HETEROARYL COMPOUNDS COMPRISING NITROGEN AND USE THEREOF
20170305861 · 2017-10-26
Assignee
Inventors
- Sung Wuk Kim (Seongnam-si, KR)
- Hong Woo Kim (Irvine, CA, US)
- SangHee Yoo (Houston, TX, US)
- Hyunwook Kim (Seoul, KR)
- Hye Jin Heo (Daejeon, KR)
- Hong Bum Lee (Daejeon, KR)
- Jiae Kook (Daejeon, KR)
- Young Woo Lee (Yangyang-gun, KR)
Cpc classification
A61K45/06
HUMAN NECESSITIES
C07D283/00
CHEMISTRY; METALLURGY
C07D227/02
CHEMISTRY; METALLURGY
International classification
C07D227/02
CHEMISTRY; METALLURGY
A61K45/06
HUMAN NECESSITIES
Abstract
The present invention relates to heteroaryl compounds comprising nitrogen and use thereof, and more specifically to compounds which exhibit a remarkable effect on inhibiting proliferation of cancer cells and metastasis and recurrence of cancer, a preparation method of the same, and a pharmaceutical composition comprising the same as an active ingredient.
The compounds according to the present invention exhibit a remarkable effect on inhibiting proliferation of cancer cells and metastasis and recurrence of cancer with a reduced dose compared to that of existing drugs. Accordingly, the compounds can be effectively used for treating various types of cancer, such as uterine cancer, breast cancer, gastric cancer, brain cancer, rectal cancer, colorectal cancer, lung cancer, skin cancer, blood cancer, pancreatic cancer, renal cancer, prostate cancer, bladder cancer, and liver cancer, and for inhibiting proliferation of cancer cells and metastasis of cancer.
Claims
1. A compound represented by Formula 1 below or a pharmaceutically acceptable salt thereof: ##STR00106## wherein, in Formula 1, refers to a single bond or double bond, and a ring of Formula 1 comprises two to three double bonds, wherein the double bonds are not adjacent to each other, X is CH, CNH.sub.2, or N, Y is CH, N, or S, n is 1 or 2, L is C.sub.1-6 alkylene or C.sub.1-6 alkenylene, R.sup.1 is C.sub.6-14 aryl, C.sub.5-20 heteroaryl, C.sub.3-8 cycloalkyl, or C.sub.3-8 heterocycloalkyl, and R.sup.2 to R.sup.4 are each independently hydrogen, amino (—NH.sub.2), substituted amino (—NHR′ or —NR′R″), nitro, halogen, cyano, oxo, hydroxy, C.sub.1-6 alkyl, C.sub.3-8 cycloalkyl, C.sub.3-8 heterocycloalkyl, C.sub.1-6 alkoxy, C.sub.1-6 haloalkyl, or C.sub.1-6 haloalkoxy; or R.sup.2 and R.sup.3 are positioned on adjacent carbon atoms and connected to each other to form a ring, wherein R′ and R″ are each independently C.sub.1-6 alkyl; or R′ and R″ are connected to each other to form a ring comprising a nitrogen atom to which R′ and R″ are bonded.
2. The compound of claim 1, wherein L is C.sub.1-6 alkylene or C.sub.1-6 alkenylene, which is unsubstituted or substituted with oxo.
3. The compound of claim 1, wherein R.sup.1 is C.sub.6-14 aryl, C.sub.5-20 heteroaryl, C.sub.3-8 cycloalkyl, or C.sub.3-8 heterocycloalkyl, which is unsubstituted or substituted with one or more substituents selected from the group consisting of hydroxy, halogen, amino, cyano, nitro, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 haloalkyl, and C.sub.1-6 haloalkoxy.
4. The compound of claim 1, wherein R.sup.2 to R.sup.4 are each independently C.sub.1-6 alkyl, C.sub.3-8 cycloalkyl, C.sub.3-8 heterocycloalkyl, C.sub.1-6 alkoxy, C.sub.1-6 haloalkyl, or C.sub.1-6 haloalkoxy, which are unsubstituted or substituted with one or more substituents selected from the group consisting of halogen, hydroxy, cyano, nitro, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 haloalkyl, and C.sub.1-6 haloalkoxy.
5. The compound of claim 1, wherein the ring formed by R.sup.2 and R.sup.3 being positioned on adjacent carbon atoms and connected to each other is substituted with one or more substituents selected from the group consisting of halogen, hydroxy, cyano, nitro, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 haloalkyl, and C.sub.1-6 haloalkoxy.
6. The compound of claim 1, wherein, R′, R″, or the ring formed when R′ and R″ are connected to each other, which comprises a nitrogen atom to which R′ and R″ are bonded, is each independently substituted with one or more substituents selected from the group consisting of hydroxy, cyano, nitro, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 haloalkyl, and C.sub.1-6 haloalkoxy.
7. The compound of claim 1, wherein the ring formed by R″ and R.sup.3 being positioned on adjacent carbon atoms and connected to each other; or the ring comprising a nitrogen atom to which R′ and R″ are bonded, formed by R′ and R″ being connected to each other, is C.sub.6-14 aryl, C.sub.5-20 heteroaryl, C.sub.3-10 cycloalkyl, or C.sub.3-10 heterocycloalkyl.
8. The compound of claim 1, wherein L is methylene, ethylene, propylene, or —CH.sub.2—C(O)—.
9. The compound of claim 1, wherein R.sup.1 is C.sub.6-8 aryl, C.sub.3-8 cycloalkyl, or C.sub.5-8 heteroaryl.
10. The compound of claim 1, wherein R.sup.1 is phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, thiophene, furan, or selenophene.
11. The compound of claim 1, wherein R.sup.1 is C.sub.6-8 aryl, C.sub.3-8 cycloalkyl, or C.sub.5-8 heteroaryl, which is unsubstituted or substituted with halogen, C.sub.1-6 haloalkoxy, or C.sub.1-6 alkyl.
12. The compound of claim 1, wherein R.sup.1 is C.sub.6-8 aryl, C.sub.3-8 cycloalkyl, or C.sub.5-8 heteroaryl, which is unsubstituted or substituted with chlorine, fluorine, trifluoromethoxy, or methyl.
13. The compound of claim 1, wherein R.sup.2 to R.sup.4 are each independently hydrogen, amino (—NH.sub.2), substituted amino (—NHR′ or —NR′R″), oxo, nitro, halogen, C.sub.3-8 cycloalkyl, C.sub.1-6 alkyl, or C.sub.1-6 alkoxy.
14. The compound of claim 1, wherein R.sup.2 to R.sup.4 are each independently hydrogen, amino (—NH.sub.2), substituted amino (—NHR′ or —NR′R″), oxo, nitro, halogen, cyclopropyl, methyl, methoxy, ethoxy, or isopropoxy.
15. The compound of claim 1, wherein R′ and R″ are each independently C.sub.1-6 alkyl; or R′ and R″ are connected to each other to form C.sub.3-10 heterocycloalkyl comprising a nitrogen atom to which R′ and R″ are bonded.
16. The compound of claim 1, wherein R′ and R″ are each independently methyl, tertiary butyl, ##STR00107##
17. The compound of claim 15, wherein C.sub.3-10 heterocycloalkyl comprising a nitrogen atom to which R′ and R″ are bonded, formed by R′ and R″ being connected to each other, is morpholinyl, azetidinyl, pyrrolidinyl, piperidinyl, or azepanyl, which is unsubstituted or substituted with one or more halogens.
18. The compound of claim 1, wherein R.sup.2 and R.sup.3 are positioned on adjacent carbon atoms and connected to each other to form C.sub.6-8 aryl, C.sub.3-10 cycloalkyl, or C.sub.3-10 heterocycloalkyl.
19. The compound of claim 18, wherein C.sub.3-10 cycloalkyl formed by R.sup.2 and R.sup.3 being positioned on adjacent carbon atoms and connected to each other is cyclohexyl.
20. The compound of claim 18, wherein C.sub.3-10 heterocycloalkyl formed by R.sup.2 and R.sup.3 being positioned on adjacent carbon atoms and connected to each other is piperidinyl or morpholinyl.
21. The compound of claim 18, wherein C.sub.6-8 aryl formed by R.sup.2 and R.sup.3 being positioned on adjacent carbon atoms and connected to each other is benzo.
22. The compound of claim 1, wherein the compound is selected from the group consisting of: 1) 4-amino-1-phenethylpyridinium chloride; 2) 4-nitro-1-phenethyl-1H-imidazole; 3) 4-nitro-1-phenethyl-1H-imidazole hydrochloride; 4) 3-nitro-1-phenethyl-1H-pyrazole; 5) 1-phenethyl-1H-pyrazol-3-amine; 6) 6-amino-3-phenethylpyrimidin-4(3H)-one; 7) 4-amino-2-bromo-1-phenethylpyridinium chloride; 8) 2,4-diamino-1-phenethylpyridinium bromide; 9) 1-phenethyl-1H-imidazole; 10) 2,6-diamino-3-phenethylpyrimidin-4(3H)-one; 11) 4-amino-1-(2-chlorophenethyl)pyridinium chloride; 12) 2,4-diamino-1-(2-chlorophenethyl)pyridinium bromide; 13) 3-phenethylthiazol-3-ium iodide; 14) 2-amino-3-phenethylthiazol-3-ium iodide; 15) 4-amino-2-cyclopropyl-1-phenethylpyridinium iodide; 16) 4-amino-1-phenethylquinolinium iodide; 17) 4-(dimethylamino)-1-phenethylpyridinium chloride; 18) 4-amino-2-fluoro-1-phenethylpyridinium chloride; 19) 4-amino-1-(3,4-dichlorophenethyl)pyridinium chloride; 20) 4-amino-1-benzylpyridinium chloride; 21) 4-amino-1-benzyl-2-fluoropyridinium chloride; 22) 1-phenethyl-5,6,7,8-tetrahydroquinolinium chloride; 23) 4-amino-1-(3-phenylpropyl)pyridinium chloride; 24) 4-amino-2-fluoro-1-(3-phenylpropyl)pyridinium chloride; 25) 4-amino-1-(2-oxo-2-(4-(trifluoromethoxy)phenyl)ethyl)pyridinium bromide; 26) 4-amino-1-(2-oxo-2-phenylethyl)pyridinium bromide 27) 4-amino-1-(2-cyclohexylethyl)pyridinium bromide; 28) 4-amino-1-(2-cyclohexylethyl)-2-fluoropyridinium bromide; 29) 2,4-diamino-1-benzylpyridinium chloride; 30) 4-amino-1-benzyl-2-chloropyridinium chloride; 31) 4-amino-1-(cyclopropylmethyl)pyridinium chloride; 32) 4-amino-2-chloro-1-phenethylpyridinium chloride; 33) 4-(methylamino)-1-phenethylpyridinium chloride; 34) 1-benzyl-4-(methylamino)pyridinium chloride; 35) 4-amino-1-(3,4-dichlorobenzyl)-2-fluoropyridinium chloride; 36) 4-amino-1-(3,4-dichlorobenzyl)pyridinium chloride; 37) 1-(3,4-dichlorobenzyl)-4-(methylamino)pyridinium chloride; 38) 1-(3,4-dichlorobenzyl)-4-(dimethylamino)pyridinium chloride; 39) 4-amino-1-(cyclopropylmethyl)-2-fluoropyridinium chloride; 40) 2,4-diamino-1-(2-cyclohexylethyl)pyridinium bromide; 41) 1-(cyclopropylmethyl)-4-(methylamino)pyridinium chloride; 42) 1-(cyclopropylmethyl)-4-(dimethylamino)pyridinium chloride; 43) 4-amino-3-methyl-1-benzylpyridinium chloride; 44) 4-amino-3-methyl-1-phenethylpyridinium chloride; 45) 4-amino-1-benzyl-2-methoxypyridinium chloride; 46) 4-amino-1-(cyclohexylmethyl)pyridinium bromide; 47) 4-amino-1-(cyclobutylmethyl)pyridinium bromide; 48) 4-amino-1-(cyclobutylmethyl)-2-fluoropyridinium bromide; 49) 4-amino-1-(4-fluorobenzyl)pyridinium bromide; 50) 1-benzyl-4-morpholinopyridinium chloride; 51) 4-morpholino-1-phenethylpyridinium chloride; 52) 4-morpholino-1-(cyclopropylmethyl)pyridinium chloride; 53) 1-(2-cyclohexylethyl)-4-morpholinopyridinium bromide; 54) 1-benzyl-4-(pyrrolidin-1-yl)pyridinium chloride; 55) 1-phenethyl-4-(pyrrolidin-1-yl)pyridinium chloride; 56) 1-(cyclopropylmethyl)-4-(pyrrolidin-1-yl)pyridinium chloride; 57) 1-(cyclohexylmethyl)-4-(pyrrolidin-1-yl)pyridinium bromide; 58) 1-(cyclobutylmethyl)-4-(pyrrolidin-1-yl)pyridinium chloride; 59) 1-benzyl-4-(piperidin-1-yl)pyridinium chloride; 60) 4-(azepan-1-yl)-1-benzylpyridinium chloride; 61) 1-benzyl-4-(neopentylamino)pyridinium chloride; 62) 4-(pyrrolidin-1-yl)-1-(thiophen-3-ylmethyl)pyridinium bromide; 63) 6-(cyclopropylmethyl)-1,2,3,4-tetrahydro-1,6-naphthyridin-6-ium chloride; 64) 6-(cyclopropylmethyl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-6-ium chloride; 65) 1-benzyl-4-(4,4-difluoropiperidin-1-yl)pyridinium chloride; 66) 4-(azetidin-1-yl)-1-benzylpyridinium chloride; 67) 1-benzyl-4-(oxetan-3-ylamino)pyridinium chloride; 68) 4-(pyrrolidin-1-yl)-1-(thiophen-2-ylmethyl)pyridinium chloride; 69) 1-benzyl-4-(tert-butylamino)pyridinium chloride; 70) 4-(azetidin-1-yl)-1-(cyclopropylmethyl)pyridinium chloride; 71) 4-(azetidin-1-yl)-1-(thiophen-3-ylmethyl)pyridinium bromide; 72) 4-(pyrrolidin-1-yl)-1-(selenophen-2-ylmethyl)pyridinium chloride; 73) 4-amino-1-(cyclopropylmethyl)pyrimidin-1-ium chloride; 74) 4-amino-1-(selenophen-2-ylmethyl)pyrimidin-1-ium chloride; 75) 4-amino-1-(selenophen-2-ylmethyl)pyridazin-1-ium chloride; 76) 4-amino-1-(selenophen-2-ylmethyl)pyridinium chloride; 77) 4-amino-1-(thiophen-2-ylmethyl)pyridinium chloride; 78) 1-(furan-2-ylmethyl)-4-(pyrrolidin-1-yl)pyridinium chloride; 79) 1-((5-methylthiophen-2-yl)methyl)-4-(pyrrolidin-1-yl)pyridinium chloride; 80) 4-(azetidin-1-yl)-1-(selenophen-3-ylmethyl)pyridinium chloride; 81) 2-amino-4-(azetidin-1-yl)-1-(cyclopropylmethyl)pyridinium chloride; 82) 2-amino-4-(azetidin-1-yl)-1-(cyclopropylmethyl)pyridinium chloride; 83) 2,4-diamino-1-(cyclopropylmethyl)pyridinium chloride; 84) 2,4-diamino-1-(4-chlorobenzyl)pyridinium chloride; 85) 2-amino-4-(azetidin-1-yl)-1-((5-methylthiophen-2-yl)methyl)pyridinium chloride; 86) 2-amino-4-(azetidin-1-yl)-1-(selenophen-2-ylmethyl)pyridinium chloride; 87) 2-amino-4-(azetidin-1-yl)-1-benzylpyridinium chloride; 88) 2-amino-1-benzyl-4-(pyrrolidin-1-yl)pyridinium chloride; 89) 2-amino-1-(cyclopropylmethyl)-4-(pyrrolidin-1-yl)pyridinium chloride; 90) 2-amino-1-((5-methylthiophen-2-yl)methyl)-4-(pyrrolidin-1-yl)pyridinium chloride; 91) 2-amino-4-(pyrrolidin-1-yl)-1-(selenophen-2-ylmethyl)pyridinium chloride; 92) 2-amino-1-(4-chlorobenzyl)-4-(pyrrolidin-1-yl)pyridinium chloride; 93) 4-amino-1-benzyl-2-ethoxypyridinium chloride; 94) 4-amino-1-benzyl-2-isopropoxypyridinium chloride; 95) 4-amino-1-benzyl-2-cyclopropylpyridinium chloride; 96) 4-(azetidin-1-yl)-1-benzyl-2-ethoxypyridinium chloride; 97) 4-(azetidin-1-yl)-1-benzyl-2-isopropoxypyridinium chloride; 98) 4-(azetidin-1-yl)-1-benzyl-2-cyclopropylpyridinium chloride; 99) 1-benzyl-2-ethoxy-4-(pyrrolidin-1-yl)pyridinium chloride; 100) 1-benzyl-2-isopropoxy-4-(pyrrolidin-1-yl)pyridinium chloride; and 101) 1-benzyl-2-cyclopropyl-4-(pyrrolidin-1-yl)pyridinium chloride.
23. A method for preparing the compound of claim 1 or a pharmaceutically acceptable salt thereof, comprising reacting a compound represented by Formula 2 below and a compound represented by Formula 3 below: ##STR00108## wherein, in Formulas 2 and 3, Z is halogen, and , X, Y, n, L, R.sup.1, R.sup.2, R.sup.3, and R.sup.4 are the same as defined in claim 1.
24. A pharmaceutical composition, comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof.
25. The composition of claim 24, wherein the composition is for anti-cancer use.
26. The composition of claim 24, wherein the composition further comprises an anti-cancer agent.
27. The composition of claim 24, wherein the cancer is one or more selected from the group consisting of uterine cancer, breast cancer, gastric cancer, brain cancer, rectal cancer, colorectal cancer, lung cancer, skin cancer, blood cancer, pancreatic cancer, renal cancer, bladder cancer, prostate cancer, and liver cancer.
28. A method for treating or preventing cancer, comprising administering a therapeutically effective amount of the pharmaceutical composition of claim 24 to a subject in need thereof.
29. The method of claim 28, wherein the method is to administer the pharmaceutical composition alone or in combination with an anti-cancer agent.
30. The method of claim 28, wherein the cancer is one or more selected from the group consisting of uterine cancer, breast cancer, gastric cancer, brain cancer, rectal cancer, colorectal cancer, lung cancer, skin cancer, blood cancer, pancreatic cancer, renal cancer, bladder cancer, prostate cancer, and liver cancer.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0060] FIG. 1 is the result of observing the volume of a tumor according to Test Example 3.
DETAILED DESCRIPTION OF THE INVENTION
[0061] Hereinafter, the present invention will be described through Examples and Comparative Examples in more detail. However, the Examples disclosed herein are only for illustrative purposes, and should not be construed as limiting the scope of the present invention.
Example 1: 4-Amino-1-phenethylpyridinium chloride
[0062] ##STR00005##
[0063] 4-Aminopyridine (0.2 g, 2.12 mmol) was dissolved in DMF (5 mL) at room temperature. 2-Chloroethylbenzene (1.392 mL, 10.6 mmol) was added thereto, and the mixture was stirred for 5 hours at 90° C. After the reaction was completed, the mixture was cooled to room temperature, diethyl ether was added, and the mixture was stirred at room temperature for 30 minutes. The solid resultant was filtered. The obtained solid was dissolved in a small amount of methanol, ethyl acetate was added, and the mixture was stirred at room temperature for 1 hour. The formed solid was filtered and dried under reduced pressure to obtain a desired compound (86 mg, 17.2%).
[0064] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.29 (s, 2H), 8.10 (d. J=7.6 Hz, 2H), 7.31 (t, J=8.4 Hz, 2H), 7.249 (d, J=7.2 Hz, 1H), 7.20 (d, J=7.2 Hz, 2H), 6.81 (d, J=7.2 Hz, 2H), 4.37 (t J=6.8 Hz, 2H), 3.09 (t, J=6.8 Hz, 2H).
[0065] LCMS: 199.1 [M].
Example 2: 4-Nitro-1-phenethyl-H-imidazole
[0066] ##STR00006##
[0067] In the same manner as in Example 1, except that 4-nitro-1H-imidazole was used instead of 4-aminopyridine, 50 mg (11.1%) of a desired compound, which is a white solid, was obtained.
[0068] .sup.1H NMR (400 MHz. DMSO-D6) δ 8.39 (d, J=1.6 Hz, 1H), 7.74 (d, J=1.2 Hz, 1H) 7.23 (m, 5H), 4.33 (t, J=7.2 Hz, 2H), 3.11 (t, J=7.6 Hz; 2H).
[0069] LCMS: 218.0 [M+H].sup.+.
Example 3: 4-Nitro-1-phenethyl-1H-imidazole hydrochloride
[0070] ##STR00007##
[0071] After dissolving the compound of Example 2 in methanol, 1 equivalent amount of 4 M HCl was added, and the mixture was stirred at room temperature for 1 hour. After concentration under reduced pressure, 50 mg of a desired compound, which is a white solid, was obtained.
[0072] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.39 (d, J=1.6 Hz, 1H), 7.74 (d, J=1.2 Hz, 1H) 7.23 (m, 5H), 4.33 (t, 0.1=7.2 Hz, 2H), 3.11 (t, J=7.6 Hz, 2H).
[0073] LCMS: 218.0 [M+H].sup.+.
Example 4: 3-Nitro-1-phenethyl-1H-pyrazole
[0074] ##STR00008##
[0075] In the same manner as in Example 1, except that 3-nitro-1H-pyrazole was used instead of 4-aminopyridine, 0.1 g (12%) of a desired compound, which is a white solid, was obtained.
[0076] .sup.1H NMR (400 MHz, DMSO-D6) δ 7.91 (d, J=2.4 Hz, 1H), 7.26 (m, 2H), 7.19 (m, 3H), 6.99 (d, J=2.0 Hz, 1H), 4.49 (t, 0.1=7.2 Hz, 2H), 3.15 (t, t, J=7.2 Hz, 2H).
[0077] LCMS: 218.0 [M+H].sup.+.
Example 5: 1-Phenethyl-1H-pyrazol-3-amine
[0078] ##STR00009##
[0079] After dissolving the compound of Example 4 in methanol, Pd/C was added, and H.sub.2 gas was added in the reactor. After stirring for 1 hour at room temperature, the mixture was filtered to remove Pd/C. The filtrate was concentrated under reduced pressure and dried under reduced pressure to obtain 0.1 g of a desired compound (12%).
[0080] .sup.1H NMR (400 MHz, DMSO-D6) δ 7.28 (m, 2H), 7.15 (m, 4H), 5.30 (s, 1H), 4.52 (s, 2H), 4.03 (t, J=7.6 Hz, 2H), 2.99 (t, J=7.2 Hz, 2H).
[0081] LCMS: 188.2 [M+H].sup.+.
Example 6: 6-Amino-3-phenethylpyrimidin-4(3H)-one
[0082] ##STR00010##
[0083] In the same manner as in Example 1, except that 6-aminopyrimidin-4(3H)-one was used instead of 4-aminopyridine, 0.2 g (34%) of a desired compound, which is a white solid, was obtained.
[0084] .sup.1H NMR (400 MHz, CD.sub.3OD) δ 6.75 (m, 1H), 6.46 (m, 5H) 4.54 (s, 1H), 3.26 (t, J=7.2 Hz, 2H), 2.15 (t, J=7.2 Hz, 2H).
[0085] LCMS: 216.1 [M+H].sup.+.
Example 7: 4-Amino-2-bromo-1-phenethylpyridinium chloride
[0086] ##STR00011##
[0087] In the same manner as in Example 1, except that 2-bromopyridin-4-amine was used instead of 4-aminopyridine, 0.12 g (18.3%) of a desired compound, which is a white solid, was obtained.
[0088] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.49 (s, 2H), 8.16 (m, 1H), 7.32 (m, 2H), 7.3 (m, 1H), 7.22 (m, 2H), 7.03 (m, 1H), 6.79 (m, 1H), 4.50 (q, J=6.4 Hz, 2H), 3.09 (t, J=6.4 Hz, 2H).
[0089] LCMS: 278.9 [M].
Example 8: 2,4-Diamino-1-phenethylpyridinium bromide
[0090] ##STR00012##
[0091] The compound of Example 7 was added to a sealed tube, and 30% of NH.sub.4OH solution was added thereto. After stirring for 12 hours at 80° C., the mixture was cooled to room temperature. After concentration under reduced pressure, the mixture was dissolved in a small amount of methanol, and ethyl acetate was added to obtain a solid. The formed solid was filtered and dried under reduced pressure to obtain 39 mg (83%) of a desired compound, which is a white solid.
[0092] .sup.1H NMR (400 MHz, DMSO-D6) δ 7.41 (s, 2H), 7.32 (m, 4H), 7.23 (m, 3H), 7.17 (m, 1H), 6.04 (d, J=7.6 Hz, 1H), 5.86 (s, 1H), 4.19 (t, J=6.8 Hz, 2H), 2.94 (t, J=6.8 Hz, 2H).
[0093] LCMS: 214.1 [M].
Example 9: 1-Phenethyl-1H-imidazole
[0094] ##STR00013##
[0095] In the same manner as in Example 1, except that 1H-imidazole was used instead of 4-aminopyridine, 0.35 g (27.7%) of a desired compound, which is a white solid, was obtained.
[0096] .sup.1H NMR (400 MHz, DMSO-D6) δ 7.74 (s, 1H), 7.21 (d, J=7.6 Hz, 1H), 7.01 (d, J=7.6 Hz, 1H), 7.21 (m, 5H), 4.33 (t, J=7.2 Hz, 2H), 3.11 (t, J=7.6 Hz, 2H).
[0097] LCMS: 173.2 [M].
Example 10: 2,6-Diamino-3-phenethylpyrimidin-4(3H)-one
[0098] ##STR00014##
[0099] In the same manner as in Example 1, except that 2,6-diaminopyrimidin-4(3H)-one was used instead of 4-aminopyridine, 0.23 g (25.2%) of a desired compound, which is a white solid, was obtained.
[0100] .sup.1H NMR (400 MHz, CD.sub.3OD) δ 7.25 (m, 5H), 5.2 (s, 1H), 4.30 (t, J=7.2 Hz, 2H), 3.00 (t, J=7.2 Hz, 2H).
[0101] LCMS: 231.0 [M+H].sup.+.
Example 11: 4-Amino-1-(2-chlorophenethyl)pyridinium chloride
[0102] ##STR00015##
[0103] In the same manner as in Example 1, except that 1-chloro-2-(2-chloroethyl) benzene was used instead of (2-chloroethyl) benzene, 20 mg (5.85%) of a desired compound, which is a white solid, was obtained.
[0104] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.07 (s, 2H), 8.02 (d, J=6.8 Hz, 1H), 7.44 (m, 1H), 7.30 (m, 3H), 6.75 (d, J=7.2 Hz, 2H), 4.40 (t, J=6.4 Hz, 2H), 3.21 (t, J=6.4 Hz, 2H).
[0105] LCMS: 233.1, 235.1 [M].
Example 12: 2,4-Diamino-1-(2-chlorophenethyl)pyridinium bromide
[0106] ##STR00016##
[0107] In the same manner as in Examples 7 and 8, except that 1-chloro-2-(2-chloroethyl) benzene was used instead of (2-chloroethyl) benzene, 21 mg (79%) of a desired compound, which is a white solid, was obtained.
[0108] .sup.1H NMR (400 MHz, DMSO-D6) δ 7.48 (s, 2H), 7.43 (m, 1H), 7.34 (m, 2H), 7.29 (m, 2H), 7.11 (d, J=7.6 Hz, 2H), 6.02 (d, J=7.6 Hz, 1H), 5.88 (s, 1H), 4.23 (t, J=6.4 Hz, 2H), 3.01 (t, J=6.4 Hz, 2H).
[0109] LCMS: 248.1, 250.1 [M].
Example 13: 3-Phenethylthiazol-3-ium iodide
[0110] ##STR00017##
[0111] In the same manner as in Example 1, except that (2-iodoethyl) benzene was used instead of 4-aminopyridine, 40 mg (63.1%) of a desired compound, which is a white solid, was obtained.
[0112] .sup.1H NMR (400 MHz, DMSO-D6) δ 10.03 (s, 1H), 8.57 (m, 1H), 8.23 (m, 1H), 7.23 (m, 5H), 4.82 (t, 0.1=7.2 Hz, 2H), 3.23 (t, J=7.2 Hz, 2H).
[0113] LCMS: 190.1 [M].
Example 14: 2-Amino-3-phenethylthiazol-3-ium iodide
[0114] ##STR00018##
[0115] In the same manner as in Example 13, except that 2-amino thiazol was used instead of thiazol, 38 mg (58%) of a desired compound, which is a white solid, was obtained.
[0116] .sup.1H NMR (400 MHz, DMSO-D6) δ 9.38 (s, 2H), 7.33 (m, 6H), 6.95 (s, 1H), 4.26 (t, J=7.6 Hz, 2H), 2.99 (t, J=7.2 Hz; 2H).
[0117] LCMS: 206.1 [M+H].sup.+.
Example 15: 4-Amino-2-cycloropyl-1-phenethylpyridinium iodide
[0118] ##STR00019##
[0119] In the same manner as in Example 1, except that 2-cyclopropylpyridine-4-amine was used instead of 4-aminopyridine, 50 mg (12.3%) of a desired compound, which is a white solid, was obtained.
[0120] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.04 (d, J=6.8 Hz, 1H), 7.91 (s, 1H), 7.77 (s, 1H), 7.27 (m, 5H), 6.63 (m, 1H), 6.51 (m, 1H), 4.53 (t, J=7.6 Hz, 2H), 3.10 (t, J=7.2 Hz, 2H), 2.14 (m, 1H), 1.17 (m, 2H), 0.81 (m, 2H).
[0121] LCMS: 239.0 [M].
Example 16: 4-Amino-1-phenethylquinolinium iodide
[0122] ##STR00020##
[0123] In the same manner as in Example 1, except that quinolin-4-amine was used instead of 4-aminopyridine, 60 mg (26.1%) of a desired compound, which is a white solid, was obtained.
[0124] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.9 (s, 1H), 8.47 (m, 1H), 8.25 (t, J=8.4 Hz, 2H), 8.05 (m, 1H) 7.70 (t, J=8.4 Hz, 1H), 7.26 (m, 3H), 7.22 (m, 2H), 6.67 (d, J=7.2 Hz, 1H), 4.80 (t, J=7.6 Hz, 2H), 3.13 (t, J=7.6 Hz, 2H).
[0125] LCMS: 249.0 [M].
Example 17: 4-(Dimethylamino)-1-phenethylpyridinium chloride
[0126] ##STR00021##
[0127] In the same manner as in Example 1, except that N,N-dimethylpyridin-4-amine was used instead of 4-aminopyridine, 50 mg (11.62%) of a desired compound, which is a white solid, was obtained.
[0128] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.26 (d, J=8.0 Hz, 2H), 7.32 (m, 2H), 7.23 (m, 3H), 7.00 (d, J=8.0 Hz, 2H), 4.44 (t, J=7.2 Hz, 2H), 3.13 (s, 6H), 3.11 (t. J=7.6 Hz, 2H).
[0129] LCMS: 227.1 [M].
Example 18: 4-Amino-2-fluor-phenethylpyridinium chloride
[0130] ##STR00022##
[0131] In the same manner as in Example 1, except that 2-fluororpyridin-4-amine was used instead of 4-aminopyridine, 0.1 g (22.18%) of a desired compound, which is a white solid, was obtained.
[0132] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.51 (m, 2H), 7.97 (t, J=6.4 Hz, 1H), 7.30 (m, 3H), 7.19 (m, 2H), 6.71 (d, J=7.6 Hz, 2H), 6.61 (d, J=7.6 Hz, 2H), 4.39 (t, J=6.8 Hz, 2H), 3.07 (t, J=6.8 Hz, 2H).
[0133] LCMS: 217.1 [M].
Example 19: 4-Amino-1-(3,4-dichlorophenethyl)pyridinium chloride
[0134] ##STR00023##
[0135] In the same manner as in Example 1, except that 1,2-dichloro-4-(2-chloroethyl) benzene was used instead of (2-chloroethyl) benzene, 0.1 g (15.5%) of a desired compound, which is a white solid, was obtained.
[0136] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.11 (s, 2H), 8.08 (d. J=7.8 Hz, 2H), 7.57 (d, J=7.8 Hz, 1H), 7.54 (s, 1H), 7.18 (d, J=7.8 Hz, 1H), 6.78 (d, J=7.8 Hz, 2H), 4.37 (t, J=7.8 Hz, 2H), 3.10 (t, J=7.8 Hz, 2H).
[0137] LCMS: 267.0, 269.0 [M].
Example 20: 4-Amino-1-benzylpyridinium chloride
[0138] ##STR00024##
[0139] In the same manner as in Example 1, except that benzyl chloride was used instead of (2-chloroethyl) benzene, 0.3 g (42.6%) of a desired compound, which is a white solid, was obtained.
[0140] .sup.1H NMR (400 MHz, DMSO-D6) S 8.41 (s, 2H), 8.31 (d, J=7.2 Hz, 2H), 7.39 (m, 5H), 6.89 (d, J=7.2 Hz, 2H), 5.37 (s, 2H).
[0141] LCMS: 185.1 [M].
Example 21: 4-Amino-1-benzyl-2-fluoropyridinium chloride
[0142] ##STR00025##
[0143] In the same manner as in Example 1, except that benzyl chloride was used instead of (2-chloroethyl) benzene and 2-fluoropyridine-4-amine was used instead of 4-amino pyridine, 0.2 g (47%) of a desired compound, which is a white solid, was obtained.
[0144] .sup.1H NMR (400 MHz, CD.sub.3OD) δ 8.09 (m, 1H), 7.43 (m, 3H), 7.40 (m, 2H), 6.80 (m, 1H), 6.63 (m, 1H), 5.37 (s, 2H).
[0145] LCMS: 203.1 [M].
Example 22: 1-Phenethyl-5,6,7,8-tetrahydroquinolinium chloride
[0146] ##STR00026##
[0147] In the same manner as in Example 1, except that 5,6,7,8-tetrahydroquinolin-4-amine was used instead of 4-aminopyridine, 0.15 g (38.5%) of a desired compound, which is a white solid, was obtained.
[0148] .sup.1H NMR (400 MHz, CD.sub.3OD) δ 8.54 (m, 1H), 8.2 (m, 1H), 7.71 (m, 1H), 7.72 (m, 3H), 7.13 (m, 2H), 4.83 (m, 2H), 3.31 (m, 2H), 3.01 (m, 4H), 1.90 (m, 2H), 1.79 (m, 2H).
[0149] LCMS: 238.1 [M].
Example 23: 4-Amino-1-(3-phenylpropyl)pyridinium chloride
[0150] ##STR00027##
[0151] In the same manner as in Example 1, except that (3-chloropropyl) benzene was used instead of (2-chloroethyl) benzene, 0.1 g (38%) of a desired compound, which is a white solid, was obtained.
[0152] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.26 (s, 2H), 8.20 (d, J=7.2 Hz, 2H), 7.28 (m, 2H), 7.21 (m, 3H), 6.86 (d, J=7.2 Hz, 2H), 4.15 (t, J=7.2 Hz, 2H), 2.57 (m, 2H), 2.08 (m, 2H).
[0153] LCMS: 213.0 [M].
Example 24: 4-Amino-2-fluoro-1-(3-phenylpropyl)pyridinium chloride
[0154] ##STR00028##
[0155] In the same manner as in Example 1, except that (3-chloropropyl) benzene was used instead of (2-chloroethyl) benzene and 2-fluoropyridine-4-amine was used instead of 4-aminopyridine, 55 mg (15.41%) of a desired compound, which is a white solid, was obtained.
[0156] .sup.1H NMR (400 MHz, CD.sub.3OD) δ 7.94 (m, 1H), 7.21 (m, 5H), 6.72 (m, 1H), 6.54 (m, 1H), 4.21 (m, 2H), 2.73 (m, 2H), 2.16 (m, 2H).
[0157] LCMS: 231.1 [M].
Example 25: 4-Amino-1-(2-oxo-2-(4-(trifluoromethoxy)phenyl)ethyl)pyridinium bromide
[0158] ##STR00029##
[0159] In the same manner as in Example 1, except that 2-bromo-1-(4-(trifluoromethoxy)phenyl)ethanone was used instead of (2-chloroethyl) benzene, 45 mg (53%) of a desired compound, which is a white solid, was obtained.
[0160] .sup.1H NMR (400 MHz, DMSO-D6) S 8.27 (s, 2H), 8.16 (m, 2H), 8.08 (d, J=7.2 Hz, 2H), 7.64 (m, 2H), 6.92 (d, J=7.6 Hz, 2H), 5.97 (s, 2H).
[0161] LCMS: 297.0 [M].
Example 26: 4-Amino-1-(2-oxo-2-phenylethyl)pyridinium bromide
[0162] ##STR00030##
[0163] In the same manner as in Example 1, except that 2-bromo-1-(phenyl)ethanone was used instead of (2-chloroethyl) benzene, 45 mg (53%) of a desired compound, which is a white solid, was obtained.
[0164] .sup.1H NMR (400 MHz, DMSO-D6) δ 7.99 (m, 4H), 7.66 (m, 1H), 7.53 (m 2H), 6.81 (m, 2H), 4.34 (s, 2H).
[0165] LCMS: 213.1 [M].
Example 27: 4-Amino-1-(2-cyclohexylethyl)pyridinium bromide
[0166] ##STR00031##
[0167] In the same manner as in Example 1, except that (2-bromoethyl)cyclohexane was used instead of (2-chloroethyl) benzene, 0.15 g (24.75%) of a desired compound, which is a white solid, was obtained.
[0168] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.10 (d. J=6.4 Hz, 2H), 6.84 (d, J=6.4 Hz, 2H), 4.17 (m, 2H), 1.77 (m, 7H), 1.24 (m, 4H), 1.00 (m, 2H).
[0169] LCMS: 205.2 [M].
Example 28: 4-Amino-1-(2-cyclohexylethyl)-2-fluoropyridinium bromide
[0170] ##STR00032##
[0171] In the same manner as in Example 1, except that (2-bromoethyl)cyclohexane was used instead of (2-chloroethyl) benzene and 2-fluoropyridine-4-amine was used instead of 4-aminopyridine, 0.35 g (64.7%) of a desired compound, which is a white solid, was obtained.
[0172] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.00 (m, 1H), 6.76 (m, 1H), 6.60 (m, 1H), 4.20 (m, 2H), 1.76 (m, 7H), 1.22 (m, 4H), 0.98 (m, 2H).
[0173] LCMS: 223.1 [M].
Example 29: 2,4-Diamino-1-benzylpyridinium chloride
[0174] ##STR00033##
[0175] In the same manner as in Example 1, except that benzyl chloride was used instead of (2-chloroethyl) benzene and pyridine-2,4-diamine was used instead of 4-aminopyridine, 0.2 g (46.3%) of a desired compound, which is a white solid, was obtained.
[0176] .sup.1H NMR (400 MHz, DMSO-D6) δ 7.72 (d, J=7.2 Hz, 1H), 7.45 (s, 2H), 7.34 (m, 5H), 7.20 (m, 2H), 6.24 (m, 1H), 5.91 (m, 1H), 5.24 (s, 2H).
[0177] LCMS: 200.1 [M].
Example 30: 4-Amino-1-benzyl-2-chloropyridinium chloride
[0178] ##STR00034##
[0179] In the same manner as in Example 1, except that benzyl chloride was used instead of (2-chloroethyl) benzene and 2-chloropyridin-4-amine was used instead of 4-aminopyridine, 0.15 g (37.8%) of a desired compound, which is a white solid, was obtained.
[0180] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.34 (d. J=13.6 Hz, 2H), 8.50 (d, J=7.2 Hz, 1H), 7.36 (m, 4H), 7.25 (m, 2H), 7.14 (m, 1H), 6.97 (m, 1H).
[0181] LCMS: 219.1 [M].
Example 31: 4-Amino-1-(cyclopropylmethyl)pyridinium chloride
[0182] ##STR00035##
[0183] In the same manner as in Example 1, except that (chloromethyl)cyclopropane was used instead of (2-chloroethyl) benzene, 0.18 g (30.6%) of a desired compound, which is a white solid, was obtained.
[0184] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.22 (d, J=7.2 Hz, 2H), 8.19 (s, 2H), 6.86 (d, J=7.2 Hz, 2H), 4.00 (d. J=7.6 Hz, 2H), 1.24 (m, 1H), 0.59 (m, 2H), 0.43 (m, 2H).
[0185] LCMS: 149.2 [M].
Example 32: 4-Amino-2-chloro-1-phenethylpyridinium chloride
[0186] ##STR00036##
[0187] In the same manner as in Example 30, except that (2-chloroethyl) benzene was used instead of benzyl chloride, 0.08 g (25.5%) of a desired compound, which is a white solid, was obtained.
[0188] .sup.1H NMR (400 MHz, CD.sub.3OD) δ 7.89 (d, J=7.6 Hz, 1H), 7.42 (m, 3H), 7.24 (m, 2H), 7.00 (m, 1H), 6.70 (m, 1H), 4.57 (t, J=7.2 Hz, 2H), 3.16 (t. J=7.6 Hz, 2H).
[0189] LCMS: 233.1, 235.1 [M, M+2].sup.+.
Example 33: 4-(Methylamino)-1-phenethylpyridinium chloride
[0190] ##STR00037##
[0191] In the same manner as in Example 1, except that N-methylpyridin-4-amine was used instead of 4-aminopyridine, 0.12 g (26.1%) of a desired compound, which is a white solid, was obtained.
[0192] .sup.1H NMR (400 MHz, DMSO-D6) δ 9.04 (m, 1H), 8.23 (m, 1H), 8.05 (m, 1H), 7.32 (m, 2H), 7.23 (m, 3H), 6.91 (m, 1H), 6.80 (m, 1H), 4.31 (t, J=7.2 Hz, 2H), 3.09 (t. J=7.2 Hz, 2H), 2.86 (d, J=4.8 Hz, 3H).
[0193] LCMS: 213.1 [M].
Example 34: 1-Benzyl-4-(methylamino)pyridinium chloride
[0194] ##STR00038##
[0195] In the same manner as in Example 1, except that N-methylpyridin-4-amine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.2 g (46.1%) of a desired compound, which is a white solid, was obtained.
[0196] .sup.1H NMR (400 MHz, DMSO-D6) δ 9.19 (m, 1H), 8.49 (d, J=7.2 Hz, 1H), 8.27 (d, J=7.2 Hz, 1H), 7.38 (m, 5H), 6.99 (m, 1H), 6.88 (m, 1H), 5.39 (s, 2H), 2.88 (d, J=6.0 Hz, 3H).
[0197] LCMS: 199.1 [M].
Example 35: 4-Amino-1-(3,4-dichlorobenzyl)-2-fluoropyridinium chloride
[0198] ##STR00039##
[0199] In the same manner as in Example 1, except that 3,4-dichlorobenzyl chloride was used instead of (2-chloroethyl) benzene and 4-amino-2-fluoropyridine was used instead of 4-aminopyridine, 50 mg (12.9%) of a desired compound, which is a white solid, was obtained.
[0200] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.79 (m, 2H), 8.26 (t, J=6.4 Hz, 1H), 7.71 (m, 2H), 7.34 (m, 1H), 6.86 (m, 1H), 6.73 (m, 1H), 5.40 (s, 2H).
[0201] LCMS: 271.0, 273.0 [M. M+2].sup.+.
Example 36: 4-Amino-1-(3,4-dichlorobenzyl)pyridinium chloride
[0202] ##STR00040##
[0203] In the same manner as in Example 1, except that 3,4-dichlorobenzyl chloride was used instead of (2-chloroethyl) benzene, 0.11 g (23.8%) of a desired compound, which is a white solid, was obtained.
[0204] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.50 (s, 2H), 8.36 (d, J=6.8 Hz, 2H), 7.80 (m, 1H), 7.71 (d, J=8.0 Hz, 1H), 7.42 (m, 1H), 6.93 (d, J=7.6 Hz, 2H), 5.41 (s, 2H).
[0205] LCMS: 253.0, 255.0 [M, M+2].sup.+.
Example 37: 1-(3,4-Dichlorobenzyl)-4-(methylamino)pyridinium chloride
[0206] ##STR00041##
[0207] In the same manner as in Example 1, except that N-methylpyridin-4-amine was used instead of 4-aminopyridine and 3,4-dichlorobenzyl chloride was used instead of (2-chloroethyl) benzene, 0.15 g (26.7%) of a desired compound, which is a white solid, was obtained.
[0208] .sup.1H NMR (400 MHz, DMSO-D6) δ 9.22 (m, 1H), 8.50 (m, 1H), 8.28 (m, 1H), 7.80 (m, 1H), 7.1 (d, J=8.0 Hz, 1H), 7.43 (m, 1H), 7.00 (m, 1H), 6.89 (m, 1H), 5.40 (s, 2H), 2.51 (d, J=5.2 Hz, 3H).
[0209] LCMS: 267.0, 269.0 [M, M+2].
Example 38: 1-(3,4-Dichlorobenzyl)-4-(dimethylamino)pyridinium chloride
[0210] ##STR00042##
[0211] In the same manner as in Example 1, except that 3,4-dimethylpyridin-4-amine was used instead of 4-aminopyridine and 3,4-dichlorobenzyl chloride was used instead of (2-chloroethyl) benzene, 0.12 g (23.08%) of a desired compound, which is a white solid, was obtained.
[0212] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.51 (d, J=8.0 Hz, 2H), 7.83 (m, 1H), 7.71 (d, J=8.4 Hz, 1H), 7.45 (m, 1H), 7.08 (d, J=7.6 Hz, 2H), 5.45 (s, 2H), 3.15 (s, 6H).
[0213] LCMS: 281.0, 283.0 [M, M+2].sup.+.
Example 39: 4-Amino-1-(cyclopropylmethyl)-2-fluoropyridinium chloride
[0214] ##STR00043##
[0215] In the same manner as in Example 1, except that 4-amino-2-fluoropyridine was used instead of 4-aminopyridine and (2-chloromethyl)cyclopropane was used instead of (2-chloroethyl) benzene, 0.07 g (25.8%) of a desired compound, which is a white solid, was obtained.
[0216] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.82 (s, 2H), 8.22 (m, 1H), 6.89 (m, 1H), 6.80 (m, 1H), 4.01 (m, 2H), 1.23 (m, 1H) 0.60 (m, 2H), 0.44 (m, 2H).
[0217] LCMS: 167.1 [M].
Example 40: 2,4-Diamino-1-(2-cyclohexylethyl)pyridinium bromide
[0218] ##STR00044##
[0219] In the same manner as in Examples 7 and 8, except that (2-chloroethyl) cyclohexane was used instead of (2-chloroethyl) benzene, 30 mg (19.96%) of a desired compound, which is a white solid, was obtained.
[0220] .sup.1H NMR (400 MHz, DMSO-D6) δ 7.72 (d, J=7.2 Hz, 1H), 7.45 (s, 2H), 7.20 (m, 1H), 6.24 (m, 1H), 4.17 (m, 2H), 1.77 (m, 7H), 1.24 (m, 4H), 1.00 (m, 2H).
[0221] LCMS: 220.1 [M].
Example 41: 1-(Cyclopropylmethyl)-4-(methylamino)pyridinium chloride
[0222] ##STR00045##
[0223] In the same manner as in Example 34, except that (2-chloromethyl)cyclopropane was used instead of benzyl chloride, 0.09 g (24.4%) of a desired compound, which is a white solid, was obtained.
[0224] .sup.1H NMR (400 MHz, DMSO-D6) δ 9.22 (m, 1H), 8.42 (m, 1H), 8.22 (m, 1H), 7.03 (m, 1H), 6.83 (m, 1H) 4.08 (d, J=7.2 Hz, 2H), 2.88 (d. J=5.2 Hz, 3H), 1.28 (m, 1H), 0.56 (m, 2H), 0.46 (m, 2H).
[0225] LCMS: 163.2 [M].
Example 42: 1-(Cyclopropylmethyl)-4-(dimethylamino)pyridinium chloride
[0226] ##STR00046##
[0227] In the same manner as in Example 1, except that N,N-dimethylpyridin-4-amine was used instead of 4-aminopyridine and (2-chloromethyl) cyclopropane was used instead of (2-chloroethyl) benzene, 90 mg (24%) of a desired compound, which is a white solid, was obtained.
[0228] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.42 (d, J=7.6 Hz, 2H), 7.07 (d, J=7.6 Hz, 2H), 4.09 (d, J=7.6 Hz, 2H), 3.13 (s, 6H), 1.28 (m, 1H), 0.56 (m, 2H), 0.48 (m, 2H).
[0229] LCMS: 177.2 [M].
Example 43: 4-Amino-3-methyl-1-benzylpyridinium chloride
[0230] ##STR00047##
[0231] In the same manner as in Example 1, except that 3-methylpyridin-4-amine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.12 g (27.6%) of a desired compound, which is a white solid, was obtained.
[0232] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.62 (s, 1H), 8.37 (s, 1H), 8.29 (m, 1H), 7.71 (s, 1H), 7.38 (m, 5H), 6.95 (d, J=6.8 Hz, 1H), 5.36 (s, 2H), 2.09 (s, 3H).
[0233] LCMS: 199.1 [M].
Example 44: 4-Amino-3-methyl-1-phenethylpyridinium chloride
[0234] ##STR00048##
[0235] In the same manner as in Example 1, except that 3-methylpyridin-4-amine was used instead of 4-aminopyridine, 0.09 g (19.5%) of a desired compound, which is a white solid, was obtained.
[0236] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.61 (s, 1H), 8.22 (s, 1H), 8.08 (m, 1H), 7.31 (m, 5H), 6.89 (d, J=7.2 Hz, 1H), 4.36 (t, J=7.2 Hz, 2H), 3.10 (t, J=6.8 Hz, 2H), 2.09 (s, 3H).
[0237] LCMS: 213.1 [M].
Example 45: 4-Amino-1-benzyl-2-methoxypyridinium chloride
[0238] ##STR00049##
[0239] In the same manner as in Example 1, except that 2-methoxylpyridin-4-amine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.08 g (26.4%) of a desired compound, which is a white solid, was obtained.
[0240] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.21 (s, 1H), 8.15 (d, J=7.2 Hz, 1H), 7.38 (m, 6H), 6.62 (m, 1H), 6.36 (m, 1H), 5.25 (s, 2H), 4.01 (s, 3H).
[0241] LCMS: 215.1 [M].
Example 46: 4-Amino-1-(cyclohexylmethyl)pyridinium bromide
[0242] ##STR00050##
[0243] In the same manner as in Example 1, except that (2-chloromethyl)cyclohexane was used instead of (2-chloroethyl) benzene, 0.1 g (23.1%) of a desired compound, which is a white solid, was obtained.
[0244] .sup.1H NMR (400 MHz; DMSO-D6) δ 8.19 (s, 2H), 8.17 (d, 0.1=5.6 Hz, 2H), 6.87 (d, J=1=6.4 Hz, 2H), 4.00 (d, J=7.6 Hz, 2H), 1.7 (m, 6H), 0.99 (m, 5H).
[0245] LCMS: 191.2 [M].
Example 47: 4-Amino-1-(cyclobutylmethyl)pyridinium bromide
[0246] ##STR00051##
[0247] In the same manner as in Example 1, except that (2-chloromethyl)cyclobutane was used instead of (2-chloroethyl) benzene, 0.045 g (35.4%) of a desired compound, which is a white solid, was obtained.
[0248] .sup.1H NMR (400 MHz, DMSO-D6) 8.19 (m, 2H), 8.09 (s, 2H), 6.84 (m, 2H), 4.16 (m, 2H), 2.69 (m, 1H), 1.83 (m, 6H).
[0249] LCMS: 163.2 [M].
Example 48: 4-Amino-1-(cyclobutylmethyl)-2-fluoropyridinium bromide
[0250] ##STR00052##
[0251] In the same manner as in Example 1, except that 4-amino-2-fluoropyridine was used instead of 4-aminopyridine and (2-chloromethyl)butane was used instead of (2-chloroethyl) benzene, 0.003 g (31.7%) of a desired compound, which is a white solid, was obtained.
[0252] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.02 (m, 2H), 6.79 (m, 1H), 6.62 (m, 1H), 4.20 (m, 2H), 2.81 (m, 1H), 1.93 (m, 6H).
[0253] LCMS: 181.2 [M].
Example 49: 4-Amino-1-(4-fluorobenzyl)pyridinium bromide
[0254] ##STR00053##
[0255] In the same manner as in Example 1, except that 4-fluorobenzyl bromide was used instead of (2-chloroethyl) benzene, 0.078 g (45.8%) of a desired compound, which is a white solid, was obtained.
[0256] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.36 (m, 2H), 8.24 (s, 2H), 7.52 (m, 2H), 7.27 (m, 2H), 6.88 (m, 2H), 5.41 (s, 2H).
[0257] LCMS: 203.1 [M].
Example 50: 1-Benzyl-4-morpholinopyridinium chloride
[0258] ##STR00054##
[0259] In the same manner as in Example 1, except that benzyl chloride was used instead of (2-chloroethyl) benzene and 4-(pyridine-4-yl)morpholine was used instead of 4-aminopyridine, 0.078 g (22%) of a desired compound, which is a white solid, was obtained.
[0260] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.55 (m, 2H), 7.45 (m, 5H), 7.38 (m, 2H), 5.45 (s, 2H), 3.71 (m, 8H).
[0261] LCMS: 255.1 [M].
Example 51: 4-Morpholino-1-phenethylpyridinium chloride
[0262] ##STR00055##
[0263] In the same manner as in Example 1, except that 4-(pyridine-4-yl)morpholine was used instead of 4-aminopyridine, 0.087 g (23.4%) of a desired compound, which is a white solid, was obtained.
[0264] .sup.1H NMR (400) MHz, DMSO-D6) δ 8.33 (m, 2H), 7.32 (m, 2H), 7.22 (m, 5H), 4.47 (m, 2H), 3.67 (m, 8H), 3.10 (m, 2H).
[0265] LCMS: 269.1 [M].
Example 52: 4-Morpholino-1-(cyclopropylmethyl)pyridinium chloride
[0266] ##STR00056##
[0267] In the same manner as in Example 1, except that 4-(pyridine-4-yl)morpholine was used instead of 4-aminopyridine and (2-chloromethyl)cyclopropane was used instead of (2-chloroethyl) benzene, 0.069 g (22.2%) of a desired compound, which is a white solid, was obtained.
[0268] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.58 (m, 2H), 7.58 (m, 2H), 4.09 (d, J=7.6 Hz, 2H), 3.71 (m, 8H), 1.28 (m, 1H), 0.56 (m, 2H), 0.48 (m, 2H).
[0269] LCMS: 219.1 [M].
Example 53: 1-(2-Cyclohexylethyl)-4-morpholinopyridinium bromide
[0270] ##STR00057##
[0271] In the same manner as in Example 1, except that 4-(pyridine-4-yl)morpholine was used instead of 4-aminopyridine and (2-chloroethyl)cyclohexane was used instead of (2-chloroethyl) benzene, 0.101 g (23.3%) of a desired compound, which is a white solid, was obtained.
[0272] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.58 (m, 2H), 7.58 (m, 2H), 4.17 (m, 2H), 3.71 (m, 8H), 1.77 (m, 7H), 1.24 (m, 4H), 1.00 (m, 2H).
[0273] LCMS: 275.1 [M].
Example 54: 1-Benzyl-4-(pyrrolidin-1-yl)pyridinium chloride
[0274] ##STR00058##
[0275] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.25 g (22.2%) of a desired compound, which is a white solid, was obtained.
[0276] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.44 (d, J=7.6 Hz, 2H), 7.38 (m, 5H), 6.92 (d, J=7.6 Hz, 2H), 5.41 (s, 2H), 3.50 (m, 4H), 2.01 (m, 4H).
[0277] LCMS: 239.3 [M].
Example 55: 1-Phenethyl-4-(pyrrolidin-1-yl)pyridinium chloride
[0278] ##STR00059##
[0279] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridine was used instead of 4-aminopyridine, 0.15 g (30.8%) of a desired compound, which is a white solid, was obtained.
[0280] .sup.1H NMR (400 MHz. DMSO-D6) δ 8.26 (d, J=8 Hz, 2H), 7.24 (m, 5H), 6.86 (d, J=8 Hz, 2H), 4.46 (t. J=7.6 Hz, 2H), 3.48 (m, 4H), 3.13 (t, J=7.6 Hz, 2H), 1.99 (m, 4H).
[0281] LCMS: 253.3 [M].
Example 56: 1-(Cyclopropylmethyl)-4-(pyrrolidin-1-yl)pyridinium chloride
[0282] ##STR00060##
[0283] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridine was used instead of 4-aminopyridine and (2-chloromethyl)cyclopropane was used instead of (2-chloroethyl) benzene, 0.2 g (49.7%) of a desired compound, which is a white solid, was obtained.
[0284] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.40 (d. J=8 Hz, 2H), 6.92 (d. J=8 Hz, 2H), 4.07 (d, J=7.2 Hz, 2H), 3.51 (m, 4H), 2.01 (m, 4H), 1.29 (m, 1H), 0.47 (m, 4H).
[0285] LCMS: 203.1 [M].
Example 57: 1-(Cyclohexylmethyl)-4-(pyrrolidin-1-yl)pyridinium bromide
[0286] ##STR00061##
[0287] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridine was used instead of 4-aminopyridine and (2-chloromethyl)cyclohexane was used instead of (2-chloroethyl) benzene, 0.21 g (47.8%) of a desired compound, which is a white solid, was obtained.
[0288] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.29 (d, J=7.6 Hz, 2H), 6.91 (d, J=7.6 Hz, 2H), 4.05 (d, J=7.2 Hz, 2H), 3.51 (m, 4H), 2.01 (m, 4H), 1.70 (m, 4H), 1.49 (m, 2H), 1.15 (m, 3H), 0.97 (m, 2H).
[0289] LCMS: 245.3 [M].
Example 58: 1-(Cyclobutylmethyl)-4-(pyrrolidin-1-yl)pyridinium chloride
[0290] ##STR00062##
[0291] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridine was used instead of 4-aminopyridine and (2-chloromethyl)cyclobutane was used instead of (2-chloroethyl) benzene, 0.13 g (38.1%) of a desired compound, which is a white solid, was obtained.
[0292] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.29 (d, J=7.6 Hz, 2H), 6.89 (d, 0.7.6 Hz, 2H), 4.20 (d, J=7.2 Hz, 2H), 3.48 (m, 4H), 2.72 (m, 1H), 2.01 (m, 4H), 1.84 (m, 6H),
[0293] LCMS: 217.2 [M].
Example 59: 1-Benzyl-4-(piperidin-1-yl)pyridinium chloride
[0294] ##STR00063##
[0295] In the same manner as in Example 1, except that 4-(piperidin-1-yl)pyridine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.078 g (32.4%) of a desired compound, which is a white solid, was obtained.
[0296] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.38 (d, J=7.6 Hz, 2H), 7.42 (m, 5H), 7.24 (d, J=7.6 Hz, 2H), 5.35 (s, 2H), 3.67 (m, 4H), 1.65 (m, 2H), 1.59 (m, 4H).
[0297] LCMS: 253.2 [M].
Example 60: 4-(Azepan-1-yl)-1-benzylpyridinium chloride
[0298] ##STR00064##
[0299] In the same manner as in Example 1, except that 4-(azepan-1-yl)pyridine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.081 g (32.1%) of a desired compound, which is a white solid, was obtained.
[0300] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.39 (d, J=7.2 Hz, 2H), 7.42 (m, 5H), 7.13 (d, J=7.2 Hz, 2H), 5.38 (s, 2H), 3.69 (m, 4H), 1.72 (m, 4H), 1.47 (m, 4H).
[0301] LCMS: 267.2 [M].
Example 61: 1-Benzyl-4-(neopentylamino)pyridinium chloride
[0302] ##STR00065##
[0303] In the same manner as in Example 1, except that 4-(neopentylamino)pyridine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.085 g (35.1%) of a desired compound, which is a white solid, was obtained.
[0304] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.69 (t, J=6.4 Hz, 2H), 8.40 (d, J=7.6 Hz, 1H), 8.22 (d, J=7.6 Hz, 1H), 7.38 (m, 5H), 7.06 (m, 2H), 5.34 (s, 2H), 3.12 (d, J=6.4 Hz, 2H), 0.93 (s, 9H).
[0305] LCMS: 255.2 [M].
Example 62: 4-(Pyrrolidin-1-yl)-1-(thiophen-3-ylmethyl)pyridinium bromide
[0306] ##STR00066##
[0307] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridine was used instead of 4-aminopyridine and 3-(bromomethyl)thiophene was used instead of (2-chloroethyl) benzene, 0.029 g (67.2%) of a desired compound, which is a white solid, was obtained.
[0308] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.39 (d, J=7.6 Hz, 2H), 7.61 (m, 2H), 7.16 (m, 1H), 6.91 (d, J=7.6 Hz, 2H), 5.38 (s, 2H), 3.46 (m, 4H), 1.99 (m, 4H).
[0309] LCMS: 245.1 [M].
Example 63: 6-(Cyclopropylmethyl)-1,2,3,4-tetrahydro-1,6-naphthyridin-6-ium chloride
[0310] ##STR00067##
[0311] In the same manner as in Example 1, except that 1,2,3,4-tetrahydro-1,6-naphthyridine was used instead of 4-aminopyridine and (2-chloromethyl)cyclopropane was used instead of (2-chloroethyl) benzene, 0.039 g (23%) of a desired compound, which is a white solid, was obtained.
[0312] .sup.1H NMR (400 MHz, DMSO-D6) δ 9.24 (s, 1H), 8.16 (s, 1H), 8.10 (m, 1H), 6.83 (d, J=7.2 Hz, 1H), 3.94 (d, J=7.6 Hz, 2H), 3.35 (m, 2H), 2.69 (m, 2H), 1.78 (m, 2H), 1.24 (m, 1H), 0.52 (m, 2H), 0.44 (m, 2H).
[0313] LCMS: 189.2 [M].
Example 64: 6-(Cyclopropylmethyl)-2,3-dihydro-1H-pyrido[3,4-b][1,4]oxazin-6-ium chloride
[0314] ##STR00068##
[0315] In the same manner as in Example 1, except that 1,2,3,4-tetrahydro-1,6-naphthyridine was used instead of 4-aminopyridine and (2-chloromethyl)cyclopropane was used instead of (2-chloroethyl) benzene, 0.019 g (23%) of a desired compound, which is a white solid, was obtained.
[0316] .sup.1H NMR (400 MHz, CD.sub.3OD) δ 8.07 (d, J=2 Hz, 2H), 7.98 (d, J=7.2 Hz, 2H), 6.87 (d, J=7.2 Hz, 2H), 4.31 (t, J=4.8 Hz, 2H), 4.12 (d, J=6.8 Hz, 2H), 3.64 (t, J=4.8 Hz, 2H), 1.35 (m, 1H), 0.72 (m, 2H), 0.53 (m, 2H).
[0317] LCMS: 191.2 [M].
Example 65: l-Benzyl-4(4,4-difluoropiperidin-1-yl)pyridinium chloride
[0318] ##STR00069##
[0319] In the same manner as in Example 1, except that 4-(4,4-difluoropiperidin-1-yl)pyridine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.018 g (16%) of a desired compound, which is a white solid, was obtained.
[0320] .sup.1H NMR (400 MHz. CD.sub.3OD) δ 8.30 (d, J=7.6 Hz, 2H), 7.41 (m, 5H), 7.40 (d, J=7.6 Hz, 2H), 5.38 (s, 2H), 3.86 (m, 4H), 2.17 (m, 4H).
[0321] LCMS: 289.2 [M].
Example 66: 4-(Azetidin-1-yl)-1-benzylpyridinium chloride
[0322] ##STR00070##
[0323] In the same manner as in Example 1, except that 4-(azetidin-1-yl)pyridine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.016 g (18%) of a desired compound, which is a white solid, was obtained.
[0324] .sup.1H NMR (400 MHz, CD.sub.3OD) δ 8.18 (d, J=7.2 Hz, 2H), 7.42 (m, 5H), 6.63 (d, J=7.2 Hz, 2H), 5.33 (s, 2H), 4.31 (t, J=8 Hz, 4H), 2.56 (m, 2H).
[0325] LCMS: 225.2 [M].
Example 67: 1-Benzyl-4-(oxetan-3-ylamino)pyridinium chloride
[0326] ##STR00071##
[0327] In the same manner as in Example 1, except that 4-(oxetan-3-ylamino)pyridine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.012 g (13%) of a desired compound, which is a white solid, was obtained.
[0328] .sup.1H NMR (400 MHz, DMSO-D6) δ 9.45 (m, 1H), 8.48 (d, J=7.6 Hz, 1H), 8.31 (d, 0.1=7.6 Hz, 1H), 7.39 (m, 5H), 6.96 (m, 1H), 6.83 (m, 1H), 5.39 (s, 2H), 4.87 (s, 2H), 4.50 (s, 2H).
[0329] LCMS: 241.2 [M].
Example 68: 4-(Pyrrolidin-1-yl)-1-(thiophen-2-ylmethyl)pyridinium chloride
[0330] ##STR00072##
[0331] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridine was used instead of 4-aminopyridine and 2-(bromomethyl)thiophene was used instead of (2-chloroethyl) benzene, 0.018 g (14.6%) of a desired compound, which is a white solid, was obtained.
[0332] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.41 (d, J=7.2 Hz, 2H), 7.60 (m, 1H), 7.31 (d, J=3.6 Hz, 2H), 7.07 (m, 1H), 6.91 (d, J=7.2 Hz, 2H), 5.61 (s, 2H), 3.49 (m, 4H), 1.98 (m, 4H),
[0333] LCMS: 245.1 [M].
Example 69: l-Benzyl-4-(tert-butylamino)pyridinium chloride
[0334] ##STR00073##
[0335] In the same manner as in Example 1, except that 4-(tert-butylamino)pyridine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl)benzene, 0.021 g (12%) of a desired compound, which is a white solid, was obtained.
[0336] .sup.1H NMR (400 MHz, DMSO-D6) δ 9.71 (s, 1H), 8.09 (d, J=6.8 Hz, 1H), 7.97 (m, 1H), 7.84 (d, J=6.8 Hz, 1H), 7.41 (m, 3H), 7.36 (m, 2H), 6.72 (m, 1H), 5.37 (s, 2H), 1.47 (s, 9H),
[0337] LCMS: 241.1 [M].
Example 70: 4-(Azetidin-1-yl)-1-(cyclopropylmethyl)pyridinium chloride
[0338] ##STR00074##
[0339] In the same manner as in Example 1, except that 4-(azetidin-1-yl)pyridine was used instead of 4-aminopyridine and (2-chloromethyl)cyclopropane was used instead of (2-chloroethyl)benzene, 0.045 g (53.7%) of a desired compound, which is a white solid, was obtained.
[0340] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.41 (d, J=8 Hz, 2H), 6.68 (d, J=8 Hz, 2H), 4.22 (d, J=7.6 Hz, 4H), 4.01 (d, J=7.6 Hz, 2H), 2.41 (m, 2H), 1.27 (m, 1H), 0.53 (m, 2H), 0.44 (m, 2H).
[0341] LCMS: 189.2 [M].
Example 71: 4-(Azetidin-1-yl)-1-(thiophen-3-ylmethyl)pyridinium bromide
[0342] ##STR00075##
[0343] In the same manner as in Example 1, except that 4-(azetidin-1-yl)pyridine was used instead of 4-aminopyridine and 3-(bromomethyl)thiophene was used instead of (2-chloroethyl)benzene, 0.068 g (58%) of a desired compound, which is a white solid, was obtained.
[0344] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.37 (d. J=6.8 Hz, 2H), 7.63 (m, 2H), 7.61 (m, 2H), 7.16 (d, J=7.2 Hz, 2H), 6.68 (d, J=6.8 Hz, 2H), 5.35 (s, 2H), 4.21 (t, J=7.6 Hz, 4H), 2.41 (m, 2H).
[0345] LCMS: 231.1 [M].
Example 72: 4-(Pyrrolidin-1-yl)-1-(selenophen-2-ylmethyl)pyridinium chloride
[0346] ##STR00076##
[0347] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridine was used instead of 4-aminopyridine and 2-(chloromethyl)selenophene was used instead of (2-chloroethyl)benzene, 0.058 g (15.8%) of a desired compound, which is a white solid, was obtained.
[0348] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.42 (d, J=7.2 Hz, 2H), 8.22 (d, J=5.6 Hz, 1H), 7.46 (d, J=3.6 Hz, 1H), 7.25 (d, J=5.6 Hz, 1H), 6.92 (d, J=7.2 Hz, 2H), 5.62 (s, 2H), 3.50 (m, 4H), 1.99 (m, 4H).
[0349] LCMS: 292.2 [M].
Example 73: 4-Amino-1-(cyclopropylmethyl)pyrimidin-1-ium chloride
[0350] ##STR00077##
[0351] In the same manner as in Example 1, except that 4-aminopyrimidine was used instead of 4-aminopyridine and (2-chloromethyl)cyclopropane was used instead of (2-chloroethyl) benzene, 0.055 g (14%) of a desired compound, which is a white solid, was obtained.
[0352] .sup.1H NMR (400 MHz, DMSO-D6) δ 9.20 (s, 1H), 9.04 (s, 1H), 8.86 (s, 1H), 8.38 (d, J=7.2 Hz, 1H), 6.87 (d, J=7.6 Hz, 1H), 3.98 (d, J=7.2 Hz, 1H), 1.30 (m, 1H), 0.53 (m, 2H), 0.47 (m, 2H).
[0353] LCMS: 150.2 [M].
Example 74: 4-Amino-1-(selenophen-2-ylmethyl)pyrimidin-1-ium chloride
[0354] ##STR00078##
[0355] In the same manner as in Example 1, except that 4-aminopyrimidine was used instead of 4-aminopyridine and 2-(chloromethyl)selenophene was used instead of (2-chloroethyl) benzene, 0.025 g (11.3%) of a desired compound, which is a white solid, was obtained.
[0356] .sup.1H NMR (400 MHz, DMSO-D6) δ 9.36 (s, 1H), 9.16 (s, 1H), 9.03 (s, 1H), 8.36 (d, =7.2 Hz, 1H), 8.26 (d, J=7.2 Hz, 1H), 7.51 (d, J=2.4 Hz, 1H), 7.28 (d, J=5.6 Hz, 1H), 6.87 (d, J=7.2 Hz, 1H),
[0357] LCMS: 240.0 [M].
Example 75: 4-Amino-1-(selenophen-2-ylmethyl)pyridazin-1-ium chloride
[0358] ##STR00079##
[0359] In the same manner as in Example 1, except that 4-aminopyridazine was used instead of 4-aminopyridine and 2-(chloromethyl)selenophene was used instead of (2-chloroethyl) benzene, 0.019 g (8.66%) of a desired compound, which is a white solid, was obtained.
[0360] .sup.1H NMR (400 MHz, DMSO-D6) δ 9.33 (s, 1H), 9.15 (s, 1H), 8.85 (s, 1H), 8.37 (d, 0.1-7.2 Hz, 1H), 8.23 (d, J=7.2 Hz, 1H), 7.50 (d, J=2.4 Hz, 1H), 7.23 (d, J=5.6 Hz, 1H), 6.89 (d, J=7.2 Hz, 1H).
[0361] LCMS: 240.0 [M].
Example 76: 4-Amino-1-(selenophen-2-ylmethyl)pyridinium chloride
[0362] ##STR00080##
[0363] In the same manner as in Example 1, except that 2-(chloromethyl)selenophene was used instead of (2-chloroethyl) benzene, 0.1 g (48%) of a desired compound, which is a white solid, was obtained.
[0364] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.33 (s, 2H), 8.31 (d, J=7.2 Hz; 2H), 8.23 (d, J=5.2 Hz, 1H), 7.44 (s, 1H), 7.27 (d, J=5.2 Hz, 1H), 6.88 (d, J=7.2 Hz, 2H), 5.59 (s, 2H).
[0365] LCMS: 239.0 [M].
Example 77: 4-Amino-1-(thiophen-2-ylmethyl)pyridinium chloride
[0366] ##STR00081##
[0367] In the same manner as in Example 1, except that 2-(chloromethyl)thiophene was used instead of (2-chloroethyl) benzene, 0.37 g (37.4%) of a desired compound, which is a white solid, was obtained.
[0368] .sup.1H NMR (400) MHz, DMSO-D6) δ 8.29 (m, 4H), 7.62 (d, J=4.8 Hz, 1H), 7.29 (s, 1H), 7.07 (m, 1H), 6.87 (d, J=7.2 Hz, 2H), 5.57 (s, 2H).
[0369] LCMS: 191.2 [M].
Example 78: 1-(Furan-2-ylmethyl)-4-(pyrrolidin-1-yl)pyridinium chloride
[0370] ##STR00082##
[0371] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridine was used instead of 4-aminopyridine and 2-(chloromethyl)furan was used instead of (2-chloroethyl) benzene, 0.028 g (2%) of a desired compound, which is a white solid, was obtained.
[0372] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.30 (d, J=7.2 Hz, 2H), 7.69 (m, 2H), 6.90 (d, J=7.2 Hz, 2H), 6.63 (d, J=3.2 Hz, 1H), 6.48 (m, 1H), 5.43 (s, 2H), 3.48 (m, 4H), 1.98 (m, 4H).
[0373] LCMS: 229.1 [M].
Example 79: 1-((5-Methylthiophen-2-yl)methyl)-4-(pyrrolidin-1-yl)pyridinium chloride
[0374] ##STR00083##
[0375] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridine was used instead of 4-aminopyridine and 2-(chloromethyl)-5-methylthiophene was used instead of (2-chloroethyl) benzene, 0.12 g (13%) of a desired compound, which is a white solid, was obtained.
[0376] .sup.1H NMR (400 MHz. DMSO-D6) δ 8.36 (d, J=7.6 Hz, 2H), 7.10 (d, J=3.2 Hz, 1H), 6.90 (d, J=7.6 Hz, 2H), 6.74 (m, 1H), 5.51 (s, 2H), 3.49 (m, 4H), 1.99 (m, 4H).
[0377] LCMS: 259.1 [M].
Example 80: 4-(Azetidin-1-yl)-1-(selenophen-3-ylmethyl)pyridinium chloride
[0378] ##STR00084##
[0379] In the same manner as in Example 1, except that 4-(azetidin-1-yl)pyridine was used instead of 4-aminopyridine and 2-(chloromethyl)selenophene was used instead of (2-chloroethyl) benzene, 0.12 g (40.7%) of a desired compound, which is a white solid, was obtained.
[0380] .sup.1H NMR (400 MHz, DMSO-D6) δ 8.37 (d. J=7.2 Hz, 2H), 8.22 (m, 1H), 7.44 (m, 1H), 7.25 (m, 1H), 6.69 (d, J=7.2 Hz, 2H), 5.58 (s, 2H), 4.22 (t, J=8 Hz, 4H), 2.41 (m, 2H),
[0381] LCMS: 279.0 [M].
Example 81: 2-Amino-4-(azetidin-1-yl)-1-(cyclopropylmethyl)pyridinium chloride
[0382] ##STR00085##
[0383] In the same manner as in Example 1, except that 4-(azetidin-1-yl)pyridin-2-amine was used instead of 4-aminopyridine and 2-(chloromethyl)thiophene was used instead of (2-chloroethyl) benzene, 0.16 g (79%) of a desired compound, which is a white solid, was obtained.
[0384] .sup.1H NMR (400 MHz, DMSO-D6) δ 7.85 (d, J=7.6 Hz, 1H), 7.59 (m, 1H), 7.45 (m, 2H), 7.05 (m, 1H), 6.16 (m, 1H), 5.49 (m, 1H), 5.18 (s, 2H), 4.05 (m, 4H), 2.39 (m, 2H).
[0385] LCMS: 246.1 [M].
Example 82: 2-Amino-4-(azetidin-1-yl)-1-(cyclopropylmethyl)pyridinium chloride
[0386] ##STR00086##
[0387] In the same manner as in Example 1, except that 4-(azetidin-1-yl)pyridin-2-amine was used instead of 4-aminopyridine and (chloromethyl)cyclopropane was used instead of (2-chloroethyl) benzene, 0.11 g (45.6%) of a desired compound, which is a white solid, was obtained.
[0388] .sup.1H NMR (300 MHz, DMSO-D6) δ 7.75 (d, J=7.5 Hz, 1H), 7.52 (s, 2H), 6.10 (m, 1H), 6.55 (m, 1H), 4.12 (m, 4H), 3.87 (d, J=7.2 Hz, 2H), 2.35 (m, 2H), 1.21 (m, 1H), 0.55 (m, 2H), 0.47 (m, 2H).
[0389] LCMS: 204.2 [M].
Example 83: 2,4-Diamino-1-(cyclopropylmethyl)pyridinium chloride
[0390] ##STR00087##
[0391] In the same manner as in Examples 7 and 8, except that (chloromethyl)cyclopropane was used instead of (2-chloroethyl) benzene, 51 mg (51.8%) of a desired compound, which is a white solid, was obtained.
[0392] .sup.1H NMR (300 MHz, DMSO-D6) δ 7.64 (d, J=7.2 Hz, 1H), 7.45 (s, 2H), 7.24 (s, 2H), 6.19 (m, 1H), 5.89 (s, 1H), 3.82 (d, J=7.2 Hz, 2H), 1.21 (m, 1H), 0.56 (m, 2H), 0.48 (m, 2H).
[0393] LCMS: 164.1 [M].
Example 84: 2,4-Diamino-1-(4-chlorobenzyl)pyridinium chloride
[0394] ##STR00088##
[0395] In the same manner as in Examples 7 and 8, except that 1-chloro-4-(chloromethyl)benzene was used instead of (2-chloroethyl) benzene, 46 mg (46.6%) of a desired compound, which is a white solid, was obtained.
[0396] .sup.1H NMR (300 MHz, DMSO-D6) δ 7.67 (d, J=7.5 Hz, 1H), 7.41 (m, 4H), 7.17 (d, J=8.4 Hz, 2H), 6.20 (m, 1H), 5.85 (s, 1H), 5.18 (s, 2H).
[0397] LCMS: 234.2 [M].
Example 85: 2-Amino-4-(azetidin-1-yl)-1-((5-methylthiophen-2-yl)methyl)pyridinium chloride
[0398] ##STR00089##
[0399] In the same manner as in Example 1, except that 4-(azetidin-1-yl)pyridin-2-amine was used instead of 4-aminopyridine and 2-(chloromethyl)-5-methylthiophene was used instead of (2-chloroethyl) benzene, 0.12 g (43%) of a desired compound, which is a white solid, was obtained.
[0400] .sup.1H NMR (400 MHz, DMSO-D6) δ 7.85 (d, J=7.6 Hz, 1H), 7.59 (m, 1H), 7.45 (m, 2H), 7.05 (m, 1H), 6.16 (m, 1H), 5.49 (m, 1H), 5.18 (s, 2H), 4.05 (m, 4H), 2.41 (s, 3H), 2.39 (m, 2H).
[0401] LCMS: 260.1 [M].
Example 86: 2-Amino-4-(azetidin-1-yl)-1-(selenophen-2-ylmethyl)pyridinium chloride
[0402] ##STR00090##
[0403] In the same manner as in Example 1, except that 4-(azetidin-1-yl)pyridin-2-amine was used instead of 4-aminopyridine and 2-(chloromethyl)selenophene was used instead of (2-chloroethyl) benzene, 0.09 g (38%) of a desired compound, which is a white solid, was obtained.
[0404] .sup.1H NMR (300 MHz, DMSO-D6) δ 8.22 (d, J=5.6 Hz, 1H), 7.75 (d. J=7.5 Hz, 1H), 7.52 (s, 2H), 7.46 (d, J=3.6 Hz, 1H), 7.25 (d, J==5.6 Hz, 1H), 6.10 (m, 1H), 6.55 (m, 1H), 5.62 (s, 2H), 4.12 (m, 4H), 2.35 (m, 2H).
[0405] LCMS: 293.1 [M].
Example 87: 2-Amino-4-(azetidin-1-yl)-1-benzylpyridinium chloride
[0406] ##STR00091##
[0407] In the same manner as in Example 1, except that 4-(azetidin-1-yl)pyridin-2-amine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.1 g (48%) of a desired compound, which is a white solid, was obtained.
[0408] .sup.1H NMR (300 MHz, DMSO-D6) δ 7.75 (d, J=7.5 Hz, 1H), 7.52 (s, 2H), 6.10 (m, 1H), 7.39 (m, 5H), 6.55 (m, 1H), 5.51 (s, 2H), 4.11 (m, 4H), 2.32 (m, 2H).
[0409] LCMS: 240.1 [M].
Example 88: 2-Amino-1-benzyl-4-(pyrrolidin-1-yl)pyridinium chloride
[0410] ##STR00092##
[0411] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridin-2-amine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.15 g (30.8%) of a desired compound, which is a white solid, was obtained.
[0412] .sup.1H NMR (300 MHz, DMSO-D6) δ 7.75 (d, J=7.5 Hz, 1H), 7.52 (s, 2H), 6.10 (m, 1H), 7.39 (m, 5H), 6.55 (m, 1H), 5.51 (s, 2H), 3.49 (m, 4H), 1.98 (m, 4H).
[0413] LCMS: 254.1 [M].
Example 89: 2-Amino-1-(cyclopropylmethyl)-4-(pyrrolidin-1-yl)pyridinium chloride
[0414] ##STR00093##
[0415] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridin-2-amine was used instead of 4-aminopyridine and (chloromethyl)cyclopropane was used instead of (2-chloroethyl) benzene, 0.18 g (14.6%) of a desired compound, which is a white solid, was obtained.
[0416] .sup.1H NMR (300 MHz, DMSO-D6) δ 7.75 (d, J=7.5 Hz, 1H), 7.52 (s, 2H), 6.10 (m, 1H), 6.55 (m, 1H), 3.82 (d, J=7.2 Hz, 2H), 3.49 (m, 4H), 1.98 (m, 4H), 1.21 (m, 1H), 0.56 (m, 2H), 0.48 (m, 2H).
[0417] LCMS: 218.1 [M].
Example 90: 2-Amino-1-((5-methylthiophen-2-yl)methyl)-4-(pyrrolidin-1-yl)pyridinium chloride
[0418] ##STR00094##
[0419] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridin-2-amine was used instead of 4-aminopyridine and 2-(chloromethyl)-5-methylthiophene was used instead of (2-chloroethyl) benzene, 0.21 g (47.8%) of a desired compound, which is a white solid, was obtained.
[0420] .sup.1H NMR (400 MHz, DMSO-D6) δ 7.85 (d, 0.1=7.6 Hz, 1H), 7.59 (m, 1H), 7.45 (m, 2H), 7.05 (m, 1H), 6.16 (m, 1H), 5.49 (m, 1H), 5.18 (s, 2H), 3.49 (m, 4H), 2.41 (s, 3H), 1.98 (m, 4H).
[0421] LCMS: 274.1 [M].
Example 91: 2-Amino-4-(pyrrolidin-1-yl)-1-(selenophen-2-ylmethyl)pyridinium chloride
[0422] ##STR00095##
[0423] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridin-2-amine was used instead of 4-aminopyridine and 2-(chloromethyl)selenophene was used instead of (2-chloroethyl) benzene, 0.17 g (38%) of a desired compound, which is a white solid, was obtained.
[0424] .sup.1H NMR (300 MHz, DMSO-D6) δ 8.22 (d, J=5.6 Hz, 1H), 7.75 (d, J=7.5 Hz, 1H), 7.52 (s, 2H), 7.46 (d, J=3.6 Hz, 1H), 7.25 (d, J=5.6 Hz, 1H), 6.10 (m, 1H), 6.55 (m, 1H), 5.62 (s, 2H), 3.49 (m, 4H), 1.98 (m, 4H).
[0425] LCMS: 307.0 [M].
Example 92: 2-Amino-1-(4-chlorobenzyl)-4-(pyrrolidin-1-yl)pyridinium chloride
[0426] ##STR00096##
[0427] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)pyridin-2-amine was used instead of 4-aminopyridine and 4-chlorobenzyl chloride was used instead of (2-chloroethyl) benzene, 0.25 g (67.4%) of a desired compound, which is a white solid, was obtained.
[0428] .sup.1H NMR (300 MHz, DMSO-D6) δ 7.75 (d. J=7.5 Hz, 1H), 7.52 (s, 2H), 7.41 (d. J=8.4 Hz, 2H), 7.17 (d, J=8.4 Hz, 2H), 6.10 (m, 1H), 6.55 (m, 1H), 5.51 (s, 2H), 3.49 (m, 4H), 1.98 (m, 4H).
[0429] LCMS: 288.1 [M].
Example 93: 4-Amino-1-benzyl-2-ethoxypyridinium chloride
[0430] ##STR00097##
[0431] In the same manner as in Example 1, except that 2-ethoxylpyridin-4-amine was used instead of 4-amino pyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.12 g (31%) of a desired compound, which is a white solid, was obtained.
[0432] .sup.1H NMR (300 MHz; DMSO-D6) δ 8.21 (s, 2H), 8.15 (d, J=7.2 Hz, 1H), 7.38 (m, 5H), 6.62 (m, 1H), 6.36 (m, 1H), 5.25 (s, 2H), 4.06 (q, J=7.2 Hz, 2H), 1.20 (t, J=7.2 Hz, 3H).
[0433] LCMS: 229.1 [M].
Example 94: 4-Amino-1-benzyl-2-isopropoxypyridinium chloride
[0434] ##STR00098##
[0435] In the same manner as in Example 1, except that 2-isopropoxylpyridin-4-amine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.14 g (28%) of a desired compound, which is a white solid, was obtained.
[0436] .sup.1H NMR (300 MHz, DMSO-D6) δ 8.20 (s, 2H), 8.13 (d, J=7.2 Hz, 1H), 7.38 (m, 5H), 6.62 (m, 1H), 6.36 (m, 1H), 5.25 (s, 2H), 4.04 (m, 1H), 1.38 (d, J=7.2 Hz, 6H).
[0437] LCMS: 243.1 [M].
Example 95: 4-Amino-1-benzyl-2-cyclopropylpyridinium chloride
[0438] ##STR00099##
[0439] In the same manner as in Example 1, except that 2-cyclopropylpyridin-4-amine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.11 g (45.6%) of a desired compound, which is a white solid, was obtained.
[0440] .sup.1H NMR (300 MHz, DMSO-D6) δ 8.21 (s, 2H), 8.11 (d, J=7.2 Hz, 1H), 7.39 (m, 5H), 6.60 (m, 1H), 6.33 (m, 1H), 5.26 (s, 2H), 1.50 (m, 1H), 0.56 (m, 2H), 0.48 (m, 2H).
[0441] LCMS: 225.1 [M].
Example 96: 4-(Azetidin-1-yl)-1-benzyl-2-ethoxypyridinium chloride
[0442] ##STR00100##
[0443] In the same manner as in Example 1, except that 4-(azetidin-1-yl)-2-ethoxypyridine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.15 g (30.8%) of a desired compound, which is a white solid, was obtained.
[0444] .sup.1H NMR (300 MHz, DMSO-D6) δ 8.17 (d, J=7.2 Hz, 1H), 7.37 (m, 5H), 6.61 (m, 1H), 6.36 (m, 1H), 5.25 (s, 2H), 4.06 (q, J=7.2 Hz, 2H), 4.12 (m, 4H), 2.35 (m, 2H), 1.20 (t, J=7.2 Hz, 3H).
[0445] LCMS: 269.1 [M].
Example 97: 4-(Azetidin-1-yl)-1-benzyl-2-isopropoxypyridinium chloride
[0446] ##STR00101##
[0447] In the same manner as in Example 1, except that 4-(azetidin-1-yl)-2-isopropoxypyridine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.07 g (21%) of a desired compound, which is a white solid, was obtained.
[0448] .sup.1H NMR (300 MHz, DMSO-D6) δ 8.15 (d. J=7.2 Hz, 1H), 7.38 (m, 5H), 6.62 (m, 1H), 6.36 (m, 1H), 5.25 (s, 2H), 4.12 (m, 4H), 4.04 (m, 1H), 2.35 (m, 2H), 1.38 (d, J=7.2 Hz, 6H).
[0449] LCMS: 283.1 [M].
Example 98: 4-(Azetidin-1-yl)-1-benzyl-2-cyclopropylpyridinium chloride
[0450] ##STR00102##
[0451] In the same manner as in Example 1, except that 4-(azetidin-1-yl)-2-cyclopropylpyridine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.05 g (42%) of a desired compound, which is a white solid, was obtained.
[0452] .sup.1H NMR (300 MHz, DMSO-D6) δ 8.13 (d. J=7.2 Hz, 1H), 7.31 (m, 5H), 6.60 (m, 1H), 6.36 (m, 1H), 5.25 (s, 2H), 4.12 (m, 4H), 2.35 (m, 2H), 1.50 (m, 1H), 0.56 (m, 2H), 0.48 (m, 2H).
[0453] LCMS: 265.1 [M].
Example 99: 1-Benzyl-2-ethoxy-4-(pyrrolidin-1-yl)pyridinium chloride
[0454] ##STR00103##
[0455] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)-2-ethoxypyridine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.18 g (14.6%) of a desired compound, which is a white solid, was obtained.
[0456] .sup.1H NMR (300 MHz, DMSO-D6) δ 8.17 (d, J=7.2 Hz, 1H), 7.37 (m, 5H), 6.61 (m, 1H), 6.36 (m, 1H), 5.25 (s, 2H), 4.06 (q, J=7.2 Hz, 2H), 3.49 (m, 4H), 1.98 (m, 4H), 1.20 (t, =7.2 Hz, 3H).
[0457] LCMS: 283.1 [M].
Example 100: 1-Benzyl-2-isopropoxy-4-(pyrrolidin-1-yl)pyridinium chloride
[0458] ##STR00104##
[0459] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)-2-isopropoxypyridine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.13 g (38.1%) of a desired compound, which is a white solid, was obtained.
[0460] .sup.1H NMR (300 MHz, DMSO-D6) δ 8.15 (d, J=7.2 Hz, 1H), 7.38 (m, 5H), 6.62 (m, 1H), 6.36 (m, 1H), 5.25 (s, 2H), 4.04 (m, 1H), 3.49 (m, 4H), 1.98 (m, 4H), 1.38 (d. J=7.2 Hz, 6H).
[0461] LCMS: 297.1 [M].
Example 101: 1-Benzyl-2-cyclopropyl-4-(pyrrolidin-1-yl)pyridinium chloride
[0462] ##STR00105##
[0463] In the same manner as in Example 1, except that 4-(pyrrolidin-1-yl)-2-cyclopropylpyridine was used instead of 4-aminopyridine and benzyl chloride was used instead of (2-chloroethyl) benzene, 0.15 g (30.8%) of a desired compound, which is a white solid, was obtained.
[0464] .sup.1H NMR (300 MHz, DMSO-D6) δ 8.13 (d, 0.1=7.2 Hz, 1H), 7.31 (m, 5H), 6.60 (m, 1H), 6.36 (m, 1H), 5.25 (s, 2H), 3.49 (m, 4H), 1.98 (m, 4H), 1.50 (m, 1H), 0.56 (m, 2H), 0.48 (m, 2H).
[0465] LCMS: 279.1 [M].
Test Example 1: Measurement of Inhibitory Effect by Oxygen Consummation Rate and Extracellular Oxidation
[0466] The compounds synthesized by the methods disclosed in the Examples of the present invention have been measured on oxygen consumption rate and extracellular oxidation by the methods disclosed in the Test Examples below.
[0467] As the synthesized drugs inhibit oxidative phosphorylation and exhibit anti-cancer effects, Oxygen Consumption Rate (OCR) of cells for the compounds was measured.
[0468] 3×10.sup.3 cells from A549 cell lines (purchased from ATCC-American Type Culture Collection), which are lung cancer cell lines, were placed on XF96 cell culture plates using RPMI1640 medium, and cultured at 37° C. in a 5% CO.sub.2 condition for 16 hours or more for attachment.
[0469] After 16 hours, the cells were treated with the drug at six different concentrations between 0 μM and 20 μM. After 24 hours, the existing medium was removed, and XF assay medium (15 mM D-Glucose, 15 mM sodium pyruvate, 4 mM L-Glutamine, pH 7.4) was added. The cells were retreated with the drug, and additionally cultured in Prep station at 37° C. in a non-CO.sub.2 condition for 1 hour. During the one-hour culture in the Prep station, a sensor cartridge was placed and calibrated for 20 minutes, and a plate with cells was placed to analyze the OCR. After the analysis was completed, XF96 plate was measured for cell viability using Cyquant assay, which measures the amount of intracellular DNA, in the following method. XF assay medium and the drug were removed, and the cells were placed in a cryogenic refrigerator (−80° C.) for at least 4 hours to be frozen. After the plate was made to be at room temperature, a solution where a lysis buffer and fluorescent GR dye were mixed was placed by 200 μL per well. After 20-minute reaction at room temperature, absorbance was measured between 480 nM to 520 nM to calculate cell viability. A measured value of a well untreated with the drug was converted to 100% by reflecting cell viability to the OCR value. Concentration of a drug which inhibits the OCR value reflecting cell viability by 50% was calculated.
TABLE-US-00002 TABLE 2 Example OCR IC.sub.50 (μM) Berberine 4.6 Example 1 1.1 Example 7 0.4 Example 8 1.2 Example 11 0.7 Example 12 0.8 Example 13 16.5 Example 15 2.3 Example 16 1.5 Example 17 4.2 Example 18 0.6 Example 19 0.5 Example 20 1.8 Example 21 0.9 Example 22 7.9 Example 23 0.9 Example 24 0.8 Example 25 1.6 Example 26 2.4 Example 27 0.4 Example 28 0.4 Example 29 5.3 Example 30 0.9 Example 31 4.3 Example 32 0.3 Example 33 1.5 Example 34 1.1 Example 35 0.5 Example 36 0.7 Example 37 1.1 Example 38 1.1 Example 39 2.7 Example 40 0.9 Example 41 2.7 Example 42 1.5 Example 43 0.9 Example 44 1.1 Example 45 5 Example 46 0.8 Example 47 5 Example 48 0.8 Example 49 2.4 Example 50 1 Example 51 3.1 Example 52 0.8 Example 53 0.5 Example 54 1.1 Example 55 3.1 Example 56 0.8 Example 57 0.5 Example 58 2.5 Example 59 2.8 Example 60 3.3 Example 61 15.7 Example 62 1 Example 63 7.1 Example 64 10.2 Example 66 5.2 Example 68 0.8 Example 70 3.4 Example 71 3.4 Example 72 1 Example 75 18.8 Example 76 1.5 Example 77 2.4 Example 78 2.1 Example 79 0.8 Example 80 1.3 Example 81 2.1
Test Example 2: Measurement of Inhibitory Effect of Cancer Cell Proliferation
[0470] The compounds prepared in the above Examples were evaluated for the inhibitory effect of cancer cell proliferation according to the method described in the following Test Example.
[0471] SK-MEL-28 cells derived from human melanoma were used, and the concentration (cell growth inhibitory concentration, IC.sub.50) at which cell growth was inhibited to 50% was measured using MTT reagent (3-(4,5-dimethylthiazole-2-yl)-2,5-ditetrazolium bromide) to confirm the inhibitory effect of cancer cell proliferation of the drugs synthesized in Examples 1 to 84.
[0472] First, SK-MEL-28 cells were cultured in 96-well plates at a cell number of about 1,250 in RPMI-1640 medium containing 11.1 mM glucose and 10% calf blood serum or 0.75 mM glucose and 10% calf blood serum, and were cultured for 16 hours. Further, in order to determine the IC.sub.50 value of each compound, the compound was added at a concentration of 1 mM, 200 μM, 40 μM, 8 μM, 1.6 μM, 0.32 μM, and 0.064 μM under the condition of 11.1 mM glucose, and 200 μM, 40 μM, 8 μM, 1.6 μM, 0.32 μM, 0.064 μM, and 0.0128 μM under the condition of 0.75 mM glucose in the well plate, and the well plate was cultured for 72 hours. After treatment of the compound, MTT was added to the culture medium to confirm living cells and further cultured for 2 hours. The resulting formazane crystal was dissolved using dimethyl sulfoxide, and the absorbance of the solution was measured at 555 nm. After culturing for 72 hours, the number of viable cells in the well plate treated with the compounds synthesized in the Examples relative to the number of cells cultured in the well plate without treatment of the compounds was expressed as cell viability (%) according to each treatment concentration. By using this, a cell viability curve graph was prepared, and the inhibitory effect of cancer cell proliferation was confirmed by calculating the concentration of the compound whose growth was inhibited to 50% (IC.sub.50).
[0473] The results of the inhibitory effect of cancer cell growth are shown in Table 3 below.
TABLE-US-00003 TABLE 3 SK-MEL-28 11.1 mM SK-MEL-28 0.75 mM glucose Cell viability glucose Cell viability Example (IC.sub.50 μM) (IC.sub.50 μM) Example 1 163.4 17.9 Example 7 16.8 20.7 Example 8 75.1 4.7 Example 11 38.5 11.9 Example 12 55.5 12.8 Example 13 319.7 129.3 Example 14 277.4 154.4 Example 15 47 6.8 Example 16 28.2 8 Example 17 75.4 6.9 Example 18 16.9 10.7 Example 19 8.8 5.9 Example 20 143.4 14.7 Example 21 27.4 12.5 Example 22 192.8 18.5 Example 23 51.1 5.4 Example 24 16.8 4.4 Example 25 40.6 15.5 Exampie 26 193.8 102.3 Example 27 19.5 1.7 Example 28 3.8 0.7 Example 29 162.5 38.5 Example 30 23 8.2 Example 31 418.5 56.2 Example 32 38.8 8.1 Example 33 101.6 7.7 Example 34 101.2 7.9 Example 35 7.5 5 Example 36 13.1 2.8 Example 37 10.8 1.5 Example 38 11.7 1.5 Example 39 76.5 45.3 Example 40 30.6 7.5 Example 41 234.3 35.1 Example 42 198.7 35 Example 43 45.5 14.4 Example 44 104.7 15.2 Example 45 81.4 16.6 Example 46 49.6 7.9 Example 47 166.6 20.3 Example 48 46.3 19.9 Example 49 54.5 10.3 Example 50 207.5 13.9 Example 51 201 29.1 Example 52 366.7 40 Example 53 76.4 8.7 Example 54 29.7 3.4 Example 55 45.9 0.1 Example 56 118 4.4 Example 57 23 1.1 Example 58 93.9 3 Example 59 33.4 2.8 Example 60 24.1 20 Example 61 42.,7 1.5 Example 62 40.1 3.1 Example 63 208.3 20.4 Example 64 187.1 21 Example 65 214.6 28.6 Example 66 77 6.1 Example 67 545.3 130.7 Example 68 38.5 1.7 Example 69 131.7 14.6 Example 70 49.5 9.4 Example 71 76.4 6.8 Example 72 37.2 2.6 Example 73 358.3 168.6 Example 74 243 74.4 Example 75 239.4 57.7 Example 76 114 14.1 Example 77 105.6 15.8 Example 78 103.5 3.55 Example 79 30.7 1.16 Example 80 45.91 3.29 Example 81 58.37 4.15
Test Example 3: Test for Observing Antitumor Effect in Mouse Kidney Cancer Cells
[0474] RENCA, which are mouse kidney cancer cells, were cultured in RPMI 1640 medium containing 10% FBS and 1% anti-anti at 37° C. and 5% CO.sub.2. 8- to 10-week-old BALB/c mice with a body weight range of 18 g to 20 g were subjected to a 7-day acclimation period, and then 1×10.sup.6/0.1 mL of RENCA cells in PBS were subcutaneously implanted on the right side of the backs of the mice. Seven days after implantation, group separation was performed based on the average of tumor volumes when the tumor volumes reached 50 mm.sup.2 to 80 mm.sup.2. A vehicle control group was intraperitoneally injected with PBS containing 2% DMSO and 2% Tween80, and an Example 62 administration group was intraperitoneally injected at a dose of 10 mg/kg, once a day for 2 weeks. Tumor volume measurements were performed twice weekly using Vemier calipers, and the volume of tumor was calculated by substituting long axis and short axis for 0.5×long axis×short axis.sup.2. The results are shown in Table 4 and FIG. 1.
TABLE-US-00004 TABLE 4 Days Post Tumor Implantation Vehicle Example 62 7 81 ± 8 81 ± 7 9 160 ± 24 146 ± 17 12 380 ± 79 227 ± 39 14 615 ± 169 364 ± 68 16 970 ± 221 545 ± 132 20 1,687 ± 358 775 ± 206
[0475] From the results of the volume measurement of tumors, it was observed that the group to which Example 62 was administered remarkably inhibited tumor growth from the 6.sup.th day of administration compared with the vehicle control group. The volume of tumor on day 14 which was the end day of observation yielded statistically significant data. Thus, Example 62 confirmed that there was a clear inhibitory effect on tumor growth in mouse kidney cancer cells.