Biguanide compounds and use thereof

Abstract

The present invention relates to biguanide compounds and use thereof, more particularly, to biguanide derivatives exhibiting excellent effects for inhibition of cancer cell proliferation and inhibition of cancer metastasis and recurrence, a method for preparing the same, a pharmaceutical composition containing the same as an active ingredient, and a method of prevention or treatment of cancer comprising the step of administering an effective amount of the composition to a subject in need thereof.

Claims

1. A compound of Chemical Formula 1, or a pharmaceutically acceptable salt thereof: ##STR00113## wherein, R.sub.1 is hydrogen, a hydroxy group, or a linear or branched C.sub.1-6 alkyl which can be substituted by hydrogen or a hydroxy group, R.sub.2 has Chemical Formula 11 ##STR00114## wherein R.sub.3 is a linear or branched C.sub.1-C.sub.6 alkyl, R.sub.4 is an amine; C.sub.1-C.sub.6 alkylamine; or amide, and, m is 1, 2 or 3.

2. A compound selected from the group consisting of: N1-pyrrolidine-N5-(benzo[1,3]dioxol-5-ylmethyl) biguanide, N1-pyrrolidine-N5-(4-methoxy)phenyl biguanide, N1-pyrrolidine-N5-(3-chloro-4-methoxy)phenyl biguanide, N1-pyrrolidine-N5-(2,6-difluoro-N5-methyl)phenyl biguanide, N1-pyrrolidine-N5-(4-dimethylamino)phenyl biguanide, N1-pyrrolidine-N5-(4-isopropyl)phenyl biguanide, N1-pyrrolidine-N5-(3-phenoxy)phenyl biguanide, N1-pyrrolidine-N5-(N-biphenyl-3-yl) biguanide, N1-pyrrolidine-N5-(N-biphenyl-4-yl) biguanide, N1-pyrrolidine-N5-(4-fluorobiphenyl-3-yl) biguanide, N1-pyrrolidine-N5-(3-fluorobiphenyl-4-yl) biguanide, N1-pyrrolidine-N5-(4-fluoro)phenethyl biguanide, N1-pyrrolidine-N5-(2-phenylpropane-2-yl) biguanide, N1-pyrrolidine-N5-(5,6,7,8-tetrahydronaphthalene-2-yl) biguanide, N1-pyrrolidine-N5-((1,2,3,4-tetrahydronaphthalene-1-yl) biguanide, N1-(S)-2-methyl pyrrolidine-N5-(4-(trifluoromethoxy)phenyl biguanide, N1-(S)-3-hydroxy pyrrolidine-N5-(4-trifluoromethoxy)phenyl biguanide, N1-(R)-3-hydroxy pyrrolidine-N5-(4-trifluoromethoxy)phenyl biguanide, N-ethyl-4-((3-(imino(pyrrolidin-1-yl)methyl)guanidino)methyl)benzamide, N-ethyl-3-((3-(imino(pyrrolidin-1-yl)methyl)guanidino)methyl)benzamide, N1-pyrrolidine-N5-(4-(2-aminoethyl)phenyl biguanide, N(N-(4-(2-oxo-2-(piperidin-1-yl)ethylthio)phenyl)carbamimidoyl)pyrrolidine-1-carboximidamide, and (S)-3-hydroxy-N(N-(4-(2-oxo-2-(piperidin-1-yl)ethylthio)phenyl)carbamimidoyl)pyrrolidine-1-carboximidaide.

3. The compound according to claim 1, wherein the pharmaceutically acceptable salt is an acid addition salt of an acid selected from the group consisting of formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, triflouroacetic acid, malic acid, maleic acid, malonic acid, fumaric acid, succinic acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, dichloroacetic acid, aminooxy acetic acid, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid and boric acid.

4. A pharmaceutical composition comprising the compound according to claim 1 or a pharmaceutical salt thereof as an active ingredient, and a pharmaceutically acceptable carrier.

Description

MODE FOR INVENTION

(1) A better understanding of the present invention may be obtained through the following examples which are set forth to illustrate, but are not to be construed as the limit of the present invention.

Example 1: Preparation of N1-pyrrolidine-N5-(benzo[1,3]dioxol-5-ylmethyl) biguanide.HCl

(2) ##STR00017##

(3) To a solution (20 mL) of piperonyl amine (0.8 ml, 6.58 mmol) dissolved in butanol was added by strong hydrochloric acid (0.6 mL, 6.58 mmol) at room temperature. The reaction mixture was added by N1-pyrrolidine cyanoguanidine (1.0 g, 7.24 mmol) and then stirred with reflux for 18 hr. The reaction mixture was concentrated under reduced pressure, separated and purified by a chromatography using MC:MeOH=9:1. The title compound was produced in white solid (1.5 g, 70%).

(4) .sup.1H NMR (400 MHz, CD.sub.3OD) 6.94 (s, 1H), 6.83 (s, 2H), 5.98 (s, 2H), 4.31 (s, 2H), 3.55 (d, 4H), 1.51 (s, 4H) LC-MS m/z 290.2 [M+1]

Example 2: Preparation of N1-pyrrolidine-N5-(4-methoxy)phenyl biguanide.HCl

(5) ##STR00018##

(6) The title compound was produced according to the substantially same method as described in Example 1, except that 4-methoxy aniline was used instead of piperonyl amine.

(7) .sup.1H NMR (400 MHz, DMSO) 9.22 (bs, 1H), 7.39 (bs, 2H), 7.27 (d, J=8.8 Hz, 2H), 6.88 (d, J=8.8 Hz, 2H), 6.72 (bs, 2H), 3.37 (m, 4H), 1.98 (m, 4H), LC-MS m/z 262.2 [M+1]

Example 3: Preparation of N1-pyrrolidine-N5-(3-chloro-4-methoxy)phenyl biguanide.HCl

(8) ##STR00019##

(9) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that 3-chloro-4-methoxy aniline was used instead of piperonyl amine

(10) .sup.1H NMR (400 MHz, DMSO) 9.22 (bs, 1H), 7.53 (d, J=2.4 Hz, 1H), 7.51 (bs, 2H), 7.25 (dd, J=8.8 Hz, 1H), 7.09 (d, J=8.8 Hz, 1H), 6.80 (bs, 2H), 3.37 (m, 4H), 1.90 (m, 4H), LC-MS m/z 296.1 [M+1]

Example 4: Preparation of N1-pyrrolidine-N5-(2,6-difluoro-N5-methyl)phenyl biguanide.HCl

(11) ##STR00020##

(12) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that 2,6-difluoro N-methylaniline was used instead of piperonyl amine

(13) .sup.1H NMR (600 MHz, CD.sub.3OD) 6.42 (s, 1H), 7.10 (m, 2H), 3.36 (m, 4H), 3.28 (s, 3H), 1.93 (m, 4H) LC-MS m/z 282.1 [M+1]

Example 5: Preparation of N1-pyrrolidine-N5-(4-dimethylamino)phenyl biguanide.HCl

(14) ##STR00021##

(15) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that 4-dimethylamino aniline was used instead of piperonyl amine.

(16) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.15 (d, J=8.8 Hz, 2H), 6.77 (d, J=8.8 Hz, 2H), 3.41 (m, 4H), 2.91 (s, 6H), 1.95 (m, 4H) LC-MS m/z 275.2 [M+1]

Example 6: Preparation of N1-pyrrolidine-N5-(4-isopropyl)phenyl biguanide.HCl

(17) ##STR00022##

(18) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that 4-isopropyl aniline was used instead of piperonyl amine.

(19) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.27 (d, J=8.8 Hz, 2H), 7.20 (d, J=8.8 Hz, 2H), 3.48 (m, 4H), 2.90 (m, 1H), 2.03 (m, 4H), 1.24 (d, J=7.2 Hz, 6H), LC-MS m/z 274.2 [M+1]

Example 7: Preparation of N1-pyrrolidine-N5-(3-phenoxy)phenyl biguanide.HCl

(20) ##STR00023##

(21) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that 3-phenoxy aniline was used instead of piperonyl amine.

(22) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.37 (m, 2H), 7.26 (m, 2H), 7.21 (m, 1H), 7.14 (m, 1H), 7.01 (m, 3H), 6.72 (m, 4H), 2.03 (m, 4H) LC-MS m/z 346.1 [M+1]

Example 8: Preparation of N1-pyrrolidine-N5-(N-biphenyl-3-yl) biguanide.HCl

(23) ##STR00024##

(24) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that N-biphenyl-3-amine was used instead of piperonyl amine.

(25) .sup.1H NMR (400 MHz, CD.sub.3OD) 7.63 (t, J=1.7 Hz, 1H), 7.59 (d, J=8.2 Hz, 2H), 7.38 (m, 6H) 3.51 (br d, 4H), 2.02 (Br d, 4H) LC-MS m/z 308.2 [M+1]

Example 9: Preparation of N1-pyrrolidine-N5-(N-biphenyl-4-yl) biguanide.HCl

(26) ##STR00025##

(27) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that N-biphenyl-4-amine was used instead of piperonyl amine.

(28) .sup.1H NMR (400 MHz, CD.sub.3OD) 7.60 (m, 4H), 7.47 (m, 4H), 7.31 (t, J=7.5 Hz, 1H), 3.52 (br d, J=41.8 Hz, 4H), 2.04 (br d, J=32.9 Hz, 4H) LC-MS m/z 308.2 [M+1]

Example 10: Preparation of N1-pyrrolidine-N5-(4-fluorobiphenyl-3-yl) biguanide.HCl

(29) ##STR00026##

(30) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that 4-fluorobiphenyl-3-amine was used instead of piperonyl amine

(31) .sup.1H NMR (400 MHz, CD.sub.3OD) 7.61 (m, 3H), 7.33 (m, 3H), 7.17 (m, 2H), 3.48 (br d, J=37.9 Hz, 4H), 2.04 (br d, J=36.3 Hz, 4H) LC-MS m/z 326.2 [M+1]

Example 11: Preparation of N1-pyrrolidine-N5-(3-fluorobiphenyl-4-yl) biguanide.HCl

(32) ##STR00027##

(33) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that 3-fluorobiphenyl-4-amine was used instead of piperonyl amine

(34) .sup.1H NMR (400 MHz, CD.sub.3OD) 7.59 (d, J=6.6 Hz, 2H), 7.48 (m, 4H), 7.35 (d, J=9.4 Hz, 1H), 7.05 (m, 1H), 3.53 (br d, J=46.1 Hz, 4H), 2.04 (br d, J=34.4 Hz, 4H) LC-MS m/z 326.2 [M+1]

Example 12: Preparation of N1-pyrrolidine-N5-(4-fluoro)phenethyl biguanide.HCl

(35) ##STR00028##

(36) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that 4-fluorophenethyl amine was used instead of piperonyl amine.

(37) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.24 (m, 2H), 7.19 (m, 2H), 3.58 (m, 6H), 2.83 (t, J=7.2 Hz, 2H), 2.08 (m, 4H) LC-MS m/z 278.2 [M+1]

Example 13: Preparation of N1-pyrrolidine-N5-(2-phenylpropane-2-yl) biguanide.HCl

(38) ##STR00029##

(39) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that 2-phenylpropane-2-amine was used instead of piperonyl amine.

(40) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.39(d, J=8.4 Hz, 2H), 7.3(t, J=8.4 Hz, 2H), 7.18 (t, J=8.4 Hz, 1H), 3.19 (m, 2H), 2.81 (m, 2H), 1.90 (m, 2H), 1.66 (s, 8H) LC-MS m/z 274.2 [M+1]

Example 14: Preparation of N1-pyrrolidine-N5-(5,6,7,8-tetrahydronaphthalene-2-yl) biguanide.HCl

(41) ##STR00030##

(42) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that 5,6,7,8-tetrahydronaphthalene-2-amine was used instead of piperonyl amine.

(43) .sup.1H (600 MHz, CD.sub.3OD) 7.03 (m, 3H), 3.48 (m, 4H), 2.73 (m, 4H), 2.02 (m, 4H), 1.79 (m, 4H) LC-MS m/z 286.2 [M+1]

Example 15: Preparation of N1-pyrrolidine-N5-(1,2,3,4-tetrahydronaphthalen-1-yl) biguanide.HCl

(44) ##STR00031##

(45) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that 1,2,3,4-tetrahydronaphthalene-1-amine was used instead of piperonyl amine.

(46) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.33 (m, 1H), 7.15 (m, 2H), 7.10 (m, 1H), 5.03 (s, 1H), 3.46 (brs, 4H), 2.85 (m, 2H), 2.051.79 (m, 10H), LC-MS m/z 286.2 [M+1]

Example 16: Preparation of N1-(S)-2-methyl pyrrolidine-N5-(4-(trifluoromethoxy)phenyl biguanide.HCl

(47) ##STR00032##

(48) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that (S)-2-methyl pyrrolidine cyanoguanidine was used instead of pyrrolidine cyanoguanidine, and 4-(trifluoromethoxy)aniline was used instead of piperonyl amine.

(49) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.37 (m, 2H), 7.31 (m, 3H), 7.22 (d, J=9.0 Hz), 7.10 (t, J=7.2 Hz, 1H), 6.99 (m, 4H), 6.80 (dd, J=1.8 Hz, 1H) LC-MS m/z 330.1 [M+1]

Example 17: Preparation of N1-(S)-3-hydroxy pyrrolidine-N5-(4-trifluoromethoxy)phenyl biguanide.HCl

(50) ##STR00033##

(51) The title compound in a white solid was produced according to the substantially same method as described in Example 1, except that (S)-3-hydroxy pyrrolidine cyanoguanidine was used instead of pyrrolidine cyanoguanidine, and 4-(trifluoromethoxy)aniline was used instead of piperonyl amine.

(52) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.47 (m, 2H), 7.22 (m, 2H), 4.42 (brs, 1H), 3.54 (m, 5H), 2.01 (m, 2H) LC-MS m/z 332.1 [M+1]

Example 18: Preparation of N1-4-hydroxy piperidine-N5-(4-trifluoromethoxy)phenyl biguanide.HCl

(53) ##STR00034##

(54) To a solution (20 mL) of 4-trifluoromethoxyphenyl(0.77 ml, 4.76 mmol) dissolved in butanol was added by strong hydrochloric acid (0.40 mL, 4.76 mmol) at room temperature. The reaction mixture was added by 4-hydroxy piperidine cyanoguanine (0.3 g, 4.76 mmol) prepared by Reaction Scheme 1 and then stirred with reflux for 12 hr. The reaction mixture was concentrated under reduced pressure, and then was stirred with addition of ethyl acetate 20 ml for 2 hr. The obtained solid was filtrated, was washed with ethyl acetate, and dried under reduced pressure. The title compound was produced in white solid (54.8 mg, 8.2%).

(55) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.46 (m, 2H), 7.26 (d, J=8.4 Hz, 2H), 3.88 (m, 3H), 3.34 (m, 3H), 1.90 (m, 2H), 1.55 (m, 2H) LC-MS m/z 346.1 [M+1]

Example 19: Preparation of N1-(R)-3-hydroxy pyrrolidine-N5-(4-trifluoromethoxy)phenyl biguanide.HCl

(56) ##STR00035##

(57) The title compound in a white solid was produced according to the substantially same method as described in Example 18, except that (R)-3-hydroxy pyrrolidine cyanoguanidine was used instead of 4-hydroxy piperidine.

(58) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.48 (d, J=9.0 Hz, 2H), 7.22 (d, J=9.0 Hz, 2H), 4.54 (brs, 1H), 3.55 (m, 4H), 1.07 (m, 2H) LC-MS m/z 332.1 [M+1]

Example 20: Preparation of N1-3-hydroxy methyl pyrrolidine-N5-(4-trifluoromethoxy)phenyl biguanide.HCl

(59) ##STR00036##

(60) The title compound in a white solid was produced according to the substantially same method as described in Example 18, except that 3-hydroxy methyl pyrrolidine cyanoguanidine was used instead of 4-hydroxy piperidine cyanoguanine.

(61) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.48 (m, 12H), 7.22 (m, 2H), 3.446 (m, 6H), 3.19 (m, 1H), 2.56 (m, 1H), 1.93 (m, 2H) LC-MS m/z 346.1 [M+1]

Example 21: Preparation of N1-4-hydroxy piperidine-N5-(4-phenoxy)phenyl biguanide.HCl

(62) ##STR00037##

(63) The title compound in a white solid was produced according to the substantially same method as described in Example 18, except that 4-phenoxyphenyl aniline was used instead of 4-trifluoromethoxy phenyl.

(64) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.33 (m, 4H), 7.09 (m, 1H), 6.96 (m, 4H), 3.85 (m, 3H), 3.27 (m, 3H), 1.88 (m, 2H), 1.50 (m, 2H) LC-MS m/z 354.2 [M+1]

Example 22: Preparation of N1-4-hydroxy piperidine-N5-(3-phenoxy)phenyl biguanide.HCl

(65) ##STR00038##

(66) The title compound in a white solid was produced according to the substantially same method as described in Example 18, except that 4-phenoxyphenyl aniline was used instead of 4-trifluoromethoxy phenyl.

(67) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.35 (t, J=7.8 Hz, 2H), 7.26 (t, J=7.8 Hz, 1H), 7.16 (m, 1H), 7.12 (t, J=7.2 Hz, 1H), 7.00 (m, 3H), 6.72 (m, 1H), 3.86 (m, 3H), 3.27 (m, 3H), 1.88 (m, 2H), 1.52(m, 2H) LC-MS m/z 354.2 [M+1]

Example 23: Preparation of N1-pyrrolidine-N5-4-benzamide biguanide.HCl

(68) ##STR00039##

(69) The title compound in a white solid was produced according to the substantially same method as described in Example 18, except that pyrrolidine cyanoguanidine was used instead of 4-hydroxy piperidine cyanoguanidine, 4-benzamide aniline was used instead of 4-trifluoromethoxy phenyl.

(70) .sup.1H NMR (400 MHz, DMSO-d6) 9.67 (s, 1H), 7.82 (s, 1H), 7.80 (d, J=8.8 Hz, 2H), 7.62 (s, 1H), 7.47 (d, J=8.8 Hz, 2H), 7.23 (s, 1H), 6.58 (s, 2H), 3.36 (m, 4H), 1.93 (m, 4H) LC-MS m/z 275.2 [M+1]

Example 24: Preparation of (E)-N(N-(4-(3-oxo-3-(piperidin-1-yl)prop-1-enyl)phenyl)carbamimidoyl)pyrrolidine-1-carboximidamide.HCl

(71) ##STR00040##

(72) The title compound in a white solid was produced according to the substantially same method as described in Example 18, except that pyrrolidine cyanoguanidine was used instead of 4-hydroxy piperidine, and (E)-3-(4-aminophenyl)-1-(piperidin-1-yl)prop-2-en-1-on was used instead of 4-trifluoromethoxy aniline.

(73) .sup.1H NMR (600 MHz, DMSO-d6) 7.62 (d, J=8.4 Hz, 2H), 7.44 (d, J=8.4 Hz, 2H), 7.41 (d, J=15.6 Hz, 1H), 7.13 (d, J=15.6 Hz, 1H), 3.62 (m, 8H), 1.94 (m, 4H), 1.61 (m, 6H) LC-MS m/z 369.2 [M+1]

Example 25: Preparation of N(N-(4-(piperidine-1-carbonyl)benzyl)carbamimidoyl)pyrrolidine-1-carboximidamide.HCl

(74) ##STR00041##

(75) The title compound in a white solid was produced according to the substantially same method as described in Example 18, except that pyrrolidine cyanoguanidine was used instead of 4-hydroxy piperidine, and (4-aminomethyl)phenyl)(piperidin-1-yl)methanone was used instead of 4-trifluoromethoxy aniline.

(76) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.43 (d, J=7.8 Hz, 2H), 7.37 (d, J=7.8 Hz, 2H), 4.57 (s, 2H), 3.59 (m, 8H), 2.04(m, 4H), 1.72 (m, 6H) LC-MS m/z 368.2 [M+2]

Example 26: Preparation of N(N-(4-(3-oxo-3-(piperidin-1-yl)propyl)phenyl)carbamimidoyl)pyrrolidine-1-carboximidamide.HCl

(77) ##STR00042##

(78) The title compound in a white solid was produced according to the substantially same method as described in Example 18, except that pyrrolidine cyanoguanidine was used instead of 4-hydroxy piperidine, and (E)-3-(4-aminophenyl)-1-(piperidin-1-yl)propyl-1-on was used instead of 4-trifluoromethoxy aniline.

(79) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.30 (d, J=8.8 Hz, 2H), 7.20 (d, J=8.8 Hz, 2H), 3.52 (m, 8H), 2.89 (t, J=7.2 Hz, 2H), 2.68(t, J=7.2 Hz, 2H), 2.05 (m, 4H), 1.61 (m, 6H) LC-MS m/z 371.3 [M+1]

Example 27: Preparation of N-ethyl-4-((3-(imino(pyrrolidin-1-yl)methyl)guanidino)methyl)benzamide.HCl

(80) ##STR00043##

(81) The title compound in a white solid was produced according to the substantially same method as described in Example 18, except that pyrrolidine cyanoguanidine was used instead of 4-hydroxy piperidine, and 4-(aminomethyl)-N-ethylbenzamide was used instead of 4-trifluoromethoxy aniline.

(82) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.77 (d, J=12 Hz, 2H), 7.397 (d, J=18 Hz, 2H), 4.46(s, 2H), 3.36 (m, 6H), 1.93 (m, 4H), 1.21 (t, J=12 Hz, 3H) LC-MS m/z 317.2 [M+1]

Example 28: Preparation of N(N-(4-(2-oxo-2-(piperidin-1-yl)ethylthio)phenyl)carbamimidoyl)pyrrolidine-1-carboximidamide.HCl

(83) ##STR00044##

(84) The title compound in a white solid was produced according to the substantially same method as described in Example 18, except that pyrrolidine cyanoguanidine was used instead of 4-hydroxy piperidine, and 2-(4-aminophenyl thio)-1-(piperidin-1-yl)ethanone was used instead of 4-trifluoromethoxy aniline.

(85) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.40 (d, J=18 Hz, 2H), 7.32 (d, J=18 Hz, 2H), 3.75 (s, 2H), 3.49 (m, 8H), 1.95 (m, 4H), 1.51 (m, 4H) LC-MS m/z 389.2 [M+1]

Example 29: Preparation of (S)-3-hydroxy-N(N-(4-(2-oxo-2-(piperidin-1-yl)ethylthio)phenyl)carbamimidoyl)pyrrolidine-1-carboximidamide.HCl

(86) ##STR00045##

(87) The title compound in a white solid was produced according to the substantially same method as described in Example 18, except that (S)-2-hydroxy pyrrolidine cyanoguanidine was used instead of 4-hydroxy piperidine, and 2-(4-aminophenyl thio)-1-(piperidin-1-yl)ethanone was used instead of 4-trifluoromethoxy aniline.

(88) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.40 (d, J=18 Hz, 2H), 7.32 (d, J=18 Hz, 2H), 4.58 (brs, 1H), 3.75(s, 2H), 3.51 (m, 3H), 2.06(m. 2H), 1.63 (m, 5H) LC-MS m/z 405.2 [M+1]

Example 30: Preparation of N-ethyl-3-((3-(imino(pyrrolidin-1-yl)methyl)guanidino)methyl)benzamide.HCl

(89) ##STR00046##

(90) The title compound in a white solid was produced according to the substantially same method as described in Example 18, except that pyrrolidine cyanoguanidine was used instead of 4-hydroxy piperidine, and 3-(aminomethyl)-N-ethylbenzamide was used instead of 4-trifluoromethoxy aniline.

(91) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.83 (m, 1H), 7.69 (d, J=7.8 Hz, 1H), 7.49 (d, J=7.8 Hz, 1H), 7.42(t, J=7.8 Hz, 1H), 4.47 (s, 2H), 3.41 (m, 4H), 3.37(t, J=6.0 Hz, 2H), 1.95 (m, 4H), 1.21 (m, 3H) LC-MS m/z 317.2 [M+1]

Example 31: Preparation of N1-4-methyl piperazine-N5-(4-bromo)phenyl biguanide.HCl

(92) ##STR00047##

(93) To a solution (10 mL) of 4-bromo aniline (154 mg, 0.897 mmol) dissolved in butanol was added by strong hydrochloric acid (0.11 mL, 0.897 mmol) at room temperature. The reaction mixture was added by methyl piperazine cyanoguanidine (150 mg, 0.897 mmol) and then stirred with reflux for 18 hr. The reaction mixture was concentrated under reduced pressure, and then was separated and purified by a chromatography using MC:MeOH=9:1. The title compound was produced in white solid phase (90.2 mg, 27%).

(94) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.47 (m, 2H), 7.29 (m, 2H), 3.58 (m, 4H), 2.51 (m, 4H), 2.35 (s, 3H); LC-MS m/z 340.0 [M+1]+; mp 227229 C.

Example 32: Preparation of N1-4-methyl piperazine-N5-(3-bromo)phenyl biguanide.HCl

(95) ##STR00048##

(96) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 3-bromo aniline was used instead of 4-bromo aniline.

(97) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.44 (m, 1H), 7.05 (m, 3H), 3.39 (m, 3H), 2.31 (m, 3H), 2.14 (s, 3H); LC-MS m/z 340.4 [M+1]+; mp 228229 C.

Example 33: Preparation of N1-4-methyl piperazine-N5-(2-bromo)phenyl biguanide.HCl

(98) ##STR00049##

(99) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 2-bromo aniline was used instead of 4-bromo aniline.

(100) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.44 (m, 1H), 7.37 (m, 1H), 7.15 (m, 1H), 6.93 (m, 1H), 3.31 (m, 4H), 2.26 (m, 4H), 2.11 (s, 3H); LC-MS m/z 340.4 [M+1]+; mp 161163 C.

Example 34: Preparation of N1-4-methyl piperazine-N5-(3-bromo)benzyl biguanide.HCl

(101) ##STR00050##

(102) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 3-bromobenzyl amine was used instead of 4-bromo aniline.

(103) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.56 (m, 1H), 7.35 (m, 2H) 7.18 (m, 1H), 4.49 (m, 2H), 3.46 (m, 4H), 2.42 (m, 4H), 2.29 (s, 3H); LC-MS m/z 354.4 [M+1]+; mp 216218 C.

Example 35: Preparation of N1-4-methyl piperazine-N5-(4-fluoro-3-trifluoromethyl)phenyl biguanide.HCl

(104) ##STR00051##

(105) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 4-fluoro-3-trifluoromethyl aniline was used instead of 4-bromo aniline.

(106) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.78 (m, 1H), 7.58 (m, 1H), 7.30 (m, 1H), 3.57 (m, 4H), 2.49 (m, 4H), 2.34 (s, 3H); LC-MS m/z 347.2 [M+1]+; mp 254256 C.

Example 36: Preparation of N1-4-methyl piperazine-N5-(3-chloro-4-trifluoromethoxy)phenyl biguanide.HCl

(107) ##STR00052##

(108) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 3-chloro-4-trifluoromethoxy aniline was used instead of 4-bromo aniline.

(109) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.72 (m, 1H), 7.37 (m, 2H), 3.60 (m, 4H), 2.53 (m, 4H), 2.35 (s, 3H); LC-MS m/z 379.4 [M+1]+; mp 229231 C.

Example 37: Preparation of N1-4-methyl piperazine-N5-(3-fluoro-4-trifluoromethoxy)phenyl biguanide.HCl

(110) ##STR00053##

(111) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 3-fluoro-4-trifluoromethoxy aniline was used instead of 4-bromo aniline.

(112) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.71 (m, 1H), 7.50 (m, 1H), 7.29 (m, 1H), 3.61 (m, 4H), 2.53 (m, 4H), 2.36 (s, 3H); LC-MS m/z 363.5 [M+1]+; mp 230232 C.

Example 38: Preparation of N1-4-methyl piperazine-N5-(4-chloro-3-trifluoromethyl)phenyl biguanide.HCl

(113) ##STR00054##

(114) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 4-chloro-3-trifluoromethyl aniline was used instead of 4-bromo aniline.

(115) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.89 (m, 1H), 7.57 (m, 2H), 3.56 (m, 4H), 2.82 (m, 4H), 2.55 (s, 3H); LC-MS m/z 363.5 [M+1]+; mp 231233 C.

Example 39: Preparation of N1-4-methyl piperazine-N5-(4-chloro-3-trifluoromethoxy)phenyl biguanide.HCl

(116) ##STR00055##

(117) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 4-chloro-3-trifluoromethoxy aniline was used instead of 4-bromo aniline.

(118) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.55 (m, 1H), 7.49 (m, 1H), 7.31 (m, 1H), 4.33 (m, 2H), 3.49 (m, 2H), 3.44 (m, 2H), 3.23 (m, 2H), 2.93 (s, 3H); LC-MS m/z 379.2 [M+1]+; mp 180182 C.

Example 40: Preparation of N1-4-methyl piperazine-N5-(4-fluoro-3-trifluoromethoxy)phenyl biguanide.HCl

(119) ##STR00056##

(120) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 4-fluoro-3-trifluoromethoxy aniline was used instead of 4-bromo aniline.

(121) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.62 (m, 1H), 7.29 (m, 2H), 4.49 (s, 2H), 3.56 (m, 4H), 2.88 (m, 4H), 2.71 (s, 3H); LC-MS m/z 363.2 [M+1]+; mp 216218 C.

Example 41: Preparation of N1-4-methyl piperazine-N5-(4-fluoro-3-trifluoromethoxy)benzyl biguanide.HCl

(122) ##STR00057##

(123) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 4-fluoro-3-trifluoromethyl benzyl amine was used instead of 4-bromo aniline.

(124) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.34 (m, 3H), 4.40 (s, 2H), 3.52 (m, 4H), 2.52 (m, 4H), 3.37 (s, 3H); LC-MS m/z 377.2 [M+1]+; mp 222224 C.

Example 42: Preparation of N1-4-methyl piperazine-N5-(3-fluoro-4-trifluoromethyl)benzyl biguanide.HCl

(125) ##STR00058##

(126) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 3-fluoro-4-trifluoromethyl benzyl amine was used instead of 4-bromo aniline.

(127) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.64 (m, 1H), 7.28 (m, 2H), 4.49 (s, 2H), 3.66 (m, 4H), 3.02 (m, 4H), 2.67 (s, 3H); LC-MS m/z 361.2 [M+1]+; mp 225227 C.

Example 43: Preparation of N1-4-methyl piperazine-N5-(3-fluoro-4-trifluoromethoxy)benzyl biguanide.HCl

(128) ##STR00059##

(129) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 3-fluoro-4-trifluoromethoxy benzyl amine was used instead of 4-bromo aniline.

(130) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.36 (m, 3H), 4.42 (s, 2H), 3.53 (m, 4H), 2.52 (m, 4H), 2.38 (s, 3H); LC-MS m/z 377.2 [M+1]+; mp 221223 C.

Example 44: Preparation of N1-4-methyl piperazine-N5-(3-fluoro-4-trifluoromethyl)phenyl biguanide.HCl

(131) ##STR00060##

(132) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 3-fluoro-4-trifluoromethyl benzyl amine was used instead of 4-bromo aniline.

(133) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.59 (m, 2H), 7.26 (m, 1H), 3.62 (m, 4H), 2.53 (m, 4H), 2.34 (s, 3H); LC-MS m/z 347.2 [M+1]+; mp 224226 C.

Example 45: Preparation of N1-4-ethyl piperazine-N5-(4-trifluoromethoxy)phenyl biguanide.HCl

(134) ##STR00061##

(135) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that ethyl piperazine cyanoguanidine was used instead of methyl piperazine cyanoguanidine, and 3-trifluoromethoxy aniline was used instead of 4-bromo aniline.

(136) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.44 (m, 2H), 7.23 (m, 2H), 3.59 (m, 4H), 3.17 (m, 1H), 2.56 (m, 4H), 2.49 (m, 1H) 1.12 (m, 3H); LC-MS m/z 359.2 [M+1]+; mp 242244 C.

Example 46: Preparation of N1-(2R,6S)-2,6-dimethyl piperidine-N5-(4-(2-aminoethyl)phenyl biguanide.HCl

(137) ##STR00062##

(138) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 2,6-dimethyl piperidine was used instead of methyl piperazine cyanoguanidine, and cyanoguanidine, 4-(2-aminoethyl) aniline was used instead of 4-bromo aniline.

(139) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.38 (d, 2H), 7.25 (d, 2H), 4.604.40 (s, 2H), 3.16 (t, 2H), 2.94 (t, 2H), 1.901.51 (m, 6H) 1.30 (s, 6H); LC-MS m/z 317.3 [M+1]; mp 230235 C.

Example 47: Preparation of N1-pyrrolidine-N5-(4-(2-aminoethyl)phenyl biguanide.HCl

(140) ##STR00063##

(141) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that pyrrolidine cyano was used instead of methyl piperazine cyanoguanidine, and guanidine, 4-(2-aminoethyl)aniline was used instead of 4-bromo aniline.

(142) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.39 (d, 2H), 7.25 (d, 2H), 3.48 (d, 4H), 3.16 (t, 2H), 2.94 (t, 2H), 2.05 (d, 4H); LC-MS m/z 275.2 [M+1]; mp 230235 C.

Example 48: Preparation of N1-2-methyl piperidine-N5-(4-(2-aminoethyl)phenyl biguanide.HCl

(143) ##STR00064##

(144) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 2-methyl piperidine cyanoguanidine was used instead of methyl piperazine cyanoguanidine, and 4-(2-aminoethyl)aniline was used instead of 4-bromo aniline.

(145) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.37 (d, 2H), 7.27 (d, 2H), 4.41 (s, 1H), 3.95 (d, 1H), 3.16 (m, 2H), 3.11 (m, 1H), 2.90 (m, 2H), 1.751.50 (m, 6H), 1.26 (d, 3H); LC-MS m/z 303.3 [M+1]; mp 230235 C.

Example 49: Preparation of N1-3,5-dimethyl piperidine-N5-(4-(2-aminoethyl)phenyl biguanide.HCl

(146) ##STR00065##

(147) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 3,5-dimethyl piperidine cyanoguanidine was used instead of methyl piperazine cyanoguanidine, and 4-(2-aminoethyl)aniline was used instead of 4-bromo aniline.

(148) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.36 (d, 2H), 7.26 (d, 2H), 4.03 (s, 2H), 3.16 (t, 2H), 2.9 (t, 2H), 2.46 (t, 2H), 1.87 (d, 1H), 1.68 (m, 2H), 0.94 (d, 6H), 0.88 (m, 1H); LC-MS m/z 317.3 [M+1]; mp 230235 C.

Example 50: Preparation of N1-3-methyl piperidine-N5-(4-(2-aminoethyl)phenyl biguanide.HCl

(149) ##STR00066##

(150) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 3-methyl piperidine cyanoguanidine was used instead of methyl piperazine cyanoguanidine, and 4-(2-aminoethyl)aniline was used instead of 4-bromo aniline.

(151) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.37 (d, 2H), 7.26 (d, 2H), 3.96 (dd, 2H), 3.16 (t, 2H), 2.99 (m, 1H), 2.94 (t, 2H), 2.67 (t, 1H), 1.86 (m, 1H), 1.74 (m, 1H), 1.67 (m, 1H), 1.57 (m, 1H), 0.94 (d, 3H); LC-MS m/z 303.2 [M+1]+; mp 230235 C.

Example 51: Preparation of N1-3,3-difluoro pyrrolidine-N5-(4-trifluoromethoxy)phenyl biguanide.HCl

(152) ##STR00067##

(153) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 3,3-difluoro pyrrolidinecyanoguanidine was used instead of methyl piperazine cyanoguanidine, and 4-(trifluoromethoxy)aniline was used instead of 4-bromo aniline.

(154) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.45 (d, J=9.0 Hz, 2H), 7.26 (d, J=8.4 Hz, 2H), 3.82 (t, J=12.6 Hz, 2H), 3.70 (t, J=6.6 Hz, 2H), 2.51 (s, 2H) LC-MS m/z 352.2 [M+1]

Example 52: Preparation of N1-4,4-difluoro piperidine-N5-(4-trifluoromethoxy)phenyl biguanide.HCl

(155) ##STR00068##

(156) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 4,4-difluoro piperidine cyanoguanidine was used instead of methyl piperazine cyanoguanidine, and 4-(trifluoromethoxy)aniline was used instead of 4-bromo aniline.

(157) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.43 (d, J=9.0 Hz, 2H), 7.25 (d, J=7.8 Hz, 2H), 3.68 (t, J=6.0 Hz, 4H), 2.06 (m, 4H) LC-MS m/z 366.2 [M+1]

Example 53: Preparation of N1-4-(methylsulfonyl)piperidine-N5-(4-trifluoromethoxy)phenyl biguanide.HCl

(158) ##STR00069##

(159) The title compound in a white solid was produced according to the substantially same method as described in Example 31, except that 4-methyl sulfonyl piperidine cyanoguanidine was used instead of methyl piperazine cyanoguanidine, and 4-(trifluoromethoxy)aniline was used instead of 4-bromo aniline.

(160) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.45 (d, J=9.0 Hz, 2H), 7.25 (d, J=7.8 Hz, 2H), 4.26 (d, J=13.8 Hz, 2H), 3.42 (m, 2H), 3.09 (t, J=12.0 Hz, 2H), 2.20 (d, J=10.8 Hz, 2H), 1.82 (m, 3H) LC-MS m/z 408.1 [M+1]

Example 54: Preparation of N5-(2,4-(dichloro)phenyl biguanide.HCl

(161) ##STR00070##

(162) To a solution (20 mL) of 2,4-dichloro aniline (0.64 g, 3.92 mmol) dissolved in butanol was added by strong hydrochloric acid (0.31 mL, 3.57 mmol) at room temperature. The reaction mixture was added by dicyandiamide (0.3 g, 3.57 mmol) and then stirred with reflux for 2 hr. The reaction mixture was concentrated under reduced pressure, and then stirred with addition of ethyl acetate 20 ml for 2 hr. The obtained solid was filtrated, was washed with ethyl acetate, and dried under reduced pressure. The title compound was produced in white solid phase (0.5 g, 54%).

(163) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.50 (d, 1H), 7.31 (d, 2H); LC-MS m/z 246.1 [M]+; mp 230235 C.

Example 55: Preparation of N5-4-fluorophenyl biguanide.HCl

(164) ##STR00071##

(165) The title compound in a white solid was produced according to the substantially same method as described in Example 54, except that 4-fluoroaniline was used instead of 2,4-dichloro aniline.

(166) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.36 (d, 2H), 7.08 (d, 2H); LC-MS m/z 196.2 [M+1]+; mp 230235 C.

Example 56: Preparation of N5-(3,4-(dichloro)phenyl biguanide.HCl

(167) ##STR00072##

(168) The title compound in a white solid was produced according to the substantially same method as described in Example 54, except that 3,4-dichloro aniline was used instead of 2,4-dichloro aniline.

(169) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.66 (s, 1H), 7.43 (d, 1H), 7.28 (d, 1H); LC-MS m/z 246.1 [M]+; mp 230235 C.

Example 57: Preparation of N5-(2-chloro-5-trifluoromethyl)phenyl biguanide.HCl

(170) ##STR00073##

(171) The title compound in a white solid was produced according to the substantially same method as described in Example 54, except that 2-chloro-5-trifluoromethyl aniline was used instead of 2,4-dichloro aniline.

(172) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.580 (s, 1H), 7.27 (s, 1H), 7.23 (t, 1H); LC-MS m/z 280.1 [M+1]+; mp 230235 C.

Example 58: Preparation of N5-(3-chloro-4-fluoro)phenyl biguanide.HCl

(173) ##STR00074##

(174) The title compound in a white solid was produced to the substantially same method as described in Example 54, except that 3-chloro-4-fluoro aniline was used instead of 2,4-dichloro aniline.

(175) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.60 (s, 1H), 7.26 (s, 1H), 7.19 (t, 1H); LC-MS m/z 230.1 [M+1]+; mp 230235 C.

Example 59: Preparation of N5-(2,3-dichloro)phenyl biguanide.HCl

(176) ##STR00075##

(177) The title compound in a white solid was produced according to the substantially same method as described in Example 54, except that 2,3-dichloro aniline was used instead of 2,4-dichloro aniline.

(178) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.58 (s, 1H), 7.34 (d, 1H), 7.28 (t, 1H); LC-MS m/z 246.0 [M]+; mp 230235 C.

Example 60: Preparation of N5-(4-trifluoromethylthio)phenyl biguanide.HCl

(179) ##STR00076##

(180) The title compound in a white solid was produced according to the substantially same method as described in Example 54, except that 4-trifluoromethylthio aniline was used instead of 2,4-dichloro aniline.

(181) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.63 (d, 2H), 7.54 (d, 2H); LC-MS m/z 278.0 [M+1]+; mp 230235 C.

Example 61: Preparation of N1,N1-dimethyl-N5-4-(2-oxo-2-(piperidin-1-yl)ethyl thio)phenyl biguanide.HCl

(182) ##STR00077##

(183) The title compound in a white solid was produced according to the substantially same method as described in Example 54, except that 2-oxo-2-(piperidine-1-yl)ethylthio aniline was used instead of 2,4-dichloro aniline.

(184) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.40 (d, J=18 Hz, 2H), 7.32 (d, J=18 Hz, 2H), 3.76(s, 2H), 3.51(m, 4H), 3.05(s. 6H), 1.63 (m, 4H) LC-MS m/z 363.2 [M+1]

Example 62: Preparation of N1-3-pyridine-N5-(4-trifluoromethyl)phenyl biguanide.HCl

(185) ##STR00078##

(186) To a solution (20 mL) of 4-trifluoromethyl aniline (0.3 g, 1.86 mmol) dissolved in butanol was added by strong hydrochloric acid (0.16 mL, 1.86 mmol) at room temperature. The reaction mixture was added by 3-pyridine cyanoguanine (0.3 g, 1.86 mmol) prepared by Reaction Scheme 1 and then stirred with reflux for 2 hr. The reaction mixture was concentrated under reduced pressure, and then was stirred by addition of ethyl acetate 20 ml for 2 hr. The obtained solid was filtrated, was washed with ethyl acetate, and dried under reduced pressure. The title compound was produced in white solid phase (54.8 mg, 8.2%).

(187) .sup.1H NMR (600 MHz, CD.sub.3OD) 8.50 (m, 3H), 7.79 (m, 2H), 7.65 (m, 1H), 7.60 (m, 1H), 7.53 (m, 1H) 7.47 (m, 1H); LC-MS m/z 323.2 [M+1]+; mp 251253 C.

Example 63: Preparation of N1-3-pyridine-N5-(4-trifluoromethoxy)phenyl biguanide.HCl

(188) ##STR00079##

(189) The title compound in a white solid was produced according to the substantially same method as described in Example 62, except that 4-trifluoromethoxy aniline was used instead of 4-trifluoromethyl aniline.

(190) .sup.1H NMR (600 MHz, CD.sub.3OD) 8.90 (m, 1H), 8.56 (m, 1H), 8.43 (m, 1H), 8.39 (m, 1H), 7.91 (m, 1H), 7.76 (m, 2H), 7.41 (m, 1H); LC-MS m/z 339.2 [M+1]+; mp 250252 C.

Example 64: Preparation of N1-3-pyridine-N5-(4-trifluoromethyl)benzyl biguanide.HCl

(191) ##STR00080##

(192) The title compound in a white solid was produced according to the substantially same method as described in Example 62, except that 4-trifluoromethyl benzyl amine was used instead of 4-trifluoromethyl aniline.

(193) .sup.1H NMR (600 MHz, DMSO) 8.49 (brs, 4H), 7.80 (m, 2H), 7.73 (m, 2H), 7.64 (m, 2H), 7.42 (m, 2H), 4.12 (s, 2H); LC-MS m/z 337.1 [M+1]+; mp 226228 C.

Example 65: Preparation of N1-3-pyridine-N5-(4-fluoro-3-trifluoromethyl)phenyl biguanide.HCl

(194) ##STR00081##

(195) The title compound in a white solid was produced according to the substantially same method as described in Example 62, except that 4-fluoro-3-trifluoromethyl aniline was used instead of 4-trifluoromethyl aniline.

(196) .sup.1H NMR (600 MHz, CD.sub.3OD) 8.51 (d, J=2.4 Hz, 1H), 8.29 (d, J=4.8 Hz, 1H), 7.81 (m, 1H), 7.66 (m, 1H), 7.65 (m, 1H), 7.39 (m, 1H), 7.29 (m, 1H); LC-MS m/z 341.1 [M+1]+; mp 254256 C.

Example 66: Preparation of N1-2-pyrazine-N5-(4-trifluoromethoxy)phenyl biguanide.HCl

(197) ##STR00082##

(198) The title compound in a white solid was produced according to the substantially same method as described in Example 62, except that 2-pyrazine cyanoguanidine was used instead of 3-pyridine cyanoguanine, and 4-trifluoromethoxy aniline was used instead of 4-trifluoromethyl aniline.

(199) .sup.1H NMR (600 MHz, CD.sub.3OD) 8.53 (m, 1H), 8.28 (m, 1H), 8.02 (m, 1H), 7.92 (m, 1H), 7.55 (m, 2H), 7.42 (m, 2H); LC-MS m/z 340.0 [M+1]+; mp 223225 C.

Example 67: Preparation of N1-2-pyrazine-N5-(4-trifluoromethyl)phenyl biguanide.HCl

(200) ##STR00083##

(201) The title compound in a white solid was produced according to the substantially same method as described in Example 62, except that 2-pyrazine cyanoguanidine was used instead of 3-pyridine cyanoguanine, and 4-trifluoromethyl aniline was used instead of 4-trifluoromethyl aniline.

(202) .sup.1H NMR (600 MHz, CD.sub.3OD) 8.55 (m, 1H), 8.32 (m, 1H), 7.83 (m, 1H), 7.60 (m, 2H), 7.50 (m, 2H), 7.39 (m, 1H); LC-MS m/z 324.0 [M+1]+; mp 223225 C.

Example 68: Preparation of N1-cyclopentyl-N5-(4-trifluoromethoxy)phenyl biguanide.HCl

(203) ##STR00084##

(204) The title compound in a white solid was produced according to the substantially same method as described in Example 62, except that cyclopentyl cyanoguanidine was used instead of 3-pyridine cyanoguanine, and 4-trifluoromethoxy aniline was used instead of 4-trifluoromethyl aniline.

(205) .sup.1H NMR (600 MHz, DMSO) 7.60 (brs, 4H), 7.37 (m, 2H), 7.35 (m, 2H), 4.11 (m, 1H) 1.64 (m, 8H); LC-MS m/z 330.3 [M+1]+; mp 271273 C.

Example 69: Preparation of N1-cyclopentyl-N5-(4-fluoro)phenyl biguanide.HCl

(206) ##STR00085##

(207) The title compound in a white solid was produced according to the substantially same method as described in Example 62, except that cyclopentyl cyanoguanidine was used instead of 3-pyridine cyanoguanine, and 4-fluoroaniline was used instead of 4-trifluoromethyl aniline.

(208) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.37 (m, 2H), 7.04 (m, 2H), 3.96 (brs, 1H), 2.00 (m, 2H), 1.70 (m, 2H), 1.58 (m, 4H); LC-MS m/z 264.2 [M+1]+; mp 272274 C.

Example 70: Preparation of N1-furan-2-ylmethyl-N5-(4-chloro)phenyl biguanide.HCl

(209) ##STR00086##

(210) The title compound in a white solid was produced according to the substantially same method as described in Example 62, except that furan-2-ylmethyl cyanoguanidine was used instead of 3-pyridine cyanoguanine, and 4-chloro aniline was used instead of 4-trifluoromethyl aniline.

(211) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.43 (m, 1H), 7.29 (m, 4H), 6.34 (m, 1H), 6.22 (m, 1H), 4.37 (s, 2H); LC-MS m/z 292.2 [M+1]+; mp 221223 C.

Example 71: Preparation of N1-furan-2-ylmethyl-N5-(4-trifluoromethoxy)phenyl biguanide.HCl

(212) ##STR00087##

(213) The title compound in a white solid was produced according to the substantially same method as described in Example 62, except that furan-2-ylmethyl cyanoguanidine was used instead of 3-pyridine cyanoguanine, and 4-trifluoromethoxy aniline was used instead of 4-trifluoromethyl aniline.

(214) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.42 (m, 3H), 7.18 (m, 2H), 6.33 (m, 2H), 4.38 (s, 2H); LC-MS m/z 342.1 [M+1]+; mp 223225 C.

Example 72: Preparation of N1-furan-2-ylmethyl-N5-(4-trifluoromethyl)phenyl biguanide.HCl

(215) ##STR00088##

(216) The title compound in a white solid was produced according to the substantially same method as described in Example 62, except that furan-2-ylmethyl cyanoguanidine was used instead of 3-pyridine cyanoguanine, and 4-trifluoromethyl aniline was used instead of 4-trifluoromethyl aniline.

(217) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.51 (m, 3H), 7.41 (m, 1H), 6.33 (m, 2H), 4.38 (s, 2H); LC-MS m/z 326.2 [M+1]+; mp 231233 C.

Example 73: Preparation of N1-dihydropyridine-N5-(4-(2-aminoethyl)phenyl biguanide.HCl

(218) ##STR00089##

(219) The title compound in a white solid was produced according to the substantially same method as described in Example 62, except that dihydropyridine cyanoguanidine was used instead of 3-pyridine cyanoguanine, and 4-(2-aminoethyl)aniline was used instead of 4-trifluoromethyl aniline.

(220) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.37 (d, 2H), 7.26 (d, 2H), 5.95 (m, 1H), 5.73 (m, 1H), 3.99 (s, 2H), 3.64 (t, 2H), 3.16 (t, 2H), 2.94 (t, 2H), 2.23 (s, 2H); LC-MS m/z 287.2 [M+1]; mp 230235 C.

Example 74: Preparation of N1-phenyl-N5-(3-trifluoromethyl)benzyl biguanide.HCl

(221) ##STR00090##

(222) The title compound in a white solid was produced according to the substantially same method as described in Example 62, except that phenyl cyanoguanidine was used instead of 3-pyridine cyanoguanine, and 3-trifluoromethylbenzyl amine was used instead of 4-trifluoromethyl aniline.

(223) .sup.1H NMR (400 MHz, CD.sub.3OD) 761 (d, J=7.8 Hz, 2H), 7.46 (d, J=7.8 Hz, 2H), 7.25 (m, 4H), 7.13 (m, 1H), 4.55 (s, 2H) LC-MS m/z 336.1 [M+1]

Example 75: Preparation of N1-(4-trifluoromethoxy)phenyl-N5-(N-biphenyl-4-yl) biguanide.HCl

(224) ##STR00091##

(225) The title compound in a white solid was produced according to the substantially same method as described in Example 62, except that 4-trifluoromethoxy phenyl cyanoguanidine was used instead of 3-pyridine cyanoguanine, and 3-trifluoromethylbenzyl amine 4-biphenyl amine was used instead of 4-trifluoromethyl aniline.

(226) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.41 (m, 4H), 7.41 (m, 4H), 7.33 (m, 12H), 7.32 (m, 2H), 7.26 (m, 2H) LC-MS m/z 415.2 [M+1]

Example 76: Preparation of N1-3-phenoxyphenyl-N5-(4-trifluoromethoxy)phenyl biguanide.HCl

(227) ##STR00092##

(228) The title compound in a white solid was produced according to the substantially same method as described in Example 62, except that 3-phenoxy phenyl cyanoguanidine was used instead of 3-pyridine cyanoguanine, and 3-trifluoromethylbenzyl amine 4-trifluoromethoxy aniline was used instead of 4-trifluoromethyl aniline.

(229) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.47 (m, 2H), 7.21 (m, 2H), 4.15 (m, 1H), 3.34 (m, 2H), 2.07 (m, 3H), 1.71 (m, 1H), 1.18 (s, 3H) LC-MS m/z 466.2 [M+1]

Example 77: Preparation of N-(4-(3-(imino(pyrrolidin-1-yl)methyl)guanidino)phenethyl)-5-(2-oxohexahydro-1H-thieno[2,3-d]imidazol-5-yl)pentaneamide

(230) ##STR00093##

(231) N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)pyrrolidine-1-carboximidamide (0.12 g, 0.48 mmol) and 2,5-dioxopyrrolidin-1-yl-5-(2-oxohexahydro-1H-thieno[2,3-d]imidazol-5-yl)pentanoate (0.20 g, 0.59 mmol) were dissolved in anhydrous DMF (5 ml), and then triethylamine (0.2 ml, 1.47 mmol) was added thereto. After stirring the reaction mixture for 24 hr, the reaction mixture was distillated under reduced pressure, separated and purified by a chromatography using MC:MeOH=10:1. The title compound was produced in white solid phase (84.0 mg, 34.2%).

(232) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.33 (d, 2H), 7.18 (d, 2H), 4.51 (m, 1H), 4.30 (m, 1H), 3.48 (m, 4H), 3.41 (m, 2H), 3.18 (m, 1H), 2.94 (dd, 1H), 2.77 (t, 2H), 2.72 (d, 1H), 2.17 (t, 2H), 2.04, 1.59 (d, 4H), 1.711.38 (m, 6H); LC-MS m/z 501.3 [M+1]; mp 230235 C.

Example 78: Preparation of N-(4-(3-(imino(2-methylpiperidin-1-yl)methyl)guanidino)phenethyl)-5-(2-oxohexahydro-1H-thieno[2,3-d]imidazol-5-yl)pentanamide

(233) ##STR00094##

(234) The title compound was produced according to the substantially same method as described in Example 92, except that N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)-2-methylpiperidine-1-carboximidamide was used instead of 4-N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)pyrrolidine-1-carboximidamide.

(235) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.29 (d, 2H), 7.19 (d, 2H), 4.51 (m, 1H), 4.40 (s, 1H), 4.30 (m, 1H), 3.95 (d, 1H), 3.41 (m, 2H), 3.19 (m, 1H), 3.09 (t, 1H), 2.94 (dd, 1H), 2.77 (t, 2H), 2.72 (d, 1H), 2.17 (t, 2H), 1.741.35 (m, 12H), 1.24 (d, 3H); LC-MS m/z 529.3 [M+1]; mp 230235 C.

Example 79: Preparation of N-(4-(3-((3,5-dimethylpiperidin-1-yl)(imino)methyl)guanidino)phenethyl)-5-(2-oxohexahydro-1H-thieno[2,3-d]imidazol-5-yl)pentanamide

(236) ##STR00095##

(237) The title compound was produced according to the substantially same method as described in Example 92, except that N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)-3,5-dimethylpiperidine-1-carboximidamide was used instead of 4-N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)pyrrolidine-1-carboximidamide.

(238) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.31 (d, 2H), 7.19 (d, 2H), 4.51 (m, 1H), 4.30 (m, 1H), 4.05 (s, 2H), 3.41 (m, 2H), 3.19 (m, 1H), 2.94 (dd, 1H), 2.77 (t, 2H), 2.72 (d, 1H), 2.45 (t, 2H), 2.17 (t, 2H), 1.86 (d, 1H), 1.731.55 (m, 6H), 1.38 (m, 2H), 0.92 (d, 6H), 0.84 (m, 1H); LC-MS m/z 543.4 [M+1]; mp 230235 C.

Example 80: Preparation of N-(4-(3-((5,6-dihydropyridin-1(2H)-yl)(imino)methyl)guanidino)phenethyl)-5-(2-oxohexahydro-1H-thieno[2,3-d]imidazol-5-yl)pentanamide

(239) ##STR00096##

(240) The title compound was produced according to the substantially same method as described in Example 92, except that N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)-5,6-dihydropyridine-1(2H)-carboximidamide was used instead of 4-N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)pyrrolidine-1-carboximidamide.

(241) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.28 (d, 2H), 7.20 (d, 2H), 5.95 (m, 1H), 5.74 (m, 1H), 4.50 (m, 1H), 4.29 (m, 1H), 3.98 (s, 2H), 3.62 (t, 2H), 3.40 (m, 2H), 3.19 (m, 1H), 2.94 (dd, 1H), 2.77 (t, 2H), 2.72 (d, 1H), 2.23 (s, 2H), 2.16 (t, 2H), 1.711.37 (m, 6H); LC-MS m/z 513.3 [M+1]; mp 230235 C.

Example 81: Preparation of N-(4-(3-(((2R,6S)-2,6-dimethylpiperidin-1-yl)(imino)methyl)guanidino)phenethyl)-5-(2-oxohexahydro-1H-thieno[2,3-d]imidazol-5-yl)pentanamide

(242) ##STR00097##

(243) The title compound was produced according to the substantially same method as described in Example 92, except that (2R,6S)N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)-2,6-dimethylpiperidine-1-carboximidamide was used instead of 4-N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)pyrrolidine-1-carboximidamide.

(244) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.27 (m, 2H), 7.14 (m, 2H), 4.47 (m, 1H), 4.26 (m, 1H), 3.37 (m, 2H), 3.18 (m, 2H), 2.91 (m, 1H) 2.75 (m, 3H), 2.15 (m, 2H), 1.67 (m, 13H) 1.26 (m, 6H); LC-MS m/z 543.2 [M+1]+

Example 82: Preparation of N-(4-(3-(imino(pyrrolidin-1-yl)methyl)guanidino)phenyl)-5-(2-oxohexahydro-1H-thieno[2,3-d]imidazol-5-yl)pentanamide

(245) ##STR00098##

(246) The title compound was produced according to the substantially same method as described in Example 92, except that N(N-(4-aminophenyl)carbamimidoyl)pyrrolidine-1-carboximidamide was used instead of 4-N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)pyrrolidine-1-carboximidamide.

(247) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.53 (d, 2H), 7.31 (d, 2H), 4.49 (m, 1H), 4.31 (m, 1H), 3.48 (s, 4H), 3.22 (m, 1H), 2.93 (dd, 1H), 2.71 (d, 1H), 2.03 (d, 4H), 1.801.47 (m, 6H); LC-MS m/z 473.3 [M+1]; mp 230235 C.

Example 83: Preparation of 2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)-N-(4-(3-(imino(pyrrolidin-1-yl)methyl)guanidino)benzyl)benzamide

(248) ##STR00099##

(249) The title compound was produced according to the substantially same method as described in Example 92, except that 2,5-dioxopyrrolidin-1-yl 2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoate was used instead of 2,5-dioxopyrrolidin-1-yl-5-(2-oxohexahydro-1H-thieno[2,3-d]imidazol-5-yl)pentanoate.

(250) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.90 (d, 1H), 7.57 (m, 2H), 7.17 (d, 2H), 7.09 (d, 1H), 6.88 (d, 2H), 6.66 (d, 2H), 6.46 (dd, 2H), 6.38 (d, 2H), 3.45 (d, 4H), 3.18 (t, 2H), 2.35 (t, 2H), 2.01 (d, 4H); LC-MS m/z 589.2 [M+1]+; mp 230235 C.

Example 84: Preparation of N-(4-(3-(((2R,6S)-2,6-dimethylpiperidin-1-yl)(imino)methyl)guanidino)benzyl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzamide

(251) ##STR00100##

(252) The title compound was produced according to the substantially same method as described in Example 92, except that N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)-2,6-dimethylpiperidine-1-carboximidamide was used instead of N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)pyrrolidine-1-carboximidamide, and 2,5-dioxopyrrolidin-1-yl 2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoate was used instead of 2,5-dioxopyrrolidin-1-yl-5-(2-oxohexahydro-1H-thieno[2,3-d]imidazol-5-yl)pentanoate.

(253) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.38 (d, 2H), 7.25 (d, 2H), 4.604.40 (s, 2H), 3.16 (t, 2H), 2.94 (t, 2H), 1.901.51 (m, 6H) 1.30 (s, 6H); LC-MS m/z 317.3 [M+1]; mp 230235 C.

Example 85: Preparation of N-(4-(3-((3,5-dimethylpiperidin-1-yl)(imino)methyl)guanidino)benzyl)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzamide

(254) ##STR00101##

(255) The title compound was produced according to the substantially same method as described in Example 92, except that N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)-3,5-dimethylpiperidine-1-carboximidamide was used instead of N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)pyrrolidine-1-carboximidamide, and 2,5-dioxopyrrolidin-1-yl 2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoate was used instead of 2,5-dioxopyrrolidin-1-yl-5-(2-oxohexahydro-1H-thieno[2,3-d]imidazol-5-yl)pentanoate.

(256) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.90 (d, 1H), 7.58 (m, 2H), 7.14 (d, 2H), 7.10 (d, 1H), 6.90 (d, 2H), 6.66 (d, 2H), 6.46 (dd, 2H), 6.39 (d, 2H), 3.98 (s, 2H), 3.18 (t, 2H), 2.41 (m, 2H), 2.36 (t, 2H), 1.83 (d, 1H), 1.60 (m, 2H), 0.89 (d, 6H), 0.81 (m, 1H); LC-MS m/z 631.4 [M+1]+; mp 230235 C.

Example 86: Preparation of 2-(6-hydroxy-3-oxo-3H-xanthene-9-yl)-N-(4-(3-(imino(3-methylpiperidine-1-yl)methyl)guanidino)benzyl)benzamide

(257) ##STR00102##

(258) The title compound was produced according to the substantially same method as described in Example 92, except that N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)-3-methylpiperidine-1-carboximidamide was used instead of N(N-(4-(2-aminoethyl)phenyl)carbamimidoyl)pyrrolidine-1-carboximidamide, and 2,5-dioxopyrrolidin-1-yl 2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoate was used instead of 2,5-dioxopyrrolidin-1-yl-5-(2-oxohexahydro-1H-thieno[2,3-d]imidazol-5-yl)pentanoate.

Example 87: Preparation of N(N-(4-(trifluoromethoxy)phenyl)carbamimidoyl)-3,4-dihydroisoquinolin-2(1H)-carboximidamide.HCl

(259) ##STR00103##

(260) The title compound was produced according to the substantially same method as described in Example 1, except that dihydro isoquinolin cyanoguanidine was used instead of pyrrolidine cyanoguanidine, and 4-(trifluoromethoxy)aniline was used instead of piperonyl amine.

(261) .sup.1H NMR (400 MHz, DMSO-d6) 9.87 (s, 1H), 7.84 (s, 2H), 7.51 (d, J=7.2 Hz, 2H), 7.29 (d, J=7.6 Hz, 2H), 7.22 (s, 4H), 7.00 (s, 2H), 4.63 (s, 2H), 3.65 (s, 2H), 2.89 (s, 2H); LC-MS m/z 378.2 [M+1]

Example 88: Preparation of N(N-(4-(trifluoromethoxy)phenyl)carbamimidoyl)indoline-1-carboximidamide.HCl

(262) ##STR00104##

(263) The title compound was produced according to the substantially same method as described in Example 1, except that indoline cyanoguanidine was used instead of pyrrolidine cyanoguanidine, and 4-(trifluoromethoxy)aniline was used instead of piperonyl amine.

(264) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.69 (d, J=8.4 Hz, 1H), 7.48 (m, 2H), 7.25 (m, 3H), 7.10 (m, 1H), 7.03 (m, 1H), 4.05 (m, 2H), 3.24 (m, 2H); LC-MS m/z 364.2 [M+1]

Example 89: Preparation of N-(imino(pyrrolidin-1-yl)methyl)-3,4-dihydroisoquinoline-2(1H)-carboximidamide HCl

(265) ##STR00105##

(266) The title compound was produced according to the substantially same method as described in Example 1, except that dihydroisoquinolin-2(1H) cyanoguanidine was used instead of pyrrolidine cyanoguanidine, and pyrrolidine was used instead of piperonyl amine.

(267) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.21 (m, 3H), 7.18 (m, 1H), 4.68, (s, 2H), 3.74 (t, J=6.6 Hz, 2H), 3.46 (s, 4H), 2.95 (t, J=6.0 Hz, 2H), 2.00 (s, 4H); LC-MS m/z 272.2 [M+1]

Example 90: Preparation of N(N-(4-phenoxyphenyl)carbamimidoyl)indoline-1-carboximidamide HCl

(268) ##STR00106##

(269) The title compound was produced according to the substantially same method as described in Example 1, except that indoline cyanoguanidine was used instead of pyrrolidine cyanoguanidine, and 4-phenoxy aniline was used instead of piperonyl amine

(270) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.73 (d, J=7.8 Hz, 1H), 7.32 (m, 4H), 7.23 (m, 1H), 7.10 (m, 2H), 7.02 (t, J=7.2 Hz, 1H), 6.97 (m, 4H), 4.03 (t, J=8.4 Hz, 2H), 3.22 (t, J=8.4 Hz, 2H); LC-MS m/z 372.2 [M+1]

Example 91: Preparation of N(N-(3-phenoxyphenyl)carbamimidoyl)indoline-1-carboximidamide HCl

(271) ##STR00107##

(272) The title compound was produced according to the substantially same method as described in Example 1, except that indoline cyanoguanidine was used instead of pyrrolidine cyanoguanidine, and 3-phenoxy aniline was used instead of piperonyl amine

(273) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.67 (m, 1H), 7.28 (m, 2H), 7.21 (m, 2H), 7.10 (m, 5H), 6.92 (m, 2H), 6.78 (m, 41H), 4.00 (m, 2H), 3.24 (m, 2H); LC-MS m/z 372.2 [M+1]

Example 92: Preparation of N-(5-(trifluoromethoxy)-1H-benzo[d]imidazol-2-yl)pyrrolidine-1-carboximidamide HCl

(274) ##STR00108##

(275) N1-pyrrolidine cyanoguanidine (72 mg, 0.52 mmol) and 4-trifluoro methoxy phenylene diamine (0.1 g, 0.52 mmol) were dissolved in distilled water (10 ml) at room temperature, and then hydrochloric acid (0.1 ml, 1.04 mmol) was added thereto, and stirred with reflux at 110 C. for 2 hr. The reaction mixture was added by 10% potassium hydroxide solution, and then the obtained solid was filtrated under reduced pressure, and was washed with distilled water. The filtrated solid was dried under reduced pressure. The title compound was produced in brown solid phase (20 mg, 12.2%).

(276) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.22 (d, J=8.4 Hz, 1H), 7.13 (s, 1H), 6.90 (d, J=8.4 Hz, 1H), 3.49 (br s, 4H), 1.92 (s, 4H); LC-MS m/z 235.1 [M+1]

Example 93: Preparation of N-(5-fluoro-1H-benzo[d]imidazol-2-yl)pyrrolidine-1-carboximidamide HCl

(277) ##STR00109##

(278) The title compound was produced according to the substantially same method as described in Example 92, except that 4-fluoro phenylene diamine was used instead of 4-trifluoro methoxy phenylene diamine

(279) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.15 (d, J=8.4 Hz, 1H), 6.95 (d, J=9.0 Hz, 1H), 6.72 (t, J=10.2 Hz, 1H), 3.48 (s, 4H), 1.98 (s, 4H); LC-MS m/z 248.0 [M+1]

Example 94: Preparation of N-(5-methyl-1H-benzo[d]imidazol-2-yl)pyrrolidine-1-carboximidamide HCl

(280) ##STR00110##

(281) The title compound was produced according to the substantially same method as described in Example 92, except that 4-methyl phenylene diamine was used instead of 4-trifluoro methoxy phenylene diamine.

(282) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.13 (d, J=7.8 Hz, 1H), 7.06 (s, 1H), 6.84 (d, J=7.2 Hz, 1H), 3.47 (s, 4H), 2.37 (s, 3H), 1.96 (s, 4H); LC-MS m/z 244.0 [M+1]

Example 95: Preparation of N-(5-methoxy-1H-benzo[d]imidazol-2-yl)pyrrolidine-1-carboximidamide.HCl

(283) ##STR00111##

(284) The title compound was produced according to the substantially same method as described in Example 92, except that 4-methoxy phenylene diamine was used instead of 4-trifluoro methoxy phenylene diamine.

(285) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.13 (d, J=7.8 Hz, 1H), 6.85 (d, J=2.4 Hz, 1H), 6.65 (dd, J=9.0 Hz, J=2.4 Hz, 1H), 3.78 (s, 3H), 3.48 (s, 4H), 1.98 (s, 4H); LC-MS m/z 260.1 [M+1]

Example 96: Preparation of N-(1H-benzo[d]imidazol-2-yl)pyrrolidine-1-carboximidamide HCl

(286) ##STR00112##

(287) The title compound was produced according to the substantially same method as described in Example 92, except that o-phenylene diamine was used instead of 4-trifluoro methoxy phenylene diamine.

(288) .sup.1H NMR (600 MHz, CD.sub.3OD) 7.25 (d, J=8.4 Hz, 2H), 7.00 (d, J=9.0 Hz, 2H), 3.48 (s, 4H), 1.98 (s, 4H); LC-MS m/z 230.1 [M+1]

Experimental Example

(289) The compounds produced in Examples 1 to 96 were evaluated for their inhibitory activities on cancer cells according to the following method.

Experimental Example 1: Evaluation of Inhibitory Activity against Cancer Cells

(290) The inhibitory activities of biguanide derivatives against cancer cells were evaluated in the cell line SK-MEL-28 derived from human Melanoma cell, Miapaka-2 cell line derived from human pancreatic cancer cell and HCT116 cell line derived from human colorectal cancer cell by measuring the concentration that inhibits 50% of the growth of the cell lines (the half maximal inhibitory concentration, IC.sub.50).

(291) Firstly, SK-MEL-28 cells were grown to be density of 5,000 cells/well in RPMI-1640 media supplemented with 10% (v/v) bovine calf serum, on 96-well plates for 16 hr. To measure the IC.sub.50 of each compound, the cells were incubated for 48 hr with the 100 mM PBS stock compounds at an amount of 10 mM, 1 mM, 200 uM, 40 uM, 8 uM, 1.6 uM, 0.32 uM or 0.064 uM; 50 mM PBS and EtOH stock compounds at an amount of 1 mM, 200 uM, 40 uM, 8 uM, 1.6 uM, 0.32 uM and 0.064 uM; and 50 mM DMSO stock compounds at an amount of 100 uM, 25 uM, 6.25 uM, 1.56 uM, 0.39 uM, 0.10 uM and 0.02 uM, respectively. To test the viable cells after incubation with each compound, the cell cultures were incubated additionally for 2 hr in the presence of MTT. The produced formazan crystals were dissolved with dimethyl sulfoxide and then measured by detecting the absorbance at 560 nm.

(292) After the 48 hr incubation, the viable cells were counted on the well plates treated with the compounds of the Examples relative to the count of the viable cells on the non-treated plates, and then was represented as the cell viability (%). By using the cell viability (%) result, the cell viability curve was plotted, and IC.sub.50 was calculated from the curve to evaluate the inhibitory activity against cancer cells.

(293) The inhibitory activities of biguanide derivatives against cancer cells of Miapaca-2 cell and HCT116 cell were evaluated according to the substantially same method as described in SK-MEL-28 cell, except that DMEM media supplemented with 10% (v/v) bovine calf serum was used instead of RPMI-1640 media supplemented with 10% (v/v) bovine calf serum for SK-MEL-28 cell.

(294) The experimental results of inhibitory activity against cancer cells are summarized in Table 1 as below.

Experimental Example 2: Evaluation of Inhibitory Activity on Oxygen Consumption Rate (OCR)

(295) The cellular metabolic activity of the compounds on Oxygen Consumption Rate (OCR) was evaluated, because the biguanide drugs have anticancer activity by inhibiting the oxidative phosphorylation of cancer cell.

(296) Compared to phenformin, the more effective compounds were selected by evaluating the inhibitory activity on the cell OCR, after lung cancer cell line A549 (ATCC-American Type Culture Collection) was treated with the compounds of Examples.

(297) A549 cells were inoculated at 510.sup.3 cells in RPMI1640 media on XF96 cell culture plate and cultured for 24 h (temp; 37 C., 5% CO.sub.2) to make the cell adhere to the plate.

(298) After 24 hr, A549 cells were treated by 10 M of the compounds or phenformin, and were washed with XF analysis media using Prep station to remove the original media. A549 cells were retreated by compounds or phenformin and incubated for 1 hr (temp; 37 C., Non-CO.sub.2) in Prep station. While incubation in Prep station, sensor cartridge was calibrated for 1 hr. Then, the calibrated sensor cartridge was inserted to the cell plate to analyze OCR.

(299) When OCR of phenformin as a reference drug was set to 100% and OCR of untreated control was set to 0%, the OCR of tested compounds was calculated by converting the measured OCR to be the relative percentage value (%). The results are summarized in Table 1, below.

(300) TABLE-US-00001 TABLE 1 OCR % to Inhibitory activity against cancer cells phenformin Test SK-MEL-28 Miapaca-2 HCT 116 (A549, 3 hrs) Compound IC.sub.50 (M) IC.sub.50 (M) IC.sub.50 (M) 10 uM Phenformin 265.8878 118.52 12.5 Example 1 106.99 36.22 >100 Example 2 >100 Example 3 >100 Example 4 >100 Example 5 >100 Example 6 28.6 44% Example 8 6.97 Example 9 7.427 Example 10 6.281 Example 11 6.464 Example 12 81.9 Example 13 >100 Example 14 29.4732 26.63 >100 Example 15 711.8 Example 16 39.810 13% Example 17 183.82 113.14 >100 Example 18 181.71 47.92 >100 57% Example 19 >100 Example 20 134.15 58.62 >100 15% Example 21 >100 Example 22 105.7 74% Example 23 >1000 2339.56 >100 Example 24 >100 >100 >100 Example 25 >100 Example 26 >100 Example 27 >100 Example 28 176.3578 208.85 >100 Example 29 >100 Example 30 >100 Example 31 93.6 Example 32 90.3 Example 33 >100 Example 34 >100 Example 35 32.4 Example 36 7.1 Example 37 32.3 Example 38 11.7 Example 39 7.5 Example 40 35.2 42.9% Example 41 >100 Example 42 >100 Example 43 >100 Example 44 42.2 Example 45 33.8 Example 46 >100 Example 47 >100 Example 48 >100 Example 49 >100 Example 50 >100 Example 51 7.828 Example 52 7.654 Example 53 >100 >100 182.1 Example 54 >100 Example 55 >100 Example 56 34.8 Example 57 >100 Example 58 >100 Example 59 >100 Example 60 49.8 Example 61 >100 Example 62 >100 Example 63 >100 Example 64 29.2 Example 65 >100 Example 66 >100 Example 67 50.7 Example 68 53.1 Example 69 68.5 Example 70 13.5 Example 71 6.9 Example 72 8.8 Example 74 6.850 Example 75 8.207 Example 76 3.121 Example 77 >100 Example 78 >100 Example 79 >100 Example 80 >100 Example 81 >100 Example 82 >100 Example 83 56.8 Example 84 27.4 Example 85 28.8 Example 86 53.2 Example 87 2.573 Example 88 7.415 Example 89 59.46 44% Example 90 27.73 Example 91 11.04 Example 92 >100 Example 93 >100 Example 94 >100 Example 95 >100 Example 96 >100