PHENANTHRENE AMPK ACTIVATOR COMPOUNDS, COMPOSITIONS, METHODS AND USES THEREOF

Abstract

The present invention relates to a compound having general formula I for use in the activation of AMPK. A composition comprising said compound for use in the activation of AMPK is also provided.

Claims

1. A method for the activation of AMPK, comprising administering a compound having the formula I ##STR00010## wherein R1, R2, R3, R4, R5, R6, R7, and R8 are each independently selected from the group consisting of H; OH; OCH3; O-glycoside; C-glycoside; acylated O-glycoside; acylated C-glycoside; sulfated O-glycoside; sulfated C-glycoside; a halogen; a primary, secondary, or tertiary alcohol; a ketone; an aldehyde; a carboxylic acid; an ester; a primary, secondary, or tertiary amine; a primary or secondary amide; a cyano; a nitro; a sulfonate; a sulfate; an optionally substituted and/or optionally branched C1 to C20 alkyl; a substituted and/or branched, C2 to C20 alkenyl; a substituted and/or branched, C4 to C20 polyalkenyl; a substituted and/or branched C2 to C20 alkynyl, or a substituted and/or branched C4 to C20 polyalkynyl, or a derivative or analogue thereof, a OCH3 group can cyclize with a neighboring OH group to form a methylene dioxy bridge.

2. A method according to claim 1 wherein R1 and R8 are each independently selected from the group consisting of H; OH; OCH3; O-glycoside; C-glycoside; acylated O-glycoside; acylated C-glycoside; sulfated O-glycoside; sulfated C-glycoside; a halogen; a primary, secondary, or tertiary alcohol; a ketone; an aldehyde; an ester; a primary, secondary, or tertiary amine; a primary or secondary amide; a cyano; a nitro; a sulfonate; a sulfate; a substituted and/or branched C1 to C20 alkyl; a substituted and/or branched, C2 to C20 alkenyl; a substituted and/or branched, C4 to C20 polyalkenyl; a substituted and/or branched C2 to C20 alkynyl, or a substituted and/or branched C4 to C20 polyalkynyl; R2 and R7 are each independently selected from the group consisting of H; OH; OCH3; O-glycoside; C-glycoside; acylated O-glycoside; acylated C-glycoside; sulfated O-glycoside; sulfated C-glycoside; a halogen; a secondary, or tertiary alcohol; a ketone; an aldehyde; a carboxylic acid; an ester; a primary, secondary, or tertiary amine; a primary or secondary amide; a cyano; a nitro; a sulfonate; a sulfate; a substituted and/or branched C1 to C20 alkyl; a substituted and/or branched, C2 to C20 alkenyl; a substituted and/or branched, C4 to C20 polyalkenyl; a substituted and/or branched C2 to C20 alkynyl, or a substituted and/or branched C4 to C20 polyalkynyl; R3 and R6 are each independently selected from the group consisting of H; OH; OCH3; O-glycoside; C-glycoside; acylated O-glycoside; acylated C-glycoside; sulfated O-glycoside; sulfated C-glycoside; a halogen; a primary, secondary, or tertiary alcohol; a ketone; an aldehyde; a carboxylic acid; an ester; a primary, secondary, or tertiary amine; a primary or secondary amide; a cyano; a nitro; a sulfonate; a sulfate; a substituted and/or branched C1 to C20 alkyl; a substituted and/or branched, C2 to C20 alkenyl; a substituted and/or branched, C4 to C20 polyalkenyl; a substituted and/or branched C2 to C20 alkynyl, or a substituted and/or branched C4 to C20 polyalkynyl; R4 and R5 are each independently selected from the group consisting of H; OH; O-glycoside; C-glycoside; acylated O-glycoside; acylated C-glycoside; sulfated O-glycoside; sulfated C-glycoside; a halogen; a primary, secondary, or tertiary alcohol; a ketone; an aldehyde; a carboxylic acid; an ester; a primary, secondary, or tertiary amine; a primary or secondary amide; a cyano; a nitro; a sulfonate; a sulfate; a substituted and/or branched C1 to C20 alkyl; a substituted and/or branched, C2 to C20 alkenyl; a substituted and/or branched, C4 to C20 polyalkenyl; a substituted and/or branched C2 to C20 alkynyl, or a substituted and/or branched C4 to C20 polyalkynyl; or a derivative or analogue thereof, a OCH3 group can cyclize with a neighboring OH group to form a methylene dioxy bridge.

3. A method according to claim 1 wherein R1 and R8 are each independently selected from the group consisting of H; CH3; OH; OCH3; O-glycoside; a sulfate; Br; CHO; CH2OH; CONH2, COCH3; CH2-COOH; CH2COOCH3; CH═CH2; CH2-CH═C(CH3)2; CH(CH3)2; CH═CH—CHO; CH(CH3)-OH; CH(CH3)-OCH3; CH(CH3)-OC2H5; CH(CH3)-O—CH2-CH═C (CH3)-(CH2)3-CH(CH3)-(CH2)3-CH(CH3)-(CH2)3-CH(CH3)2; (CH2)8-CH═CH2; CH2-CO—CH2-CO—CH2-C(OCH3)-(CH2)4-CH3; C≡C—(CH2)2-CO—CH3; (CH2)2-NH2; (CH2)2-NH—CO—CH3; CHOH—CH2-N(CH3)2; (CH2)2-N(CH3)2; (CH2)2-NH—CH3; (CH2)2-N(OH)—CH3; (CH2)2-N(CH3)2=O; (CH2)2-N+(CH3)3; (CH2)2-N(CH3)-CO—CH3; NH—CO—CH3; NH—CH═CH2; 4-hydroxybenzyl; 3,4-dihydroxybenzyl; 4-hydroxy-3-methoxybenzyl; 2-bromo-3,4-dihydroxybenzyl; R2 and R7 are each independently selected from the group consisting of H; CH3; OH; OCH3; O-glycoside; a sulfate; Br; CHO; COOH, CONH2, COCH3; CH2-COOH; CH2COOCH3; CH═CH2; CH2-CH═C(CH3)2; CH(CH3)2; CH═CH—CHO; CH(CH3)-OH; CH(CH3)-OCH3; CH(CH3)-OC2H5; CH(CH3)-O—CH2-CH═C(CH3)-(CH2)3-CH(CH3)-(CH2)3-CH(CH3)-(CH2)3-CH(CH3)2; (CH2)8-CH═CH2; CH2-CO—CH2-CO—CH2-C(OCH3)-(CH2)4-CH3; C≡C—(CH2)2-CO—CH3; (CH2)2-NH2; (CH2)2-NH—CO—CH3; CHOH—CH2-N(CH3)2; (CH2)2-N(CH3)2; (CH2)2-NH—CH3; (CH2)2-N(OH)—CH3; (CH2)2-N(CH3)2=O; (CH2)2-N+(CH3)3; (CH2)2-N(CH3)-CO—CH3; NH—CO—CH3; NH—CH═CH2; 4-hydroxybenzyl; 3,4-dihydroxybenzyl; 4-hydroxy-3-methoxybenzyl; 2-bromo-3,4-dihydroxybenzyl; R4 and R5 are each independently selected from the group consisting of H; CH3; OH; O-glycoside; a sulfate; Br; CHO; CH2OH; COOH, CONH2, COCH3; CH2-COOH; CH2COOCH3; CH═CH2; CH2-CH═C(CH3)2; CH(CH3)2; CH═CH—CHO; CH(CH3)-OH; CH(CH3)-OCH3; CH(CH3)-OC2H5; CH(CH3)-O—CH2-CH═C(CH3)-(CH2)3-CH(CH3)-(CH2)3-CH(CH3)-(CH2)3-CH(CH3)2; (CH2)8-CH═CH2; CH2-CO—CH2-CO—CH2-C(OCH3)-(CH2)4-CH3; C≡C—(CH2)2-CO—CH3; (CH2)2-NH2; (CH2)2-NH—CO—CH3; CHOH—CH2-N(CH3)2; (CH2)2-N(CH3)2; (CH2)2-NH—CH3; (CH2)2-N(OH)—CH3; (CH2)2-N(CH3)2=O; (CH2)2-N+(CH3)3; (CH2)2-N(CH3)-CO—CH3; NH—CO—CH3; NH—CH═CH2; 4-hydroxybenzyl; 3,4-dihydroxybenzyl; 4-hydroxy-3-methoxybenzyl; 2-bromo-3,4-dihydroxybenzyl; R3 and R6, are each independently selected from the group consisting of H; CH3; OH; OCH3; O-glycoside; a sulfate; Br; CHO; CH2OH; COOH, CONH2, COCH3; CH2-COOH; CH2COOCH3; CH═CH2; CH2-CH═C(CH3)2; CH(CH3)2; CH═CH—CHO; CH(CH3)-OH; CH(CH3)-OCH3; CH(CH3)-OC2H5; CH(CH3)-O—CH2-CH═C (CH3)-(CH2)3-CH(CH3)-(CH2)3-CH(CH3)-(CH2)3-CH(CH3)2; (CH2)8-CH═CH2; CH2-CO—CH2-CO—CH2-C(OCH3)-(CH2)4-CH3; C≡C—(CH2)2-CO—CH3; (CH2)2-NH2; (CH2)2-NH—CO—CH3; CHOH—CH2-N(CH3)2; (CH2)2-N(CH3)2; (CH2)2-NH—CH3; (CH2)2-N(OH)—CH3; (CH2)2-N(CH3)2=0; (CH2)2-N+(CH3)3; (CH2)2-N(CH3)-CO—CH3; NH—CO—CH3; NH—CH═CH2; 4-hydroxybenzyl; 3,4-dihydroxybenzyl; 4-hydroxy-3-methoxybenzyl; 2-bromo-3,4-dihydroxybenzyl, or a derivative or analogue thereof, and a OCH3 group can cyclize with a neighboring OH group to form a methylene dioxy bridge.

4. A method according to claim 1 wherein R1, R2, R3, R6, R7, and R8 are each independently selected from the group consisting of H; CH3; OH; OCH3; O-glycoside; a sulfate; CH2-CH═C(CH3)2; R4 and R5 are each independently selected from the group consisting of H; CH3; OH; O-glycoside; a sulfate; CH2-CH═C(CH3)2 or a derivative or analogue thereof, optionally and a OCH3 group can cyclize with a neighboring OH group to form a methylene dioxy bridge.

5. A method according to claim 1 wherein R1 and R3 are each independently selected from the group consisting of H; CH3; OH; OCH3; O-glycoside; a sulfate; CH2-CH═C (CH3)2; 4-hydroxybenzyl; 3,4-dihydroxybenzyl; or 4-hydroxy-3-methoxybenzyl; R2, R4 and R7 are each independently selected from the group consisting of OH; OCH3; O-glycoside; or a sulfate; R5 is H; OH; OCH3; O-glycoside; or a sulfate; R6 and R8 are each independently selected from the group consisting of H; CH3; OH; OCH3; O-glycoside; a sulfate; CH2-CH═C (CH3)2; 4-hydroxybenzyl; 3,4-dihydroxybenzyl; or 4-hydroxy-3-methoxybenzyl, or a derivative or analogue thereof, optionally and a OCH3 group can cyclize with a neighboring OH group to form a methylene dioxy bridge.

6. A method according to claim 1 wherein R1 and R3 are H; R2 and R4 are each independently selected from the group consisting of OH; OCH3; O—CH═CH2; O-glycoside; or a sulfate; R5, R6, R7; and R8 are each independently selected from the group consisting of H; OH; OCH3; O-glycoside; or a sulfate, or a derivative or analogue thereof, optionally and a OCH3 group can cyclize with a neighboring OH group to form a methylene dioxy bridge.

7. A compound method according to claim 1, wherein the compound is Lusianthrin also known as 7-Methoxyphenanthrene-2,5-diol, 7-Methoxy-2,5-phenanthrenediol, 2,5-Phenanthrenediol, 7-methoxy, CAS number 126767-85-9.

8. A method according to claim 1, wherein the compound is 2-Methoxyphenanthrene-4,5-diol, 4,5-Phenanthrenediol, 2-methoxy, CAS 874659-27-5.

9. A method according to claim 1 for the activation of AMPK to treat or prevent a condition, disorder, or disease related to cardiometabolic health, obesity, type 2 diabetes, non-alcoholic fatty liver disease, cardiovascular disease, and/or cancer.

10. A method according to claim 9, wherein the subject is a human.

11. A method according to claim 1, wherein the activation of AMPK is through a direct activation mechanism.

12. A method according to claim 1, wherein the activation of AMPK is in muscle and liver tissues.

13. A method according to claim 1, wherein the activation of AMPK is AMPK which comprises an α2 subunit, a β1 subunit, and a γ1 subunit.

14. A method according to claim 1, wherein the compound is obtained from a plant or plant extract.

15-16. (canceled)

17. A method according to claim 1, wherein the composition is a food, beverage, or dietary supplement.

18. A method according to claim 1, wherein the composition further comprises a pharmaceutically acceptable carrier.

19-21. (canceled)

22. An in vitro method of activating AMPK, comprising contacting a compound of general formula I ##STR00011## wherein R1, R2, R3, R4, R5, R6, R7, and R8 are each independently selected from the group consisting of H; OH; OCH3; O-glycoside; C-glycoside; acylated O-glycoside; acylated C-glycoside; sulfated O-glycoside; sulfated C-glycoside; a halogen; a primary, secondary, or tertiary alcohol; a ketone; an aldehyde; a carboxylic acid; an ester; a primary, secondary, or tertiary amine; a primary or secondary amide; a cyano; a nitro; a sulfonate; a sulfate; an optionally substituted and/or branched C1 to C20 alkyl; a substituted and/or branched, C2 to C20 alkenyl; a substituted and/or branched, C4 to C20 polyalkenyl; a substituted and/or branched C2 to C20 alkynyl, or a substituted and/or branched C4 to C20 polyalkynyl, or a derivative or analogue thereof, a OCH3 group can cyclize with a neighboring OH group to form a methylene dioxy bridge with AMPK.

23. A method of treatment or prevention of a condition, disorder, or disease related to cardiometabolic health, obesity, type 2 diabetes, non-alcoholic fatty liver disease, cardiovascular disease, and/or cancer comprising administration of a composition comprising administering a compound having the formula I ##STR00012## wherein R1, R2, R3, R4, R5, R6, R7, and R8 are each independently selected from the group consisting of H; OH; OCH3; O-glycoside; C-glycoside; acylated O-glycoside; acylated C-glycoside; sulfated O-glycoside; sulfated C-glycoside; a halogen; a primary, secondary, or tertiary alcohol; a ketone; an aldehyde; a carboxylic acid; an ester; a primary, secondary, or tertiary amine; a primary or secondary amide; a cyano; a nitro; a sulfonate; a sulfate; an optionally substituted and/or branched C1 to C20 alkyl; a substituted and/or branched, C2 to C20 alkenyl; a substituted and/or branched, C4 to C20 polyalkenyl; a substituted and/or branched C2 to C20 alkynyl, or a substituted and/or branched C4 to C20 polyalkynyl, or a derivative or analogue thereof, a OCH3 group can cyclize with a neighboring OH group to form a methylene dioxy bridge to an individual in need of same.

Description

BRIEF DESCRIPTION OF FIGURES

[0184] FIG. 1. Compound 1 and 2 activation of bacterially-expressed AMPKα2β1γ1.

[0185] Compound 1 is Lusianthrin, also known as 7-Methoxyphenanthrene-2,5-diol, 7-Methoxy-2,5-phenanthrenediol, 2,5-Phenanthrenediol, 7-methoxy, CAS number 126767-85-9.

[0186] Compound 2 is 2-Methoxyphenanthrene-4,5-diol, 4,5-Phenanthrenediol, 2-methoxy, CAS 874659-27-5.

[0187] FIG. 2. Compound 1 and 2 increases the phosphorylation of the AMPK substrate, acetyl-CoA carboxylase (ACC), in U2OS Flp-In T-REx mammalian cells.

[0188] Compound 1 is Lusianthrin, also known as 7-Methoxyphenanthrene-2,5-diol, 7-Methoxy-2,5-phenanthrenediol, 2,5-Phenanthrenediol, 7-methoxy, CAS number 126767-85-9.

[0189] Compound 2 is 2-Methoxyphenanthrene-4,5-diol, 4,5-Phenanthrenediol, 2-methoxy, CAS 874659-27-5.

EXAMPLES

Example 1: Synthesis of Lusianthrin from (3-(benzyloxy)-5-methoxybenzyl)triphenylphosphonium Bromide and 5-(benzyloxy)-2-iodobenzaldehyde

Part 1: Synthesis of (3-(benzyloxy)-5-methoxybenzyl)triphenylphosphonium bromide

[0190] After suitable protection, 3,5-dihydroxybenzoic acid methyl ester was reduced to a primary alcohol, and converted to its corresponding alkyl halide before reaction with triphenylphosphine to give the desired triphenylphosphonium ylide reagent (Scheme 1).

##STR00007##

[0191] Step a. To a solution of methyl 3,5-dihydroxybenzoate 1 (300 g, 1784.12 mmol) in acetone (7200 mL) was added potassium carbonate (271.22 g, 1962.53 mmol). The suspension was stirred at room temperature for 10 min. Benzyl bromide (222.50 mL, 1873.32 mmol) was added, and the resultant suspension was heated at 60° C. for 12 h. After cooling to room temperature, the suspension was filtered, the filter cake washed with acetone, and the filtrate was concentrated to a residue. The residue was purified by automated normal-phase chromatography and eluted with ethyl acetate/hexanes to give methyl 3-(benzyloxy)-5-hydroxybenzoate 2 as an off-white solid. (144 g, 31% yield). 1H NMR (300 MHz, DMSO-d6) δ ppm: 9.89 (s, 1H), 7.33-7.46 (m, 5H), 7.01 (dd, J=6.30, 0.90 Hz, 2H), 6.67 (t, J=2.40 Hz, 1H), 5.11 (s, 2H), 3.82 (s, 3H); MS (ES+) m/z 257.1 [M−H]+; HPLC-UV analysis: retention time=13.35 min; detection: 190-400 nm: peak area, 99.81%; eluent A, 0.1% TFA in water; eluent B, Acetonitrile; isocratic/gradient over 30 min with a flow rate of 1.0 mL min-1.

[0192] Step b. To a solution of methyl 3-(benzyloxy)-5-hydroxybenzoate 2 (140 g, 542.06 mmol) in acetone (7000 mL) was added potassium carbonate (224.74 g, 1626.20 mmol). The suspension was stirred at room temperature for 10 min. Iodomethane (168.73 mL, 2710.34 mmol) was added, and the resultant suspension was stirred at room temperature for 16 h. The suspension was filtered, the filter cake washed with acetone, and the filtrate was concentrated to a residue. The residue was purified by automated normal-phase chromatography and eluted with ethyl acetate/hexanes to give methyl 3-(benzyloxy)-5-methoxybenzoate 3 as liquid. (125 g, 94% yield). 1H NMR (300 MHz, DMSO-d6) δ ppm: 7.33-7.48 (m, 6H), 7.16 (t, J=2.10 Hz, 1H), 7.08 (d, J=1.20 Hz, 1H), 6.87 (t, J=2.40 Hz, 1H), 5.15 (s, 2H), 3.84 (s, 3H), 3.79 (s, 3H); MS (ES+) m/z 273.1 [M+H]+; HPLC-UV analysis: retention time=15.31 min; detection: 190-400 nm: peak area, 99.78%; eluent A, 0.1% TFA in water; eluent B, Acetonitrile; isocratic/gradient over 30 min with a flow rate of 1.0 mL min-1.

[0193] Step c. Lithium aluminium hydride (16.86 g, 444.36 mmol) in THF (605 mL) was added to methyl 3-(benzyloxy)-5-methoxybenzoate 3 (121 g, 444.36 mmol) in THF (1600 mL) at 0° C. The suspension was stirred at 0° C. for 20 min, at room temperature for 1 h. The reaction mixture was diluted with THF and quenched by addition of water. The resultant mixture was filtered through a pad of celite, and washed with ethyl acetate. The filtrate was concentrated in vacuo to give (3-(benzyloxy)-5-methoxyphenyl)methanol 4 as liquid. (100 g, 92% yield). 1H NMR (300 MHz, DMSO-d6) δ ppm: 7.30-7.46 (m, 5H), 6.59 (d, J=0.60 Hz, 1H), 6.51 (s, 1H), 6.45 (d, J=2.40 Hz, 1H), 5.19 (t, J=5.70 Hz, 1H), 5.07 (s, 2H), 4.44 (d, J=5.70 Hz, 2H), 3.72 (s, 3H); MS (ES+) m/z 245.1 [M+H]+; HPLC-UV analysis: retention time=12.86 min; detection: 190-400 nm: peak area, 99.64%; eluent A, 0.1% TFA in water; eluent B, Acetonitrile; isocratic/gradient over 30 min with a flow rate of 1.0 mL min-1.

[0194] Step d. To a solution of (3-(benzyloxy)-5-methoxyphenyl)methanol 4 (100 g, 409.34 mmol) in 1,4-dioxane (1000 mL) was added phosphorous tribromide (50.54 mL, 532.15 mmol). The reaction mixture was stirred at 40° C. for 1 h and quenched by addition of water. The aqueous phase was extracted with ethyl acetate, and the combined organic extracts were washed with water, brine and concentrated to give 1-(benzyloxy)-3-(bromomethyl)-5-methoxybenzene 5 as pale yellow solid. (100 g, 80% yield). 1H NMR (300 MHz, DMSO-d6) δ ppm: 7.33-7.46 (m, 5H), 6.72 (s, 1H), 6.63 (s, 1H), 6.54 (d, J=1.80 Hz, 1H), 5.09 (d, J=5.40 Hz, 2H), 4.62 (d, J=5.70 Hz, 2H), 3.74 (s, 3H); MS (ES+) m/z 309 [M+2H]+; HPLC-UV analysis: retention time=15.77 min; detection: 190-400 nm: peak area, 99.71%; eluent A, 0.1% TFA in water; eluent B, Acetonitrile; isocratic/gradient over 30 min with a flow rate of 1.0 mL min-1.

[0195] Step e. To a solution of 1-(benzyloxy)-3-(bromomethyl)-5-methoxybenzene 5 (100 g, 325.53 mmol) in toluene (2488 mL) was added triphenylphosphine (85.38 g, 325.53 mmol). The reaction mixture was stirred at 100° C. for 6 h, then allowed to cool to room temperature. The solid was collected by filtration, washed with ether, and dried under vacuum to give (3-(benzyloxy)-5-methoxybenzyl)triphenylphosphonium bromide 6 as an off-white solid. (150 g, 82% yield). 1H NMR (400 MHz, DMSO-d6) δ ppm: 7.89-7.91 (m, 3H), 7.65-7.75 (m, 12H), 7.28-7.37 (m, 5H), 6.51 (s, 1H), 6.23 (s, 1H), 6.12 (s, 1H) 5.07 (d, J=15.60 Hz, 2H), 4.82 (s, 2H), 3.48 (s, 3H); MS (ES+) m/z 489.2 [M−HBr]+; HPLC-UV analysis: retention time=14.19 min; detection: 190-400 nm: peak area, 95.51%; eluent A, 0.1% TFA in water; eluent B, Acetonitrile; isocratic/gradient over 30 min with a flow rate of 1.0 mL min-1.

Part 2: Synthesis of 5-(benzyloxy)-2-iodobenzaldehyde

[0196] 3-Hydroxybenzaldehyde was protected before ortho iodination, as displayed in Scheme 2.

##STR00008##

[0197] Step a. To a solution of 3-hydroxybenzaldehyde 7 (25 g, 204.85 mmol) in acetone (250 mL) was added potassium carbonate (42.46 g, 307.27 mmol). The suspension was stirred at room temperature for 10 min. Benzyl bromide (31.38 mL, 264.25 mmol) was added, and the resultant suspension was heated at 60° C. for 12 h. After cooling to room temperature, the suspension was filtered, the filter cake washed with acetone, and filtrate concentrated to a residue. The residue was purified by automated normal-phase chromatography and eluted with ethyl acetate/hexanes to give 3-(benzyloxy)benzaldehyde 8 as an off-white solid. (42 g, 96% yield). 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.98 (s, 1H), 7.27-7.50 (m, 5H), 7.25-7.26 (m, 2H), 5.14 (s, 2H); GCMS: m/z 212.1: (GCMS condition: column: HP-5 (30 m×320 μm×0.25 μm); gradient: 120° C.-300° C., 40° C. min-1; HPLC-UV analysis: retention time=14.37 min; detection: 190-400 nm: peak area, 99.58%; eluent A, 0.1% TFA in water; eluent B, Acetonitrile; isocratic/gradient over 30 min with a flow rate of 1.0 mL min-1.

[0198] Step b. To a solution of 3-(benzyloxy)benzaldehyde 8 (42 g, 197.87 mmol) in chloroform (1050 mL) was added Silver trifluoroacetate (65.56 g, 296.81 mmol). The suspension was stirred at 0° C. for 10 min. Iodine (32.43 g, 126.90 mmol) was added at 0° C. and the resultant suspension was stirred at room temperature for 12 h and quenched by addition of water. The resultant mixture was filtered through a pad of celite, washed with dichloromethane. The aqueous phase was extracted dichloromethane, and the combined organic extracts were washed with water, brine and concentrated to a residue. The residue was purified by automated normal-phase chromatography and eluted with ethyl acetate/hexanes to give 5-(benzyloxy)-2-iodobenzaldehyde 9 as an off-white solid. (40 g, 59% yield). 1H NMR (400 MHz, CDCl3) δ ppm: 10.04 (s, 1H), 7.83 (d, J=8.40 Hz, 1H), 7.54 (s, 1H), 7.37-7.53 (m, 5H), 7.01 (dd, J=8.80, 3.20 Hz, 1H), 5.12 (s, 2H); GCMS m/z 338: (GCMS condition: column: ZB1MS (10 m×100 μm×0.1 μm); gradient: 120° C.-300° C., 40° C. min-1; HPLC-UV analysis: retention time=16.04 min; detection: 190-400 nm: peak area, 99.84%; eluent A, 0.1% TFA in water; eluent B, Acetonitrile; isocratic/gradient over 30 min with a flow rate of 1.0 mL min-1.

Part 3: Synthesis of Lusianthrin

[0199] Lusianthrin was prepared through a Wittig reaction between (3-(benzyloxy)-5-methoxybenzyl)triphenylphosphonium bromide and 5-(benzyloxy)-2-iodobenzaldehyde, followed by cyclization and deprotection, as shown in Scheme 3.

##STR00009##

[0200] Step a. To a solution of 5-(benzyloxy)-2-iodobenzaldehyde 9 (36 g, 106.46 mmol) in THF (3600 mL) was added (3-(benzyloxy)-5-methoxybenzyl)triphenylphosphonium bromide 6 (127.32 g, 223.57 mmol). The suspension was stirred at 0° C. Potassium tert-butoxide (26.28 g, 234.08 mmol) was added at 0° C. and the resultant suspension was stirred at room temperature for 12 h. The reaction mixture was concentrated to a residue and the residue was purified by automated normal-phase chromatography and eluted with ethyl acetate/hexanes to give (Z)-4-(benzyloxy)-2-(3-(benzyloxy)-5-methoxystyryl)-1-iodobenzene 10 as an off-white solid. (50 g, 86% yield). 1H NMR (400 MHz, CDCl3) δ ppm: 7.74 (d, J=8.80 Hz, 1H), 7.26-7.42 (m, 8H), 6.89 (d, J=3.20 Hz, 1H), 6.49-6.65 (m, 4H), 6.40 (s, 3H), 6.33 (t, J=1.60 Hz, 1H), 4.85 (s, 4H), 3.62 (s, 3H); MS (ES+) m/z 549.1 [M+H]+; HPLC-UV analysis: retention time=18.74 min; detection: 190-400 nm: peak area, 89.98%; eluent A, 0.1% TFA in water; eluent B, Acetonitrile; isocratic/gradient over 30 min with a flow rate of 1.0 mL min-1.

[0201] Step b. To a solution of (Z)-4-(benzyloxy)-2-(3-(benzyloxy)-5-methoxystyryl)-1-iodobenzene 10 (50 g, 91.17 mmol) in toluene (1250 mL) was added tributyltin hydride (49.14 mL, 182.34 mmol) and azobisisobutyronitrile (7.48 g, 45.58 mmol). The reaction mixture was sparged with nitrogen for 5 min and heated at 100° C. for 16 h. The reaction mixture was concentrated to a residue and the residue was purified by automated normal-phase chromatography and eluted with ethyl acetate/hexanes to give 1:1 mixture of 4,7-bis(benzyloxy)-2-methoxyphenanthrene 11a and 2,7-bis(benzyloxy)-4-methoxyphenanthrene 1 lb as an off-white solid. (25 g, 65% yield). MS (ES+) m/z 421.3 [M+H]+; HPLC-UV analysis: retention time=(6.59 & 6.70) min; detection: 190-400 nm: peak area, 99.28%; eluent A, 0.1% TFA in water; eluent B, 0.1% TFA in Acetonitrile; isocratic/gradient over 10 min with a flow rate of 2.0 mL min-1.

[0202] Step c. To a solution of 4,7-bis(benzyloxy)-2-methoxyphenanthrene 11a and 2,7-bis(benzyloxy)-4-methoxyphenanthrene 1 lb (22 g, 52.31 mmol) in THF (660 mL) and 2-propanol (220 mL) was added 10% Pd/C (22 g) and Pd(OH)2 (22 g). The reaction mixture was stirred at room temperature 1 day under hydrogen balloon. The resultant mixture was filtered through a pad of Celite, washed with ethyl acetate and the filtrate was concentrated to give 1:1 mixture of regioisomers. (10.12 g, 80% yield). This crude product (10.12 g, 1:1 mixture) was purified by SFC (SFC condition: column: YMC Amylose-C; retention time=3.76 min; detection: 210 nm: co-solvent: 0.5% isopropyl amine in methanol; flow rate of 4.0 mL min-1) to give Lusianthrin as a pale yellow solid (1.1 g). 1H NMR (400 MHz, DMSO-d6) δ ppm: 9.59 (bs, 2H), 9.42 (d, J=9.20 Hz, 1H), 7.54-7.60 (m, 2H), 7.15 (d, J=2.60 Hz, 1H), 7.07-7.10 (m, 1H), 6.90 (d, J=2.40 Hz, 1H), 6.74 (d, J=2.50 Hz, 1H), 3.83 (s, 3H); 13C NMR (100 MHz, DMSO-d6) δ ppm: 157.20, 157.01, 154.78, 134.34, 133.27, 129.29, 127.69, 127.56, 124.12, 117.00, 114.69, 111.59, 102.77, 101.41, 55.43; MS (ES+) m/z 241.2 [M+H]+; Elemental analysis: Calculated (%) for C15H12O3+0.1CH3OH: C 74.49, H 5.13. Found: C 74.50, H 5.11; HPLC-UV analysis: retention time=12.04 min; detection: 190-400 nm: peak area, 99.49%; eluent A, 0.1% TFA in water; eluent B, Acetonitrile; isocratic/gradient over 30 min with a flow rate of 1.0 mL min-1.

Example 2: Compound 1 and 2 Activate Bacterially-Expressed AMPKα2β1γ1 in In Vitro

[0203] The AMPK heterotrimers were expressed in bacteria and purified through the His-α subunit by nickel purification before further purification through gel filtration. After being phosphorylated by incubation with CaMKKβ, AMPK complexes were further purified with a final gel filtration purification step. Phosphorylated purified AMPK was incubated with varying concentrations of Compound 1 or 2 for 30 mins using substrate and reagents from the HTRF-KinEASE Cisbio assay kit (STK S1 Kit). Phosphorylation of the substrate was measured by incubating with donor and acceptor antibodies for 2 h at room temperature as per the manufacturer's protocol (Coulerie et al., (2016) PMID: 27792327) and phosphorylated peptide detected by performing HTRF. The 665 nm/620 nm ratio was determined and plotted vs the log of the concentration of ligand.

[0204] FIG. 1 shows that Compound 1 and Compound 2 directly activate AMPK complexes in vitro.

Example 3: Compound 1 and 2 Increase the Phosphorylation of the AMPK Substrate, Acetyl-CoA Carboxylase (ACC), in U2OS Flp-In T-REx Mammalian Cells

[0205] U2OS Flp-In T-REx cells were first seeded at 50 K in a 96-well plate and left overnight at 37 C in DMEM GlutaMAX (Thermo Fisher Scientific) supplemented with 10% (vol/vol) FBS and 100 U/ml penicillin G, and 100 μg/ml streptomycin. Cells were treated for 1 h with varying concentrations of Lusianthridin in media lacking FBS and then cells were lysed in 50 μl of Cisbio lysis buffer #1 supplemented with blocking solution as per the manufacturer's protocol (Cisbio). Cells were lysed for 30 mins at room temperature before 16 μl of lysate was incubated with 4 μl of the HTRF antibodies (1:40 dilution of the acceptor and donor (p)ACC antibodies, as per the manufacturers protocol). Lysates were incubated overnight with the antibodies before 665 nm/620 nm ratio was determined using a MolecularDevices i3 plate reader (with a HTRF cartridge add-on).

[0206] FIG. 2 shows that using this pACC HTRF assay kit (Cisbio), Compound 1 and compound 2 increase the phosphorylation of the AMPK substrate, ACC, in a dose-dependent manner in U2OS Flp-In T-REx mammalian cells. Phosphorylation of ACC is widely used as a cellular indicator of AMPK activity.