Aryl naphthyl methanone oxime(s) and process for preparation thereof
09994515 ยท 2018-06-12
Assignee
Inventors
- Sabyasachi Sanyal (Lucknow, IN)
- Atul Kumar (Lucknow, IN)
- Naibedya Chattopadhyay (Lucknow, IN)
- Jawahar Lal (Lucknow, IN)
- Arun Kumar Trivedi (Lucknow, IN)
- Dipak Datta (Lucknow, IN)
- Srikanta Kumar RATH (Lucknow, IN)
- Tahseen Akhtar (Lucknow, IN)
- Shailendra Kumar Dhar Dwivedi (Lucknow, IN)
- Manisha Yadav (Lucknow, IN)
- Bandana Chakravarti (Lucknow, IN)
- Abhishek Kumar Singh (Lucknow, IN)
- Jay Sharan Mishra (Lucknow, IN)
- Nidhi Singh (Lucknow, IN)
- Anil Kumar Tripathi (Lucknow, IN)
Cpc classification
C07C251/48
CHEMISTRY; METALLURGY
C07C249/08
CHEMISTRY; METALLURGY
C07C251/58
CHEMISTRY; METALLURGY
International classification
C07C251/48
CHEMISTRY; METALLURGY
C07C249/08
CHEMISTRY; METALLURGY
C07C251/58
CHEMISTRY; METALLURGY
C07C251/60
CHEMISTRY; METALLURGY
Abstract
The present invention relates to substituted aryl naphthyl methanone oximes of general formula (I), their process for preparation and their derivatives, salts, pharmaceutical composition thereof and their use in treatment of chronic myelogenous leukemia, acute myelogenous leukemia, lymphoma, multiple myeloma, solid tumor forming cell-lines including such as breast cancer, endometrial cancer colon cancer, prostate cancer and killing of drug resistant cancer stem cells, as subject in need thereof. ##STR00001##
Claims
1. A compound of formula I or a pharmaceutically acceptable salt thereof, ##STR00056## R.sup.1=H, Halogen, alkyl, alkoxy, or nitro R.sup.2=H, Halogen, alkyl, alkoxy, hydroxy, or nitro R.sup.3=H, alkyl, alkoxy, nitro, or halogen X=O, or S R.sup.4=hydrogen, alkyl group (C.sub.1-C.sub.6), alkylamino group (C.sub.1-C.sub.6), cyclic, or open chain amines R.sup.5=alkyl group (C.sub.1-C.sub.6), alkylepoxy, alkylhydroxyamino group (C.sub.1-C.sub.6), alkylamino group (C.sub.1-C.sub.6), or cyclic or open chain amines, ester and amide derivatives selected from the group consisting of ##STR00057## ##STR00058##
2. The compound of claim 1, wherein the compound is selected from the group consisting of: (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(piperidin-1-yl)ethyl oxime oxalate (5a ), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(pyrrolidin-1-yl)ethyl oxime oxalate (5b), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(dimethylamino)ethyl oxime oxalate (5c), (4-(methylthio)phenyl)(naphthalen-1-yl methanone O-2-(diethylamino)ethyl oxime oxalate (5d), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(diisopropylamino)ethyl oxime (5e), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-morpholinoethyl oxime (5f), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-1-(dimethylamino)propan-2-yl oxime oxalate (5g), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(azepan-1-yl)ethyl oxime oxalate (5h), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(diethylamino)ethyl oxime citrate (5i), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(diethylamino)ethyl oxime fumarate (5j), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(diethylamino)ethyl oxime tartrate (5k), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(diethylamino)ethyl oxime (5l), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-3-(dimethylamino)propyl oxime oxalate (5m), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-3-chloropropyl oxime (5n), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-4-chlorobutyl oxime (5o), Ethyl 2-(4-(methylthio)phenyl)(naphthalen-1-yl)methyleneaminooxy)acetate (5p), N,N-diethyl-2-(4-(methylthio)phenyl)(naphthalen-1-yl)methyleneaminooxy) acetamide (5q), (4-methoxyphenyl)(naphthalen-1-yl)methanone O-2-(piperidin-1-yl)ethyl oxime oxalate (6a), (4-methoxyphenyl)(naphthalen-1-yl)methanone O-2-(pyrrolidin-1-yl)ethyl oxime oxalate (6b), (4-methoxyphenyl)(naphthalen-1-yl)methanone O-2-(dimethylamino)ethyl oxime oxalate (6c), (4-methoxyphenyl)(naphthalen-1-yl)methanone O-2-(diethylamino)ethyl oxime oxalate (6d), (4-methoxyphenyl)(naphthalen-1-yl)methanone O-2-(diisopropylamino)ethyl oxime oxalate (6e), (4-methoxyphenyl)(naphthalen-1-yl)methanone O-1-(dimethylamino)propan-2-yl oxime oxalate (6f), (4-methoxyphenyl)(naphthalen-1-yl)methanone O-2-(azepan-1-yl)ethyl oxime oxalate (6g), (4-methoxyphenyl)(naphthalen-1-yl)methanone O-2-motpholinoethyl oxime (6h), (4-methoxyphenyl)(naphthalen-1-yl)methanone O-3-(dimethylamino)propyl oxime oxalate (6i), (4-methoxyphenyl)(naphthalen-1-yl)methanone O-3-chloropropyl oxime (6j), (4-methoxyphenyl)(naphthalen-1-yl)methanone O-3-(piperidin-1-yl)propyl oxime (6k), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-3-(piperidin-1-yl)propyl oxime oxalate (7a), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-3-morpholinopropyl oxime (7b), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-3-(4-methylpiperidin-1-yl)propyl oxime oxalate (7c), (4-(methylthio)phenyl) (naphthalen-1-yl)methanone O-3-(butyl(methyl)amino) propyl oxime (7d), (4-methoxyphenyl)(naphthalen-1-yl)methanone O-3-(2-ethylhexylamino)propyl oxime (7e), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-4-(piperidin-1-yl)butyl oxime (8a), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-4-(pyrrolidin-1-yl)butyl oxime (8b), (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-4-(phenethylamino)butyl oxime (8c), and (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-4-(benzyl(ethyl)amino) butyl oxime (8d).
3. The compound of claim 1, wherein the structural formula of the compound is: ##STR00059## ##STR00060## ##STR00061## ##STR00062## ##STR00063## ##STR00064## ##STR00065##
4. The compound of claim 1, wherein the pharmaceutically acceptable salt is selected from the group consisting of hydrochlorides, citrates, oxalates, fumarates, malates and tartrates.
5. A pharmaceutical composition comprising the compound of claim 1.
6. The pharmaceutical composition of claim 5, further comprising a pharmaceutically acceptable carrier or diluent.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
(1)
(2)
(3)
(4)
(5)
(6)
(7)
(8)
(9)
ABBREVIATIONS
(10) Centans: Abbreviation used for compounds based on the IUPAC name of the compound, prefixed with Cent; a Central Drug Research Institute signature. CML: Chronic myelogenous leukemia AML: Acute myelogenous leukemia PBMC: Peripheral blood mononuclear cells BCR: Break point cluster region gene ABL: Abelson murine leukemia viral oncogene homolog 1 BCR-ABL: a fusion protein of BCR and ABL APOP: Apoptosis Stat5: Signal transducer and activator of transcription 5 Crkl: v-CRK avian sarcoma virus CT10-homolog-like eIF4E: Eukaryotic translation initiation factor 4E GATA-2: GATA binding protein 2 ER: Estrogen Receptor MTT: 3-(4,5-dimethythiazol-. 2-yl)-2,5-diphenyl tetrazolium bromide PI: Propidium Iodide
DETAILED DESCRIPTION OF THE INVENTION
(11) Accordingly, the present invention provides the process for preparation of substituted aryl naphthyl methanone oximes of general formula (I) and their derivatives, salts, pharmaceutical composition thereof and their use in treatment of chronic myelogenous leukemia, acute myelogenous leukemia, lymphoma, multiple myeloma, breast cancer, endometrial cancer prostate cancer and colon cancer as subject in need thereof.
(12) The present invention provides substituted aryl naphthyl methanone oxime (s) of formula (I), for use in the treatment of hematological malignancy, particularly for use in treatment of leukemia, acute leukemia, lymphoma and multiple myeloma and solid tumors such as breast cancer, endometrial cancer, prostate cancer and colon cancer.
(13) ##STR00012##
(14) R.sup.1=H, halogen, alkyl, alkoxy, nitro
(15) R.sup.2=H, halogen, alkyl, alkoxy, hydroxy, nitro
(16) R.sup.3=H, alkyl, alkoxy, nitro, halogen
(17) X=O, S
(18) R.sup.4=hydrogen, alkyl group (c.sub.1-c.sub.6), alkylamino group (c.sub.1-c.sub.6), cyclic or open chain amines
(19) R.sup.5=hydrogen, alkyl group (c.sub.1-c.sub.6) alkylepoxy, alkylhydroxyamino group (c.sub.1-c.sub.6), alkylamino group (c.sub.1-c.sub.6), cyclic or open chain amines, alkyl ester and amides derivatives selected from the group consisting of
(20) ##STR00013## ##STR00014##
(21) This invention also relates to a process for the preparation of Substituted Aryl naphthyl methanone oxime (s). Reacting the substituted Naphthalene-1-carboxylic acid (1) and suitably substituted phenol or thiophenol (2) in the presence of PPA at 90 C. to yield compound 3b. (Formula II) [Atul Kumar, S. R. Pathak, Pervez Ahmad, S. Ray, P. Tewari, A. K. Srivastava Bioorganic & Medicinal Chemistry Letters 16 (2006) 2719-2723].
(22) ##STR00015##
(23) The compound 3 on treatment with a hydroxylamine hydrochloride in the presence of base like pyridine, sodium acetate in dry ethanol yields compound 4. (Formula I)
(24) Compound 4 on treatment with a number of alkyl halides in the presence of base, like K.sub.2CO.sub.3, CsCO.sub.3, KOH, NaH etc. in dry DMF or acetone, yields compound 5, 6 of formula I.
(25) Compound 4 on treatment with various dihaloalkanes in the presence of base, like K.sub.2CO.sub.3, CsCO.sub.3 etc. in dry DMF/acetone also yields compound 5n, o (formula I), which on reaction with a number of amines in dry DMF/Methanol yields compound 7 & 8 of formula I (Scheme 2).
(26) Compound 4b on treatment with HBr/acetic acid, HBr/H.sub.2O, BBr.sub.3 or pyridine hydrochloride yields compound 4c (formula I).
(27) Following examples are given by way of illustrations and therefore, should not be construed to limit the scope of the present invention:
EXAMPLES
Synthesis of Compounds
(28) The following examples are given by way of illustrating the present invention and should not be construed to limit the scope of the invention:
Example 1
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone oxime (Compound 4a)
(29) ##STR00016##
(30) Into a 50 ml round bottom flask, (4-Methylsulfanyl-phenyl)-naphthalen-1-yl-methanone (1 mmole, 278 mg), hydroxylamine hydrochloride (2 mmole, 140 mg), dry pyridine (0.5 ml) and absolute ethanol (7.0 ml) were taken to obtain a reaction mixture. The reaction mixture was refluxed at 80 C. under anhydrous conditions for 8 hours. The reaction was followed by TLC (thin layer chromatography) monitoring. After completion of the reaction, ethanol was evaporated under reduced pressure. The resulting mixture was poured onto crushed ice and extracted with ethyl acetate (350 ml), washed with water until pH=7, dried over anhydrous Na.sub.2SO.sub.4 (8-9g) and concentrated. The crude product was purified by silica gel column chromatography using ethyl acetate and distilled Hexane as an eluent (8%) to yield the pure product It was then recrystellize from ethyl acetate and Hexane. Yield: 215 mg, 73.4%.
(31) M.P.=135 C., ESI MS (m/z)=294 (M+H); IR (KBr, Cm.sup.1): 3235.8, 2921.5, 1899.5, 1591.0, 1491.1, 1434.8, 1390.9, 1352.4, 1313.1, 1093.0, 1074.0, 969.6, 922.0, 770.1, 695.6; .sup.1H NMR(CDCl.sub.3, 300MHz): =7.99 (t, J=8.7 Hz, 2H, ArH), 7.73(d, J=8.1 Hz, 1H, ArH), 7.64-7.33(m, 6H, ArH), 7.16(d, J=7.2 Hz, 2H, ArH), 2.46(s, 3H, SCH.sub.3); .sup.13C NMR (CDCl3, 50 MHz): =157.43, 141.26, 133.89, 132.75, 131.73, 130.80, 129.61, 128.89, 127.84(2), 127.14, 126.74, 126.54 (2), 126.23, 126.09, 125.73, 15.67; Analysis calculated for C.sub.18H.sub.15NOS: C, 73.69; H, 5.15; N, 4.77. found: C, 73.71; H, 5.16; N, 4.80.
Example 2
Synthesis of (4-methoxyphenyl)(naphthalen-1-yl)methanone oxime (Compound 4b)
(32) ##STR00017##
(33) Into a 50 ml round bottom flask, (4-Methoxy phenyl)-naphthalen-1-yl-methanone (1 mmole, 262 mg), hydroxylamine hydrochloride (2 mmole, 140 mg), dry pyridine (0.5 ml) and absolute ethanol (7.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 8 hours. The reaction was followed by TLC monitoring. After completion of the reaction, ethanol was evaporated under reduced pressure. The resulting mixture was poured onto crushed ice and extracted with ethyl acetate (350 ml), washed with water until pH=7, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by silica gel column chromatography using ethyl acetate and distilled Hexane as an eluent (6%) to yield the pure product. It was then recrystallized from ethyl acetate and Hexane. Yield: 209 mg, 75.45%.
(34) M.P.=132 C.; ESI MS(m/z)=278(M+H); IR (KBr, Cm.sup.1): 3254.2, 3056.7, 2924.9, 1603.6, 1510.5, 1460.0, 1252.1, 1176.3, 1025.4, 970.4, 928.8, 835.0, 771.8; .sup.1H NMR(CDCl.sub.3, 300MHz): =7.95(t, J=6.6 Hz, 2H, ArH), 7.76(d, J=8.1 Hz, 1H, ArH), 7.63-7.39(m, 6H, ArH), 6.83(d, J=8.9 Hz, 2H, ArH), 3.79(s, 3H, OCH.sub.3); .sup.13C NMR (CDCl.sub.3, 75MHz): 160.82, 156.91, 133.51, 132.14, 131.87, 130.47, 129.11, 128.65, 128.48, 128.39, 126.72, 126.32, 126.13, 125.87, 125.38, 125.20, 113.91, 55.32; Analysis calculated for C.sub.18H.sub.15NO.sub.2: C, 77.96; H, 5.45; N, 5.05, found: C, 77.97; H, 5.43; N, 5.07.
Example 3
Synthesis of (4-hydroxyphenyl)(naphthalen-1-yl)methanone oxime (Compound 4c)
(35) ##STR00018##
(36) Into a 50 ml round bottom flask, (4-Hydroxy-phenyl)-naphthalen-1-yl-methanone (1 mmole, 248 mg), hydroxylamine hydrochloride (2 mmole, 140 mg), dry pyridine (0.5 ml) and absolute ethanol (7.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 7 hours. The reaction was followed by TLC monitoring. After completion of the reaction, ethanol was evaporated under reduced pressure. The resulting mixture was poured onto crushed ice and extracted with ethyl acetate (350 ml), washed with water until pH=7, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by silica gel column chromatography using chloroform as eluent to yield the pure product. It was then recrystellized with ethyl acetate and hexane. Yield: 221 mg, 84.03%. ESI MS (m/z)=264 (M+H); .sup.1H NMR (200 MHz, CDCl.sub.3): =7.91(t, J=8.0 Hz, 2H, ArH), 7.71(d, J=8.0 Hz, 1H, ArH), 7.55-7.31(m, 5H, ArH), 7.26(m, 1H, ArH), 6.52-6.47(m, 2H, ArH); .sup.13C NMR(CDCl.sub.3+CD.sub.3OD, 75MHz): 159.61, 157.99, 134.92, 134.07, 131.88, 129.84, 129.78(2), 129.51, 129.37, 127.50, 127.27, 127.11, 126.94, 126.48, 116.41; Analysis calculated for C.sub.17H.sub.13NO.sub.2: C, 77.55; H, 4.98; N, 5.32, found: C, 77.50; H, 5.06; N, 5.26.
Example 4
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(piperidin-1-yl)ethyl oxime oxalate (Compound 5a)
(37) ##STR00019##
(38) Into a 50 ml round bottom flask, 4-(methylthiophenyl)(naphthalen-1-yl)methanone oxime (1 mmole, 293 mg), 1-(2-chloroethyl)piperidine hydrochloride (1.2 mmole, 220.8 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (349 mg, 86.6%). The product obtained was oily, so a salt of the compound was prepared.
(39) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (109.1 mg) and oily product (349 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 390 mg, 85.2%.
(40) M.P. (Oxalate Salt of compound)=155 C., ESI MS (m/z)=404; IR (KBr, Cm.sup.1)=3020.4, 2970.1, 2360.8, 1758.4, 1630.1, 1522.3, 1216.4, 761.8, 670.3; .sup.1H NMR(CDCl.sub.3, 200MHz): =7.94 (d, J=8.0 Hz, 2H, ArH), 7.63-7.41(m, 5H, ArH), 7.29-7.27(m, 2H, ArH), 7.18 (d, J=8.2 Hz, 2H, ArH), 4.46(m, 2H, OCH.sub.2), 3.21(m, 4H, NCH.sub.2)2.47-2.46(m, 5H, SCH.sub.3 & NCH.sub.2), 1.51(m, 5H, CH.sub.2). Analysis calculated for C.sub.25H.sub.28N.sub.2OS: C, 74.22; H, 6.98; N, 6.92; found: C, 74.25; H, 6.95; N, 6.96.
Example 5
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(pyrrolidin-1-yl)ethyl oxime oxalate (Compound 5b)
(41) ##STR00020##
(42) Into a 50 ml round bottom flask, 4-(methylthiophenyl)(naphthalen-1-yl)methanone oxime (1 mmole, 293 mg), 1-(2-chloroethyl)pyrrolidine hydrochloride (1.2 mmole, 204 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (250 mg, 64.26%). The product obtained was oily, so a salt of the compound was prepared.
(43) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (108.9 mg) and oily product (250 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 390 mg, 85.2%.
(44) M.P. (Oxalate Salt of compound)=153 C., ESI MS (m/z)=390 (M+H); IR (KBr, Cm.sup.1)=3021.01, 2359.2, 1757.5, 1629.5, 1216.6, 761.0, 671.4; .sup.1H NMR (CDCl.sub.3, 200MHz): =7.93 (d, J=7.6 Hz, 2H, ArH), 7.62-7.41(m, 5H, ArH), 7.29-7.27(m, 2H, ArH), 7.17 (d, J=8.4 Hz, 2H, ArH), 4.61-4.45(m, 2H, OCH.sub.2), 3.29(m, 2H, NCH.sub.2), 2.82-2.80(m, 4H, NCH.sub.2), 2.47(s, 3H, SCH.sub.3), 1.70(m, 4H, CH.sub.2). Analysis calculated for C.sub.24H.sub.26N.sub.2OS (free base): C, 73.81; H, 6.71; N, 7.17, found: C, 73.79; H, 6.68; N, 7.15.
Example 6
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(dimethylamino)ethyl oxime oxalate (Compound 5c)
(45) ##STR00021##
(46) Into a 50 ml round bottom flask, 4-(methylthiophenyl) (naphthalen-1-yl)methanone oxime (1 mmole, 293 mg), (2-Chloro-ethyl)-dimethyl-amine hydrochloride (1.2 mmole, 172.8 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (280 mg, 76.9%). The product obtained was oily, so a salt of the compound was prepared.
(47) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (96.9 mg) and oily product (280 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 310 mg, 63.27%.
(48) M. P. (Oxalate Salt of compound)=105 C.; ESI MS (m/z)=365 (M+H), IR (KBr, Cm.sup.1): 3020.5, 2970.8, 2362.1, 1757.3, 1629.1, 1216.4, 1044.6, 760.9, 670.2; .sup.1H NMR (CDCl3, 200MHz): =7.93 (d, J=7.7 Hz, 2H, ArH), 7.61-7.41(m, 5H, ArH), 7.28-7.25(m, 2H, ArH), 7.17(d, J=8.4 Hz, 2H, ArH), 4.65-4.49(m, 2H, OCH.sub.2), 3.47(s, 6H, NCH.sub.3), 2.45(s, 3H, SCH.sub.3), 2.40(m, 2H, NCH.sub.2). Analysis calculated for C.sub.22H.sub.24N.sub.2OS (free base): C, 72.49; H, 6.64; N, 7.69; Found: C, 72.46; H, 6.63; N, 7.67.
Example 7
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(diethylamino)ethyl oxime oxalate (Compound 5d)
(49) ##STR00022##
(50) Method A: Into a 50 ml round bottom flask, 4-(methylthiophenyl)(naphthalen-1-yl)methanone oxime (1 mmole, 293 mg), 2-chloro-N,N-diethylethanamine hydrochloride (1.2 mmole, 205 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (250 mg, 63.6%). The product obtained was oily so a salt of the compound was prepared.
(51) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (80.15 mg) and the oily product (250 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 259 mg, 48.3%.
(52) M.P. (Oxalate Salt of compound)=115 C.; ESI MS (m/z)=393 (M+H); .sup.1H NMR(CDCl.sub.3, 200MHz): =7.93(d, J=8.0 Hz, 2H, ArH), 7.63-7.41(m, 5H, ArH), 7.27-7.25(m, 2H, ArH), 7.17-7.13(d, J=8.0 Hz, 2H, ArH), 4.48(m, 2H, OCH.sub.2), 3.26(m, 2H, NCH.sub.2), 2.72(m, 4H, NCH.sub.2), 2.46(s, 3H, SCH.sub.3), 0.81(m, 6H, CH.sub.3); Analysis calculated for C.sub.26H.sub.30N.sub.2O.sub.5S: C, 64.71; H, 6.27; N, 5.80 found: C, 64.76; H, 6.26; N, 5.85.
(53) Method B: Into a 50 ml round bottom flask, 4-(methylthiophenyl)(naphthalen-1-yl)methanone oxime (1 mmole, 293 mg), 2-chloro-N,N-diethylethanamine hydrochloride (1.2 mmole, 205 mg), Cs.sub.2CO.sub.3 (5 mmole) and dry DMF (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 5 hours. The reaction was followed by TLC monitoring. After completion of the reaction, filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (62.6%).
Example 8
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(diisopropylamino)ethyl oxime (Compound 5e)
(54) ##STR00023##
(55) Into a 50 ml round bottom flask, 4-(methylthiophenyl) (naphthalen-1-yl)methanone oxime (1 mmole, 293 mg), (2-Chloro-ethyl)-diisopropyl-amine hydrochloride (1.2 mmole, 240 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (325 mg, 77.38%). Oily Compound; ESI MS (m/z)=421(M+H); .sup.1H NMR(CDCl.sub.3, 200MHz): 7.92(d, J=7.6 Hz, 2H, ArH),7.57-7.39(m, 5H, ArH), 7.32-7.26(m, 2H, ArH), 7.15 (d, J=8.5 Hz, 2H, ArH), 4.09 (t, J=7.2 Hz, 2H, OCH.sub.2), 2.94-2.88(m, 2H, NCH), 2.68(m, 2H, NCH.sub.2), 2.45(s, 3H, SCH.sub.3), 0.93-0.89(d, J=6.5 Hz, 12H, CH.sub.3). Analysis calculated for C.sub.26H.sub.32N.sub.2OS: C, 74.24; H, 7.67; N, 6.66, found: C, 74.20; H, 7.65; N, 6.65.
Example 9
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-morpholinoethyl oxime (Compound 5f)
(56) ##STR00024##
(57) Into a 50 ml round bottom flask, 4-(methylthiophenyl) (naphthalen-1-yl)methanone oxime (1 mmole, 293 mg), 4-(2-Chloro-ethyl)-morpholine hydrochloride (1.2 mmole, 223.2 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (298 mg, 73.22%). Oily compound, ESI MS (m/z)=407 (M+H); IR(Neat, Cm.sup.1): 4300.2, 3020.6, 1757.4, 1593.6, 1216.5, 1042.5, 762.0, 671.3; .sup.1H (300MHz, CDCl.sub.3): =7.93 (d, J=8.3 Hz, 2H, ArH), 7.68 (d, J=8.2 Hz, 1H, ArH), 7.58-7.40(m, 5H, ArH), 7.32-7.28(m, 1H, ArH), 7.18(d, J=8.4 Hz, 2H, ArH), 4.33(t, J=5.6 Hz, 2H, OCH.sub.2), 3.77-3.51(m, 4H, OCH.sub.2), 2.63(t, J=5.6 Hz, 2H, NCH.sub.2), 2.47(s, 311, SCH.sub.3), 2.27(t, J=4.3 Hz, 4H, NCH.sub.2); Analysis calculated for C.sub.24H.sub.26N.sub.2O.sub.2S: C, 70.90; H, 6.45; N, 6.89, found: C, 70.89; H, 6.40; N, 6.86.
Example 10
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-1-(dimethylamino)propan-2-yl oxime oxalate (Compound 5g)
(58) ##STR00025##
(59) Method A: Into a 50 ml round bottom flask, 4-(methylthio)phenyl (naphthalen-1-yl)methanone oxime (1 mmole, 293 mg), (2-Chloro-propyl)-dimethyl-amine hydrochloride (1.2 mmole, 189.6 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry DMF (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 5 hours. The reaction was followed by TLC monitoring. After completion of the reaction, the mixture was quenched into water (100 ml), extracted with ethyl acetate (375 ml) and dried over anhydrous Na.sub.2SO.sub.4. The filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (67.13%). The product obtained was oily so a salt of the compound was prepared.
(60) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (83.3 mg) and oily product (250 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 260 mg, 64.35%.
(61) M.P. (Oxalate Salt of compound)=101 C., ESI MS (m/z)=379 (M+H); IR (KBr, Cm.sup.1)=3020.7, 2361.4, 1761.8, 1631.9, 1525.6, 1426.5, 1216.1, 761.0, 670.3; .sup.1H NMR(300MHz, CDCl.sub.3): =7.9240(d, J=7.8 Hz, 2H, ArH) 7.6788(d, J=8.2 Hz, 1H, ArH), 7.5763-7.3997(m, 5H, ArH), 7.3223-7.2806(m, 1H, ArH), 7.1766(d, J=8.4Hz, 2H, ArH), 4.0737-4.0187(m, 1H, OCH), 3.0861-2.8784(m, 2H, NCH.sub.2), 2.4656(s, 3H, SCH.sub.3), 2.1236(s, 6H, NCH.sub.3), 0.9154 (d, J=6.0 Hz, 3H, CHCH.sub.3); Analysis calculated for C.sub.23H.sub.26N.sub.2OS(free base): C, 72.98; H, 6.92; N, 7.40, found: C, 72.93; H, 6.88; N, 7.37.
(62) Method B: Into a 50 ml round bottom flask, 4-(methylthio)phenyl (naphthalen-1-yl)methanone oxime (1 mmole, 293 mg), (2-Chloro-propyl)-dimethyl-amine hydrochloride (1.2 mmole, 189.6 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (250 mg, 66.13%).
Example 11
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(azepan-1-yl)ethyl oxime oxalate (Compound 5h)
(63) ##STR00026##
(64) Into a 50 ml round bottom flask, 4-(methylthio)phenyl (naphthalen-1-yl)methanone oxime (1 mmole, 293 mg), 1-(2-Chloro-ethyl)-azepane hydrochloride (1.2 mmole, 237.6 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (350 mg, 83.7%). The product obtained was oily so a salt of the compound was prepared.
(65) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (105.5 mg) and oily product (350 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 427 mg, 78.49%.
(66) M.P. (Oxalate Salt of compound)=145 C.; ESI MS(m/z)=419 (M+H);); IR (KBr, Cm.sup.1): 3444.7, 2931.2, 2604.7, 1921.2, 1741.9, 1430.3, 1246.1, 1095.6, 1066.0, 961.0, 777.3; .sup.1H NMR(300MHz, CDCl.sub.3): =7.93 (d, J=8.2 Hz, 2H, ArH), 7.68(d, J=8.2 Hz, 1H, ArH), 7.58-7.39(m, 5H, ArH), 7.33-7.28(m, 1H, ArH), 7.18(d, J=8.6 Hz, 2H, ArH), 4.28(t, J=6.1 Hz, 2H, OCH.sub.2), 2.78(t, J=0.5 Hz, 2H, NCH.sub.2), 2.51(m, 4H, NCH.sub.2), 2.47(s, 3H, SCH.sub.3), 1.49(m, 8H, CH.sub.2); Analysis Calculated for C.sub.26H.sub.30N.sub.2OS (free base): C, 74.60; H, 7.22; N, 6.69, found: C, 74.62; H, 7.20; N, 6.68.
Example 12
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(diethylamino)ethyl oxime citrate (Compound 5i)
(67) ##STR00027##
(68) Procedure for citrate salt formation: citric acid, 1 mmole/1 mmole of compound, (122.45 mg) and (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(diethylamino)ethyl oxime (250 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly and kept in refrigerator for overnight after adding dry diethyl ether. The salt was filtered, washed with dry diethyl ether and collected. Yield: 49%. Analysis calculated for C.sub.30H.sub.36N.sub.2O.sub.8SC, 61.63; H, 6.21; N, 4.79; found: C, 61.62; H, 6.17; N, 4.75.
Example 13
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(diethylamino)ethyl oxime fumarate (Compound 5j)
(69) ##STR00028##
(70) Procedure for fumarate salt formation: fumaric acid, 1 mmole/1 mmole of compound, (73.98 mg) and (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(diethylamino)ethyl oxime (250 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly and kept in refrigerator for overnight after adding dry diethyl ether. The salt was filtered, washed with dry diethyl ether and collected. Yield: 48.3%. Analysis calculated for C.sub.28H.sub.32N.sub.2O.sub.5S, C, 66.12; H, 6.34; N, 5.51; found: C, 66.13; H, 6.37; N, 5.55.
Example 14
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(diethylamino)ethyl oxime tartrate (Compound 5k)
(71) ##STR00029##
Procedure for tartrate salt formation: tartric acid, 1 mmole/1 mmole of compound, (95.66 mg) and (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(diethylamino)ethyl oxime (250 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly and kept in refrigerator for overnight after adding dry diethyl ether. The salt was filtered, washed with dry diethyl ether and collected. Yield: 51%. Analysis calculated for C.sub.28H.sub.34N.sub.2O.sub.7S, C, 61.97; H, 6.32; N, 5.16; found: C, 61.93; H, 6.30; N, 5.11.
Example 15
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-2-(diethylamino)ethyl oxime (Compound 5l)
(72) ##STR00030##
(73) Into a 50 ml round bottom flask, 4-(methylthiophenyl)(naphthalen-1-yl)methanone oxime (1 mmole, 293 mg), 2-chloro-N,N-diethylethanamine hydrochloride (1.2 mmole, 205 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (250 mg, 63.6%). The product obtained was oily so a salt of the compound was prepared. ESI MS (m/z)=393 (M+H); IR (KBr, Cm.sup.1): 3445.9, 2939.4, 2362.7, 1742.4, 1645.1, 1400.5, 1235.1, 964.2, 718.3; NMR(CDCl.sub.3, 200MHz): =7.93(d, J=8.0 Hz, 2H, ArH), 7.63-7.41(m, 5H, ArH), 7.27-7.25(m, 2H, ArH), 7.17-7.13(d, J=8.0 Hz, 2H, ArH), 4.48(m, 2H, OCH.sub.2), 3.26(m, 2H, NCH.sub.2), 2.72(m, 4H, NCH.sub.2), 2.46(s, 3H, SCH.sub.3), 0.81(m, 6H, CH.sub.3); Analysis calculated for C.sub.24H.sub.28N.sub.2OS: C, 73.43; H, 7.19; N, 7.14, found: C, 73.41; H, 7.15; N, 7.10.
Example 16
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-3-(dimethylamino)propyl oxime oxalate (Compound 5m)
(74) ##STR00031##
(75) Into a 50 ml round bottom flask, 4-(methylthiophenyl)(naphthalen-1-yl)methanone oxime (1 mmole, 293 mg), 3-chloro-N,N-dimethylpropan-1-amine hydrochloride (1.2 mmole, 190 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (240 mg, 63.5%). The product obtained was oily so a salt of the compound was prepared.
(76) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (80 mg) and oily product (240 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 251 mg, 49.8%.
(77) M.P. (Oxalate Salt of compound)=110 C.; ESI MS (m/z)=379 (M+H); IR (KBr, Cm.sup.1): 3448.1, 2938.1, 2682.5, 1621.9, 1471.4, 1240.3, 1048.1, 719.1; .sup.1H NMR (CD.sub.3OD, 200MHz): =7.97 (d, J=8.2 Hz, 2H, ArH), 7.60-7.44(m, 5H, ArH), 7.39-7.26(m, 2H, ArH), 7.16 (d, J=8.0 Hz, 2H, ArH), 4.21-4.16(t, J=5.6 Hz 2H, OCH.sub.2), 2.90-2.85(m, 2H, NCH.sub.2), 2.63(s, 6H, NCH.sub.3), 2.43(s, 3H, SCH.sub.3), 2.02-1.96(m, 2H, CH.sub.2); Analysis calculated for C.sub.23H.sub.26N.sub.2OS(Free base): C, 72.98; H, 6.92; N, 7.40, found: C, 72.95; H, 6.89; N, 7.38.
Example 17
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-3-chloropropyl oxime (Compound 5n)
(78) ##STR00032##
(79) Into a 50 ml round bottom flask, 4-(methylthiophenyl) (naphthalen-1-yl)methanone oxime (2 mmole, 586 mg), 1-bromo-3-chloropropane (4 mmole, 0.39 ml), baked K.sub.2CO.sub.3 (10 mmole, 1380 mg) and dry acetone (15.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 8 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 nil). Filtrate was concentrated and the crude product was purified by silica gel column chromatography using distilled Hexane to yield the pure product (510 mg, 69.11%).
(80) Oily compound, ESI MS(m/z)=370 (M+H); IR(Neat, Cm.sup.1): 3464.5, 3055.8, 1594.0, 1493.4, 1437.8, 1314.6, 1217.4, 1095.5, 1042.5, 964.0, 758.3; (200 MHz, CDCl.sub.3): =7.92 (d, J=6 Hz, 2H, ArH), 7.66-7.41(m, 6H, ArH), 7.32-7.26(m, 1H, ArH), 7.17 (d, J=8 Hz, 2H, ArH), 4.28(t, J=4.9 Hz, 2H, OCH.sub.2), 3.42(t, J=6.7 Hz, 2H, CH.sub.2Cl), 2.46(s, 3H, SCH.sub.3), 2.10-2.04(m, 2H, CH.sub.2); Analysis calculated for C.sub.21H.sub.20ClNOS: C, 68.19; H, 5.45; N, 3.79, found: C, 68.21; H, 5.44; N, 3.80.
Example 18
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-4-chlorobutyl oxime (Compound 5o)
(81) ##STR00033##
(82) Into a 50 ml round bottom flask 4-(methylthiophenyl) (naphthalen-1-yl)methanone oxime (2 mmole, 586 mg), 1-bromo-4-chlorobutane (4 mmole, 0.46 ml), baked K.sub.2CO.sub.3 (10 mmole, 1380 mg) and dry acetone (15.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 8 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 nil). Filtrate was concentrated and the crude product was purified by silica gel column chromatography using distilled Hexane to yield the pure product (547 mg, 71.22%).
(83) Oily compound, ESI MS (m/z)=385 (M+H); .sup.1H (200MHz, CDCl.sub.3): =7.92 (d, J=7.6 Hz, 2H, ArH), 7.55-7.39(m, 6H, ArH), 7.32-7.20(m, 1H, ArH), 7.17(d, J=8.5 Hz, 2H, ArH), 4.19(t, J=5.7 Hz, 2H, OCH.sub.2), 3.42(t, J=6.5 Hz, 2H, CH.sub.2Cl), 2.46(s, 3H, SCH.sub.3), 1.79-1.61(m, 4H, CH.sub.2); Analysis calculated for C.sub.22H.sub.22ClNOS: C, 68.82; H, 5.78; N, 3.65; found: C, 68.80; H, 5.76; N, 3.64.
Example 19
Synthesis of Ethyl 2-((4-(methylthio)phenyl) (naphthalen-1-yl) methyleneaminooxy) acetate (Compound 5p)
(84) ##STR00034##
(85) Into a 50 ml round bottom flask, 4-(methylthiophenyl)(naphthalen-1-yl)methanone oxime (1 mmole, 293 mg), ethyl-2-bromoacetate (1.2 mmole), baked K.sub.2CO.sub.3 (5 mmole) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (65.0%).
(86) ESI MS (m/z)=393 (M+H); .sup.1H NMR(CDCl.sub.3, 300MHz): =7.93(d, J=6.0 Hz, 2H, ArH), 7.64-7.43(m, 7H, ArH), 7.16(d, J=6.0 Hz, 2H, ArH), 4.62(s, 1H, OCH.sub.2), 4.16(q, J=12.0 Hz, 2H, OCH.sub.2), 2.45(s, 3H, SCH.sub.3), 1.34 (t, J=12.0 Hz, 3H, CH.sub.3); Analysis calculated for C.sub.22H.sub.21NO.sub.3S: C, 69.63; H, 5.58; N, 3.69; found: C, 69.60; H, 5.53; N, 3.66.
Example 20
Synthesis of N,N-diethyl-2-((4-(methylthio)phenyl) (naphthalen-1-yl)methyleneaminooxy)acetamide (Compound 5q)
(87) ##STR00035##
(88) Into a microwave vial (2-5 cm), ethyl 2-((4-(methylthio)phenyl)(naphthalen-1-yl)methyleneaminooxy)acetate (379 mg, 1 mmole) and potassium tertiary butoxide (1 mmole) and diethylamine (1 mmole) was taken. The reaction mixture was then irradiated to microwave for 3 min. On completion of the reaction, the mixture was diluted with 15 ml of dichloromethane and extracted. It was then washed with brine, then with water and dried over anhydrous Na.sub.2SO.sub.4. The crude product was purified by silica gel column chromatography using 1% MeOH/CHCl.sub.3 as eluent to yield the pure product. Yield: 54.3%. .sup.1H NMR (CDCl.sub.3, 300 MHz): =7.92(d, J=8.0 Hz, 2H, ArH), 7.70(d, J=9.0 Hz, 1H, ArH), 7.62-7.46(m, 6H, ArH), 7.36-7.31(m, 1H, ArH), 7.20(d, J=9.0 Hz, 2H, ArH), 4.71(s, 2H, OCH.sub.2), 4.02(q, J=6.0 Hz, 4H, NCH.sub.2), 2.50(s, 3H, SCH.sub.3), 1.86(t, 6H, CH.sub.3); Analysis calculated for C.sub.24H.sub.26N.sub.2O.sub.2S: C, 70.90; H, 6.45; N, 6.89; found: C, 70.95; H, 6.43; N, 6.84.
Example 21
Synthesis of (4-methoxyphenyl)(naphthalen-1-yl)methanone O-2-(piperidin-1-yl)ethyl oxime oxalate (Compound 6a)
(89) ##STR00036##
(90) Into a 50 ml round bottom flask, (4-(methoxy phenyl)(naphthalen-1-yl)methanone oxime (1 mmole, 277 mg), 1-(2-chloroethyl)piperidine hydrochloride (1.2 mmole, 220.8 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). The filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (312 mg, 80.41%). The product obtained was oily so a salt of the compound was prepared.
(91) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (101.3 mg) and oily product (312 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 364 mg, 70.82%.
(92) M. P. (Oxalate Salt of compound)=171 C.; ESI MS (m/z): 389 (M+H), IR (KBr, Cm.sup.1): 3409.7, 3020.1, 2959.6, 2361.4, 1771.7, 1609.9, 1513.2, 1216.1, 1030.0, 761.0, 670.1. .sup.1H NMR(CDCl.sub.3, 300MHz): =7.92(d, J=8.0 Hz, 2H, ArH), 7.71(d, J=8.2 Hz, 1H, ArH), 7.58-7.42(m, 5H, ArH), 7.33-7.28(m, 1H, ArH), 6.84(d, J=8.8 Hz, 2H, ArH), 4.30(t, J=6.0 Hz, 2H, OCH.sub.2), 3.80(s, 3H, OCH.sub.3), 2.62(t, J=5.4 Hz, 2H, NCH.sub.2), 2.25-2.23(m, 4H, NCH.sub.2), 1.48-1.43(m, 4H, CH.sub.2), 1.34-1.28(m, 2H, CH.sub.2); Analysis calculated for C.sub.25H.sub.28N.sub.2O.sub.2 (free base): C, 77.29; H, 7.26; N, 7.21, found: C, 77.31; H, 7.30; N, 7.20.
Example 22
Synthesis of (4-methoxyphenyl)(naphthalen-1-yl)methanone O-2-(pyrrolidin-1-yl)ethyl oxime oxalate (Compound 6b)
(93) ##STR00037##
(94) Into a 50 ml round bottom flask, (4-(methoxy phenyl)(naphthalen-1-yl)methanone oxime (1 mmole, 277 mg), 1-(2-chloroethyl)pyrrolidine hydrochloride (1.2 mmole, 204 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). The filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (262 mg, 69.87%). The product obtained was oily so a salt of the compound was prepared.
(95) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (88.02 mg) and oily product (262 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 325 mg, 64.87%.
(96) M. P. (Oxalate Salt of compound)=158 C.; ESI MS(m/z)=375 (M+H); IR (KBr, Cm.sup.1): 3450.2, 3020.0, 1758.1, 1629.6, 1216.0, 1045.5, 761.6, 669.7; .sup.1H NMR(CDCl.sub.3, 300MHz): =7.93(d, J=8.0 Hz, 2H, ArH), 7.73(d, J=9.8 Hz, 1H, ArH), 7.58-7.42(m, 5H, ArH), 6.84(d, J=8.7 Hz, 2H, ArH), 4.32(t, J=6.1 Hz, 2H, OCH.sub.2), 3.75(s, 3H, OCH.sub.3), 2.75-2.68(m, 2H, NCH.sub.2), 2.47-2.44 (m, 4H, NCH.sub.2), 1.66-1.57. (m, 4H, CH.sub.2); Analysis calculated for C.sub.24H.sub.26N.sub.2O.sub.2 (free base): C, 76.98; H, 7.00; N, 7.48, found: C, 76.95; H, 7.03; N, 7.45.
Example 23
Synthesis of (4-methoxyphenyl)(naphthalen-1-yl)methanone O-2-(dimethylamino)ethyl oxime oxalate (Compound 6c)
(97) ##STR00038##
(98) Into a 50 ml round bottom flask, 4-(methylthio)phenyl (naphthalen-1-yl)methanone oxime (1 mmole, 277 mg), (2-Chloro-ethyl)-dimethyl-amine hydrochloride (1.2 mmole, 172.8 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). The filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (281 mg, 80.8%). The product obtained was oily so a salt of the compound was prepared.
(99) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (101.8 mg) and oily product (281 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 329 mg, 69.4%.
(100) M. P. (Oxalate Salt of compound)=141 C.; ESI MS (m/z)=349 (M+H), IR (KBr, Cm.sup.1): 3443.3, 3019.8, 2962.4, 2361.6, 1756.8, 1627.5, 1510.3, 1466.8, 1216.5, 1032.0, 761.1, 670.0; .sup.1H NMR(CDCl.sub.3, 200 MHz): =7.92(m, 2H, ArH), 7.61-7.37(m, 6H, ArH), 7.31-7.26(m, 1H, ArH), 6.84(d, J=6.8 Hz, 2H, ArH), 4.49(t, J=4.6 Hz, 2H, OCH.sub.2), 3.78(s, 3H, OCH.sub.3), 3.29-3.27(m, 2H, NCH.sub.2), 2.46(s, 6H, NCH.sub.3). Analysis calculated for C.sub.22H.sub.24N.sub.2O.sub.2 (free base): C, 75.83; H, 6.94; N, 8.04, Found: C, 75.80; H, 6.90; N, 8.00.
Example 24
Synthesis of (4-methoxyphenyl)(naphthalen-1-yl)methanone O-2-(diethylamino)ethyl oxime oxalate (Compound 6d)
(101) ##STR00039##
(102) Into a 50 ml round bottom flask, (4-(methoxy phenyl)(naphthalen-1-yl)methanone oxime (1 mmole, 277 mg), 2-chloro-N,N-diethylethanamine hydrochloride (1.2 mmole, 205 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). The filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (289 mg, 76.86%). The product obtained was oily so a salt of the compound was prepared.
(103) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (96.85 mg) and oily product (289 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 295 mg, 58.78%.
(104) M.P. (Oxalate Salt of compound)=137 C.; ESI MS(m/z)=377 (M+H), IR (KBr, Cm.sup.1): 3445.4, 2940.8, 2645.9, 1900.0, 1607.6, 1510.1, 1313.4, 1249.4, 1177.5, 1030.3, 963.3, 717.1; NMR(CDCl.sub.3, 200MHz): =7.91(d, J=8.1 Hz, 2H, ArH), 7.69(d, J=8.2 Hz, 1H, ArH), 7.57-7.39(m, 5H, ArH), 7.32-7.26(m, 1H, ArH), 6.83(d, J=7.1 Hz, 2H, ArH), 4.25(t, J=6.3 Hz, 2H, OCH.sub.2), 3.78(s, 3H, OCH.sub.3), 2.74(t, J=5.9 Hz, 2H, NCH.sub.2), 2.45-2.34(q, J=7.2 Hz, 4H, CH.sub.2), 0.88(t, J=7.2 Hz, 6H, CH.sub.3); Analysis calculated for C.sub.24H.sub.28N.sub.2O.sub.2: C, 76.56; H, 7.50; N, 7.44, found: C, 76.53; H, 7.52; N, 7.42.
Example 25
Synthesis of (4-methoxyphenyl)(naphthalen-1-yl)methanone O-2-(diisopropylamino)ethyl oxime oxalate (Compound 6e)
(105) ##STR00040##
(106) Into a 50 ml round bottom flask, 4-(methoxy phenyl (naphthalen-1-yl)methanone oxime (1 mmole, 277 mg), (2-Chloro-ethyl)-diisopropyl-amine hydrochloride (1.2 mmole, 240 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). The filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (318 mg, 78.71%). The product obtained was oily so a salt of the compound was prepared.
(107) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (99.18 mg) and oily product (318 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 331 mg, 62.45%.
(108) M.P. (Oxalate Salt of compound)=130 C.; ESI MS(m/z)=405 (M+H), IR (KBr, Cm.sup.1): 3445.3, 2977.4, 2691.6, 1748.4, 1638.8, 1508.0, 1394.7, 1252.6, 1178.0, 966.1, 777.6; .sup.1H NMR(CDCl.sub.3, 200MHz): =7.92(d, J=8.1 Hz, 2H, ArH), 7.69 (d, J=8.0 Hz, 1H, ArH), 7.58-7.40(m, 5H, ArH), 7.33-7.29(m, 1H, ArH), 6.83(d, J=6.9 Hz, 2H, ArH) 4.09(t, J=7.3 Hz, 2H, OCH.sub.2), 3.79(s, 3H, OCH.sub.3), 2.98-2.85(m, 2H, NCH), 2.69-2.60(m, 2H, NCH.sub.2), 0.94(d, J=6.5 Hz, 12H, OHCH.sub.3); Analysis calculated for C.sub.26H.sub.32N.sub.2O.sub.2: C, 77.19; H, 7.97; N, 6.92, found: C, 77.24; H, 7.99; N, 6.90.
Example 26
Synthesis of (4-methoxyphenyl)(naphthalen-1-yl)methanone O-1-(dimethylamino)propan-2-yl oxime oxalate (Compound 6f)
(109) ##STR00041##
(110) Into a 50 ml round bottom flask, 4-(methoxy phenyl (naphthalen-1-yl)methanone oxime (1 mmole, 277 mg), (2-Chloro-propyl)-dimethyl-amine hydrochloride (1.2 mmole, 189.6 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (270 mg, 74.59%). The product obtained was oily so a salt of the compound was prepared.
(111) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (93.98 mg) and oily product (270 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 250 mg, 51.23%.
(112) M.P. (Oxalate Salt of compound)=105 C.; ESI MS(m/z)=363 (M+H), IR (KBr, Cm.sup.1): 3746.4, 2931.5, 1754.4, 1694.1, 1629.4, 1559.5, 1511.4, 1462.6, 1252.0, 1176.7, 962.8, 739.5; .sup.1H NMR (200MHz, CDCl.sub.3): =7.91(d, J=8.8 Hz, 2H, ArH), 7.69(d, J=8.1 Hz, 1H, ArH), 7.54-7.39(m, 5H, ArH), 7.32-7.26(m, 2H, ArH & CHCl.sub.3), 6.83(m, 2H, ArH), 4.00-3.97(m, 1H, OCH), 3.80(s, 3H, OCH.sub.3), 2.09(s, 6H, NCH.sub.3), 0.85(d, J=6.6 Hz, 3H, CHCH.sub.3); Analysis calculated for C.sub.23H.sub.26N.sub.2O.sub.2: C, 76.21; H, 7.23; N, 7.73, found: C, 76.21; H, 7.23; N, 7.73.
Example 27
Synthesis of (4-methoxyphenyl)(naphthalen-1-yl)methanone O-2-(azepan-1-yl)ethyl oxime oxalate (Compound 6g)
(113) ##STR00042##
(114) Into a 50 ml round bottom flask, 4-(methoxy phenyl (naphthalen-1-yl)methanone oxime (1 mmole, 277 mg), 1-(2-Chloro-ethyl)-azepane hydrochloride (1.2 mmole, 237.6 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (312 mg, 77.61%). The product obtained was oily so a salt of the compound was prepared.
(115) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (97.79 mg) and oily product (312 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 331 mg, 62.69%.
(116) M.P. (Oxalate Salt of compound)=122 C.; ESI MS(m/z)=403 (M+H), IR (KBr, Cm.sup.1): 3446.5, 2933.9, 2599.9, 1721.6, 1607.9, 1510.2, 1459.5, 1251.3, 1177.9, 1026.7, 960.1, 719.2; .sup.1H NMR (300MHz, CDCl.sub.3): =7.92(d, J=8.0 Hz, 2H, ArH), 7.69(d, J=8.2 Hz, 1H, ArH), 7.58-7.39(m, 5H, ArH), 7.33-7.28(m, 1H, ArH), 6.84(d, 3=8.5 Hz, 2H, ArH), 4.27(t, J=6.1 Hz, 2H, OCH.sub.2), 3.80(s, 3H, OCH.sub.3), 2.78-2.67(m, 2H NCH.sub.2), 2.51(m, 4H NCH.sub.2), 1.49(m, 8H, CH.sub.2); Analysis calculated for C.sub.26H.sub.30N.sub.2O.sub.2: C, 77.58; H, 7.51; N, 6.96; found: C, 77.55; H, 7.53; N, 6.95.
Example 28
Synthesis of (4-methoxyphenyl)(naphthalen-1-yl)methanone O-2-morpholinoethyl oxime (Compound 6h)
(117) ##STR00043##
(118) Into a 50 ml round bottom flask, 4-(methoxy phenyl (naphthalen-1-yl)methanone oxime (1 mmole), 1-(2-Chloro-ethyl)-morpholine hydrochloride (1.2 mmole), baked K.sub.2CO.sub.3 (5 mmole) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 4 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (78.5%).
(119) ESI MS(m/z)=391 (M+H), NMR (300MHz, CDCl.sub.3): =7.94 (d, J=8.22Hz, 2H, ArH), 7.66-7.43 (m, 6H, ArH), 7.31(m, 1H, ArH), 6.86(d, J=8.9 Hz, 2H, ArH), 4.47(br, s, 2H, OCH.sub.3), 3.81(s, 3H, OCH.sub.3), 3.60-3.53(m, 4H, OCH.sub.2), 3.25-3.24(m, 2H, OCH.sub.2), 2.71(broad, 6H NCH.sub.2); Analysis calculated for: C.sub.24H.sub.26N.sub.2O.sub.3: C, 73.85; H, 6.73; N, 7.20; found: C, 73.82; H, 6.71; N, 7.17.
Example 29
Synthesis of (4-methoxyphenyl)(naphthalen-1-yl)methanone O-3-(dimethylamino)propyl oxime oxalate (Compound 6i)
(120) ##STR00044##
(121) Into a 50 ml round bottom flask (4-(methoxy phenyl)(naphthalen-1-yl)methanone oxime (1 mmole, 277 mg), 3-chloro-N,N-dimethylpropan-1-amine hydrochloride (1.2 mmole, 190 mg), baked K.sub.2CO.sub.3 (5 mmole, 690 mg) and dry acetone (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (253 mg, 69.89%). The product obtained was oily so the salt of the compound was prepared.
(122) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (88.06 mg) and oily product (253 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 250 mg, 65.96%.
(123) M.P. (Oxalate Salt of compound)=155 C.; ESI MS(m/z)=363 (M+H), IR (KBr, Cm.sup.1): 3446.3, 3045.6, 2957.4, 2680.0, 2470.8, 1741.9, 1602.8, 1510.0, 1251.9, 1177.4, 1043.6, 719.1; .sup.1H NMR (200MHz, CDCl.sub.3): =7.92(d, J=7.7 Hz, 2H, ArH), 7.69(d, J=8.0 Hz, 1H, ArH), 7.63-7.44(m, 5H, ArH), 7.32-7.28(m, 2H, ArH), 6.83(d, J=8.4 Hz, 2H, ArH), 4.17(t, J=6.0 Hz, 2H, OCH.sub.2), 3.80(s, 3H, OCH.sub.3), 2.65(m, 2H, NCH.sub.2), 2.12(s, 2H, NCH.sub.3), 1.78(m, 2H, CH.sub.2). Analysis calculated for C.sub.23H.sub.26N.sub.2O.sub.2: C, 76.21; H, 7.23; N, 7.73; Found: C, 76.20; H, 7.25; N, 7.70.
Example 30
Synthesis of (4-methoxyphenyl)(naphthalen-1-yl)methanone O-3-chloropropyl oxime (Compound 6j)
(124) ##STR00045##
(125) Into a 50 ml round bottom flask, (4-Methoxy-phenyl)-naphthalen-1-yl-methanone oxime (2 mmole, 526.58 mg), 1-bromo-3-chloropropane (4 mmole, 0.39 ml), baked K.sub.2CO.sub.3 (10 mmole, 1380 mg) and dry acetone (15.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 8 hours. The reaction was followed by TLC monitoring. After completion of the reaction, K.sub.2CO.sub.3 was filtered off and washed with acetone (210 ml). Filtrate was concentrated and the crude product was purified by silica gel column chromatography using 30% chloroform/Hexane to yield the pure product (614 mg, 86.7%).
(126) Oily compound, ESI MS(m/z)=354 (M+H); .sup.1H NMR (200MHz, CDCl.sub.3): =7.93(d, J=8.0 Hz, 2H, ArH), 7.70(m, 1H, ArH), 7.64-7.45(m, 5H, ArH), 7.33-7.29(m, 2H, ArH), 6.84(d, J=4.0 Hz, 2H, ArH), 4.29(t, J=5.7 Hz, 2H, OCH.sub.2), 3.42-3.35(m, 2H, CH.sub.2Cl), 3.79(s, 3H, OCH.sub.3), 2.14-2.01(m, 2H,CH.sub.2); Analysis calculated for C.sub.21H.sub.20ClNO.sub.2: C, 71.28; H, 5.70; N, 3.96; found: C, 71.32; H, 5.75; N, 3.90.
Example 31
Synthesis of (4-methoxyphenyl)(naphthalen-1-yl)methanone O-3-(piperidin-1-yl)propyl oxime (Compound 6k)
(127) ##STR00046##
(128) Into a 50 ml round bottom flask, (4-Methoxy-phenyl)-naphthalen-1-yl-methanone O-(3-chloro-propyl)-oxime (1 mmole, 353 mg), Piperidine (5 mmole, 0.49 ml) and dry methanol (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, methanol was evaporated under reduced pressure. The resulting mixture was poured onto water and extracted with ethyl acetate (350 ml), washed with water, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (289 mg, 69.1%).
(129) Oily Compound; ESI MS (m/z)=403 (M+H); .sup.1H NMR (200MHz, CDCl.sub.3): =7.93(d, J=8 Hz, 2H, ArH), 7.70(d, J=4 Hz, 1H, ArH), 7.64-7.45(m, 5H, ArH), 7.33-7.29(m, 1H, ArH), 6.84(d, J=4 Hz, 2H, ArH), 4.13-4.15(m, 2H, OCH.sub.2), 3.8(s, 3H, OCH.sub.3), 3.75(m, 2H, NCH.sub.2), 3.34(m, 2H, NCH.sub.2), 2.70-2.58(m, 2H, NCH.sub.2), 2.10-1.71(m, 8H, CH.sub.2). Analysis calculated for C.sub.21H.sub.20ClNO.sub.2: C, 77.58; H, 7.51; N, 6.96; found: C, 77.55; H, 7.50; N, 6.89.
Example 32
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-3-(piperidin-1-yl)propyl oxime oxalate (Compound 7a)
(130) ##STR00047##
(131) Into a 50 ml round bottom flask, 4-(methylthiophenyl)(naphthalen-1-yl)methanone O-3-chloropropyl oxime (1 mmole, 369 mg), piperidine (5 mmole, 0.49 ml) and dry methanol (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, methanol was evaporated under reduced pressure. The resulting mixture was poured onto water and extracted with ethyl acetate (350 ml), washed with water, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (335 mg, 75.59%). The product obtained was oily so a salt of the compound was prepared.
(132) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (100.7 mg) and oily product (335 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 395 mg, 90.65%.
(133) M.P. (Oxalate Salt of compound)=137 C.; ESI MS (m/z)=419 (M+H); IR (KBr, Cm.sup.1): 3854, 3740, 3443, 2925, 2358, 1753, 1633, 1553, 1220, 760; .sup.1H NMR (300MHz, CDCl.sub.3): =7.93(d, J=8.2 Hz, 2H, ArH), 7.64-7.38(m, 6H, ArH), 7.30(m, 1H, ArH), 7.16(d, J=8.0 Hz, 2H, ArH), 4.13(m, 2H, OCH.sub.2), 3.75(m, 2H, NCH.sub.2), 3.34(m, 2H, NCH.sub.2), 2.70-2.58(m, 2H, NCH.sub.2), 2.45(s, 3H, SCH.sub.3), 2.10-1.71(m, 8H, CH.sub.2). Analysis calculated for C.sub.26H.sub.30N.sub.2OS: C, 74.60; H, 7.22; N, 6.69; found: C, 74.58; H, 7.20; N, 6.67.
Example 33
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-3-morpholinopropyl oxime (Compound 7b)
(134) ##STR00048##
(135) Into a 50 ml round bottom flask, 4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-3-chloropropyl oxime (1 mmole, 369 mg), Morpholine (5 mmole, 0.44 ml) and dry methanol (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, methanol was evaporated under reduced pressure. The resulting mixture was poured onto water and extracted with ethyl acetate (350 ml), washed with water, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (265 mg, 60%).
(136) M.P.=89 C., ESI MS(m/z)=421(M+H); IR (KBr, Cm.sup.1): 3440.1, 2953.6, 2858.6, 2807.4, 2689.8, 1556.0, 1590.3, 1485.7, 1437.4, 1394.3, 1369.8, 1310.1, 1252.3, 1113.2, 1058.5, 585.8, 799.7, 777.7; (300MHz, CDCl.sub.3): =7.93(d, J=7.9 Hz, 2H, ArH), 7.54-7.39(m, 6H, ArH), 7.30 (d, J=8.1 Hz, 1H, ArH), 7.15(d, J=8.0 Hz, 2H, ArH), 4.19(t, J=6.3 Hz, 2H, OCH.sub.2), 3.70-3.60(m, 4H, OCH.sub.2), 2.44(s, 3H, SCH.sub.3), 2.30-2.15(m, 6H, NCH.sub.2), 1.26(m, 2H, CH.sub.2). Analysis calculated for C.sub.25H.sub.28N.sub.2O.sub.2S: C, 71.40; H, 6.71; N, 6.66. found: C, 71.38; H, 6.69; N, 6.64.
Example 34
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-3-(4-methylpiperidin-1-yl)propyl oxime oxalate (Compound 7c)
(137) ##STR00049##
(138) Into a 50 ml round bottom flask, 4-(methylthiophenyl)(naphthalen-1-yl)methanone O-3-chloropropyl oxime (1 mmole, 369 mg), 4-methylpiperidine (5 mmole, 0.59 ml) and dry methanol (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, methanol was evaporated under reduced pressure. The resulting mixture was poured onto water and extracted with ethyl acetate (350 ml), washed with water, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (350 mg, 81.1%). The product obtained was oily so a salt of the compound was prepared.
(139) Procedure for oxalate salt formation: Oxalic acid, 1 mmole/1 mmole of compound, (102.1 mg) and oily product (350 mg) were dissolved in dry methanol separately into two round bottom flasks. The acid and the compound were mixed and shaken thoroughly. The salt was precipitated using dry diethyl ether, filtered, washed with the same and collected. Yield: 400 mg, 71.7%.
(140) M.P. (oxalate salt of compound): 145 C., ESI MS(m/z)=433 (M+H), IR Spectra: (KBr, Cm.sup.1): 3445.4, 2928.8, 1648.1, 1451.6, 1246.1, 779.6. .sup.1H NMR (300MHz, CDCl.sub.3): =7.93(d, J=8.0 Hz, 2H, ArH), 7.56-7.38(m, 6H, ArH), 7.30(d, J=8.1 Hz, 1H, ArH), 7.16(d, J=8.5 Hz, 2H, ArH), 4.13(m, 2H, OCH.sub.2), 3.49-3.42(m, 2H, NCH.sub.2), 2.63(m, 2H, NCH.sub.2), 2.45(s, 3H, SCH.sub.3), 2.00(m, 2H, NCH.sub.2), 1.63(m, 5H, CH.sub.2 & CH), 1.24(m, 211, CH.sub.2), 0.96(d, J=5.6 Hz, 3H, CHCH.sub.3). Analysis calculated for C.sub.27H.sub.32N.sub.2OS (free base): C, 74.96; H, 7.46; N, 6.48; found: C, 74.92; H, 7.43; N, 6.47.
Example 35
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-3-(butyl(methyl)amino)propyl oxime (Compound 7d)
(141) ##STR00050##
(142) Into a 50 ml round bottom flask, (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-3-chloropropyl oxime (1 mmole, 369 mg), N-methylbutan-1-amine (5 mmole, 0.59 ml) and dry methanol (15.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, methanol was evaporated under reduced pressure. The resulting mixture was poured onto water and extracted with ethyl acetate (350 ml), washed with water, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (287 mg, 68.3%).
(143) Oily Compound; ESI MS (m/z)=421 (M+H); .sup.1H NMR (300MHz, CDCl.sub.3): =7.93(d, J=8 Hz, 2H, ArH), 7.62-7.39(m, 6H, ArH), 7.31-7.26(m, 1H, ArH), 7.15-7.11(m, 2H, ArH), 4.19-4.10(m, 2H, OCH.sub.2), 2.46(s, 3H, SCH.sub.3), 2.44(t, J=6.7 Hz, 4H, NCH.sub.2), 2.24(s, 3H, NCH.sub.2), 1.34-1.22(m, 6H, CH.sub.2), 0.91(t, J=6.9 Hz, 3H, CH.sub.3). Analysis calculated for C.sub.26H.sub.32N.sub.2OS: C, 74.24; H, 7.67; N, 6.66; found: C, 74.22; H, 7.65; N, 6.63.
Example 36
Synthesis of (4-methoxyphenyl)(naphthalen-1-yl)methanone O-3-(2-ethylhexylamino)propyl oxime (Compound 7e)
(144) ##STR00051##
(145) Into a 50 ml round bottom flask, (4-Methoxy-phenyl)-naphthalen-1-yl-methanone O-(3-chloro-propyl)-oxime (1 mmole, 353 mg), 2-ethylhexan-1-amine (5 mmole, 0.82 ml) and dry methanol (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, methanol was evaporated under reduced pressure. The resulting mixture was poured onto water and extracted with ethyl acetate (350 ml), washed with water, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (321 mg, 71.9%).
(146) Oily Compound; ESI MS (m/z)=447 (M+H); .sup.1H NMR (300MHz, CDCl.sub.3): =8.07(d, J=9 Hz, 1H, ArH), 7.94-7.91(m, 1H, ArH), 7.67-7.61(m, 2H, ArH), 7.59-7.54(m, 3H, ArH), 7.48-7.45 (m, 1H, ArH), 7.41-7.39 (d, 1H, ArH), 7.25-7.22(d, 2H, ArH), 3.48-3.28(m, 1H, OCH.sub.2), 3.25-3.23 (m, 1H, OCH.sub.2), 2.47 (s, 3H, SCH.sub.3), 1.93-1.87 (m, 2H, NCH.sub.2), 1.50-1.48(m, 2H, NCH.sub.2), 1.12-0.92(m, 5H, CH.sub.2CH.sub.2, CH), 0.81-0.60(m, 12H, 3CH.sub.2, 2CH.sub.3); Analysis calculated for C.sub.29H.sub.38N.sub.2O.sub.2: C, 77.99; H, 8.58; N, 6.27; found: C, 78.03; H, 8.61; N, 6.30.
Example 37
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-4-(piperidin-1-yl)butyl oxime (Compound 8a)
(147) ##STR00052##
(148) Into a 50 ml round bottom flask, 4-(methylthiophenyl)(naphthalen-1-yl)methanone O-4-chlorobutyl oxime (1 mmole, 383 mg), piperidine (5 mmole, 0.49 ml) and dry methanol (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 7 hours. The reaction was followed by TLC monitoring. After completion of the reaction, methanol was evaporated under reduced pressure. The resulting mixture was poured onto water and extracted with ethyl acetate (350 ml), washed with water, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product. Yield: 361 mg, 83.6%.
(149) Oily Compound, ESI MS (m/z)=433 (M+H); IR Spectra: (Neat, Cm.sup.1): 3020.7, 2934.8, 1595.0, 1216.0, 761.4; .sup.1H NMR (CDCl.sub.3, 300MHz): =7.93 (d, J=8.04 Hz, 2H, ArH), 7.67 (d, J=8.6 Hz, 1H, ArH), 7.58-7.41(m, 5H, ArH), 7.39-7.28(m, 1H, ArH), 7.17(d, J=8.6 Hz, 2H, ArH), 4.17-4.13(t, J=6.8 Hz, 2H, OCH.sub.2), 2.46(s, 3H, SCH.sub.3), 2.24-2.16(m, 6H, NCH.sub.2), 1.64-1.28(m, 10H, CH.sub.2); Analysis calculated for C.sub.27H.sub.32N.sub.2OS (free base)=C, 74.96; H, 7.46; N, 6.48; found: C, 74.92; H, 7.51; N, 6.50.
Example 38
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-4-(pyrrolidin-1-yl)butyl oxime (Compound 8b)
(150) ##STR00053##
(151) Into a 50 ml round bottom flask, 4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-3-chloropropyl oxime (1 mmole, 369 mg), Pyrrolidine (5 mmole, 0.42 ml) and dry methanol (10.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 7 hours. The reaction was followed by TLC monitoring. After completion of the reaction, methanol was evaporated under reduced pressure. The resulting mixture was poured onto water and extracted with ethyl acetate (350 ml), washed with water, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (289 mg, 69.1%).
(152) Oily Compound; ESI MS(m/z)=419(M+H); IR (Neat, Cm.sup.1): 3379.8, 3020.8, 1635.5, 1514.8, 1216.4, 761.7; NMR (300MHz, CDCl.sub.3): =7.93(d, J=8.1 Hz, 2H, ArH), 7.56-7.42(m, 6H, ArH), 7.33-7.28(m, 1H, ArH), 7.16(d, J=8.6 Hz, 2H, ArH), 4.18(t, J=6.5 Hz, 2H, OCH.sub.2), 2.46(s, 3H, SCH.sub.3), 2.37-2.31(m, 4H, NCH.sub.2), 1.77-1.43(m, 8H, CH.sub.2). Analysis calculated for C.sub.26H.sub.30N.sub.2OS: C, 74.60; H, 7.22; N, 6.69; found: C, 74.55; H, 7.18; N, 6.67.
Example 39
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-4-(phenethylamino)butyl oxime (Compound 8c)
(153) ##STR00054##
(154) Into a 50 ml round bottom flask, 4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-4-chlorobutyl oxime (1 mmole, 383 mg), 2-phenylethanamine (5 mmole, 0.63 ml) and dry methanol (15.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 6 hours. The reaction was followed by TLC monitoring. After completion of the reaction, methanol was evaporated under reduced pressure. The resulting mixture was poured onto water and extracted with ethyl acetate (350 ml), washed with water, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (329 mg, 72.5%).
(155) Oily Compound; ESI MS (m/z)=455 (M+H); .sup.1H NMR (300MHz, CDCl.sub.3): =7.90-7.85(m, 2H, ArH), 7.63(d, J=9 Hz, 1H, ArH), 7.54-7.49(m, 2H, ArH), 7.45-7.36(m, 5H, ArH), 7.29(m, 1H, ArH), 7.23-7.14(m, 5H, ArH), 4.13(t, J=6 Hz, 2H, OCH.sub.2), 2.50-2.45(m, 2H, NCH.sub.2), 2.45(s, 3H, SCH.sub.3), 1.60-1.55(m, 4H, NCH.sub.2, ArCH.sub.2), 0.91-0.85(m, 4H, CH.sub.2CH.sub.2). Analysis calculated for C.sub.29H.sub.30N.sub.2OS: C, 76.61; H, 6.65; N, 6.16; found: C, 76.56; H, 6.64; N, 6.15.
Example 40
Synthesis of (4-(methylthio)phenyl)(naphthalen-1-yl) methanone O-4-(benzyl(ethyl)amino)butyl oxime (Compound 8d)
(156) ##STR00055##
(157) Into a 50 ml round bottom flask, 4-(methylthio)phenyl)(naphthalen-1-yl)methanone O-4-chlorobutyl oxime (1 mmole, 383 mg), N-benzylethanamine (5 mmole) and dry methanol (15.0 ml) were taken. The reaction mixture was refluxed under anhydrous conditions for 7 hours. The reaction was followed by TLC monitoring. After completion of the reaction, methanol was evaporated under reduced pressure. The resulting mixture was poured onto water and extracted with ethyl acetate (350 ml), washed with water, dried over anhydrous Na.sub.2SO.sub.4 and concentrated. The crude product was purified by basic alumina column chromatography using distilled Hexane to yield the pure product (329 mg, 72.5%). Oily Compound; ESI MS (m/z)=483 (M+H); .sup.1H NMR (300MHz, CDCl.sub.3): =7.93-7.91(m, 3H, ArH), 7.64-7.14(m, 13H, ArH), 4.17(t, J=6 Hz, 2H, OCH.sub.2), 3.82(s, 2H, NCH.sub.2), 2.75(q, J=4.0 Hz, 2H, NCH.sub.2), 2.54-2.47(m, 5H, SCH.sub.3, NCH.sub.2), 1.77-1.66(m, 4H, CH.sub.2), 1.19-1.14(t, J=4.0 Hz, 3H, CH.sub.3). Analysis calculated for C.sub.31H.sub.34N.sub.2OS: C, 77.14; H, 7.10; N, 5.80; found: C, 77.17; H, 7.12; N, 5.85.
(158) Primary Evaluation of Aryl Naphthyl Methanone Oxime Derivative Compounds in Leukemia Cell Lines K562 and HL-60
(159) All the compounds synthesized were screened for cytotoxicity on leukemia cell lines K562, HL-60 and mouse fibroblast cell line NIH3T3 using (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay kit from Sigma according to manufacturer's instructions and trypan blue exclusion method. Activity of these compounds is given in Table-1. Cytotoxicity of five most active compounds (5a, 5d, 5j, 5m and 7c) was further checked in Kasumil and U937 cells. Compounds 5a, 5d, 5m and 7c exhibited higher efficacy than imatinib (IC.sub.50 values are shown in Table 2) in K562.
(160) TABLE-US-00001 TABLE 1 IC.sub.50 (M) values of compounds in K562, HL60 and NIH3T3 cell lines as tested using MTT assay after 48 h treatment. Results were calculated from three independent experiments performed in triplicate. Compounds with IC.sub.50 more than 15 M were considered inactive. S. No. Compound K562 HL60 NIH3T3 1 4a >50 M >50 M >50 M 2 4b >50 M >50 M >50 M 3 5a 5.8 M >20 M 15 M 4 5b 25 M 19 M 20 M 5 5c >50 M >50 M >50 M 6 5d 3.61 M 5.99 M >20 M 7 5e 7 M 14 M 16 M 8 5f >25 M >50 M 26 M 9 5g >50 M >50 M >50 M 10 5h >50 M >50 M >50 M 11 5i >30 M >50 M 40 M 12 5j 8.5 M 10 M 25 M 13 5k >50 M >50 M >50 M 14 5l >50 M >50 M >50 M 15 5m 4.8 M 7.78 M >25 M 16 5p >50 M 15 M >50 M 17 5q >50 M >50 M >50 M 18 6a >50 M >50 M >50 M 19 6b >50 M >50 M >50 M 20 6c >50 M >50 M >50 M 21 6d >50 M >50 M >50 M 22 6e >50 M >50 M >50 M 23 6f >50 M >50 M >50 M 24 6g 6.66 M >50 M 16 M 25 6h 19 M >50 M 26 M 26 6i >50 M >50 M >50 M 27 7a >20 >50 M >50 M 28 7b >50 M >50 M >50 M 29 7c 5.73 M >50 M 21 M 30 7d >50 M 13 M >50 M 31 8a 6.34 M 16 M 20 M 32 8b >50 M >50 M >50 M 33 8c >30 M >50 M >50 M 34 8d 16.3 M >50 M >50 M
Evaluation of Hit Compounds in Multiple Leukemia Cell-Lines:
(161) To consider a molecule as a candidate compound in cancer drug therapy, the test compound must have specific cytotoxicity towards the cancer cells without affecting normal cell growth. Therefore, all active compounds were assessed for toxicity in non-cancer cell-lines: mouse fibroblast cell line (NIH3T3) and monkey kidney epithelial cells (Vero). 5d and 5m exhibited much higher IC.sub.50 values in the normal cells compared to leukemia cells. Therefore, based on safety index (IC.sub.50/CC.sub.50), 5d [Centans; based on IUPAC name prefixed with Central Drug Research Institute's signature (Cent)] was selected as a lead compound.
(162) TABLE-US-00002 TABLE 2 IC.sub.50 of Active compounds in Leukemia and normal cell lines. Results were calculated from three independent experiments performed in triplicates. Compound K562 HL60 Kasumi 1 U937 Vero NIH3T3 5a 5.8 M >20 M 27 M >18 M >25 M 15 M 5d 3.61 M 5.99 M 6.78 M 8.12 M >25 M >20 M (Centans) 5j 8.5 M 10 M 20 M >24 M >25 M 25 M 5m 4.8 M 7.78 M >10 M >15 M >25 M >25 M 7c 5.73 M >50 M 19 M >25 M >25 M 21 M IMT 7.53 M 9.63 M ND ND ND ND
(163) As described in table 2, among all the compounds tested compound 5d (Centans) showed strong cytotoxicity against BCR-ABL-positive CML model K562, acute myeloblastic leukemia model HL60, Kasumil (Acute Myeloid Leukemia with t(8; 21) translocation) and U937 (monoblast), while it showed no toxicity against non-cancerous fibroblastic cell-lines Vero and NIH3T3. ND; not done.
(164) Evaluation of the Efficacy of Lead Compound Centans in Comparison with Imatinib and Dasatinib:
(165) The efficacy of Centans in comparison to marketed drugs imatinib and dasatinib were determined in K562 and HL-60 cells that were used as examples of CML and AML respectively. As shown in table 3, Centans exhibited a much lower IC50 in comparison to both imatinib and dasatinib, indicating its higher efficacy.
(166) TABLE-US-00003 TABLE 3 IC.sub.50 of Centans, imatinib and dasatinib in K562 and HL60 cell lines after 48 h of treatment. Following 48 h treatment of indicated cell lines with Centans, imatinib or dasatinib (10 point dose response) MTT assays were performed and IC-50 for the compounds were determined. Results were calculated from three independent experiments performed in triplicates. Cell-line IC 50(M) Centans HL 60 5.99 K562 3.61 Imatinib HL 60 9.63 K562 5.16 Dasatinib HL 60 6.85 K562 6.8
(167) Centans shows higher efficacy than imatinib and dasatinib in peripheral blood mononuclear cells (PBMC) isolated from CML patients.
(168) Next PBMCs isolated from freshly diagnosed, imatinib sensitive, imatinib resistant CML (BCR-ABL+ve) patient samples or healthy controls were treated for 48h with Centans, imatinib and dasatinib and following treatment the efficacy of each drug in each group was determined by MTT assay. As shown in Table 4. Centans showed most robust cytotoxic effect in PBMCs from imatinib-resistant CML patients (72%) compared to imatinib (40%) and dasatinib (59%). Centans also showed higher cytotoxicity than imatinib or dasatinib in PBMCs from freshly diagnosed or imatinib sensitive patients, while all three drugs showed comparative activity on PBMCs from healthy controls.
(169) TABLE-US-00004 TABLE 4 PBMCs isolated from indicated groups of BCR-ABL-positive CML patients or healthy individuals were treated with 1 M Centans, imatinib or dasatinib for 48 h and the cytotoxicity was determined by MTT assay in triplicates. Data represents % cell death. Centans showed better efficacy than both imatinib and dasatinib in all the groups, but this efficacy was highest in imatinib-resistant patient samples (p < 0.01). P value P value Patient Sample Centans vs Centans vs Type ID Centans Imatinib Imatinib Dasatinib Dasatinib Healthy C1 14.01 0.76978 10.51 0.30456 2.39 control C2 20.65 17.91 12.03 C3 16.93 19.93 18.93 Average 17.19667 16.11667 11.11667 SEM 1.921435 2.863339 4.796476 Freshly P1 68.82 0.102813 61.58 0.017607 55.46 Diagnosed P10 73.32 53.34 43.59 P11 55.61 49.51 32.75 P15 71.19 64.1 52.79 Average 67.235 57.1325 46.1475 SEM 3.982484 3.42554 5.138736 Imatinib P2 69.46 0.007977 32.81 0.0074 56.61 Resistant P13 70.03 34.05 53.82 P14 77.68 48.35 68.84 P17 78.93 47.84 56.41 P18 64.49 36.67 57.84 Average 72.118 39.944 58.704 SEM 3.030576 3.786158 2.926022 Imatinib P3 50.92 0.32157 49.79 0.510364 73.22 treated/ P12 62.41 48.39 43.65 Responder P16 76.59 62.39 47.26 Average 63.30667 53.52333 54.71 SEM 6.429235 3.855299 8.065716
(170) Next, comparative cytotoxic efficacy of centans with imatinib and dasatinib was assessed in CML patient samples harbouring different BCR-ABL mutations by MTT assay. PBMCs isolated from patients harbouring indicated mutations in BCR-ABL were treated with 5 M centans, imatinib or dasatinib for 48h, followed by MTT assay. As shown in Table 5. Centans showed much stronger cytotoxic activity in comparison to imatinib or dasatinib. Importantly centans showed appreciable cytotoxicity in patient sample harbouring multidrug resistant T315I BCR-ABL mutation.
(171) TABLE-US-00005 TABLE 5 Centans induces cell death in PBMCs from patients with BCR-ABL mutations including multi-drug resistant T315I mutation with higher efficacy than imatinib and dasatinib. % Cell Death Mutation centans Imatinib Dasatinib T315I 34.60 4.81 8.91 H396R 40.78 8.66 27.82 Y253H 42.93 19.36 32.96 E255V 48.18 23.10 44.40
(172) Centans-Induced Cell-Death in K562 and HL-60 is Apoptotic in Nature.
(173) To determine whether cell death caused by Centans is specifically associated with apoptosis: phosphatidylserine exposure (Annexine V/PI; Flow cytometry) was performed in K562 and HL-60 cell-lines. As apparent form
(174)
(175) Centans Induces G0/G1 Growth Arrest in K562 and HL-60 Cells.
(176) To evaluate the cytostatic effect of Centans, K562 and HL-60 cells were analysed for cell cycle progression in presence of Centans in and as shown in
(177)
(178) Propidium iodide (PI) staining of K562 cells (A) and HL-60 cells (B) treated with vehicle (DMSO) or 0.5 M Centans, followed by flow cytometry analysis to find % distribution of cells in different phases of the cell cycle. Data has been shown as percent of total cells from data pooled from three independent experiments. (C and D). mRNA expression of cell cycle regulatory factorsK562 (C) or HL-60 (D) cells were treated with Vehicle or Centans, followed by mRNA isolation and cDNA preparation and quantitative real-time PCR (p21, p27 and cyclin D1). Graphs represented pooled data from three independent experiments. *P<0.05, **P<0.01,***P<0.001 compared to vehicle treated control.
(179) Centans Induces Dephosphorylation of BCR-ABL and SRC:
(180) Kinase activity of Bcr-Abl is the major cause of CML, and since Centans causes growth arrest and apoptosis in Bcr-Abl positive K562 cells, phosphorylation status of Bcr-Abl which is representative of its kinase activity was measured by immunoblotting. As shown in
(181) Effect of Centans was also observed in terms of SRC kinase activation. SRC kinases activated by BCR-ABL remain fully active in imatinib-treated mouse leukemic cells, suggesting that imatinib does not inactivate all BCR-ABL activated signalling pathways. This SRC pathway is responsible for survival of leukemic cells that becomes imatinib resistant and for CML transition to lymphoid blast crisis. Effect of Centans and Imatinib was compared on its dephosphorylation by immunoblotting and we found that Centans dephosphorylating SRC-1 in a dose dependent manner, while imatinib failed to do so (
(182)
(183) Centans Causes Differentiation in K562 Cells:
(184) In order to clarify whether the inhibitory effects of Centans on the growth and cell cycle of K562 cells was followed by differentiation, Centans treated cells was stained with Wright-Giemsa stain.
(185)
(186) Centans Induces Granulocytic/Monocytic Differentiation in HL-60:
(187) Differentiation potential of Centans on HL-60 cells was evaluated by the effect of this compound on Nuclear/cytosolic ratio and change in nuclear morphology, which are reliable markers of differentiation. For this, HL60 cell treated with Centans were cyto-spinned on microscope slides and stained with Wright-Giemsa and micro-photographed. As shown in
(188) Further confirm of granulocytic/monocytic differentiation of HL-60 cells by Centans, was obtained by QRT-PCR analysis of expression of lineage specific differentiation markers following Centans treatment. mRNA level of CD11b, a marker for granulocytic differentiation was significantly increased (
(189)
(190) Evaluation of Centans in Solid Tumor:
(191) Efficacy of Centans and related series of compounds were evaluated in solid tumor cell-lines MCF-7 (estrogen receptor positive, non-metastatic breast tumor), MDA-MB-231 (estrogen receptor negative, metastatic breast tumor), DU-145 (prostrate tumor), Ishikawa (endometrial adenocarcinoma) cells by MTT assay following 24h treatment of indicated compounds. As depicted in table 6. Centans showed selective cytotoxicity in all the cancer cells tested but not in non-cancer cell-lines Vero and HEK-293. Efficacy of Centans was comparable to that of the positive control compound 4-hydroxy-tamoxifen (4-OHT).
(192) TABLE-US-00006 TABLE 6 IC.sub.50 of Centans and related compounds in cell line models of breast, prostate and endometrial cancers after 48 h of treatment. Following 48 h treatment of indicated cell lines with Centans, Centans analogs or 4-OHT (10 point dose response) MTT assays were performed and IC-50 for the compounds were determined. Results were calculated from three independent experiments performed in triplicates. Compound MCF-7 MDA-MB231 DU-145 Ishikawa 4a 18 20 >50 >50 5a 13.5 8.8 10 10 5b 6.6 5.01 9.5 6 5c 14 12.78 15 9 5d (Centans) 7.5 7.2 9 12 5f >50 >50 >50 >50 5m 8 7.2 9.5 7 6a 18 >50 >50 >50 6b 25 >50 >50 >50 6c 15 >50 >50 >50 6d 20 >50 >50 >50 6e 23 >50 >50 >50 6i 20.5 >50 >50 >50 7a 5 20 15 12 7b >50 >50 >50 >50 7c 6.5 20 10 >50 8a 8 18 7.5 18 8b >50 >50 >50 >50 OH-TAM 8.8 8 10 20
(193) Efficacy of centans in colon cancer cell-lines were also assessed using sulphorhodamine B assay and as shown in table 7, centans exhibited robust cytotoxicity in four different colon cancer cell lines compared to the marketted drug for colon cancer 5-fluoro uracil (5-FU).
(194) TABLE-US-00007 TABLE 7 IC50 of Centans in colon cancer cell lines as determined by Sulphorhodamine B (SRB) assay. Cancer cell-line SW620 DLD1 HCT116 Colo205 Centans 8.78 M 15.6 M 7.7 M 4.2 M 5-FU >50 M 16.38 M >50 M 38.06 M
Centans Induces Apoptosis in MCF-7 and MDA-MB-231 Cell-Lines.
(195) To assess if the cell death induced by centans in MCF-7 and MDA-MB231 lines is apoptotic in nature, apoptosis was measured by flow cytometry using annexin V/PI staining kit from Sigma according to manufacturer's instructions. A concentration-dependent increase in apoptotic cell population was observed upon treatment with Centans and the effect was stronger in MDA-MB231 than MCF-7 cells (
(196)
(197) Centans Inhibits EGFR and Her2 Phosphorylation
(198) Since EGFR and Her2 play major roles in breast cancer development and metastatis, the effect of Centans on regulation of these two factors was evaluated in MCF-7 and MDA-MB-231 cells by enzyme-linked immunosorbant assays (ELISA). As depicted in
(199)
(200) Centans Reduces Tumor Volume in Mouse Xenograft Model:
(201) The effect of Centans on the growth of MCF-7 xenograft tumor was studied. Injection of MCF-7 cells in the peritoneal cavity of NIH-III strain of nude mice resulted in the appearance of palpable tumors that rapidly grew from 9 d to 25 d. Beyond 25 d, vehicle treated mice bearing tumors (control) show significant mortality (data not shown). Administration of Centans at 16 mg.Math.kg.sup.1.Math.day.sup.1 dose resulted in nearly 50% reduction in relative tumor volume compared to vehicle treated mice on days 21 and 25 following the inoculation of MCF-7 cells (
(202) Histological assessment showed that tumors from vehicle-treated mice were primarily composed of tumor epithelial cells with small amounts of mouse-derived stroma and frequent blood vessels. Tumors from mice treated with Centans presented with large areas of stroma where deletion of epithelial cells had occurred as evident from few mitotic figures (
(203)
(204) Centans Reduces CD133+Colon Cancer Stem Cells
(205) Since centans showed cytotoxicity in drug resistant leukemic cells, the efficacy of centans in cancer stem cells (CD133+DLD1 colon cancer cells) was assessed. As shown in
(206)
(207) Advantages of the Present Invention
(208) 1. The present compound shows better efficacy than existing drugs imatinib and dasatinib both in leukemia cell-lines and patient samples. 2. The present compound causes robust apoptosis in imatinib resistant cancer samples. 3. The present compound induces cell death in broader range of leukemic cells than imatinib or dasatinib. 4. The present compound induces differentiation in blast cells which is indicative of restoring normal blood cell functions. 5. The present compound is also efficacious in solid tumor forming cell-lines including, breast, prostate and endometrial cancers.