1-carboxylate
(35) δ.sub.H (500 MHz, DMSO) 7.88-7.97 (2 H, m) 7.30 (1 H, d) 4.78-4.89 (1 H, m) 2.77-2.89 (1 H, m) 1.64-1.74 (1 H, m) 1.33 (6 H, d) 1.14-1.25 (2 H, m). Tr=4.03 min m/z (ES.sup.+) (M+H.sup.+) 283, 285.
(36) The following compounds were prepared substantially as described above.
Sodium (1S,2S)-2-{[3-chloro-4-(trifluoromethyl)phenyl]carbonyl}cyclopropane-1-carboxylate
(37) δ.sub.H (500 MHz, DMSO) 7.99-8.16 (3 H, m) 2.86-2.98 (1 H, m) 1.71-1.84 (1 H, m) 1.14-1.39 (2H, m). Tr=4.19 min m/z (ES.sup.−) (M−H.sup.−) 291.
Sodium (1S,2S)-2-[(3-chloro-4-methylphenyl)carbonyl]cyclopropane-1-carboxylate
(38) δ.sub.H (500 MHz, DMSO) 7.94 (1 H, d) 7.79 (1 H, dd) 7.58 (1 H, d) 2.83-2.93 (1 H, m) 2.41 (3 H, s) 1.68-1.78 (1 H, m) 1.14-1.31 (2 H, m). Tr=3.98 min m/z (ES.sup.+) (M+H.sup.+) 239.
Sodium (1S,2S)-2-[(3-chloro-4-fluorophenyl)carbonyl]cyclopropane-1-carboxylate
(39) δ.sub.H (500 MHz, DMSO) 8.10 (2 H, d) 7.97-8.05 (2 H, m) 7.57 (2 H, t) 2.82-2.94 (2 H, m) 1.67-1.80 (2 H, m) 1.17-1.34 (4 H, m). Tr=3.79 min m/z (ES.sup.−) (M−H.sup.−) 241.
Example 4
(40) ##STR00008##
Step 1, (1S,2S)-2-[(3-Chloro-4-fluorophenyl)carbonyl]cyclopropane-1-carboxamide
(41) Oxalyl chloride (2.37 mL, 27.2 mmol) was added dropwise to a stirred solution of (1S,2S)-2-{[3-Chloro-4fluorophenyl]carbonyl}cyclopropane-1-carboxylic acid (2.2 g, 9.07 mmol) in DCM (44 mL) containing DMF (0.07 mL) and the mixture was stirred at room temperature for 1 hour. After this time, the reaction mixture was concentrated and the resulting residue was co-evaporated with DCM (3×30 mL). The residue was dissolved in THF (44 mL), cooled to 0° C. and treated with a saturated solution of aqueous ammonia (10.0 mL, 516.7 mmol) added dropwise over 10 minutes and the reaction mixture was allowed to warm to room temperature under a nitrogen atmosphere and stirred for 1 hour. After this time the reaction mixture was concentrated and the resulting residue was suspended in water (50 mL) and extracted with ethyl acetate (3×100 mL). The organic layers were combined, dried (Na.sub.2SO.sub.4), filtered and concentrated. The resulting residue was then purified by using a Biotage Isolera (100 g silica gel cartridge, elution; 0% EtOAc, 100% heptanes to 100% EtOAc) to give the title compound (1.8 g, 80% yield) as a white solid. Tr=1.59 min m/z (ES.sup.+) (M+H.sup.+) 242.
Intermediate 2, Step 1, (1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropane-1-carboxamide
(42) Tr=1.72 min (3.5 minute method) m/z (ES.sup.+) (M+H.sup.+) 258, 260.
Step 2, Method 4: (1S,2S)-2-[(3-Chloro-4-fluorophenyl)carbonyl]cyclopropane-1-carbonitrile
(43) See, e.g., WO2009/98144.
(44) 2,4,6-Trichloro-1,3,5-triazine (1.92 g, 10.4 mmol) was added portionwise to a stirred, cooled (0° C.) solution of (1S,2S)-2-[(3-chloro-4-fluorophenyl)carbonyl]cyclopropane-1-carboxamide (1.8 g, 7.45 mmol) in DMF (19 mL) and the mixture was stirred for 1 hour before being warmed to room temperature over 30 minutes. After this time, the reaction mixture was poured onto sodium hydroxide (0.5M solution, 50 mL) and extracted with ethyl acetate (2×50 mL). The organic layers were combined, washed consecutively with water (2×50 mL) and brine (30 mL) before being dried (Na.sub.2SO.sub.4), filtered and concentrated. The resulting residue was then purified by using a Biotage Isolera (100 g silica gel cartridge, elution; 0% EtOAc, 100% heptanes to 50% EtOAc, 50% heptanes) to give the title compound (1.6 g, 93% yield) as a colourless oil. Tr=1.94 min m/z (ES.sup.+) (M+H.sup.+) 222.
Intermediate 2, Step 2, (1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropane-1-carbonitrile
(45) δ.sub.H (500 MHz, DMSO) 8.36 (1 H, d) 8.05 (1 H, dd) 7.87 (1 H, d) 3.79 (1 H, ddd) 2.35 (1 H, ddd) 1.69 (1 H, ddd) 1.52 (1 H, ddd). Tr=4.27 min m/z (ES.sup.+) (M+H.sup.+) 238, 240.
Step 3, Method 4: (1S,2S)-2-[(3-Chloro-4-fluorophenyl)carbonyl]-N-hydroxycyclopropane-1-carboximidamide
(46) See, e.g., Tkachev et al, Synthesis, 2000, 1148-1159.
(47) Diisopropylethyl amine (0.59 mL, 3.47 mmol) was added dropwise to a stirred solution of (1S,2S)-2-[(3-chloro-4-fluorophenyl)carbonyl]cyclopropane-1-carbonitrile (0.8 g, 3.47 mmol) and hydroxylamine hydrochloride (0.25 g, 3.47 mmol) in ethanol (50 mL) and the mixture was stirred at room temperature under a nitrogen atmosphere for 18 hours. After this time, the reaction mixture was concentrated and the resulting residue suspended in water (50 mL) and extracted with ethyl acetate (3×50 mL). The organic layers were combined, dried (Na.sub.2SO.sub.4), filtered and concentrated. The resulting residue was then purified by using a Biotage Isolera (100 g silica gel cartridge, elution; 0% EtOAc, 100% heptanes to 100% EtOAc) to give the title compound (0.81 g, 89% yield) as a white solid. Tr=1.18 min m/z (ES.sup.+) (M+H.sup.+) 258.
Intermediate 2, Step 3, (Z,1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]-N′-hydroxycycloprop-1-carboximidamide
(48) Tr=2.86 min (3.5 minute method) m/z (ES.sup.+) (M+H.sup.+) 273, 275.
Step 4, 3-[(1S,2S)-2-[(3-Chloro-4-fluorophenyl)carbonyl]cyclopropyl]-2,5-dihydro-1,2,4-oxadiazol-5-one
(49) Carbonyl diimidazole (0.77 g, 4.76 mmol) and DBU (0.54 mL, 3.65 mmol) were added sequentially to a stirred solution of (1S,2S)-2-[(3-chloro-4-fluorophenyl)carbonyl]-N-hydroxycyclopropane-1-carboximidamide (0.8 g, 3.17 mmol) in dioxane (22 mL) and the mixture was heated to 105° C. under a nitrogen atmosphere for 18 hours. After this time, the reaction mixture was allowed to cool to room temperature before being concentrated. The resulting residue was suspended in water (20 mL) and extracted with ethyl acetate (1×10 mL). The aqueous layer was removed, acidifed to pH 1 with HCl (2M solution) and extracted with ethyl acetate (4×30 mL). The organic layers were combined, dried (Na.sub.2SO.sub.4), filtered and concentrated. The resulting residue was then purified by using a Biotage Isolera (50 g silica gel cartridge, elution; 0% EtOAc, 100% heptanes to 100% EtOAc). The resulting residue was then purified further by prep HPLC to give the title compound (0.02 g, 3% yield) as a white solid.
3-[(1S,2S)-2-[(3-Chloro-4-fluorophenyl)carbonyl]cyclopropyl]-2,5-dihydro-1,2,4-oxadiazol-5-one
(50) δ.sub.H (500 MHz, DMSO) 8.32 (dd, 2.05 Hz, 1 H) 8.11 (ddd, 4.77, 2.05 Hz, 1 H) 7.64 (t, 1 H) 6.53 (s, 1 H) 3.42-3.49 (m, 1H) 2.42-2.47 (m, 1 H) 1.59-1.72 (m, 2 H). Tr=3.89 min m/z (ES.sup.+) (M+H.sup.+) 281, 283.
(51) The following compounds were prepared substantially as described above.
3-[(1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropyl]-2,5-dihydro-1,2,4-oxadiazol-5-one
(52) δ.sub.H (500 MHz, DMSO) 12.29 (1 H, br. s.) 8.32 (1 H, d) 8.02 (1 H, dd) 7.87 (1 H, d) 3.47 (1 H, ddd) 2.44-2.49 (1 H, m) 1.69 (1 H, ddd) 1.63 (1 H, ddd). Tr=4.22 min m/z (ES.sup.+) (M+H.sup.+) 297, 299.
Example 5
(53) ##STR00009##
Step 1, Method 5: 5-[(1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropyl]-2H-1,2,3,4-tetrazole
(54) See, e.g., Curran et al, Tetrahedron, 1999, 8997-9006.
(55) Trimethylsilyl azide (0.44 mL, 3.33 mmol) and dibutyl(oxo) stannane (0.042 g, 0.17 mmol) were added sequentially to a stirred solution of (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carbonitrile (0.4 g, 1.37 mmol) in toluene (10 mL) and the reaction mixture was heated to 115° C. and stirred at this temperature under a nitrogen atmosphere for 18 hours. After this time the reaction mixture was concentrated, the resulting residue dissolved in ethyl acetate (50 mL) and extracted with saturated sodium bicarbonate (3×10 mL). The combined aqueous extracts were combined and acidified to pH 1 with HCl (6M solution) before being extracted with ethyl acetate (3×10 mL). The organic layers were combined, dried (MgSO.sub.4), filtered and concentrated. The resulting residue was purified by prep HPLC to give the title compound as the ammonium salt which was dissolved in water (2 mL), to pH 1 with HCl (6M solution) before being extracted with ethyl acetate (3×10 mL). The organic layers were combined, dried (MgSO.sub.4), filtered and concentrated to give the title compound (0.26 g, 55% yield) as a white solid.
5-[(1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropyl]-2H-1,2,3,4-tetrazole
(56) δ.sub.H (500 MHz, DMSO) 16.29 (1 H, br. s.) 8.32 (1 H, d) 8.04 (1 H, dd) 7.85 (1 H, d) 3.49-3.55 (1 H, m) 2.87 (1 H, ddd) 1.75-1.85 (2 H, m). Tr=3.87 min m/z (ES.sup.+) (M+H.sup.+) 283, 285.
Example 6
(57) ##STR00010##
Step 1, (1S,2S)-2-[(3-chloro-4-cyclopropoxyphenyl)carbonyl]cyclopropane-1-carboxamide
(58) EDC (0.07 g, 0.36 mmol) and HOBt (0.048 g, 0.36 mmol) were added sequentially to a stirred solution of (1S,2S)-2-{[3-chloro-4-cyclopropoxyphenyl]carbonyl}cyclopropane-1-carboxylic acid (0.1 g, 0.36 mmol) in DMF (2 mL) and the mixture was stirred at room temperature under a nitrogen atmosphere for 15 minutes. After this time, saturated aqueous ammonia (1 mL) was added and stirring was continued for a further 2 hours. After this time, water (5 mL) was added and the mixture was extracted with DCM (2×10 mL). The organic layers were combined, washed consecutively with saturated sodium bicarbonate (50 mL) and brine (30 mL) before being dried (Na.sub.2SO.sub.4), filtered and concentrated. The resulting residue was then purified by prep HPLC to give the title compound (0.02 g, 22% yield) as a white solid.
(1S,2S)-2-[(3-chloro-4-cyclopropoxyphenyl)carbonyl]cyclopropane-1-carboxamide
(59) δ.sub.H (500 MHz, DMSO) 8.07 (dd, J=8.67, 2.21 Hz, 1 H) 8.00 (d, J=2.05 Hz, 1 H) 7.73 (br. s., 1 H) 7.53-7.60 (m, 1 H) 7.06 (br. s., 1 H) 4.09 (tt, J=5.99, 2.92 Hz, 1 H) 3.04 (ddd, J=8.51, 5.04, 3.94 Hz, 1 H) 2.16-2.24 (m, 1 H) 1.35 (dddd, J=18.70, 8.45, 5.64, 2.84 Hz, 2 H) 0.69-0.96 (m, 4 H). Tr=3.87 min m/z (ES.sup.+) (M+H.sup.+) 280, 281.
Example 7
(60) ##STR00011##
Step 1, [(5R,6S,6aR)-6-Hydroxy-tetrahydro-2H-furo[2,3-d][1,3]dioxol-5-yl]methyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(61) See, e.g., Smith et al, Journal of organic Chemistry, 1980, 5000-5002.
(62) Triethylamine (0.08 mL, 0.61 mmol) and 2,2-dimethylpropanoyl chloride (0.07 mL, 0.61 mmol) were added dropwise sequentially to a stirred solution of (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylic acid (0.16 g, 0.61 mmol) in THF (5 mL) and the mixture was stirred at room temperature for 1 hour. After this time the mixture was added portion wise to a solution of (1R)-1-[(3aR,5R,6S,6aR)-6-hydroxy-2,2-dimethyl-tetrahydro-2H-furo[2,3-d][1,3]dioxol-5-yl]ethane-1,2-diol (0.4 g, 1.82 mmol) in pyridine (5 mL) and the reaction mixture was stirred at room temperature under a nitrogen atmosphere for 18 hours. The resulting mixture was concentrated and the residue partitioned between DCM (50 ml) and water (10 ml). The organic layer was separated, dried (MgSO.sub.4), filtered and concentrated. The resulting residue was then purified by flash column chromatography (elution: 100% ethyl acetate) to give the title compound (0.3 g, 99% yield) as a white solid. Tr=2.03 min m/z (ES.sup.+) (M+Na) 485.
Step 1, [(2R,3S,4S,5R)-3,4,5,6-Tetrahydroxyoxan-2-yl]methyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(63) 4M HCl in dioxane solution (5 ml) was added in one portion to a stirred solution of [(5R,6S,6aR)-6-hydroxy-tetrahydro-2H-furo[2,3-d][1,3]dioxol-5-yl]methyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate (0.3 g, 0.6 mmol) in dioxane (5 ml) and the mixture was stirred at room temperature overnight. The resulting mixture was concentrated and the resulting residue was then purified by prep HPLC to give the title compound (0.06 g, 14% yield) as a pale yellow solid.
[(2R,3S,4S,5R)-3,4,5,6-tetrahydroxyoxan-2-yl]methyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(64) δ.sub.H (500 MHz, MeOD) 8.15 (d, J=2.05 Hz, 1 H) 7.94-8.03 (m, 1 H) 7.71 (dd, J=8.43, 2.29 Hz, 1 H) 5.09 (d, J=3.63 Hz, 0.5 H) 4.49 (d, J=7.88 Hz, 0.5 H) 4.37-4.47 (m, 1 H) 4.23-4.32 (m, 1 H) 3.95-4.03 (m, 0.5 H) 3.67 (dd, J=9.30 Hz, 0.5H) 3.51 (ddd, J=9.46, 5.91, 2.13 Hz, 0.5 H) 3.33-3.40 (m, 2 H) 3.22-3.28 (m, 1H) 3.14 (dd, J=8.91, 7.96 Hz, 0.5H) 2.27-2.39 (m, 1 H) 1.61-1.68 (m, 1 H) 1.54-1.61 (m, 1H). Tr=3.44 min m/z (ES.sup.+) (M+H.sup.+) 443, 445.
(65) The following compound was prepared substantially as described above.
[(2R,3R,4S,5R,6S)-3,4,5,6-tetrahydroxyoxan-2-yl]methyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(66) Tr=1.62 min m/z (ES.sup.+) (M+Na.sup.+) 443.
Example 8
(67) ##STR00012##
Step 1, 2-Methylpropyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(68) Oxalyl chloride (0.33 mL, 3.4 mmol) was added dropwise to a stirred solution of (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylic acid (0.3 g, 1.09 mmol) in DCM (9 mL) with a drop of DMF and the mixture was stirred at room temperature for 1 hour. After this time the mixture was concentrated, re-dissolved in DCM and butan-1-ol (0.05 ml, 0.56 mmol) was added in one portion and stirring was continued for a further 72 hours. The resulting mixture was concentrated and purified by flash column chromatography (elution: 10% ethyl acetate, 90% heptanes) to give the title compound (0.16 g, 58% yield) as a colourless oil.
2-Methylpropyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(69) δ.sub.H (500 MHz, CDCl.sub.3) 8.10 (d, J=2.05 Hz, 1 H) 7.85 (dd, J=8.35, 2.05 Hz, 1 H) 7.59 (d, J=8.35 Hz, 1 H) 3.89-3.97 (m, 2 H) 3.06-3.13 (m, 1 H) 2.39-2.46 (m, 1 H) 1.92-2.03 (m, 1 H) 1.60-1.68 (m, 2 H) 0.96 (d, J=6.62 Hz, 6 H). Tr=5.49 min m/z (ES.sup.+) (M+H.sup.+) 313, 315.
(70) The following compounds were prepared substantially as described above.
Butyl (1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(71) δ.sub.H (500 MHz, DMSO) 8.26 (d, J=2.05 Hz, 1 H), 8.02 (dd, J=2.05, 8.35 Hz, 1 H), 7.83 (d, J=8.35 Hz, 1 H), 4.08 (t, J=6.62 Hz, 2 H), 3.30 (ddd, J=3.94, 5.44, 8.91 Hz, 1 H), 2.22 (ddd, J=3.86, 5.91, 8.67 Hz, 1 H), 1.48-1.61 (m, 4 H), 1.34 (qd, J=7.40, 14.99 Hz, 2 H), 0.89 (t, J=7.41 Hz, 3 H). Tr=5.66 min m/z (ES.sup.+) (M+H.sup.+) 315, 317.
Propan-2-yl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(72) δ.sub.H (500 MHz, CDCl.sub.3) 8.10 (d, J=2.05 Hz, 1H) 7.85 (dd, J=8.35, 2.05 Hz, 1 H) 7.59 (d, J=8.35 Hz, 1 H) 5.01-5.10 (m, 1H) 3.04-3.11 (m, 1 H) 2.34-2.41 (m, 1 H) 1.59-1.65 (m, 2H) 1.28 (m, J=6.00, 6.00 Hz, 6H). Tr=5.25 min m/z (ES.sup.+) (M+H.sup.+) 299, 301.
Propan-2-yl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(73) δ.sub.H (500 MHz, CDCl.sub.3) 8.09 (d, J=2.05 Hz, 1 H, 7.81-7.88 (m, 1 H, 7.58 (d, J=8.35 Hz, 1 H, 4.16-4.26 (m, 2 H, 3.07-3.15 (m, 1 H, 2.74 (t, J=6.07 Hz, 2 H, 2.60 (q, J=7.09 Hz, 4 H, 2.39-2.46 (m, 1 H, 1.55-1.71 (m, 2 H, 1.05 (t, J=7.09 Hz, 6 H,. Tr=3.20 min m/z (ES.sup.+) (M+H.sup.+) 358, 360.
4-tert-Butylphenyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(74) δ.sub.H (500 MHz, CDCl.sub.3) 8.14 (d, J=1.9 Hz, 1 H,, 7.89 (dd, J=2.0, 8.4 Hz, 1 H,, 7.61 (d, J=8.4 Hz, 1 H,, 7.43-7.37 (m, 2 H,, 7.08-7.01 (m, 2 H,, 3.27-3.20 (m, 1 H,, 2.65 (ddd, J=3.8, 6.5, 8.2 Hz, 1 H,, 1.81-1.73 (m, 2 H,, 1.33 (s, 9 H,. Tr=5.86 min m/z (ES.sup.+) (M+Na.sup.+) 413, 415.
Example 9
(75) ##STR00013##
Step 1, (2E)-4-(3,4-dichlorophenyl)-4-oxobut-2-enoic acid
(76) See, e.g., U.S. Pat. No. 6,323,240.
(77) AlCl.sub.3 (20.4 g, 152.9 mmol) was added portion wise to a stirred solution of 2,5-dihydrofuran-2,5-dione (5.0 g, 51.0 mmol) in 1,2-dichlorobenzene (34.6 ml, 305.9 mmol) at room temperature under a nitrogen atmosphere and the reaction mixture was heated to 50° C. for 4 hours. After this time the mixture was allowed to cool to room temperature and poured slowly onto cold HCl (6M solution, 120 ml) maintaining the temperature below 30° C. The quenched mixture was stirred for 15 minutes before the addition of hexane (120 ml) and stirring was continued for a further 30 minutes. The resulting solid was then collected by filtration and dried in a vacuum oven for 4 hours before being heated to reflux in diethyl ether (50 ml) and allowed to cool to room temperature. The solid was then filtered, washed with diethyl ether (10 ml) and air dried to to give the title compound (10.0 g, 80% yield) as a pale yellow solid. Tr=1.95 min m/z (ES.sup.+) (M+H.sup.+) 243, 245.
Step 2, 4-(3,4-Dichloro-phenyl)-4-oxo-but-2-enoic acid methyl ester
(78) A mixture of 4-(3,4-dichloro-phenyl)-4-oxo-but-2-enoic acid (4.0 g, 16.3 mmol), NaHCO.sub.3 (2.74 g, 32.6 mmol) and dimethyl sulfate (3.1 ml, 32.6 mmol) in acetone (50 ml) were stirred at reflux under a nitrogen atmosphere for 6 hours. After this time the reaction mixture was allowed to cool to room temperature and the remaining solid was removed by filtration. The solid was washed with acetone (10 ml) and the filtrate concentrated under reduced pressure. The resulting residue was dissolved in DCM (15 ml), washed sequentially with saturated NaHCO.sub.3 (2×10 ml) followed by water (10 ml). The organic layer was dried (MgSO.sub.4), filtered and concentrated. The resulting residue was purified by flash column chromatography (elution: 10% ethyl acetate, 90% heptane) to give the title compound (4.1 g, 97% yield) as a yellow solid. Tr=2.25 min m/z (ES.sup.+) (M+H.sup.+) 259, 261.
Step 3, 3-(3,4-Dichloro-benzoyl)-aziridine-2-carboxylic acid
(79) See, e.g., Wincewicz et al, Organic Letters, 2007, 351-353.
(80) 4-Methylmorpholine (0.14 ml, 1.29 mmol) was added dropwise to a stirred solution of amino diphenylphosphinate (0.3 g, 1.29 mmol) in DCM (20 ml) and the white mixture was stirred at room temperature under a nitrogen atmosphere for 30 minutes before the sequential addition of NaOH (0.15 g, 3.86 mmol) and methyl (2E)-4-(3,4-dichlorophenyl)-4-oxobut-2-enoate (0.5 g, 1.93 mmol). The resulting mixture was stirred at room temperature for a further 20 hours. After this time, water (30 ml) was added and the mixture was acidified to pH 2-3 with HCl (1M solution). The phases were separated and the aqueous phase was extracted with DCM (3×15 ml). The combined organic extracts were dried (MgSO.sub.4), filtered and concentrated. The resulting residue was triturated with diethyl ether (30 ml) and the solvent was decanted off. The residue was then purified by prep HPLC to give the title compound (0.021 g, 4% yield) as a pale yellow solid.
3-(3,4-Dichloro-benzoyl)-aziridine-2-carboxylic acid
(81) δ.sub.H (500 MHz, DMSO) 13.12 (br. s., 1 H, 8.27 (d, J=1.42 Hz, 1 H, 8.02 (dd, J=8.35, 1.89 Hz, 1 H, 7.85 (d, J=8.35 Hz, 1 H, 3.84 (br. s., 1 H, 2.79 (br. s., 1 H, 2.69 (br. s., 1H). Tr=3.49 min m/z (ES.sup.+) (M+H.sup.+) 260, 262.
Example 10
(82) ##STR00014##
(83) See, e.g., International Journal of Pharmaceutics, 2010, 39.
Step 1, Method 11: Methyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-6-{[(1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropyl]formamido}hexanoate
(84) Diisopropylethylamine (0.22 mL, 1.3 mmol) was added in one portion to a stirred solution of (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylic acid (0.11 g, 0.43 mmol), methyl (2R)-6-amino-2-{[(tert-butoxy)carbonyl]amino}hexanoate (0.11 g, 0.43 mmol) and HATU (0.16 g, 0.43 mmol) in DMF (8 ml) and the mixture was stirred at room temperature for 16 hours. After this time the resulting mixture was concentrated, and purified by flash column chromatography (elution: 30% ethyl acetate, 60% heptanes to 50% ethyl acetate, 50% heptanes) to give the title compound (0.16 g, 64% yield) as a white gum. δ.sub.H (500 MHz, CDCl.sub.3) 8.11 (d, J=1.89 Hz, 1H), 7.87 (dd, J=1.89, 8.35 Hz, 1H), 7.58 (d, J=8.35 Hz, 1H), 5.98 (br. s., 1H), 5.15 (d, J=7.57 Hz, 1H), 4.20-4.37 (m, 1H), 3.74 (s, 3H), 3.31 (q, J=6.36 Hz, 2H), 3.04-3.21 (m, 1H), 2.11-2.27 (m, 1H), 1.75-1.89 (m, 1H), 1.62-1.73 (m, 2H), 1.49-1.62 (m, 3H), 1.34-1.49 (m, 11H). Tr=2.45 min m/z (ES.sup.+) (M+Na.sup.+) 524, 526.
Step 2, Methyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-6-{[(1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropyl]formamido}hexanoate
(85) NaOH (2M solution, 2 mL, 4.3 mmol) was added in one portion to a stirred solution of methyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-6-{[(1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropyl]formamido}hexanoate
(86) (0.15 g, 0.3 mmol) in THF (2 mL) and the mixture was stirred at room temperature for 1 hour. After this time the resulting mixture was concentrated, and added in one portion to a stirred solution of HCl in dioxane (4M solution, 4 mL) and stirred at room temperature for 72 hours. After this time the resulting mixture was concentrated, suspended in acetonitrile/water (1:1, 3 mL) and the resulting precipitate was collected by filtration to give the title compound (0.05 g, 40% yield) as a white powder.
(25)-2-amino-6-{[(1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropyl]formamido}hexanoic acid
(87) δ.sub.H (500 MHz, DMSO) 8.32 (t, J=5.36 Hz, 1H), 8.19 (d, J=1.73 Hz, 1H), 8.16-8.20 (m, 1H), 8.19 (br. s, 3H), 7.99 (dd, J=1.81, 8.43 Hz, 1H), 7.83 (d, J=8.51 Hz, 1H), 3.85 (t, J=5.75 Hz, 1H), 2.99-3.15 (m, 3H), 2.23-2.32 (m, 1H), 1.66-1.87 (m, 2H), 1.21-1.50 (m, 6H). Tr=3.12 min m/z (ES.sup.+) (M+H.sup.+) 387, 389.
Example 11
(88) ##STR00015##
Step 1, 1-(3,4-Dichlorophenyl)prop-2-en-1-ol)
(89) See, e.g., Toshiyuki et al, Tetrahedron Letters, 2007, 7774-7777.
(90) Vinylmagnesium bromide (1.0M solution in THF, 27.4 ml, 27.4 mmol) was added dropwise to a cold (−78° C.), stirred solution of 3,4-dichlorobenzaldehyde (4.00 g, 22.9 mmol) in dry THF (35 ml) under a nitrogen atmosphere. The reaction mixture was allowed to warm to 0° C. and stirred at this temperature for 1 hour before being re-cooled to −78° C. and quenched by the addition of saturated ammonium chloride (50 ml). Water (40 ml) and brine (30 ml) were added and the mixture extracted with ethyl acetate (2×100 ml). The combined organic layers were dried (Na.sub.2SO.sub.4), filtered and concentrated to give the title product (4.66 g, 96% yield) as a colourless oil. Tr=1.90 min m/z (ES.sup.+) (M+H.sup.+) 185, 187.
Step 2, 1-(3,4-dichlorophenyl)prop-2-en-1-one
(91) IBX (15.3 g, 24.7 mmol) was added portionwise to a stirred solution of 143,4-dichloro-phenyl)-prop-2-en-1-ol (3.34 g, 16.5 mmol) in DMSO (130 ml) and the mixture was stirred at room temperature under a nitrogen atmosphere for 4 days. After this time the mixture was partitioned between water (250 ml) extracted with DCM (3×100 ml) and the organic layers combined, dried (Na.sub.2SO.sub.4), filtered and concentrated. The resulting residue was purified using a Biotage Isolera (340 g silica column eluting with 1% ethyl acetate/99% heptane to 20% ethyl acetate, 80% heptane) to give the title compound (2.00 g, 55% yield) as a cream oil. Tr=2.15 min m/z (ES.sup.+) (M+H.sup.+) 201, 203.
Step 3, 1-[(3,4-dichlorophenyl)carbonyl]-5-oxaspiro[2.4]heptan-4-one
(92) See, e.g., Trost et al, Journal of organic Chemistry, 1973, 3140.
(93) Dimethyl-(2-oxo-tetrahydro-furan-3-yl)-sulfonium tetrafluoroborate (0.67 g, 2.89 mmol) was added portionwise to a stirred solution of 1-(3,4-dichlorophenyl)prop-2-en-1-one (0.58 g, 2.89 mmol) in THF (12 ml) and the mixture was stirred at 0° C. for 10 minutes, after which time sodium hydride (0.13 g of a 60% dispersion in oil, 3.17 mmol) was added portion wise and stirring was continued for a further 5 hours under a nitrogen atmosphere. After this time the mixture was partitioned between saturated ammonium chloride (50 ml) extracted with ethyl acetate (3×100 ml) and the organic layers combined, dried (Na.sub.2SO.sub.4), filtered and concentrated. The resulting residue was purified using a Biotage Isolera (100 g silica column eluting with 1% ethyl acetate/99% heptane to 40% ethyl acetate, 60% heptane) to give the title compound (0.06 g, 7% yield) as a white crystalline solid.
1-[(3,4-dichlorophenyl)carbonyl]-5-oxaspiro[2.4]heptan-4-one
(94) δ.sub.H (500 MHz, CDCl.sub.3) 8.09 (d, J=2.05 Hz, 1 H, 7.84 (dd, J=8.35, 2.05 Hz, 1 H, 7.60 (d, J=8.35 Hz, 1 H, 4.30-4.52 (m, 2 H, 3.26 (dd, J=8.59, 6.23 Hz, 1 H, 2.23-2.55 (m, 2 H, 1.76-1.88 (m, 2 H,. Tr=4.40 min m/z (ES.sup.+) (M+Na.sup.+) 307, 309.
Example 12
(95) ##STR00016##
Step 1, (1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]-N-phenylcyclopropane-1-carboxamide
(96) Triethylamine (0.005 mL, 0.04 mmol) was added in one portion to a stirred solution of (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylic acid (0.01 g, 0.04 mmol), aniline (0.004 mL, 0.04 mmol), EDC (0.007 g, 0.04 mmol) and HOBt (0.001 g, 0.004 mmol) in DMF (8 ml) and the mixture was stirred at room temperature for 16 hours. After this time the resulting mixture was concentrated, partitioned between water (5 ml) extracted with ethyl acetate (3×10 ml) and the organic layers combined, dried (MgSO.sub.4), filtered and concentrated to give the title compound (0.01 g, 82% yield) as a white solid.
(1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]-N-phenylcyclopropane-1-carboxamide
(97) Tr=4.84 min m/z (ES.sup.+) (M+H.sup.+) 334, 336.
(98) The following compounds were prepared substantially as described above.
(1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]-N-(2-hydroxyethyl)cyclopropane-1-carboxamide
(99) δ.sub.H (500 MHz, CDCl.sub.3) 8.02-8.18 (1 H, m), 7.77-7.95 (1 H, m), 7.49-7.65 (1 H, m), 6.30 (1 H, br. s.), 3.69-3.83 (2 H, m), 3.43-3.55 (2 H, m), 3.08-3.20 (1 H, m), 2.38 (1 H, br. s.), 2.20-2.29 (1 H, m), 1.65-1.74 (1 H, m), 1.51-1.58 (1H, m). Tr=3.49 min m/z (ES.sup.+) (M+H.sup.+) 302.
4-{[(1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropyl]carbonyl}piperazin-2-one
(100) δ.sub.H (250 MHz, DMSO) 8.15 (1 H, d, J=1.98 Hz), 7.98 (1 H, dd, J=8.38, 1.98 Hz), 7.57-7.84 (2 H, m), 3.90-4.24 (2 H, m), 3.60-3.83 (2 H, m), 3.20-3.34 (2 H, m), 3.08-3.20 (1 H, m), 2.54-2.62 (1 H, m), 1.42-1.60 (2 H, m). Tr=3.52 min m/z (ES.sup.+) (M+H.sup.+) 341, 343.
Example 13
(101) ##STR00017##
Step 1, 2-(Oxan-2-yloxy)ethyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(102) 2-(Tetrahydro-2H-pyran-2-yloxy) ethanol (0.07 mL, 0.46 mmol) was added in one portion to a stirred solution of (+)-(1S,2S)-cyclopropane-1,2-dicarboxylic acid monomethyl ester (0.12 g, 0.46 mmol) in chloroform (6 mL) and the reaction mixture was stirred at room temperature under a nitrogen atmosphere for 72 hours. After this time, the reaction mixture was concentrated and the resulting residue was purified by flash column chromatography (elution: 20% ethyl acetate, 80% heptanes) to give the title compound (0.14 g, 78% yield) as a white solid. Tr=2.39 min m/z (ES.sup.+) (M+Na.sup.+) 409.
Step 2, 2-Hydroxyethyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(103) Concentrated HCl (5 drops) was added dropwise to a stirred solution of 2-(oxan-2-yloxy)ethyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate (0.14 g, 0.36 mmol) and the reaction mixture was stirred at room temperature under a nitrogen atmosphere for 18 hours. After this time, the reaction mixture was concentrated and the resulting residue was purified by flash column chromatography (elution: 1% methanol, 99% DCM) to give the title compound (0.03 g, 29% yield) as a white solid.
2-Hydroxyethyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(104) δ.sub.H (250 MHz, CDCl.sub.3) 8.24-8.18 (1 H, m),7.87-7.83 (1 H, m), 7.62-7.56 (1 H, m), 4.37 4.26 (2 H, m), 3.93-3.84 (2 H, m), 3.22-3.06 (1H, m), 2.54-2.43 (1H, m), 1.92-1.87 (1H, m), 1.71-1.63 (2H, m). Tr=3.49 min m/z (ES.sup.+) (M+H.sup.+) Ion not seen.
Example 14
(105) ##STR00018##
Step 1, {[(1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropyl]carbonyloxy}methyl 2,2-dimethylpropanoate
(106) A solution of 2,2-dimethyl-propionic acid iodomethyl ester (0.26 g, 1.07 mmol) in dry DMF (4 mL) was added dropwise over 10 minutes to a cool (0° C.), stirred solution of sodium (1S,2S)-2-(3,4-dichloro-benzoyl)-cyclopropanecarboxylate (0.2 g, 0.71 mmol) in dry DMF (4 mL) and the reaction was warmed to room temperature and stirring continued for 22 hours. After this time a solution of 2,2-dimethyl-propionic acid iodomethyl ester (0.4 g, 1.65 mmol) in DMF (1 mL) was added and stirring continued for a further 1 hour. After this time potassium carbonate (0.3 g, 2.14 mmol) was added portion wise and stirring continued for 14 hours. After this time the reaction mixture was filtered and the filtrate concentrated. The resulting residue was treated with brine (30 mL), extracted with ethyl acetate (3×30 mL) and the combined organic extracts were dried (Na.sub.2SO.sub.4), filtered and concentrated. The resulting residue was purified on a Biotage Isolera (25 g column, elution; 2% ethyl acetate, 98% heptanes to 20% ethyl acetate, 80% heptanes) to give the title product (0.08 g, 29% yield) as a white crystalline solid.
{[(1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropyl]carbonyloxy}methyl 2,2-dimethylpropanoate
(107) δ.sub.H (500 MHz, CDCl.sub.3) 8.09 (d, J=2.05 Hz, 1 H) 7.79-7.91 (m, 1 H) 7.59 (d, J=8.35 Hz, 1 H) 5.80 (s, 2 H) 3.06-3.21 (m, 1H) 2.30-2.52 (m, 1 H) 1.62-1.76 (m, 2 H) 1.23 (s, 9 H). Tr=5.34 min m/z (ES.sup.+) (M+Na.sup.+) 395, 397.
Example 15
(108) ##STR00019##
Step 1, Method 17: 1-{[(1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropyl]carbonyloxy}ethyl acetate
(109) 1-Chloroethyl acetate (0.14 g, 1.18 mmol) was added in one portion to a stirred suspension of sodium (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate (0.15 g, 0.54 mmol) in dry DMF (3 mL) and the mixture was heated to 70° C. and stirred at this temperature under nitrogen atmosphere for 6 hours. After this time the reaction mixture was added to water (5 mL) and extracted with ethyl acetate (4×5 ml). The combined organic extracts were washed with a 1:1 mixture of water and brine (3×20 mL), the aqueous layers were back extracted with EtOAc (2×5 mL). The combined organic layers were washed with brine (5 mL), dried (MgSO.sub.4), filtered and the concentrated. The resulting residue was dry loaded onto silica gel using DCM and purified by dry flash chromatography (elution; 100% heptanes to 20% ethyl acetate, 80% heptanes) to give the title compound (0.08 g, 45% yield) as a light brown solid.
1-{[(1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropyl]carbonyloxy}ethyl acetate
(110) δ.sub.H (500 MHz, CDCl.sub.3) 8.07-8.13 (m, 1 H), 7.81-7.89 (m, 1 H), 7.56-7.63 (m, 1H), 6.85-6.93 (m, 1 H), 3.07-3.16 (m, 1 H), 2.35-2.43 (m, 1 H), 2.05-2.15 (m, 3 H), 1.61-1.71 (m, 2 H), 1.48-1.55 (m, 3 H). Tr=3.29 min m/z (ES.sup.+) (M+H.sup.+) 397, 399.
(111) The following compounds were prepared substantially as described above.
1-{[(Propan-2-yloxy)carbonyl]oxy}ethyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(112) δ.sub.H (500 MHz, CDCl.sub.3) 8.03-8.13 (m, 1 H) 7.81-7.89 (m, 1 H) 7.59 (m, J=8.40, 1.30 Hz, 1 H) 6.75-6.86 (m, 1 H) 4.83-4.98 (m, 1 H) 3.07-3.17 (m, 1 H) 2.36-2.44 (m, 1 H) 1.62-1.73 (m, 2 H) 1.51-1.60 (m, 3 H) 1.24-1.38 (m, 6H). Tr=5.33 min m/z (ES.sup.+) (M+Na.sup.+) 411, 413.
1-[(Ethoxycarbonyl)oxy]ethyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(113) δ.sub.H (500 MHz, CDCl.sub.3) 8.07-8.13 (m, 1 H) 7.82-7.89 (m, 1 H) 7.56-7.63 (m, 1 H) 6.75-6.84 (m, 1 H) 4.16-4.30 (m, 2 H) 3.08-3.18 (m, 1 H) 2.37-2.45 (m, 1 H) 1.62-1.73 (m, 2 H) 1.53-1.61 (m, 3 H) 1.27-1.38 (m, 3H). Tr=5.17 min m/z (ES.sup.+) (M+Na.sup.+) 397, 399.
1-{[(1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropyl]carbonyloxy}ethyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate
(114) δ.sub.H (500 MHz, CDCl.sub.3) 8.06-8.13 (m, 2 H) 7.80-7.88 (m, 2 H) 7.55-7.63 (m, 2 H) 6.88-6.96 (m, 1 H) 3.06-3.17 (m, 2 H) 2.36-2.46 (m, 2 H) 1.61-1.71 (m, 4 H) 1.52-1.59 (m, 3 H). Tr=5.10 min m/z (ES.sup.+) (M+Na.sup.+) 565, 567, 569.
Example 16
(115) See, e.g., Molecules, 2009, 3268-3274.
(116) ##STR00020##
Step 1, (3-Carboxy-2-{[(1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropyl]carbonyloxy}propyl)trimethylazanium
(117) Diisopropylethylamine (0.31 mL, 1.80 mmol) was added in one portion to a stirred solution of (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylic acid (0.16 g, 0.60 mmol), L-Carnitine (0.11 g, 0.66 mmol) and HATU (0.23 g, 0.60 mmol) in DMF (5 mL) and the mixture was stirred at room temperature for 45 hours before being heated to 50° C. and stirred at this temperature for a further 8 hours. After this time the resulting mixture was concentrated and the resulting residue purified by prep HPLC to give the title compound (0.01 g, 4% yield) as a brown gum.
(3-Carboxy-2-{[(1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropyl]carbonyloxy}propyl)trimethylazanium
(118) δ.sub.H (500 MHz, DMSO-d.sub.6) 8.29 (br. s., 1H), 8.25 (dd, J=0.32, 1.73 Hz, 1H), 8.00-8.05 (m, 1H), 7.84 (d, J=8.35 Hz, 1H), 5.36-5.51 (m, 1H), 3.55-3.80 (part. obsc. m, 2H), 3.36 (ddd, J=4.02, 5.28, 8.83 Hz, 1H), 3.11 (s, 9H), 2.54-2.62 (m, 1H), 2.39-2.48 (m, 1H), 2.22-2.29 (m, 1H), 1.60 (ddd, J=3.70, 5.71, 8.87 Hz, 1H), 1.52 (ddd, J=3.63, 5.44, 8.75 Hz, 1H). Tr=3.12 min m/z (ES.sup.+) (M+H.sup.+) 402, 404.
Example 17
(1S,2S)-2-(7-chloro-2,3-dihydro-1-benzofuran-5-carbonyl)cyclopropane-1-carboxylic acid
(119) ##STR00021##
Step 1: 2-Chloro-6-(2-hydroxyethyl)phenol
(120) Sodium periodate (8.9 g, 41.5 mmol) was added portion wise to a stirred solution of 2-chloro-6-(prop-2-en-1-yl)phenol (3.72 g, 20.7 mmol) in THF (100 mL) and water (100 mL) at 0° C. After 5 minutes, osmium tetroxide (0.1 g, 0.41 mmol) was added and stirring continued for 1.5 hours. After this time, the mixture was poured into ice and brine (100 mL) and extracted with ethyl acetate (3×100 mL). The combined organic extracts were dried (Na.sub.2SO.sub.4), filtered and concentrated. The resulting residue was dissolved in methanol (100 mL) and cooled to 0° C. before being treated with sodium tetrahydroborate (1.57 g, 41.5 mmol) in small portions over 30 minutes. After this time, the reaction mixture was warmed to room temperature and stirred overnight. The resulting mixture was concentrated, treated with aqueous 1M hydrochloric acid (80 mL) and extracted with ethyl acetate (3×100 mL). The combined organic extracts were dried (Na.sub.2SO.sub.4), filtered and concentrated. The resulting residue was purified by flash column chromatography (elution; 0-40% ethyl acetate in heptanes) to give the title compound (1.41 g, 36% yield) as a yellow oil. δ.sub.H (250 MHz, CDCl.sub.3) 7.23 (dd, J=8.0, 1.6 Hz, 1H), 7.03 (dd, J=7.5, 1.6 Hz, 1H), 6.81 (t, J=7.8 Hz, 1H), 3.95 (t, J=5.9 Hz, 2H), 2.95 (t, J=5.8 Hz, 2H).
Step 2: 7-Chloro-2,3-dihydro-1-benzofuran
(121) Diisopropyldiazene-1,2-dicarboxylate (1.97 ml, 9.9 mmol) was added portionwise to a stirred, cooled (0° C.) solution of 2-chloro-6-(2-hydroxyethyl)phenol (1.4 g, 7.7 mmol) and triphenylphosphine (2.62 g, 9.9 mmol) in anhydrous THF (25 ml) and the reaction mixture was stirred a nitrogen atmosphere for 15 hours. After this time the reaction mixture was concentrated and the resulting residue purified by Biotage (Isolera snap 50 g cartridge, eluting with 0-20% EtOAc in Heptanes) to give the title compound (1.46 g, 92% yield) as an orange oil. δ.sub.H (250 MHz, CDCl.sub.3) 7.16-7.02 (m, 2H), 6.85-6.71 (m, 1H), 4.67 (t, J=8.8 Hz, 2H), 3.29 (t, J=8.8 Hz, 2H).
Step 3: Methyl (1S,2S)-2-(7-chloro-2,3-dihydro-1-benzofuran-5-carbonyl)cyclopropane-1-carboxylate
(122) Methyl (1S,2S)-2-(carbonochloridoyl)cyclopropane-1-carboxylate (0.39 g, 2.43 mmol) in DCE (2 mL) was added portionwise to a cooled (0° C.), stirred solution of aluminium trichloride (0.65 g, 4.85 mmol) in DCE (4 mL) under a nitrogen atmosphere. 7-Chloro-2,3-dihydro-1-benzofuran (0.5 g, 2.43 mmol) was added dropwise over 5 minutes and the reaction mixture was stirred at 0° C. for a further 1 hour. After this time, the mixture was allowed to warm to room temperature before being stirred overnight. The reaction mixture was then cooled to 0° C. before being added portionwise to a mixture of concentrated HCl (4 mL) and ice (20 g). The resulting mixture was then extracted with DCM (3×50 mL), the combined organic extracts were washed with brine (30 mL), before being dried (MgSO.sub.4), filtered and concentrated. The resulting residue was purified using a Biotage Isolera, (Snap 50 g cartridge, eluting in 0-35% EtOAc in Heptanes) to give the title compound (0.14 g, 21% yield) as a white solid. δ.sub.H (250 MHz, CDCl.sub.3) 7.88 (d, J=1.6 Hz, 1H), 7.78 (d, J=1.5 Hz, 1H), 4.79 (t, J=8.9 Hz, 2H), 3.74 (s, 3H), 3.35 (t, J=8.8 Hz, 2H), 3.07 (ddd, J=8.6, 5.9, 3.9 Hz, 1H), 2.37 (ddd, J=8.6, 6.0, 3.8 Hz, 1H), 1.58 (ddt, J=12.0, 5.9, 2.9 Hz, 2H). Tr=2.08 min m/z (ES.sup.+) (M+H.sup.+) 281.
Step 4: (1S,2S)-2-(7-chloro-2,3-dihydro-1-benzofuran-5-carbonyl)cyclopropane-1-carboxylic acid
(123) 2M NaOH (0.25 mL, 0.5 mmol) was added in one portion to a stirred solution of methyl (1S,2S)-2-[(7-chloro-2,3-dihydro-1-benzofuran-6-yl)carbonyl]cyclopropane-1-carboxylate (0.07 g, 0.25 mmol) in dioxane (5 mL) and the resulting solution was stirred at room temperature for 18 hours. After this time the reaction mixture was acidified with 1M HCl, and the resulting suspension was extracted with EtOAc (2×10 mL). The combined organic extracts were washed with brine (10 mL), before being dried (MgSO4), filtered and concentrated. The resulting residue was part-dissolved in TBME (2 mL), sonicated and the resulting precipitate was collected by filtration and dried under vacuum to give the title compound (0.04 g, 54% yield) as a white powder.
(1S,2S)-2-(7-chloro-2,3-dihydro-1-benzofuran-5-carbonyl)cyclopropane-1-carboxylic acid
(124) δ.sub.H (500 MHz, CDCl.sub.3) 7.88 (s, 1H), 7.78 (s, 1H), 4.79 (t, J=8.9 Hz, 2H), 3.36 (t, J=8.8 Hz, 2H), 3.12 (ddd, J=9.4, 5.8, 3.9 Hz, 1H), 2.37 (ddd, J=9.3, 5.7, 3.9 Hz, 1H), 1.70-1.60 (m, 2H). Tr=2.61 min (7 minute method, low pH) m/z (ES+) (M+H+) 267.
(1S,2S)-2-(2,3-dihydro-1-benzofuran-5-carbonyl)cyclopropane-1-carboxylic acid
(125) δ.sub.H (500 MHz, CDCl.sub.3) 7.96-7.83 (m, 2H), 6.84 (d, J=8.78 Hz, 1H), 4.68 (t, J=8.78 Hz, 2H), 3.27 (t, J=8.76 Hz, 2H), 3.18 (ddd, J=3.84, 5.89, 9.44 Hz, 1H), 2.35 (ddd, J=3.83, 5.65, 9.20 Hz, 1H), 1.67 (ddd, J=3.50, 5.90, 9.02 Hz, 1H), 1.59 (ddd, J=3.51, 5.66, 8.96 Hz, 1H). Tr=2.21 min, m/z (ES+) (M+H+) 233.
(126) TABLE-US-00006 Structure IUPAC Name Molecular Weight LCMS Data
Example 18
(1S,2S)-2-(8-Chloro-3,4-dihydro-2H-1-benzopyran-6-carbonyl)cyclopropane-1-carboxylic acid
(127) ##STR00024##
Step 1: 1-Chloro-2-(prop-2-en-1-yloxy)benzene
(128) Sodium hydride (60%, 5.6 g, 140.02 mmol) was suspended in anhydrous DMF (80 mL) in an ice-bath before 2-chlorophenol (11.9 mL, 116.7 mmol) as a solution DMF (20 mL) was added dropwise over 30 minutes. Once the addition was complete, the reaction mixture was allowed to stir for 45 minutes. After this time 3-bromoprop-1-ene (12.12 mL, 140.0 mmol) was added dropwise and the reaction was allowed to warm to room temperature and stirring was continued for a further 15 hours. After this time, aqueous saturated ammonium chloride (50 mL) was added and the mixture was extracted with ethyl acetate (3×200 mL). The combined organic extracts were washed with brine (50 mL) dried (Na.sub.2SO.sub.4), filtered, and concentrated, The resulting residue was dissolved in ethyl acetate (200 mL) and washed with water (3×200 mL) and brine solution (50 mL). The organics were dried (Na.sub.2SO.sub.4), filtered and concentrated to give the title compound (17.5 g, 89% yield) as an orange oil which was taken on directly without further purification.
Step 2: 2-Chloro-6-(prop-2-en-1-yl)phenol
(129) A solution of 1-chloro-2-(prop-2-en-1-yloxy)benzene (17.5 g, 93.4 mmol) in mesitylene (150 mL) was heated under nitrogen for 48 h at 190° C. with stirring. After this time, the reaction was cooled to room temperature and concentrated. The resulting residue was purified Biotage (Snap Isolera 340 g, eluting with 100% heptanes). The purified material was treated with 2M NaOH (6 mL), diluted water (50 mL) and the starting material was extracted with TBME (100 mL). The basic aqueous layer was acidified with 6M HCl (6 mL). The aqueous layer was extracted with TBME (2×100 mL), the organic extracts were combined, dried (Na.sub.2SO.sub.4), filtered and concentrated to give the title compound (6.5 g, 41% yield) as a brown oil. δ.sub.H (250 MHz, CDCl.sub.3) 7.34 (s, 1H), 7.17 (s, 1H), 6.08-5.82 (m, 1H), 5.60 (s, 1H), 5.15 (d, J=1.2 Hz, 1H), 5.12-5.05 (m, 1H), 3.40 (d, J=6.7 Hz, 2H).
Step 3: 2-Chloro-6-(3-hydroxypropyl)phenol
(130) 2-Chloro-6-(prop-2-en-1-yl)phenol (1.37 g, 0.01 mol) in anhydrous THF (20 mL) at room temperature was treated dropwise with 1M borane-tetrahydrofuran (1:1 solution, 7.3 mL) and stirring was continued for 15 hours. After this time, water (0.13 mL) was added carefully over 5 minutes, before 2M NaOH (1.68 mL) was added dropwise over 15 minutes. Hydrogen peroxide (0.2 mL, 0.01 mol) was then added dropwise and the mixture stirred at room temperature for a further 1.5 hours. After this time, the mixture was treated with water (20 mL) and then partitioned between ethyl acetate (100 mL) and water (50 mL). The organic layer was separated and the aqueous extracted with ethyl acetate (2×100 mL), the combined organic extracts were washed with brine (50 mL), before being dried (MgSO.sub.4) filtered and concentrated. The resulting residue was purified by flash column chromatography (elution: 0-80% EtOAc, in heptanes) to give the title compound (0.69 g, 43% yield) as a pale orange oil which was used directly.
Step 4: 8-Chloro-3,4-dihydro-2H-1-benzopyran
(131) Diisopropyldiazene-1,2-dicarboxylate (0.64 ml, 3 mmol) was added portionwise to a stirred, cooled (0° C.) solution of 2-chloro-6-(3-hydroxypropyl)phenol (0.49 g, 2.0 mmol) and triphenylphosphine (0.85 g, 3.0 mmol) in anhydrous THF (15 ml) and the reaction mixture was stirred a nitrogen atmosphere for 15 hours. After this time the reaction mixture was concentrated and the resulting residue purified by Biotage (Isolera snap 50 g cartridge, eluting with 0-20% EtOAc in Heptanes) to give the title compound (0.36 g, 83% yield) as a pale pink oil. δ.sub.H (250 MHz, CDCl.sub.3) 7.17 (d, J=7.9 Hz, 1H), 6.98-6.88 (m, 1H), 6.76 (t, J=7.7 Hz, 1H), 4.35-4.25 (m, 2H), 2.81 (t, J=6.5 Hz, 2H), 2.11-1.93 (m, 2H).
Step 5: Methyl (1S,2S)-2-(8-chloro-3,4-dihydro-2H-1-benzopyran-6-carbonyl)cyclopropane-1-carboxylate
(132) Methyl (1S,2S)-2-(carbonochloridoyl)cyclopropane-1-carboxylate (0.19 g, 1.19 mmol) in DCE (2 mL) was added portionwise to a cooled (0° C.), stirred solution of aluminium trichloride (0.32 g, 2.37 mmol) in DCE (4 mL) under a nitrogen atmosphere. 8-Chloro-3,4-dihydro-2H-1-benzopyran (0.2 g, 1.19 mmol) was added dropwise over 5 minutes and the reaction mixture was stirred at 0° C. for a further 1 hour. After this time, the mixture was allowed to warm to room temperature before being stirred overnight. The reaction mixture was then cooled to 0° C. before being added portionwise to a mixture of concentrated HCl (4 mL) and ice (20 g). The resulting mixture was then extracted with DCM (3×50 mL), the combined organic extracts were washed with brine (30 mL), before being dried (MgSO.sub.4), filtered and concentrated. The resulting residue was purified using a Biotage Isolera, (Snap 50 g cartridge, eluting in 0-45% EtOAc in Heptanes) to give the title compound (0.15 g, 43% yield) as a pale yellow oil. δ.sub.H (250 MHz, CDCl.sub.3) 7.88 (d, J=2.1 Hz, 1H), 7.69-7.62 (m, 1H), 4.43-4.31 (m, 2H), 3.73 (s, 3H), 3.08 (ddd, J=8.5, 5.9, 3.8 Hz, 1H), 2.86 (t, J=6.4 Hz, 2H), 2.36 (ddd, J=8.5, 6.1, 3.8 Hz, 1H), 2.16-1.96 (m, 2H), 1.58 (tdd, J=7.6, 5.9, 3.4 Hz, 2H). Tr=2.00 min m/z (ES.sup.+) (M+H.sup.+) 295
Step 6: (1S,2S)-2-(8-chloro-3,4-dihydro-2H-1-benzopyran-6-carbonyl)cyclopropane-1-carboxylic acid
(133) 2M NaOH (0.6 mL, 1.02 mmol) was added in one portion to a stirred solution of methyl (1S,2S)-2-[(8-chloro-3,4-dihydro-2H-1-benzopyran-6-yl)carbonyl]cyclopropane-1-carboxylate (0.15 g, 0.51 mmol) in dioxane (5 mL) and the resulting solution was stirred at room temperature for 18 hours. After this time the reaction mixture was acidified with 1M HCl, and the resulting suspension was extracted with EtOAc (2×10 mL). The combined organic extracts were washed with brine (10 mL), before being dried (MgSO4), filtered and concentrated. The resulting residue was part-dissolved in TBME (2 mL), sonicated and the resulting precipitate was collected by filtration and dried under vacuum to give the title compound (0.01 g, 10% yield) as a white powder.
(1S,2S)-2-(8-Chloro-3,4-dihydro-2H-1-benzopyran-6-carbonyl)cyclopropane-1-carboxylic acid
(134) δ.sub.H(500 MHz, DMSO-d6) 12.63 (s, 1H), 7.95-7.79 (m, 2H), 4.45-4.23 (m, 2H), 3.23-3.16 (m, 1H), 2.87 (t, J=6.35 Hz, 2H), 2.06 (ddd, J=3.86, 5.82, 9.44 Hz, 1H), 1.97 (p, J=6.20 Hz, 2H), 1.44 (ddd, J=3.25, 5.85, 8.81 Hz, 1H), 1.40 (ddd, J=3.26, 5.58, 8.70 Hz, 1H). Tr=2.81 min (7 minute method, low pH) m/z (ES+) (M+H+) 281, 283.
(135) TABLE-US-00007 Structure IUPAC Name Molecular Weight LCMS Data
Example 19
(1S,2S)-2-(4-chloro-3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-6-carbonyl)cyclopropane-1-carboxylic acid
(136) ##STR00026##
Step 1: 2-Amino-5-bromo-3-chlorophenol
(137) Bromine (1.08 ml, 20.9 mmol) was added dropwise to an ice-cooled solution of 2-amino-3-chlorophenol (2.00 g, 13.9 mmol) in DCM (100 ml). The reaction mixture was stirred for 1 h after addition was complete, then filtered. Collected grey solid was washed with DCM (4 ×10 ml) and dried under suction to afford the crude product as a grey-black powder. The powder was partitioned between sat. aq. NaHCO.sub.3 (20 ml) and DCM (50 ml). The layers were separated and the aqueous extracted with DCM (3×50 ml). Combined DCM extracts were washed with water (25 ml) and brine (25 ml), then dried (MgSO.sub.4), filtered and concentrated to afford the title compound as a dark red-brown powder (1.7 g, 27% at ca. 50% NMR purity). δ.sub.H (500 MHz, DMSO) δ 10.13 (s, 1H), 6.90 (d, J=2.16 Hz, 1H), 6.76 (d, J=2.17 Hz, 1H), 4.81 (s, 2H). Tr=1.74 min 67% m/z 222, 224, 226 (M+H).sup.+.
Step 2: 6-Bromo-4-chloro-2,3-dihydro-1,3-benzoxazol-2-one
(138) 2-Amino-5-bromo-3-chlorophenol (1.55 g, 3.5 mmol) was dissolved in THF (20 ml). CDI (2.73 g, 16.9 mmol) was added and the reaction was stirred at 65° C. After 2 h the reaction was cooled to rt and concentrated to give an orange solid. The residue was redissolved in EtOAc (100 mL) and the organic phase was washed with water (50 mL), 2M HCl (3×50 mL), water (100 mL) and brine (20 mL) and dried (MgSO.sub.4). Filtration and concentration afforded the title compound (1.7 g, 97% yield) as a red-brown powder. δ.sub.H (500 MHz, DMSO-d.sub.6) 12.28 (s, 1H), 7.61 (d, J=1.60 Hz, 1H), 7.51 (d, J=1.61 Hz, 1H). Tr (3 min)=1.87 min m/z (ES.sup.−) 246, 248 (M−H).sup.−.
(139) Intermediate 2, Step 2: (5-Bromo-7-chloro-2,3-dihydro-1,3-benzoxazol-2-one) δ.sub.H (500 MHz, DMSO-d.sub.6) 12.01 (br. s., 1 H) 7.44 (d, J=1.73 Hz, 1 H) 7.26 (d, J=1.73 Hz, 1 H). Tr (3 min)=1.87 min m/z (ES.sup.−) 246, 248 (M−H).sup.−.
Step 3: 6-Bromo-4-chloro-3-methyl-2,3-dihydro-1,3-benzoxazol-2-one
(140) 6-Bromo-4-chloro-2,3-dihydro-1,3-benzoxazol-2-one (1.26 g, 3.1 mmol) was dissolved in anhydrous DMF (20 mL) and the reaction was cooled in an ice bath. Sodium hydride (60% in oil, 0.31 g, 7.7 mmol) was added portionwise and the reaction was stirred in the ice bath for 1 h. Methyl iodide (0.4 ml, 6.5 mmol) was added and the reaction was stirred at rt for 2 hours. The reaction was cooled in an ice-water bath. Water (30 mL) was added cautiously followed by EtOAc (50 mL). The layers were separated and the aqueous layer re-extracted with EtOAc (2×50 mL). The combined organic layers were washed with water (4×30 mL) and brine (2×30 mL) and dried (MgSO.sub.4). Filtration and concentration gave the title compound (1.3 g, 75% yield) as a brown powder. δ.sub.H (500 MHz, DMSO-d.sub.6) 7.68 (d, J=1.71 Hz, 1H), 7.53 (d, J=1.74 Hz, 1H), 3.54 (s, 3H).
Intermediate 2, Step 3: (5-bromo-7-chloro-3-methyl-2,3-dihydro-1,3-benzoxazol-2-one)
(141) δ.sub.H (500 MHz, CDCl.sub.3) 7.30 (d, J=1.73 Hz, 1 H) 7.03 (d, J=1.73 Hz, 1 H) 3.41 (s, 3 H); Tr (3 min)=1.97 min m/z (ES.sup.+) No ionisation.
Step 4, Method 7: 4-Chloro-3-methyl-6-(trimethylstannyl)-2,3-dihydro-1,3-benzoxazol-2-one
(142) 6-Bromo-4-chloro-3-methyl-2,3-dihydro-1,3-benzoxazol-2-one (0.65 g, 1.19 mmol) and lithium chloride (55 mg, 1.31 mmol) were dissolved in anhydrous dioxane (25 ml) and deoxygenated with nitrogen for 1 min. Hexamethyldistannane (246 μl, 1.19 mmol) and Pd(PPh.sub.3).sub.4 (137 mg, 0.12 mmol), were added and the reaction was stirred at 100° C. for 18 h. The reaction mixture was cooled to rt and concentrated. The resulting residue was purified by flash column chromatography (elution: 10% ethyl acetate, 90% heptanes) to give the title compound (210 mg, 44% yield) as a red-orange solid. δ.sub.H (500 MHz, CDCl.sub.3) 7.23-7.17 (m, 1H), 7.16-7.10 (m, 1H), 3.69 (s, 3H), 0.41-0.25 (m, 9H).
(143) Intermediate 2, Step 4: 7-Chloro-3-methyl-5-(trimethylstannyl)-2,3-dihydro-1,3-benzoxazol-2-one δ.sub.H (500 MHz, CDCl.sub.3) 7.17 (s, 1H), 6.91 (s, 1H), 3.43 (s, 3H), 0.35 (s, 9H); Tr (3 min)=2.61 min m/z (ES.sup.+) 344, 346, 348.
Step 5: Methyl (1S,2S)-2-(4-chloro-3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-6-carbonyl)cyclopropane-1-carboxylate
(144) 4-Chloro-3-methyl-6-(trimethylstannyl)-2,3-dihydro-1,3-benzoxazol-2-one (210 mg, 0.52 mmol) and methyl (1S,2S)-2-(carbonochloridoyl)cyclopropane-1-carboxylate (85 mg, 0.52 mmol) were dissolved in anhydrous toluene (5 ml) and de-oxygenated with a stream of nitrogen for 1 min. PdCl.sub.2(PPh.sub.3).sub.2 (18 mg, 0.02 mmol) was added and the reaction was stirred at 110° C. under nitrogen for 1 h. The reaction mixture was cooled to rt, concentrated and the resulting residue purified by flash column chromatography (elution: 25% ethyl acetate, 75% heptanes) to give the title compound (101 mg, 59% yield) as a light orange powder. δ.sub.H (500 MHz, CDCl.sub.3) 7.23-7.17 (m, 1H), 7.16-7.10 (m, 1H), 3.69 (s, 3H), 0.41-0.25 (m, 9H). Tr (3 min)=2.11 min m/z (ES.sup.+) (M+H.sup.+) 310, 312.
(145) Intermediate 2, Step 5: (1S,2S)-2-(7-chloro-3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-carbonyl)cyclopropane-1-carboxylate δ.sub.H (500 MHz, DMSO-d.sub.6) 8.00 (d, J=1.53 Hz, 1H), 3.71-3.65 (m, 3H), 3.47-3.38 (m, 4H), 2.27 (ddd, J=8.6, 5.9, 3.8 Hz, 1H), 1.57 (ddd, J=8.9, 5.8, 3.4 Hz, 1H), 1.50 (ddd, J=8.7, 5.5, 3.5 Hz, 1H); Tr (3 min)=1.94 min m/z (ES.sup.+) (M+H.sup.+) 310, 312.
Steps 6 & 7: (1S,2S)-2-[3-Chloro-5-hydroxy-4-(methylamino)benzoyl]cyclopropane-1-carboxylic acid
(146) Methyl (1S,2S)-2-(4-chloro-3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-6-carbonyl)cyclopropane-1-carboxylate (80 mg, 0.25 mmol) was dissolved in dioxane (5 ml) and treated with 2M NaOH (0.5 ml, 1.0 mmol). The reaction mixture was stirred at room temperature for 2 h. Further 2M NaOH (0.5 ml, 1.0 mmol) was added and the reaction mixture stirred at room temperature for 64 h. The reaction mixture was neutralised with 1M HCl and extracted with DCM (2×10 ml). The aqueous layer was adjusted to pH3 with 1M HCl and extracted with IPA-CHCl.sub.3 (1:1; 2×10 ml). IPA-CHCl.sub.3 extracts were dried (MgSO.sub.4), filtered and concentrated to afford a yellow oil. The aqueous was adjusted to pH7 with 2M NaOH and again extracted with IPA-CHCl.sub.3 (1:1; 2×10 ml). IPA-CHCl.sub.3 extracts were dried (MgSO.sub.4), filtered and concentrated. The resulting residue was re-dissolved in THF (5 ml), CDI (73 mg, 0.46 mmol) was added and the mixture heated to 65° C. for 2 h. The reaction mixture was cooled to room temperature and concentrated to give a dark red solid. The crude product was purified by reverse phase acidic preparative HPLC (H.sub.2O/MeCN/0.1% formic acid) to afford the title compound (24 mg, 54% yield) as a white powder.
(1S,2S)-2-(4-Chloro-3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-6-carbonyl)cyclopropane-1-carboxylic acid
(147) δ.sub.H (500 MHz, DMSO) 12.62 (br. s, 1H), 8.01 (d, J=1.41 Hz, 1H), 7.98 (d, J=1.42 Hz, 1H), 3.61 (s, 3H), 3.29-3.25 (part. obsc. m, 1H), 2.11 (ddd, J=3.87, 5.92, 9.55 Hz, 1H), 1.49 (ddd, J=3.29, 5.95, 8.93 Hz, 1H), 1.43 (ddd, J=3.31, 5.50, 8.70 Hz, 1H). Tr=2.40 min 97% m/z 296, 298 (M+H).sup.+.
(1S,2S)-2-(7-Chloro-3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5-carbonyl)cyclopropane-1-carboxylic acid
(148) 1H NMR (500 MHz, DMSO-d6) δ 12.73 (s, 1H), 7.99 (s, 1H), 7.93 (s, 1H), 3.50-3.41 (part. obsc. m, 4H), 2.19-2.09 (m, 1H), 1.47 (dt, J=36.9, 8.8 Hz, 2H). Tr=2.45 min 100% m/z (M+H).sup.+ 296, 298.
(149) TABLE-US-00008 Molecular Structure IUPAC Name Weight LCMS Data
Example 20
(150) ##STR00029##
Step 1: 2,3-Dihydro-1,4-benzodioxin-6-yltrimethylstannane
(151) n-Butyllithium (3.1 ml of a 1.6M solution in hexane, 4.98 mmol) was added dropwise under nitrogen to a stirred solution of 6-bromo-2,3-dihydro-1,4-benzodioxine (1.00 g, 4.65 mmol) in dry THF (20 ml) at −78° C. After 45 min, trimethyltin chloride (5.0 ml of a 1.0M solution in THF, 5 mmol) was added dropwise over 5 min. After 20 min the reaction mixture was allowed to warm to room temperature and left overnight. The reaction mixture was poured into brine (100 ml), extracted with ethyl acetate (3×80 ml) and the combined, dried (Na.sub.2SO.sub.4) organic extracts were evaporated in vacuo to give the title compound (1.363 g, 98%). as a colourless oil. δ.sub.H (500 MHz, CDCl.sub.3) 6.78 (d, J=1.1 Hz, 1H), 6.74 (dd, J=7.7, 1.1 Hz, 1H), 6.67 (d, J=7.7 Hz, 1H), 4.06 (s, 4H), 0.06 (s, 9H). Tr=2.42 min; no ionisation.
Step 2: Methyl (1S,2S)-2-(2,3-dihydro-1,4-benzodioxine-6-carbonyl)cyclopropane-1-carboxylate
(152) A mixture of 2,3-dihydro-1,4-benzodioxin-6-yltrimethylstannane (700 mg, 2.34 mmol), (1S,2S)-2-(methoxycarbonyl)cyclopropane-1-carboxylic acid (571 mg, 3.51 mmol), PdCl.sub.2(PPh.sub.3).sub.2 (82 mg, 0.12 mmol), and toluene (8 mL) was de-gassed by bubbling a stream of nitrogen through the mixture for 15 min, and then stirred at 110° C. for 2 h. The reaction was cooled, and then absorbed onto silica gel (Merck 9385, 8 mL). The resultant silica was purified on a Biotage machine (100 g cartridge of silica gel) eluting with ethyl acetate-heptane (5% EtOAc, 1CV; 5% to 40% EtOAc 10 CV; 40% EtOAc, 2 CV), to give the desired product (373 mg, 58%) as pale yellow oil. δ.sub.H (500 MHz, CDCl.sub.3) 7.61-7.57 (m, 2H), 7.00-6.86 (m, 1H), 4.33 (ddd, J=20.0, 5.8, 2.6 Hz, 4H), 3.13 (ddd, J=9.4, 5.8, 3.9 Hz, 1H), 2.36 (ddd, J=9.5, 5.8, 3.9 Hz, 1H), 1.59 (dddd, J=25.1, 9.1, 5.8, 3.4 Hz, 2H).). Tr=1.86 min; 100% m/z (ES+) 263 (M+H.sup.+).
Step 3: (1S,2S)-2-(2,3-Dihydro-1,4-benzodioxine-6-carbonyl)cyclopropane-1-carboxylic acid
(153) A solution of methyl (1S,2S)-2-(2,3-dihydro-1,4-benzodioxine-6-carbonyl)cyclopropane-1-carboxylate (347 mg, 1.32 mmol) in 1,4-dioxane (8 ml) was treated with aqueous 2M sodium hydroxide (595 μl, 1.19 mmol) at room temperature, and stirred for 22 h under nitrogen. The reaction mixture was evaporated in vacuo, treated with water (25 ml), extracted with ether (3×30 ml), and the ethereal extracts were discarded. The aqueous phase was filtered through a PTFE frit (0.45 μM). The aqueous solution was freeze-dried to give a foam (300 mg). A solution of the foam in DMSO (3 ml) was treated with aqueous 2M hydrochloric acid (0.6 ml) and purified by low pH HPLC. The resultant gum was further dried in vacuo at 40° C. to give the title compound (104 mg, 31%) as a colourless gum.
(154) (1S,2S)-2-(2,3-Dihydro-1,4-benzodioxine-6-carbonyl)cyclopropane-1-carboxylic acid δ.sub.H (500 MHz, CDCl.sub.3) 7.62-7.56 (m, 2H), 6.99-6.93 (m, 1H), 4.33 (ddd, J=20.6, 5.8, 2.6 Hz, 4H), 3.18 (ddd, J=9.5, 5.9, 3.8 Hz, 1H), 2.37 (ddd, J=9.3, 5.7, 3.8 Hz, 1H), 1.65 (dddd, J=33.1, 9.1, 5.8, 3.5 Hz, 2 Hz). LCMS Tr=2.16 min; 99% m/z (ES+) 249 (M+H.sup.+).
(155) TABLE-US-00009 Molecular Structure IUPAC Name Weight LCMS Data
Example 21
(156) ##STR00031##
Step 1: 4-Bromo-2-chloro-1-cyclopropoxy-benzene
(157) Bromocyclopropane (87.5 g, 0.723 mol) was added portionwise over 10 minutes to a stirred solution of 4-bromo-2-chlorophenol (30.0 g, 0.145 mol) and caesium dicarbonate (118 g, 0.362 mol) in dimethyl acetamide (450 mL). The mixture was heated to 150° C. and stirred at this temperature for 16 hours. After this time, the reaction was cooled to room temperature, poured onto ice-water (600 mL) and extracted with TBME (3×400 mL). The organic layers were combined and washed with water (2×200 mL) before being dried (MgSO.sub.4), filtered and concentrated to give intermediate 1 (31.2 g, 72% yield) as a pale orange oil which was used directly without purification. .sup.1H NMR (500 MHz, CDCl.sub.3) δ ppm 7.51-7.45 (m, 1H) 7.34 (dd, J=8.8, 2.4 Hz, 1H) 7.21-7.13 (m, 1H) 3.84-3.74 (m, 1H) 0.93-0.77 (m, 4H). Tr=2.46 min (3.5 Minute method) m/z (ES.sup.+) (M+H.sup.+) No ionisation observed.
Step 2: (3-Chloro-4-cyclopropoxyphenyl)trimethylstannane
(158) To a solution of 4-bromo-2-chloro-1-cyclopropoxy-benzene (111 g, 417 mmol) in dioxane (1.1 L, previously degassed with N.sub.2) was added lithium chloride (19.45 g, 458.7 mol), Pd(PPh.sub.3).sub.4 (24.1 g, 20.8 mmol) and hexamethyl ditin (150 g, 458.7 mmol). The reaction mixture was heated to 100° C. for 90 mins, then cooled and the volatiles evaporated. The residue was purified by dry flash chromatography on 1 kg of silica using 10% EtOAc-heptane to afford the product as light yellow oil, 151 g (115%) yield. This was re-purified by dry flash chromatography on 1.5 kg of silica using a gradient of heptane to 10% EtOAc-heptane to give the desired product (85 g, 65% yield) as a faint yellow oil. Tr=2.75 min (3.5 Minute method) m/z (ES.sup.+) (M+Na.sup.+) 352.
Step 3: Methyl (1S,2S)-2-(3-chloro-4-cyclopropoxybenzoyl)cyclopropane-1-carboxylate
(159) To a solution of (3-chloro-4-cyclopropoxyphenyl)trimethylstannane (20.7 g, 62 mmol) in toluene (400 ml, previously degassed with N.sub.2) was added a solution of freshly prepared methyl (1S,2S)-2-(carbonchloridoyl) cyclopropane-1-carboxylate (10.1 g, 62 mmol) in toluene (100 ml, degassed) and PdCl.sub.2(PPh.sub.3).sub.2 (2.19 g, 3 mmol). The reaction mixture was degassed for a further 5 minutes, before being heated to 110° C. for 90 mins. The reaction mixture was allowed to cool overnight before the volatiles were evaporated. The residue was purified by dry flash chromatography on 500 g of silica using neat heptane to 20% THF-heptane to afford the product as a sticky off-white solid (10.9 g, 59% yield). Trituration with ether (100 ml) and heptanes (50 ml) gave a white solid (4.54 g) in 98% UV purity. δ.sub.H (250 MHz, DMSO-d6) 8.39-7.78 (m, 2H), 7.56 (d, J=8.47 Hz, 1H), 4.09 (dt, J=3.05, 5.97 Hz, 1H), 3.66 (s, 3H), 3.28 (ddd, J=3.90, 5.79, 8.68 Hz, 1H), 2.19 (ddd, J=3.86, 5.90, 8.57 Hz, 1H), 1.74-1.26 (m, 2H), 1.00-0.64 (m, 4H). Tr=2.14 min m/z (ES.sup.+) (M+H.sup.+) 295.
Step 4: (1S,2S)-2-(3-Chloro-4-cyclopropoxybenzoyl)cyclopropane-1-carboxylic acid
(160) To a solution of methyl (1S,2S)-2-(3-chloro-4-cyclopropoxybenzoyl)cyclopropane-1-carboxylate (either pure, or impure) in dioxane (7-10 volumes) was added aq NaOH (2M, 0.9 eq, adjusted for the purity of the methyl ester) and the mixture stirred overnight. The dioxane was then carefully removed on a rotary evaporator. The gummy residue was dissolved in water/EtOAc (c. 10 vol of each) and the EtOAc layer was separated and the aqueous layer further extracted with EtOAc (5 vol). The aqueous layer was carefully evaporated on a rotary evaporator to give a gummy foamy solid. This gum was redissolved in water and freeze-dried overnight to afford the desired material as a free-flowing off-white powder. 31.67 g.
(161) There were significant aliphatic impurities present at this point, so the material was treated with dilute HCl to afford the free acid (95% wt recovery). Dry flash chromatography using acetone and column chromatography using THF-heptane 1:1 failed to substantially improve the quality of the material to hand. Finally, recrystallisation from heptanes using a minimum of EtOAc (c. 20%) gave the acid in 2 crops of sufficiently high purity to re-convert to the sodium salt. This was freeze dried to give 21.45 g of white powder, of which 21.007 g remained after analytical submissions.
(1S,2S)-2-(3-Chloro-4-cyclopropoxybenzoyl)cyclopropane-1-carboxylic acid
(162) δ.sub.H (500 MHz, D.sub.2O) 7.99 (d, J=7.68 Hz, 2H), 7.55 (d, J=9.03 Hz, 1H), 4.03 (tt, J=2.82, 6.02 Hz, 1H), 3.11 (dt, J=4.84, 8.99 Hz, 1H), 2.14 (ddd, J=3.94, 6.36, 9.68 Hz, 1H), 1.54 (ddt, J=3.75, 6.19, 10.26 Hz, 2H), 1.28-0.45 (m, 4H). Tr=3.0 min; 99%; m/z 281, 283 (M+H.sup.+).
(163) TABLE-US-00010 Molecular Structure IUPAC Name Weight LCMS Data
Example 22
(164) ##STR00033##
Step 1: Ethyl (1S,2S)-2-2[(benzyloxy)methyl]-1-methylcyclopropane-1-carboxylate (Ref. Chem. Comm., 2010, 46, 5867-5869)
(165) n-Butyllithium (2.5M solution in hexanes, 80 mL, 200.0 mmol) was added dropwise to a stirred solution of 2-(diethoxy-phosphoryl)-propionic acid ethyl ester (47.6 g, 200.0 mmol) in dry DME (80 mL) at room temperature under a nitrogen atmosphere. After 5 minutes (2S)-2-[(benzyloxy)methyl]oxirane (3.28 g, 20.00 mmol) was added portion wise, followed by heptanes (140 mL) and DME (40 mL) and the reaction mixture was heated at 130° C. in a sealed autoclave for 20 hours. After this time, the reaction was cooled to room temperature and poured into aqueous saturated ammonium chloride (400 mL). The mixture was extracted with ethyl acetate (3×250 mL), the combined organic extracts were dried (Na.sub.2SO.sub.4), filtered and concentrated. The resulting residue was purified on a Biotage column (HPSIL, 100 g, elution with ethyl acetate-heptane 20% EtOAc to 98% EtOAc gradient in heptanes) to give the title compound (2.28 g, 46% yield) as a colourless oil. δ.sub.H (500 MHz, CDCl.sub.3) 7.40-7.29 (m, 5H), 4.55 (d, J=14.3 Hz, 2H), 4.11 (dd, J=7.1, 2.7 Hz, 2H), 3.68 (dd, J=10.6, 5.8 Hz, 1H), 3.38 (dd, J=10.6, 8.5 Hz, 1H), 1.95-1.77 (m, 1H), 1.42 (dd, J=9.4, 4.2 Hz, 1H), 1.32 (s, 3H), 1.26 (t, J=7.1 Hz, 3H), 0.56 (dd, J=6.5, 4.2 Hz, 1H). Tr=2.24 min m/z (ES.sup.+) (M+H.sup.+) 249.
Step 2: Ethyl (1S,2S)-2-(hydroxymethyl)-1-methylcyclopropane-1-carboxylate (Ref. J. Org. Chem., 2002, 67, 4520-4525)
(166) 10% Palladium-on-carbon (2.00 g, 200.0 mmol) was added to a stirred solution of (1S,2S)-2-benzyloxymethyl-1-methyl-cyclopropanecarboxylic acid ethyl ester (2.3 g, 9.18 mmol) in ethanol (130 mL), the mixture was purged twice with nitrogen before being evacuated and refilled with hydrogen. The resulting mixture was then stirred at room temperature for 18 hours. After this time the mixture was filtered and the filtrate concentrated to give the title compound (1.45 g, 99% yield) as a colourless oil. δ.sub.H (500 MHz, CDCl.sub.3) 4.04 (qd, J=7.1, 3.0 Hz, 2H), 3.78 (dd, J=11.7, 5.9 Hz, 1H), 3.47 (dd, J=11.7, 8.8 Hz, 1H), 1.75 (tt, J=9.0, 6.3 Hz, 1H), 1.33 (dd, J=9.3, 4.2 Hz, 2H), 1.29 (s, 3H), 1.18 (t, J=7.1 Hz, 3H), 0.49 (dd, J=6.5, 4.3 Hz, 1H).
Step 3: (1S,2S)-2-(ethoxycarbonyl)-2-methylcyclopropane-1-carboxylic acid (Ref. Org. Lett., 2003, 5, 4669)
(167) Sodium bicarbonate (4.83 g, 55.6 mmol), sodium periodate (10.75 g, 50.2 mmol) and ruthenium trichloride (0.19 g, 0.91 mmol) were added sequentially to a stirred solution of (1S,2S)-2-hydroxymethyl-1-methyl-cyclopropanecarboxylic acid ethyl ester (1.44 g, 9.13 mmol) in chloroform (90 mL), acetonitrile (90 mL), and water (135 mL) and the mixture was stirred at room temperature for 4 hours. After this time, the mixture was acidified to pH 1 by the addition of 6M hydrochloric acid (10 mL) and the mixture extracted with DCM (4×200 mL). The combined organic extracts were dried (Na.sub.2SO.sub.4), filtered and concentrated. The resulting residue was stirred in dichloromethane (20 mL) and then filtered. The resultant black oil was filtered through silica gel (25 g) eluting with ethyl acetate-heptane-acetic acid 30:69:1 to give the title compound (1.42 g, 90% yield) as a colourless oil. δ.sub.H (500 MHz, CDCl.sub.3) 12.3-10.1 (br s, 1H), 4.08 (q, J=7.1 Hz, 2H), 2.28 (dd, J=8.6, 6.5 Hz, 1H), 1.57 (dd, J=8.6, 4.3 Hz, 1H), 1.38 (s, 3H), 1.27 (dd, J=6.5, 4.3 Hz, 1H), 1.20 (t, J=7.1 Hz, 3H). Tr=1.48 min m/z (ES.sup.+) (M+H.sup.+) 173.
Step 4: Ethyl (1S,2S)-2-(3,4-dichlorobenzoyl)-1-methylcyclopropane-1-carboxylate
(168) Oxalyl chloride (0.46 mL, 5.23 mmol) was added dropwise under nitrogen to a stirred solution of (1S,2S)-1-methyl-cyclopropane-1,2-dicarboxylic acid 1-ethyl ester (0.3 g, 1.74 mmol) and DMF (14 μl, 0.17 mmol) in DCM (11 mL). After 1 hour, the solution was concentrated and subsequently co-evaporated with DCM (3×10 mL). The resulting residue was dissolved in toluene (1 mL) and added portion wise to a stirred solution of (3,4-dichloro-phenyl)-trimethyl-stannane (0.53 g, 1.71 mmol) in toluene (6 mL). The solution was de-gasses under a stream of nitrogen for 15 minutes, and then stirred at 110° C. under a nitrogen atmosphere for 20 hours. After this time, the mixture was cooled to room temperature and concentrated. The resulting residue was absorbed from ethyl acetate onto silica gel (Merck9385, 8 mL). The resultant silica was purified on a Biotage machine (100 g column, elution; 2% EtOAc to 20% EtOAc gradient with heptanes) to give the title compound (0.21 g, 35% yield) as a colourless oil. δ.sub.H (500 MHz, CDCl.sub.3) 7.96 (d, J=2.0 Hz, 1H), 7.71 (dd, J=8.3, 2.0 Hz, 1H), 7.49 (d, J=8.3 Hz, 1H), 4.17 (q, J=7.1 Hz, 2H), 3.13 (dd, J=8.1, 6.7 Hz, 1H), 1.63-1.55 (m, 2H), 1.25 (t, J=7.1 Hz, 3H), 1.19 (s, 3H). Tr=2.35 min m/z (ES.sup.+) (M+H.sup.+) 301.
Step 5: (1S,2S)-2-(3,4-Dichlorobenzoyl)-1-methylcyclopropane-1-carboxylic acid
(169) 2M NaOH (0.15 mL, 0.31 mmol) was added in one portion to a stirred solution of ethyl (1S,2S)-2-(3,4-dichlorobenzoyl)-1-methylcyclopropane-1-carboxylate (0.08 g, 0.28 mmol) in dioxane (5 mL) and the resulting solution was stirred at room temperature for 18 hours. After this time the reaction mixture was acidified with 1M HCl, and the resulting suspension was extracted with EtOAc (2×10 mL). The combined organic extracts were washed with brine (10 mL), before being dried (MgSO4), filtered and concentrated. The resulting residue was purified by prep HPLC to give the title compound (0.03 g, 39% yield) as a white solid.
(1S,2S)-2-(8-Chloro-3,4-dihydro-2H-1-benzopyran-6-carbonyl)cyclopropane-1-carboxylic acid
(170) δ.sub.H (500 MHz, CDCl.sub.3) 9.94-12.01 (m, 1H) 8.06 (d, J=1.89 Hz, 1H) 7.81 (dd, J=8.35, 2.05 Hz, 1H) 7.59 (d, J=8.35 Hz, 1H) 3.29 (dd, J=8.20, 6.62 Hz, 1H) 1.72-1.81 (m, 2H) 1.30 (s, 3H). Tr=3.29 min (7 minute method, low pH) m/z (ES+) (M+H+) 273/275.
(171) TABLE-US-00011 Molecular Structure IUPAC Name Weight LCMS Data
Example 23
(172) ##STR00035##
Step 1: 2-(Methoxycarbonyl)-3-methylcyclopropane-1-carboxylic acid
(173) A solution of potassium hydroxide (0.54 g, 9.58 mmol) in methanol (4 mL) was added dropwise over 10 minutes to a stirred solution of dimethyl 3-methyl-trans-1,2-cyclopropanedicarboxylate (1.5 g, 8.7 mmol) and the reaction mixture was heated to 65° C. and stirred at this temperature for 3 hours. After this time, the reaction was cooled to room temperature and concentrated. The resulting residue was dissolved in water (20 mL) and the mixture extracted with ethyl acetate (2×50 mL). The organic layers were discarded and the aqueous layer acidified to pH 1 with 1M HCl. The acidic aqueous layer was then extracted with ethyl acetate (3×30 mL), the combined organic extracts were washed with brine (50 mL), before being dried (MgSO.sub.4), filtered and concentrated to give the title compound (1.14 g, 83% yield) as a colourless oil. Compound was isolated as a mixture of diastereoisomers. Tr=1.14 min m/z (ES.sup.+) (M+H.sup.+) No ionisation; 1.10 min m/z (ES.sup.+) (M+H.sup.+) No ionisation.
Step 2: Methyl 2-(carbonochloridoyl)-3-methylcyclopropane-1-carboxylate
(174) Oxalyl chloride (1.7 mL, 18.9 mmol) was added dropwise over 10 minutes to a stirred solution of 2-(methoxycarbonyl)-3-methylcyclopropane-1-carboxylic acid (1.0 g, 6.3 mmol) in DCM (20 mL) containing 1 drop of DMF and the mixture was stirred at room temperature for 3 hours under a nitrogen atmosphere. After this time, the reaction mixture was concentrated to give the title compound (1.12 g, 99% yield) as a pale yellow oil which was taken on directly without further purification.
Step 3: Methyl 2-[methoxy(methyl)carbamoyl]-3-methylcyclopropane-1-carboxylate
(175) N-Methoxymethanamine hydrochloride (0.68 g, 6.98 mmol) was added in one portion to a cooled (0° C.), stirred solution of methyl 2-(carbonochloridoyl)-3-methylcyclopropane-1-carboxylate (1.12 g, 6.34 mmol). Pyridine (1.13 mL, 13.9 mmol) was then added dropwise over 5 minutes and the reaction mixture was allowed to warm to room temperature before being stirred under a nitrogen atmosphere for 2 hours. After this time, the reaction mixture was washed sequentially with saturated sodium bicarbonate solution (50 mL), 10% citric acid (300 mL), water (50 mL) and brine (20 mL). The organic layer was dried (MgSO.sub.4), filtered and concentrated to give the title compound (1.1 g, 85% yield) as an orange oil. Tr=1.36 min m/z (ES.sup.+) (M+H.sup.+) 202 (76% purity); 1.33 min m/z (ES.sup.+) (M+H.sup.+) 202 (24% purity).
Step 4: Methyl 2-[(3,4-dichlorophenyl)carbonyl]-3-methylcyclopropane-1-carboxylate
(176) 3,4-Dichlorophenylmagnesium bromide (0.5M solution in THF, 16.2 mL, 8.13 mmol) was added dropwise over 20 minutes to a cold (−50° C.), stirred solution of methyl 2-[methoxy(methyl)carbamoyl]-3-methylcyclopropane-1-carboxylate (1.1 g, 5.4 mmol) in THF and the reaction was stirred at −50° C. under a nitrogen atmosphere for 1 hour before being allowed to warm to 0° C. over 1 hour. After this time, the reaction mixture was warmed to room temperature and quenched by the addition of saturated ammonium chloride solution (20 mL). The mixture was extracted with ethyl acetate (2×50 mL) and the combined organic layer washed sequentially with water (50 mL) and brine (50 mL), before being dried (MgSO.sub.4), filtered and concentrated. The resulting residue was purified on a Biotage isolera (elution: 8% ethyl acetate, 92% heptanes, to 15% ethyl acetate, 85% heptanes). Purification led to the isolation of two sets of diastereosiomers (combined total: 0.82 g, 53% yield): Diastereoisomer 1 (as a mixture of (1S,2S,3S) and (1R,2R,3R) enantiomers) and diastereoisomer 2 (as a mixture of (1S,2S,3R) and (1R,2R,3S) enantiomers).
(177) Diastereosiomer 1 (1S,2S,3S) and (1R,2R,3R): δ.sub.H(500 MHz, CDCl.sub.3) 8.08 (d, J=2.05 Hz, 1 H), 7.84 (dd, J=8.35, 2.05 Hz, 1 H), 7.53-7.63 (m, 1 H), 3.67-3.81 (m, 3 H), 2.97-3.08 (m, 1 H), 2.56 (dd, J=9.62, 4.41 Hz, 1 H), 2.04 (dquin, J=9.52, 6.17, 6.17, 6.17, 6.17 Hz, 1 H), 1.32-1.42 (m, 3 H).
(178) Diastereosiomer 2 (1S,2S,3R) and (1R,2R,3S): δ.sub.H (500 MHz, CDCl.sub.3) 8.10 (d, J=2.05 Hz, 1 H), 7.85 (dd, J=8.35, 2.05 Hz, 1 H), 7.53-7.63 (m, 1 H), 3.71-3.76 (m, 3 H), 3.18 (dd, J=9.85, 4.65 Hz, 1 H), 2.49 (dd, J=5.60, 4.81 Hz, 1 H), 2.08-2.20 (m, 1 H), 1.15 (d, J=6.31 Hz, 3 H).
(179) Each diastereoisomeric mixture was taken on separately as described in Step 5 below.
Step 5: 2-[(3,4-Dichlorophenyl)carbonyl]-3-methylcyclopropane-1-carboxylic acid (1S,2S,3S and 1R,2R,3R)
(180) NaOH (2M solution, 0.41 mL, 0.83 mmol) was added in one portion to a stirred solution of methyl 2-[(3,4-dichlorophenyl)carbonyl]-3-methylcyclopropane-1-carboxylate (0.28 g, 0.92 mmol) in THF (5 mL) and the mixture was stirred at room temperature for 72 hours. After this time, the reaction mixture was concentrated and the resulting residue was dissolved in water (10 mL). The aqueous layer was washed with diethyl ether (2×25 mL) and DCM (2×50 mL) before being acidified to pH 1 with 2M HCl. The mixture was then extracted with ethyl acetate (3×30 mL) and the combined organic extracts were washed sequentially with water (10 mL) and brine (10 mL) before being dried (MgSO.sub.4), filtered and concentrated to give the title compound (0.16 g, 64% yield) as a white solid. The resulting enantiomers were then separated by chiral HPLC.
(1S,2S,3S or 1R,2R,3R)-2-(3,4-Dichlorobenzoyl)-3-methylcyclopropane-1-carboxylic acid
(181) δ.sub.H (500 MHz, DMSO-d.sub.6) 12.39-12.86 (m, 1 H), 8.19 (d, J=1.89 Hz, 1 H), 7.95-8.04 (m, 1 H), 7.84 (d, J=8.35 Hz, 1 H), 3.08-3.17 (m, 1 H), 2.26-2.35 (m, 1 H), 1.83-1.95 (m, 1 H), 1.29 (d, J=6.31 Hz, 3 H). Tr=4.20 min (7 minute method) m/z (ES.sup.+) (M+H.sup.+) 273, 275.
(1S,2S,3S or 1R,2R,3R)-2-(3,4-Dichlorobenzoyl)-3-methylcyclopropane-1-carboxylic acid
(182) δ.sub.H (500 MHz, DMSO-d.sub.6) 12.43-12.79 (m, 1 H), 8.18 (d, J=2.05 Hz, 1 H), 7.98 (dd, J=8.35, 2.05 Hz, 1 H), 7.83 (d, J=8.35 Hz, 1 H), 3.08-3.17 (m, 1 H), 2.30 (dd, J=9.62, 4.57 Hz, 1 H), 1.82-1.94 (m, 1 H), 1.28 (d, J=6.31 Hz, 3 H). Tr=4.20 min (7 minute method) m/z (ES.sup.+) (M+H.sup.+) 273, 275.
(1R,2R,3S or 1S,2S,3R)-2-(3,4-Dichlorobenzoyl)-3-methylcyclopropane-1-carboxylic acid
(183) δ.sub.H (500 MHz, CDCl.sub.3) 8.10 (d, J=2.05 Hz, 1H), 7.85 (dd, J=8.35 2.05 Hz, 1H), 7.59 (d, J=8.35 Hz, 1H), 3.22 (dd, J=9.77, 4.57 Hz 1H), 2.50 (t, J=5.12 Hz, 1H), 2.19 (dt, J=9.77, 6.07 Hz, 1H), 1.16 (d, J=6.31 Hz, 3H). Tr=3.27 min (7 minute method) m/z (ES.sup.+) (M+H.sup.+) 273/275.
(1S,2S,3R or 1R,2R,3S)-2-(3,4-Dichlorobenzoyl)-3-methylcyclopropane-1-carboxylic acid
(184) δ.sub.H (500 MHz, CDCl.sub.3) 8.10 (d, J=1.89 Hz, 1H), 7.85 (dd, J=8.35, 2.05 Hz, 1H), 7.59 (d, J=8.35 Hz, 1H), 3.22 (dd, J=9.85, 4.49 Hz, 1H), 2.50 (t, J=5.12 Hz, 1H), 2.19 (dt, J=9.93, 6.07 Hz, 1H), 1.16 (d, J=6.31 Hz, 3H). Tr=3.27 min (7 minute method) m/z (ES.sup.+) (M+H.sup.+) 273/275.
(1S,2S)-2-(3-chloro-4-cyclopropoxybenzoyl)cyclopropane-1-carboxylic acid
(185) δ.sub.H (500 MHz, DMSO) 12.58 (br. s., 1H), 8.11 (d, J=2.05 Hz, 1H), 7.90-7.97 (m, 1H), 7.83 (d, J=8.35 Hz, 1H), 3.14 (d, J=5.67 Hz, 1H), 2.32 (d, J=5.67 Hz, 1H), 1.37-1.48 (m, 3H), 1.05 (s, 3H). Tr=3.47 min (7 minute method) m/z (ES.sup.+) (M+H.sup.+) 287.
(1R,2R)-2-(3-chloro-4-cyclopropoxybenzoyl)cyclopropane-1-carboxylic acid
(186) δ.sub.H (500 MHz, DMSO) 12.58 (br. s., 1H), 8.11 (d, J=2.05 Hz, 1H), 7.91-7.99 (m, 1H), 7.83 (d, J=8.35 Hz, 1H), 3.14 (d, J=5.67 Hz, 1H), 2.32 (d, J=5.67 Hz, 1H), 1.42 (s, 3H), 1.05 (s, 3H). Tr=3.47 min (7 minute method) m/z (ES.sup.+) (M+H.sup.+) 287.
(187) TABLE-US-00012 Molecular Structure IUPAC Name Weight LCMS Data
Example 24
(188) ##STR00042##
(189) The following examples may be prepared according to the method described above.
(190) TABLE-US-00013 Molecular Structure IUPAC Name weight
Example 25
(191) ##STR00074##
(192) The following examples may be prepared according to the methods described above.
(193) TABLE-US-00014 Molecular Structure IUPAC Name weight
Example 26
(194) ##STR00107##
(195) The following examples may be prepared according to the methods described above.
(196) TABLE-US-00015 Molecular Structure IUPAC Name weight
Example 27
(197) The following routes (A, B, C, D, E & F) describe methods for the synthesis of compounds described in the table below:
(198) Route A:
(199) ##STR00140##
Route B:
(200) ##STR00141##
Route C:
(201) ##STR00142##
Route D:
(202) ##STR00143##
Route E:
(203) ##STR00144##
Route F:
(204) ##STR00145##
(205) TABLE-US-00016
Example 28
(206) The following route (G) describes a generic method for the synthesis of compounds described in the table below:
(207) Route G:
(208) ##STR00154##
(209) TABLE-US-00017
Example 29
(210) The following route (H) describes a generic method for the synthesis of compounds described in the table below:
(211) Route H:
(212) ##STR00162##
(213) TABLE-US-00018
Example 30
(214) The following examples may be prepared according to the methods described above.
(215) TABLE-US-00019 Structure IUPAC Name
Example 31
(216) A generalized procedure for monitoring L-Kynurenine (KYN) hydroxylation to form product 3-Hydroxy-Kynurenine (3OH-KYN) by LC/MS is described below. Product is quantified by multiple reaction monitoring using MS.
(217) Key Reagents:
(218) Compound: Stock concentrations: 10 mM in 100% DMSO Cell line: CHO GST HIS KMO cell line, 1E4 cells/well/100 μl in 96 well cell plate Substrate: L-Kynurenine (Sigma: Cat# K3750, stock concentration: 10 mM in 100 mM potassium phosphate buffer, pH 7.4)
Assay Conditions: Medium: OptiMem (Reduced Serum Medium 1×, +L-Glutamine+HEPES−Phenol Red; GIBCO: Cat#11058) Assay Volume: 200 μl Plate Format: 96 well plate, transparent (Corning) Read-Out: product (3OH-KYN) quantification using product specific MRM Reader: LC/MS/MS
Assay Protocol: prepare serial dilution (factor 3) of compound in 100% DMSO (top concentration=6.67 mM, 100% DMSO) [8 points: 6.67 mM; 2.22 mM; 0.74 mM; 0.247 mM; 0.082 mM; 0.027 mM; 0.009 mM; 0.003 mM] prepare 300-fold concentrated solution of each compound concentration (top concentration 22.22 μM, 0.3% DMSO) in OptiMem medium [22.2 μM; 7.41 μM; 2.47 μM; 0.82 μM; 0.27 μM; 0.09 μM; 0.03 μM; 0.01 μM] prepare substrate (10 mM) at concentration of 1.1 mM in medium medium of cell plate is drawed off cells are washed with OptiMem (100 μl/well) and drawed off again assay mix: 90 μl OptiMem/well+90 μl compound/well of each concentration [final compound top concentration: 10 μM; 0.15% DMSO] [final compound bottom concentration: 0.004 μM; 0.15% DMSO] pre-incubation: 30 min at 37° C. add 20 μl/well of the 1.1 mM substrate solution (final assay concentration: 100 μM) positive control: 200 μl OptiMem negative control: 180 μl OptiMem+20 μ1.1 mM substrate incubate ˜24 h at 37° C. transfer 100 μl of each well in a transparent 96 well plate (Corning) add 100 μl/well 10% trichloro acetic acid (TCA) in water centrifugate plate for 3 min at 4000 rpm detect product by LC/MS (injection of 50 μl/well; 2.5 fold overfill of the 20 μl sample loop)
Data Analysis: IC.sub.50's are calculated using automated fitting algorithm (A+ Analysis).
Example 32
(219) A method of monitoring L-Kynurenine (KYN) hydroxylation to form product 3-Hydroxy-Kynurenine (3OH-KYN) by LC/MS is described below. Product is quantified by multiple reaction monitoring.
(220) Key Reagents:
(221) Compound: Stock concentrations: 10 mM in 100% DMSO Enzyme: KMO enzyme prepared at Evotec via mitochondria isolation from CHO-GST HIS KMO cells Substrate: L-Kynurenine (Sigma: Cat# K3750) [stock concentration: 10 mM in 100 mM potassium phosphate buffer, pH 7.4]
Assay Conditions: Buffer: 100 mM potassium phosphate, pH 7.4, 200 μM NADPH, 0.4 U/ml G6P-DH (Glucose 6-phosphate dehydrogenase), 3 mM G6P (D-Glucose 6-phosphate) Assay Volume: 40 μl Plate Format: 384 well plate, transparent (Matrix) Read-Out: product (3OH-KYN) quantification using product specific MRM Reader: LC/MS/MS
Assay Protocol: prepare serial dilution (factor 3) of compound in 100% DMSO (top concentration=10 mM, 100% DMSO) [8 points: 10 mM; 3.33 mM; 1.11 mM; 0.37 mM; 0.12 mM; 0.04 mM; 0.0137 mM; 0.0045 mM, 0.0015 mM] prepare 3.33-fold concentrated solution of each compound concentration (top concentration 300 μM, 3% DMSO) in assay buffer [concentrations: 300 μM; 100 μM; 33.3 μM; 11.1 μM; 3.70 μM; 1.23 μM; 0.41 μM; 0.137 μM] prepare substrate (10 mM) at concentration of 1 mM in assay buffer assay mix: 4 μl compound/well of each concentration+24 μl assay buffer/well+8 μl KMO human enzyme+4 μl 1 mM substrate (final concentration=100 μM) [final compound top concentration: 30 μM; 0.3% DMSO] [final compound bottom concentration: 0.0137 μM; 0.3% DMSO] positive control: 4 μl 50 μM FCE28833 in assay buffer [0.5% DMSO] (final assay concentration=5 μM)+24 μl assay buffer/well+8 μl KMO human enzyme+4 μl 1 mM substrate (final concentration=100 μM) negative control: 28 μl assay buffer/well+8 μl KMO human enzyme+4 μl 1 mM substrate (final concentration=100 μM) incubate 400 min at RT add 40 μl/well 10% trichloro acetic acid in water to stop the assay and precipitate protein centrifuge plate for 3 min at 4000 rpm product detection by LC/MS (injection of 50 μl/well; 2.5 fold overfill of the 20 μl sample loop)
Data Analysis: IC.sub.50's are calculated using automated fitting algorithm (A+ Analysis).
Example 33
(222) A method of monitoring L-Kynurenine (KYN) hydroxylation to form 3-Hydroxy-Kynurenine (3OH-KYN) by LC/MS is described. Product is quantified by multiple reaction monitoring (MRM method).
(223) Key Reagents:
(224) Compound: Stock concentrations: 10 mM in 100% DMSO Enzyme: KMO enzyme prepared at Evotec from mouse liver (4-6 weeks old) via mitochondria isolation as described in the literature Substrate: L-Kynurenine (Sigma: Cat# K3750, stock concentration: 10 mM in 100 mM potassium phosphate buffer, pH 7.4)
Assay Conditions: Buffer: 100 mM potassium phosphate, pH 7.4, 200 μM NADPH, 0.4 U/ml G6P-DH (Glucose 6-phosphate Dehydrogenase), 3 mM G6P (D-Glucose 6-phosphate) Assay Volume: 40 μl Plate Format: 384 well plate, transparent (Matrix) Read-Out: product (3OH-KYN) quantification using product specific MRM Reader: LC/MS/MS
Assay Protocol: prepare serial dilution (factor 3) of compound in 100% DMSO (top concentration=10 mM, 100% DMSO) [8 points: 10 mM; 3.33 mM; 1.11 mM; 0.37 mM; 0.12 mM; 0.04 mM; 0.0137 mM; 0.0045 mM, 0.0015 mM] prepare 3.33-fold concentrated solution of each compound concentration (top concentration 300 μM, 3% DMSO) in assay buffer [concentrations: 300 μM; 100 μM; 33.3 μM; 11.1 μM; 3.70 μM; 1.23 μM; 0.41 μM; 0.137 μM] prepare substrate (10 mM) at concentration of 1 mM in assay buffer assay mix: 4 μl compound/well of each concentration+24 μl assay buffer/well+8 μl KMO mouse enzyme+4 μl 1 mM substrate (final concentration=100 μM) [final compound top concentration: 30 μM; 0.3% DMSO] [final compound bottom concentration: 0.0137 μM; 0.3% DMSO] positive control: 4 μl 50 μM FCE28833 in assay buffer, 0.5% DMSO [final assay concentration=5 μM]+24 μl assay buffer/well+8 μl KMO mouse enzyme+4 μl 1 mM substrate [final concentration=100 μM] negative control: 28 μl assay buffer/well+8 μl KMO mouse enzyme+4 μl 1 mM substrate [final concentration=100 μM] incubate 40 min at RT add 40 μl/well 10% trichloro acetic acid in water to stop the assay and precipitate protein centrifuge plate for 3 min at 4000 rpm product detection by LC/MS (injection of 20 μl/well, 2 fold overfill of the 10 μl sample loop)
Data Analysis: IC.sub.50's are calculated using automated fitting algorithm (A+ Analysis).
Example 34
(225) Using the assay protocols substantially similar to those of Example 33, the following compounds were tested.
(226) TABLE-US-00020 % Inhibition IUPAC Name at 30 mM (1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropane-1-carboxylic acid 106 (1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]-N-phenylcyclopropane-1-carboxamide 86 Methyl (1S,2S)-2-[(3,5-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate 104 (1S,2S)-2-[(3,5-Dichlorophenyl)carbonyl]cyclopropane-1-carboxylic acid 103 (1S,2S)-2-[(3-Chlorophenyl)carbonyl]cyclopropane-1-carboxylic acid 105 [(2R,3R,4S,5R,6S)-3,4,5,6-Tetrahydroxyoxan-2-yl]methyl (1S,2S)-2-[(3,4- 102 dichlorophenyl)carbonyl]cyclopropane-1-carboxylate 2-Hydroxyethyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1- 102 carboxylate (1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]-N-(2-hydroxyethyl)cyclopropane-1- 94 carboxamide 4-{[(1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropyl]carbonyl}piperazin-2- 33 one (1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropane-1-carbonitrile 68 3-[(1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropyl]-4,5-dihydro-1,2,4- 101 oxadiazol-5-one (1S)-2-[(3-Chloro-4-fluorophenyl)carbonyl]cyclopropane-1-carboxylic acid 68 (1S,2S)-2-{[3-Chloro-4-(trifluoromethyl)phenyl]carbonyl}cyclopropane-1- 102 carboxylic acid (1S,2S)-2-[(3,4-Difluorophenyl)carbonyl]cyclopropane-1-carboxylic acid 88 (1S,2S)-2-{[3-Chloro-4-(propan-2-yloxy)phenyl]carbonyl}cyclopropane-1- 98 carboxylic acid 5-[(1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropyl]-1H-1,2,3,4-tetrazole 101 (1S,2S)-2-[(3-Chloro-4-methylphenyl)carbonyl]cyclopropane-1-carboxylic acid 100 Sodium (1S,2S)-2-{[3-chloro-4-(propan-2-yloxy)phenyl]carbonyl}cyclopropane- 102 1-carboxylate Sodium (1S,2S)-2-{[3-chloro-4-(trifluoromethyl)phenyl]carbonyl}cyclopropane- 102 1-carboxylate Sodium (1S,2S)-2-[(3-chloro-4-methylphenyl)carbonyl]cyclopropane-1- 100 carboxylate (1S,2S)-2-[(4-Chloro-3-fluorophenyl)carbonyl]cyclopropane-1-carboxylic acid 103 Sodium (1S,2S)-2-[(3-chloro-4-fluorophenyl)carbonyl]cyclopropane-1- 102 carboxylate 3-[(1S,2S)-2-[(3-Chloro-4-fluorophenyl)carbonyl]cyclopropyl]-4,5-dihydro-1,2,4- 101 oxadiazol-5-one (1S,2S)-2-[(3-Chloro-4-cyclopropoxyphenyl)carbonyl]cyclopropane-1-carboxylic 101 acid (1S,2S)-2-[(3-Chloro-4-cyclopropoxyphenyl)carbonyl]cyclopropane-1- 42 carboxamide [(2R,3S,4S,5R)-3,4,5,6-Tetrahydroxyoxan-2-yl]methyl (1S,2S)-2- 100 [(3,4dichlorophenyl)carbonyl]cyclopropane-1-carboxylate (2S,3S)-3-[(3,4-Dichlorophenyl)carbonyl]aziridine-2-carboxylic acid 101 1-[(3,4-Dichlorophenyl)carbonyl]-5-oxaspiro[2.4]heptan-4-one 38 2-Methylpropyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1- 101 carboxylate Butyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate 101 Propan-2-yl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate 101 2-(Diethylamino)ethyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1- 102 carboxylate (2S)-2-Amino-6-{[(1S,2S)-2-[(3,4- 98 dichlorophenyl)carbonyl]cyclopropyl]formamide}hexanoic acid {[(1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropyl]carbonyloxy}methyl 2,2- 102 dimethylpropanoate 1-{[(1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropyl]carbonyloxy}ethyl 103 acetate (3-Carboxy-2-{[(1S,2S)-2-[(3,4- 99 dichlorophenyl)carbonyl]cyclopropyl]carbonyloxy}propyl)trimethylazanium 1-{[(Propan-2-yloxy)carbonyl]oxy}ethyl (1S,2S)-2-[(3,4- 103 dichlorophenyl)carbonyl]cyclopropane-1-carboxylate 1-[(Ethoxycarbonyl)oxy]ethyl (1S,2S)-2-[(3,4- 102 dichlorophenyl)carbonyl]cyclopropane-1-carboxylate 4-tert-Butylphenyl (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1- 101 carboxylate 1-{[(1S,2S)-2-[(3,4-Dichlorophenyl)carbonyl]cyclopropyl]carbonyloxy}ethyl 98 (1S,2S)-2-[(3,4-dichlorophenyl)carbonyl]cyclopropane-1-carboxylate (1S,2S)-2-(7-chloro-2,3-dihydro-1-benzofuran-5-carbonyl)cyclopropane-1- 100.3 carboxylic acid (1S,2S)-2-(2,3-dihydro-1-benzofuran-5-carbonyl)cyclopropane-1-carboxylic acid 98.7 (1S,2S)-2-(8-chloro-3,4-dihydro-2H-1-benzopyran-6-carbonyl)cyclopropane-1- 100.3 carboxylic acid (1S,2S)-2-(4-chloro-3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-6- 93.5 carbonyl)cyclopropane-1-carboxylic acid (1S,2S)-2-(7-chloro-3-methyl-2-oxo-2,3-dihydro-1,3-benzoxazole-5- 101.1 carbonyl)cyclopropane-1-carboxylic acid (1S,2S)-2-(2,3-dihydro-1,4-benzodioxine-6- 101.6 carbonyl)cyclopropane-1-carboxylic acid 1S,2S)-2-(3-chloro-4- 100.5 cyclopropoxybenzoyl)cyclopropane-1-carboxylic acid (1S,2S)-2-(3,4-dichlorobenzoyl)-1-methylcyclopropane-1-carboxylic acid 98.6 (1S,2S,3S)-2-(3,4-dichlorobenzoyl)-3-methylcyclopropane-1-carboxylic acid 52.5 (1R,2R,3R)-2-(3,4-dichlorobenzoyl)-3-methylcyclopropane-1-carboxylic acid 100.6 (1R,2R,3S)-2-(3,4-dichlorobenzoyl)-3-methylcyclopropane-1-carboxylic acid 96.7 (1S,2S,3R)-2-(3,4-dichlorobenzoyl)-3-methylcyclopropane-1-carboxylic acid 100.9 (1R,3R)-3-(3,4-dichlorobenzoyl)-2,2-dimethylcyclopropane-1-carboxylic acid 54 (1S,3S)-3-(3,4-dichlorobenzoyl)-2,2-dimethylcyclopropane-1-carboxylic acid 15.4
(227) While some embodiments have been shown and described, various modifications and substitutions may be made thereto without departing from the spirit and scope of the invention. For example, for claim construction purposes, it is not intended that the claims set forth hereinafter be construed in any way narrower than the literal language thereof, and it is thus not intended that exemplary embodiments from the specification be read into the claims. Accordingly, it is to be understood that the present invention has been described by way of illustration and not limitations on the scope of the claims.