Biguanide derivative, a preparation method thereof, and a pharmaceutical composition containing the biguanide derivative as an active ingredient

Abstract

A biguanide derivative compound with N1-N5 substitution, which is represented by Formula 1, or a pharmaceutically acceptable salt thereof, a method of preparing the same, and a pharmaceutical composition containing the same as an active ingredient are provided. The biguanide derivative may exhibit excellent effect on activation of AMPK and inhibition of cancer cell proliferation in a low dose, compared to conventional drugs, and thus, may be useful to treat diabetes mellitus, obesity, hyperlipidemia, hypercholesterolemia, fatty liver, coronary artery disease, osteoporosis, polycystic ovarian syndrome, metabolic syndrome, cancer, etc.

Claims

1. A method of activating AMPK(5-AMP-activated protein kinase alpha) in cells, the method comprising: administering a therapeutically effective amount of a compound of Formula 1 below or a pharmaceutically acceptable salt to a subject in need thereof: ##STR00044## wherein R.sub.1is C.sub.5-12aryl, or C.sub.1-12 alkyl unsubstituted or substituted with C.sub.5-12 aryl, and the aryl is unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, and halogen; R.sub.2 is hydrogen, or C.sub.1-12 alkyl unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.3-10 cycloalkyl, C.sub.5-12 aryl, C.sub.5-12 heteroaryl, hydroxyl, halogen, and C.sub.1-4 alkoxycarbonyl, and the aryl and heteroaryl are unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, and halogen; R.sub.3is hydrogen, C.sub.5-12 aryl, or C.sub.1-12 alkyl unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.3-10 cycloalkyl, C.sub.5-12 aryl, C.sub.5-12 heteroaryl, hydroxyl, halogen, and C.sub.1-4 alkoxycarbonyl, and the aryl and heteroaryl are unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, and halogen; and R.sub.4 is C.sub.3-10 cycloalkyl, C.sub.5-12 alkoxy, C.sub.5-12 aryl, C.sub.5-12 heteroaryl, hydroxyl, halogen, or C.sub.1-12 alkyl unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.3-10 cycloalkyl, C.sub.5-12 aryl, C.sub.5-12 heteroaryl, hydroxyl, halogen, and C.sub.1-4 alkoxycarbonyl, and the aryl and heteroaryl are unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, and halogen.

2. The method of claim 1, wherein R.sub.1 is C.sub.5-12 aryl, unsubstituted C.sub.1-7 alkyl, or C.sub.1-6 alkyl substituted with C.sub.5-12 aryl, and the aryl is unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, and halogen; R.sub.2 is hydrogen, unsubstituted C.sub.1-7 alkyl, or C.sub.1-6 alkyl substituted with at least one non-hydrogen substituent selected from the group consisting of unsubstituted C.sub.3-7 cycloalkyl, C.sub.5-12 aryl, C.sub.5-12 heteroaryl, hydroxyl, halogen, and C.sub.1-4 alkoxycarbonyl, and the aryl and heteroaryl are unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, and halogen; R.sub.3 is hydrogen, C.sub.5-12 aryl, unsubstituted C.sub.1-7 alkyl, or C.sub.1-6 alkyl substituted with at least one non-hydrogen substituent selected from the group consisting of unsubstituted C.sub.3-7 cycloalkyl, C.sub.5-12 aryl, C.sub.5-12 heteroaryl, hydroxyl, halogen, and C.sub.1-4 alkoxycarbonyl, and the aryl and heteroaryl are unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, and halogen; and R.sub.4 is unsubstituted C.sub.3-7 cycloalkyl, alkoxy, C.sub.5-12 aryl, C.sub.5-12 heteroaryl, hydroxyl, halogen, unsubstituted C.sub.1-7 alkyl, or C.sub.1-6 alkyl substituted with at least one non-hydrogen substituent selected from the group consisting of unsubstituted C.sub.3-7 cycloalkyl, C.sub.5-12 aryl, C.sub.5-12 heteroaryl, hydroxyl, halogen, and C.sub.1-4 alkoxycarbonyl, and the aryl and heteroaryl are unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, and halogen.

3. The method of claim 2, wherein R.sub.1 is C.sub.5-12 aryl, unsubstituted C.sub.1-7 alkyl, or C.sub.1-6 alkyl substituted with C.sub.5-12 aryl, and the aryl is unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, and halogen; R.sub.2 is hydrogen, unsubstituted C.sub.1-7 alkyl, or C.sub.1-6 alkyl substituted with C.sub.5-12 heteroaryl, and the heteroaryl is unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, and halogen; R.sub.3 is hydrogen, C.sub.5-12 aryl, unsubstituted C.sub.1-7 alkyl, or C.sub.1-6 alkyl substituted with C.sub.5-12 heteroaryl, and the aryl and heteroaryl are unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, and halogen; and R.sub.4 a is unsubstituted C.sub.3-7 cycloalkyl, C.sub.5-12 aryl, C.sub.5-12 heteroaryl, unsubstituted C.sub.1-7 alkyl, or C.sub.1-6 alkyl substituted with at least one non-hydrogen substituent selected from the group consisting of unsubstituted C.sub.3-7 cycloalkyl, C.sub.5-12 aryl, C.sub.5-12 heteroaryl, and C.sub.1-4 alkoxycarbonyl, and the aryl and heteroaryl are unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, and halogen.

4. The method of claim 3, wherein R.sub.1 is C.sub.5-12 aryl, unsubstituted C.sub.1-7 alkyl, or C.sub.1-4 alkyl substituted with C.sub.5-12 aryl, and the aryl is unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, and halogen; R.sub.2 is hydrogen, unsubstituted C.sub.1-7 alkyl, or C.sub.1-4 alkyl substituted with C.sub.5-12 heteroaryl, and the heteroaryl may be unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkoxy, hydroxyl, and halogen, and the heteroaryl is selected from the group consisting of pyridinyl, furanyl, and isoquinolinyl; R.sub.3 is hydrogen, C.sub.5-12 aryl, unsubstituted C.sub.1-7 alkyl, or C.sub.1-4 alkyl substituted with C.sub.5-12 heteroaryl, and the aryl and heteroaryl may be unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkoxy, hydroxyl, and halogen, and the aryl and heteroaryl is selected from the group consisting of phenyl, pyridinyl, furanyl, and isoquinolinyl; and R.sub.4 is unsubstituted C.sub.3-7 cycloalkyl, C.sub.5-12 aryl, C.sub.5-12 heteroaryl, unsubstituted C.sub.1-7 alkyl, or C.sub.1-4 alkyl substituted with at least one non-hydrogen substituent selected from the group consisting of unsubstituted C.sub.3-7 cycloalkyl, C.sub.5-12 aryl,C.sub.5-12 heteroaryl, and C.sub.1-4 alkoxycarbonyl, and the aryl and heteroaryl are unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkyl, C.sub.1-4 alkoxy, hydroxyl, and halogen.

5. The method of claim 4, wherein R.sub.1 is C.sub.5-12 aryl, unsubstituted C.sub.1-7 alkyl, or C.sub.1-4 alkyl substituted with C.sub.5-12 aryl; R.sub.2 is hydrogen or unsubstituted C.sub.1-7 alkyl; R.sub.3 is hydrogen, C.sub.5-12 aryl, or unsubstituted C.sub.1-7 alkyl; and R.sub.4 is unsubstituted C.sub.3-7 cycloalkyl,C.sub.5-12 aryl, C.sub.5-12 heteroaryl, unsubstituted C.sub.1-7 alkyl, or C.sub.1-4 alkyl substituted with at least one non-hydrogen substituent selected from the group consisting of unsubstituted C.sub.3-7 cycloalkvl, C.sub.5-12 aryl, C.sub.5-12 heteroaryl, and C.sub.1-4 alkoxycarbonyl; and the aryl and heteroaryl may be unsubstituted or substituted with at least one non-hydrogen substituent selected from the group consisting of C.sub.1-4 alkoxy, hydroxyl, and halogen, and the aryl and heteroaryl is selected from the group consisting of phenyl, pyridinyl, furanyl, and isoquinolinyl.

6. The method of claim 1, wherein the compound of Formula 1 is: N1 -hexyl-N5-propyl biguanide; N1 -hexyl-N5-cyclopropylmethyl biguanide; N1 -hexyl -N5-cyclohexylmethyl biguanide; N1 -hexyl-N5-benzyl big uanide; N1 ,N5-bis(4-chlorophenyl) biguanide; N1 ,N5-bis(3-chlorophenyl) biguanide; N1 -(4-chlorophenyl-N5-(4-methoxy)phenyl biguanide; N1 ,N5-bis(3-chloro-4-methoxyphenyl) biguanide; N1 ,N5 bis(3,4-dichlorophenyl) biguanide; N1 ,N5-bis(3,5-dichlorophenyl) biguanide; N1 ,N5-bis(4-bromophenyl) biguanide; N1 -benzyl-N5-(pyridine-3-yl)methyl biguanide; N1 -(phenethyl)-N5-propyl biguanide; N1 -(phenethyl)-N5-cyclopropylmethyl biguanide; N1 -(phenethyl)-N5-cycloheptyl biguanide; N1 ,N5-bis(phenethyl) biguanide; N1 ,N1 , N5-trimethyl biguanide; N1 ,N1-dimethyl-N5-butyl biguanide; N1 ,N1-dimethyl-N5-(butan-2-yl) biguanide; N1 ,N1-dimethyl-N5-t-butyl biguanide; N1 ,N1-dimethyl-N5-pentyl biguanide; N1 ,N1-dimethyl-N5-methoxycarbonylethyl biguanide; N1 ,N1-dimethyl-N5-cycloheptyl biguanide; N1 ,N1-dimethyl-N5-cyclopropylmethyl biguanide; N1 ,N1-dimethyl-N5-(4-bromo)phenyl biguanide; N1 ,N1-dimethyl-N5-(furan-2-yl)methyl biguanide; N1 ,N1-dimethyl-N5-(pyridine-3-yl)methyl biguanide; N1 ,N1-dimethyl-N5-benzyl biguanide; N1 ,N1-dimethyl-N5-(phenethyl) biguanide; N1 ,N1-diethyl-N5-(3-chloro)phenyl biguanide; N1 ,N1-dipropyl-N5-(3-chloro)phenyl biguanide; N1 ,N1-(ethyl)(propyl)-N5-(4-chloro)phenyl biguanide; N1 ,N1-dipropyl-N5-(isoquionline-5-yl) biguanide; N1 ,N1-dihexyl-N5-(3-chloro)phenyl biguanide; N1 ,N1, N5, N5-tetraethyl biguanide; N1 ,N1-diethyl-N5, N.5-(cyclohexyl)(methyl) biguanide; N1 ,N1-dipropyl-N5, N5-diethyl biguanide; N1 ,N1-dipropyl-N5, N5-(methyl)(phenethyl) biguanide; N1 ,N1-dipropyl-N5, N5-(4-hydroxylphenyl)(phenyl) biguanide; or N1 ,N1,N5, N5-bis((benzyl)(methyl)) biguanide.

Description

BEST MODE

(1) The advantages and features of the present invention and the method of revealing them will be explicit from the following examples described in detail. However, it is to be distinctly understood that the present invention is not limited thereto but may be otherwise variously embodied and practiced. It is obvious that the following examples are to complete the disclosure of the invention and to indicate the scope of the present invention to a skilled artisan completely, and the present invention will be defined only by the scope of the claims.

EXAMPLES

Example 1

Preparation of N1-hexyl-N5-propyl Biguanide Hydrochloride

(2) ##STR00004##

(1-1) Synthesis of 1-hexyl-3-cyanoguanidine

(3) While a solution prepared by dissolving 1-hexyl amine (3.6 g, 35.9 mmol) in n-butanol (30 ml) was stirred, sodium dictanoamide (3.5 g, 39.5 mmol) and concentrated hydrochloric acid (3.4 ml, 39.5 mmol) were added thereto at room temperature. The mixed solution was refluxed with stirring for 24 hours. After confirming the completion of the reaction, the generated sodium chloride was removed by filtering the reaction mixture, and then the filtered solution was concentrated under reduced pressure. After the concentrate was filtered, the filter-cake was washed with distilled water (30 ml). The filter-cake was vacuum-dried, thereby obtaining a target compound as a white solid (3.4 g, 58%). The compound was used in a subsequent reaction without another step of purification.

(1-2) Preparation of N1-hexyl-N5-propyl Biguanide Hydrochloride

(4) Concentrated hydrochloric acid (0.47 ml, 5.31 mmol) was added to a solution prepared by dissolving 1-propyl amine (0.58 g, 5.84 mmol) in n-butanol (10 ml), and the mixed solution was stirred for 30 minutes at room temperature. The compound obtained in the previous step (1-1) (0.89 g, 5.31 mmol) was added to the reaction mixture and refluxed with stirring for 24 hours. The mixture was concentrated under reduced pressure, and the concentrate was purified using flash column chromatography (dichloromethane:methanol=9:1). A 6N methanol hydrochloride solution (1 ml) was added to the compound to dissolve, and the mixture was concentrated under reduced pressure, thereby obtaining a target compound as a white solid (0.92 g, 65%).

(5) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 7.46 (br s, 2H), 6.81 (br s, 2H), 3.05 (m, 4H), 1.44 (m, 4H) 1.25 (m, 6H), 0.85 (t, 3H, J=7.2 Hz); mp 148-149 C.

(6) Target compounds of the following Examples 2 to 40 were prepared by the same method as described in Example 1, except that an amine compound corresponding to the target compound was used instead of 1-hexyl amine and 1-propyl amine respectively used in steps (1-1) and (1-2) of Example 1.

Example 2

N1-propyl-N5-cyclopropylmethyl Biguanide Hydrochloride

(7) ##STR00005##

(8) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 7.50 (br s, 2H), 6.83 (br s, 3H), 3.07 (m, 2H), 2.97 (m, 2H), 1.42 (m, 2H), 1.26 (m, 6H), 0.95 (m, 1H), 0.85 (t, 3H, J=6.6 Hz), 0.42 (m, 2H), 0.17 (d, 2H, J=4.8 Hz); mp 162-163 C.

Example 3

N1-hexyl-N5-cyclohexylmethyl Biguanide Hydrochloride

(9) ##STR00006##

(10) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 8.22 (br s, 2H), 3.12 (m, 2H), 2.73 (dd, 2H, J=7.5, 7.2 Hz), 1.96 (m, 2H), 1.68 (m, 2H), 1.50-1.59 (m, 7H), 1.46 (m, 2H), 1.38 (m, 2H), 1.23-1.36 (m, 4H), 0.87 (t, 3H, J=7.2 Hz)

Example 4

N1-hexyl-N5-benzyl Biguanide Hydrochloride

(11) ##STR00007##

(12) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 8.45 (br s, 1H), 7.24-7.36 (m, 5H), 6.92 (br s, 2H), 4.33 (d, 2H, J=6.0 Hz), 3.08 (m, 2H), 1.21-1.44 (m, 8H), 0.86 (m, 3H): mp 121-123 C.

Example 5

N1,N5-bis(4-chlorophenyl) Biguanide Hydrochloride

(13) ##STR00008##

(14) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 10.02 (br s, 2H), 7.54 (br s, 2H), 7.38 (d, 4H, J=9.0 Hz), 7.32 (d, 4H, J=9.0 Hz); mp 263-264 C.

Example 6

N1,N5-bis(3-chlorophenyl) Biguanide Hydrochloride

(15) ##STR00009##

(16) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.23 (br s, 2H), 7.54 (dd, 2H, J=2.0, 2.0 Hz), 7.32 (dd, 2H, J=8.2, 8.2 Hz), 7.23 (ddd, 2H, J=8.2, 2.0, 0.8 Hz), 7.15 (br s, 2H), 7.11 (ddd, 2H, J=8.2, 2.0, 0.8 Hz); mp 129-131 C.

Example 7

N1-(4-chloro)phenyl-N5-(4-methoxy)phenyl Biguanide Hydrochloride

(17) ##STR00010##

(18) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 9.87 (br s, 1H), 9.75 (br s, 1H), 7.49 (br s, 1H), 7.35 (d, 2H, J=9.0 Hz), 7.33 (d, 2H, J=9.0 Hz), 7.30 (br s, 1H), 7.20 (d, 2H, 9.0 Hz), 6.91 (d, 2H, 9.0 Hz), 3.72 (s, 3H); mp 247-249 C.

Example 8

N1,N5-bis(3-chloro-4-methoxyphenyl) Biguanide Hydrochloride

(19) ##STR00011##

(20) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 9.11 (br s, 1H), 7.46 (d, 2H, J=2.4 Hz), 7.20 (dd, 2H, J=9.0, 2.4 Hz), 7.09 (d, 2H, J=9.0 Hz), 7.05 (br s, 1H), 3.83 (s, 6H); mp 203-204 C.

Example 9

N1,N5-bis(3,4-dichlorophenyl) Biguanide Hydrochloride

(21) ##STR00012##

(22) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 10.3 (br s, 1H), 7.73 (br s, 1H), 7.63 (s, 2H), 7.59 (d, 2H, J=9.0 Hz), 7.28 (d, 2H, J=9.0 Hz); mp 255-257 C.

Example 10

N1,N5-bis(3,5-dichlorophenyl) Biguanide Hydrochloride

(23) ##STR00013##

(24) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 10.48 (br s, 1H), 7.85 (br s, 1H), 7.40 (d, 4H, J=1.8 Hz), 7.35 (d, 2H, J=1.8 Hz); mp 250-251 C.

Example 11

N1,N5-bis(4-bromophenyl) Biguanide Hydrochloride

(25) ##STR00014##

(26) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 10.17 (br s, 1H), 7.59 (br s, 1H), 7.49 (d, 4H, J=7.2 Hz), 7.27 (d, 4H, J=7.2 Hz); mp 242-243 C.

Example 12

N1-benzyl-N5-(pyridine-3-yl)methyl Biguanide Hydrochloride

(27) ##STR00015##

(28) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.67 (s, 1H), 8.57 (dd, 1H, J=4.8, 1.6 Hz), 8.45 (br s, 2H), 7.93 (dd, 1H, J=7.6, 1.6 Hz), 7.90 (br a, 1H), 7.45 (dd, 1H, J=7.6, 4.8 Hz), 7.18-7.34 (m, 5H), 7.21 (br s, 1H), 4.32 (m, 2H), 4.06 (s, 2H); mp 138-140 C.

Example 13

N1-(phenethyl)-N5-propyl Biguanide Hydrochloride

(29) ##STR00016##

(30) .sup.1H NMR (400 MHz, DMSO-ds) 7.33 (br s, 2H), 7.21-7.37 (m, 5H), 6.87 (br s, 2H), 3.33 (m, 2H), 2.79 (m, 2H), 1.46 (m, 2H), 0.87 (t, 3H, J=7.2 Hz); mp 126-128 C.

Example 14

N1-(phenethyl)-N5-cyclopropylmethyl Biguanide Hydrochloride

(31) ##STR00017##

(32) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 7.20-7.32 (m, 5H), 3.32 (m, 2H), 2.98 (t, 2H, J=6.0 Hz), 2.76 (m, 2H), 0.96 (m, 1H), 0.41 (dd, 2H, J=7.8, 1.8 Hz), 0.19 (dd, 2H, J=4.8, 1.8 Hz); mp 143-146 C.

Example 15

N1-(phenethyl)-N5-cycloheptyl Biguanide Hydrochloride

(33) ##STR00018##

(34) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.08 (br s, 2H), 7.10-7.38 (m, 5H), 6.86 (br s, 2H), 3.66 (m, 1H), 3.33 (m, 4H), 3.15 (m, 2H), 2.77 (m, 2H), 1.81 (m, 2H), 1.36-1.57 (m, 6H); mp 135-137 C.

Example 16

N1,N5-bis(phenethyl) Biguanide Hydrochloride

(35) ##STR00019##

(36) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 8.29 (br s, 2H), 7.34 (m, 2H), 7.26 (m, 3H), 3.00 (t, 2H, J=9.0 Hz), 2.92 (t, 2H, J=9.0 Hz); mp 204-205 C.

Example 17

N1,N1,N5-trimethyl Biguanide Hydrochloride

(37) ##STR00020##

(38) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 7.98 (br s, 2H), 6.91 (br s, 1H), 2.82 (s, 9H); mp 175-177 C.

Example 18

N1,N1-dimethyl-N5-butyl Biguanide Hydrochloride

(39) ##STR00021##

(40) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 7.95 (br s, 2H), 6.93 (br s, 1H), 2.85 (s, 6H), 2.72 (t, 2H, J=7.2 Hz), 1.49 (m, 2H), 1.31 (m, 2H), 0.86 (t, 3H, J=7.8 Hz); mp 131-133 C.

Example 19

N1,N1-dimethyl-N5-(butane-2-yl) Biguanide Hydrochloride

(41) ##STR00022##

(42) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 7.92 (br s, 2H), 6.94 (br s, 1H), 3.05 (m, 1H), 2.87 (s, 6H), 1.60 (m, 1H), 1.43 (m, 1H), 1.15 (d, 3H, J=5.4 Hz), 0.88 (t, 3H), J=7.2 Hz); mp 110-112 C.

Example 20

N1,N1-dimethyl-N5-t-butyl Biguanide Hydrochloride

(43) ##STR00023##

(44) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 8.05 (br s, 2H), 6.93 (br s, 1H), 2.85 (s, 6H), 1.23 (s, 9H); mp 186-187 C.

Example 21

N1,N1-dimethyl-N5-pentyl Biguanide Hydrochloride

(45) ##STR00024##

(46) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 7.94 (br s, 2H), 6.95 (br s, 1H), 2.86 (s, 6H), 2.73 (t, 2H, J=7.2 Hz), 1.54 (m, 2H), 1.27 (m, 4H), 0.86 (t, 3H, J=7.2 Hz); mp 131-133 C.

Example 22

N1,N1-dimethyl-N5-(methoxycarbonylethyl)biguanide Hydrochloride

(47) ##STR00025##

(48) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.12 (br s, 2H), 6.93 (br s, 1H), 3.61 (s, 3H), 2.97 (t, 2H, J=4.8 Hz), 2.69 (t, 2H, J=4.8 Hz); mp 100-102 C.

Example 23

N1,N1-dimethyl-N5-cycloheptyl Biguanide Hydrochloride

(49) ##STR00026##

(50) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 8.01 (br s, 2H), 6.93 (br s, 1H), 3.10 (m, 1H), 2.86 (s, 6H), 1.91 (m, 2H), 1.63 (m, 2H), 1.38-1.53 (m, 6H), 1.34 (m, 2H); mp 132-133 C.

Example 24

N1,N1-dimethyl-N5-cyclopropylmethyl Biguanide Hydrochloride

(51) ##STR00027##

(52) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 8.08 (br s, 2H), 6.94 (br s, 1H), 2.86 (s, 6H), 2.63 (d, 2H, J=4.8 Hz), 1.01 (m, 1H), 0.51 (m, 2H), 0.31 (m, 2H); mp 117-118 C.

Example 25

N1,N1-dimethyl-N5-(4-bromo)phenyl Biguanide Hydrochloride

(53) ##STR00028##

(54) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 10.07 (br s, 1H), 7.68 (br s, 2H), 7.40 (d, 2H, J=9.0 Hz), 7.35 (d, 2H, J=9.0 Hz), 2.92 (s, 6H); mp 272-273 C.

Example 26

N1,N1-dimethyl-N5-(furan-2-yl)methyl Biguanide Hydrochloride

(55) ##STR00029##

(56) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 7.59 (m, 1H), 7.38 (br s, 2H), 6.77 (br s, 1H), 6.40 (dd, 1H, J=2.7, 1.5 Hz), 6.31 (s, 1H), 4.31 (d, 2H, J=6.0 Hz), 2.92 (s, 6H); mp 176-177 C.

Example 27

N1,N1-dimethyl-N5-(pyridine-3-yl)methyl Biguanide Hydrochloride

(57) ##STR00030##

(58) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 8.65 (d, 1H, J=1.8 Hz), 8.62 (br s, 2H), 8.52 (dd, 1H, J=4.8, 1.8 Hz), 7.93 (ddd, 1H, J=7.8, 1.8, 1.8 Hz), 7.40 (dd, 1H, J=7.8, 4.8 Hz), 4.01 (s, 2H), 3.34 (s, 6H); mp 112-114 C.

Example 28

N1,N1-dimethyl-N5-benzyl Biguanide Hydrochloride

(59) ##STR00031##

(60) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 8.51 (br s, 2H), 7.46 (m, 2H), 7.36 (m, 3H), 6.91 (br s, 1H), 3.95 (s, 2H), 2.83 (s, 6H); mp 151-152 C.

Example 29

N1,N1-dimethyl-N5-(phenethyl) Biguanide Hydrochloride

(61) ##STR00032##

(62) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 8.24 (br s, 2H), 7.20-7.34 (m, 6H), 3.00 (t, 2H, J=7.8 Hz), 2.92 (s, 6H), 2.91 (t, 2H, J=7.8 Hz); mp 155-157 C.

Example 30

N1,N1-diethyl-N5-(3-chloro)phenyl Biguanide Hydrochloride

(63) ##STR00033##

(64) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.91 (br s, 1H), 7.64 (br s, 2H), 7.60 (dd, 1H, J=2.0, 2.0 Hz), 7.29 (d, 1H, J=8.0 Hz), 7.26 (ddd, H, 1H, J=80, J=8.0, 1.6, 1.6 Hz), 7.05 (ddd, 1H, J=8.0, 1.6, 1.6 Hz), 6.92 (br s, 1H), 3.31 (m, 4H), 1.08 (m, 6H); mp 219-220 C.

Example 31

N1,N1-dipropyl-N5-(3-chloro)phenyl Biguanide Hydrochloride

(65) ##STR00034##

(66) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 10.20 (br s, 1H), 7.70 (br s, 2H), 7.63 (d, 1H, J=1.8 Hz), 7.30 (m, 2H), 7.05 (d, 1H, J=7.8 Hz), 3.27 (m, 4H), 1.56 (m, 4H), 0.85 (m, 6H); mp 201-202 C.

Example 32

N1,N1-(ethyl)(propyl)-N5-(4-chloro)phenyl Biguanide Hydrochloride

(67) ##STR00035##

(68) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 9.71 (br s, 1H), 8.69 (br s, 1H), 7.42 (d, 2H, J=6.6 Hz), 7.35 (d, 2H, J=6.6 Hz), 7.23 (br s, 2H), 2.90 (q, 2H, J=7.2 Hz), 2.80 (t, 2H, J=7.2 Hz), 1.61 (m, 2H), 1.18 (t, 3H, J=7.2 Hz), 0.91 (t, 3H, J=7.2 Hz); mp 110-112 C.

Example 33

N1,N1-dipropyl-N5-(isoquinoline-5-yl) Biguanide Hydrochloride

(69) ##STR00036##

(70) .sup.1H NMR (400 MHz, DMSO-d.sub.6) 9.27 (br s, 2H), 8.64 (d, 1H, J=8.4 Hz), 7.97 (m, 2H), 7.78 (ddd, 1H, J=8.4, 6.0, 2.4 Hz), 7.71 (d, 1H, J=7.2 Hz), 7.38 (br s, 1H), 7.23 (d, 1H, J=7.2 Hz), 3.21 (t, 2H, J=7.6 Hz), 0.85 (t, 3H, J=7.2 Hz); mp 191-192 C.

Example 34

N1,N1-dihexyl-N5-(3-chloro)phenyl Biguanide Hydrochloride

(71) ##STR00037##

(72) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 10.07 (br s, 1H), 7.67 (br s, 1H), 7.62 (dd, 1H, J=2.4, 1.8 Hz), 7.29 (dd, 1H, J=7.8, 7.8 Hz), 7.25 (ddd, 1H, J=7.8, 1.8, 1.2 Hz), 7.05 (ddd, 1H, J=7.8, 2.4, 1.2 Hz), 3.27 (t, 4H, J=7.8 Hz), 1.52 (m, 4H), 1.18-1.34 (m, 12H), 0.82 (m, 6H); mp 187-188 C.

Example 35

N1,N1,N5,N5-tetraethyl Biguanide Hydrochloride

(73) ##STR00038##

(74) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 8.76 (br s, 2H), 6.90 (br s, 1H), 2.87 (q, 8H, J=7.2 Hz), 1.17 (t, 12H, J=7.2 Hz); mp 140-141 C.

Example 36

N1,N1-diethyl-N5,N5-(cyclohexyl)(methyl) Biguanide Hydrochloride

(75) ##STR00039##

(76) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 8.73 (br s, 2H), 6.90 (br s, 1H), 3.26 (q, 4H, J=7.2 Hz), 2.86 (m, 1H), 2.49 (s, 3H), 1.99 (m, 2H), 1.74 (m, 2H), 1.20-1.30 (m, 6H), 1.02 (t, 6H, J=7.2 Hz); mp 115-117 C.

Example 37

N1,N1-dipropyl-N5,N5-diethyl Biguanide Hydrochloride

(77) ##STR00040##

(78) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 6.92 (br s, 1H), 6.89 (br s, 1H), 3.30 (m, 4H), 3.21 (m, 4H), 1.51 (m, 4H), 1.08 (t, 6H, J=6.0 Hz), 0.84 (t, 6H, J=6.6 Hz); mp 151-152 C.

Example 38

N1,N1-dipropyl-N5,N5-(methyl)(phenethyl) Biguanide Hydrochloride

(79) ##STR00041##

(80) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 7.16-7.30 (m, 5H), 6.90 (br s, 2H), 3.47 (t, 2H, J=7.8 Hz), 3.18 (t, 4H, J=7.2 Hz), 2.84 (s, 3H), 2.76 (t, 2H, J=7.8 Hz), 1.49 (m, 4H), 0.83 (m, 6H); mp 110-111 C.

Example 39

N1,N1-dipropyl-N5,N5-(4-hydroxylphenyl)(phenyl) Biguanide Hydrochloride

(81) ##STR00042##

(82) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 9.86 (br s, 1H), 7.20-7.38 (m, 5H), 7.16 (d, 2H, J=8.4 Hz), 6.84 (br s, 2H), 6.81 (d, 2H, J=8.4 Hz), 3.10 (t, 4H, J=5.4 Hz), 1.54 (m, 4H), 0.89 (t, 6H, J=7.2 Hz); mp 189-190 C.

Example 40

N1,N1,N5,N5-bis((benzyl)(methyl)) Biguanide Hydrochloride

(83) ##STR00043##

(84) .sup.1H NMR (600 MHz, DMSO-d.sub.6) 7.59 (m, 4H), 7.41 (m, 6H), 7.00 (br s, 2H), 4.07 (s, 4H), 2.47 (s, 6H); mp 141-142 C.

EXPERIMENTAL EXAMPLES

(85) The compounds synthesized by the method described in the examples of the present invention were treated to cancer cells, according to a method to be described in the following Experimental Examples, to measure an effect on inhibition of cancer cell proliferation. A simple experimental method is as follows:

Experimental Example 1

Measurement of Effect on Inhibition of Cancer Cell Proliferation

(86) HCT116 cells derived from human colorectal cancer were used, and an effect of a biguanide derivative on the inhibition of cancer cell proliferation was confirmed by measuring a concentration value (cell growth inhibition concentration, GIC50) at which 50% of the cell growth was inhibited using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenytetrazolium bromide (MTT) reagent.

(87) First, HCT116 cells were put on a 96-well plate and cultured in a DMEM medium containing 10% bovine serum for 24 hours to each have cell count of approximately 5000. Subsequently, to obtain the GIC50 value of each compound, 100 M (or 200 M), 25 M, 6.25 M, 1.56 M or 0.39 M of the compound was treated to each culture medium and then incubated for 48 hours. To confirm living cells after treatment with the compound, MTT was added to each culture medium and further incubated for 3 hours. Generated formazane crystal was dissolved using dimethyl sulfoxide (DMSO) and absorbance of the solution was measured at 560 nm. After the 48-hour incubation, a ratio of a cell count cultured on a well plate not treated with the compound to a cell count on a well plate treated with compounds synthesized in the examples was indicated as cell viability (%) according to each administered concentration. A cell viability curve was plotted using the cell viability (%), and the calculated concentration value (GIC50) of the compound, at which 50% of the growth was inhibited was, to confirm an effect on the inhibition of cancer cell proliferation.

(88) Results of cancer cell growth inhibition effect are shown in Table 1.

(89) TABLE-US-00001 TABLE 1 Example GIC50 (uM) @ HCT116 Metformin HCl 2172 1 23.8 2 20.7 3 >100 4 8.4 5 3.2 6 >100 7 18.3 8 17.0 9 5.9 10 6.3 11 3.1 12 >200 13 47.6 14 47.0 15 17.2 16 >100 17 >200 18 >200 19 >200 20 >200 21 >200 22 >200 23 >200 24 >200 25 >100 26 >200 27 >200 28 >200 29 >200 30 28.3 31 7.6 32 >200 33 >100 34 7.8 35 >200 36 >200 37 >100 38 36.0 39 >100 40 >100

Experimental Example 2

Measurement of Effect on AMPK Activation

(90) MCF7 cells derived from human breast cancer cells were used, and an effect of a biguanide derivative on 5-AMP-activated protein kinase (AMPK) activation was confirmed using an AMPK immunoassay kit (Invitrogen, catalog No. KHO0651).

(91) The MCF7 cells were put on a 6-well plate and incubated in a DMEM medium containing 10% fetal bovine serum in an incubator to which 5% CO.sub.2 was supplied to have a cell count of approximately 510.sup.5. 50 M of derivatives synthesized in the examples were treated to the each culture medium, and the cells were incubated for 24 hours. Subsequently, the cells were lysed by a method presented in the operation manual of the AMPK immunoassay kit, and 20 g of cell lysates were yielded through protein assay. A degree of phosphorylation of an AMPK threonine 172.sup.nd residue (Thr172) from the cell lysates were confirmed according to a method presented in the operation manual of the AMPK immunoassay kit to thereby obtain results. A degree of the AMPK activation by a biguanide derivative was exhibited as a degree of phosphorylated AMPK in the cells cultured in the presence of the compounds synthesized in the examples based on phosphorylated AMPK in cells cultured without treating the biguanide derivative.

(92) In addition, an experiment was performed in the same manner as described in Experimental Example 2 using metformin as a control group, and the results of an effect on AMPK activation were compared to the effect on AMPK activation when 1 mM metformin was treated.

(93) The results are shown in Table 2.

(94) TABLE-US-00002 TABLE 2 AMPK Activation Example 0 50 M fold Metformin HCl 6.8 21.5 (@ 1 mM) 3.2 1 5.3 35.6 6.7 2 5.3 29.3 5.5 3 6.8 13.1 1.9 4 2.5 15.5 6.2 5 N.D 6 6.8 4.6 0.7 7 N.D 8 N.D 9 N.D 10 N.D 11 N.D 12 5.3 7.8 1.5 13 5.3 36.2 6.8 14 5.3 31.2 5.9 15 4.6 7.0 1.5 16 6.8 4.7 0.7 17 4.9 5.5 1.1 18 5.3 4.6 0.9 19 5.3 3.6 0.7 20 5.3 4.6 0.9 21 5.3 5.6 1.1 22 5.3 3.2 0.6 23 5.3 5.4 1.0 24 5.3 5.1 1.0 25 5.3 23.1 4.4 26 5.3 8.9 1.7 27 5.3 4.6 0.9 28 4.9 5.9 1.2 29 5.3 14.7 2.8 30 2.5 19.3 7.7 31 N.D 32 5.3 5.2 1.0 33 6.8 11.3 1.7 34 N.D 35 5.3 3.8 0.7 36 5.3 4.5 0.8 37 5.3 33.3 6.3 38 5.3 35.8 6.8 39 6.8 28.8 4.2 40 6.8 11.3 1.7

(95) Consequently, it was seen that the derivatives synthesized in the examples effectively inhibited the viability of cancer cells, particularly, colorectal cancer cells in terms of the effect on inhibition of cancer cell proliferation. In addition, it could be observed that the compounds exhibiting a greater effect on AMPK activation at a concentration 20 times lower than the control group, metformin, may have an effect at least 20 times greater than the control group.