COMPOSITION FOR TREATMENT, PREVENTION, OR AMELIORATION OF LYMPHEDEMA

Abstract

Provided are a novel compound exhibiting the effect of treating, preventing or ameliorating lymphedema, and a composition including the same as an active ingredient, and may be applied as a medicament product, functional food or food supplement, or cosmetics. The compound used as an active ingredient compound according to the disclosure exhibits a remarkable ameliorating effect on lymphedema in various aspects such as TNFα production inhibitory activity, metabolic stability, solubility, and blood exposure.

Claims

1. A compound of Formula 1 or Formula 2, or a pharmaceutically, food, or cosmetically acceptable salt thereof: ##STR00028## wherein, in this formula, R1 and R2 are each independently one or more selected from a group consisting of —H, —OH, —OR3, —OCOR3, —NO.sub.2, —CN, —SH, —SR3, —COR4, —COOR4, —CONHR4, —CON(R4).sub.2, —NH.sub.2, —NHR4, —N(R4).sub.2, —NHCOR4, —N(R5)COR.sub.4, a C.sub.1-C.sub.6 alkyl group, a C.sub.2-C.sub.6 alkenyl group, a C.sub.2-C.sub.6 alkynyl group, halogen, an allyloxy group, a C.sub.3-C.sub.7 cycloalkyl group, an allyl group, a heteroallyl group having at least one hetero atom and consisting of 3-7 atoms, and a heterocycloalkyl group having at least one hetero atom and consisting of 3-7 atoms, R3 are each independently selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, a C.sub.3-C.sub.10 aryl group, ##STR00029## R6 are each independently one or more selected from the group consisting of —H, —OH, —OR7, —NO.sub.2, —CN, a C.sub.1-C.sub.6 alkyl group, a C.sub.1-C.sub.6 alkoxy group, halogen, —NH.sub.2, —NHR7, and —N(R7).sub.2, R7 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, R4 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, R5 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, provided that butein is excluded from the compound of Formula 1; ##STR00030## wherein, in this formula, R8 and R9 are each independently one or more selected from the group consisting of —H, —OH, —OR10, —NO.sub.2, —CN, —SH, —SR10, —COR11, —COOR11, —CONHR11, —CON(R11).sub.2, —NH.sub.2, —NHR11, —N(R11).sub.2, —NHCOR11, —N(R12)COR11, a C.sub.1-C.sub.6 alkyl group, a C.sub.2-C.sub.6 alkenyl group, a C.sub.2-C.sub.6 alkynyl group, halogen, an allyloxy group, a C.sub.3-C.sub.7 cycloalkyl group, an allyl group, a heteroallyl group having at least one hetero atom and consisting of 3-7 atoms, and a heterocycloalkyl group having at least one hetero atom and consisting of 3-7 atoms, R10 is each independently selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, a C.sub.3-C.sub.10 aryl group, ##STR00031## R13 is each independently one or more selected from the group consisting of —H, —OH, —NO.sub.2, —CN, a C.sub.1-C.sub.6 alkyl group, a C.sub.1-C.sub.6 alkoxy group, halogen, —NH.sub.2, —NHR14, and —N(R14).sub.2, R14 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, R11 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, and R12 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group.

2. The compound or pharmaceutically, food, or cosmetically acceptable salt thereof of claim 1, wherein the hetero atom is at least one compound selected from the group consisting of O, N and S.

3. The compound or pharmaceutically, food, or cosmetically acceptable salt thereof of claim 1, wherein the compound or the pharmaceutically, food, or cosmetically acceptable salt thereof is selected from the group consisting of: (E)-1-(2,4-dihydroxyphenyl)-3-(4-hydroxy-3-methoxyphenyl)prop-2-en-1-one; (E)-1-(2,4-dihydroxyphenyl)-3-(3-hydroxy-4-methoxyphenyl)prop-2-en-1-one; (E)-1-(2,4-dihydroxyphenyl)-3-(3-fluoro-4-hydroxyphenyl)prop-2-en-1-one; (E)-1-(2,4-dihydroxyphenyl)-3-(4-fluoro-3-hydroxyphenyl)prop-2-en-1-one; (E)-1-(2,4-dihydroxyphenyl)-3-(4-hydroxyphenyl)prop-2-en-1-one; (E)-1-(2,4-dihydroxyphenyl)-3-(3-hydroxyphenyl)prop-2-en-1-one; (E)-3-(3,4-dihydroxyphenyl)-1-(2-fluoro-4-hydroxyphenyl)prop-2-en-1-one; (E)-3-(3,4-dihydroxyphenyl)-1-(4-hydroxy-2-methylphenyl)prop-2-en-1-one; (E)-3-(3,4-dihydroxyphenyl)-1-(4-hydroxy-2-methoxyphenyl)prop-2-en-1-one; (E)-3-(3,4-dihydroxyphenyl)-1-(4-hydroxy-2-isopropoxyphenyl)prop-2-en-1-one; (E)-3-(3,4-dihydroxyphenyl)-1-(2,4-dimethoxyphenyl)prop-2-en-1-one; (E)-3-(3,4-dihydroxyphenyl)-1-(p-tolyl)prop-2-en-1-one; (E)-1-(4-chlorophenyl)-3-(3,4-dihydroxyphenyl)prop-2-en-1-one; (E)-4-(3-(2,4-diacetoxyphenyl)-3-oxoprop-1-en-1-yl)-1,2-phenylene diacetate; (E)-4-(3-(2,4-bis(benzoyloxy)phenyl)-3-oxoprop-1-en-1-yl)-1,2-phenylene dibenzoate; (E)-2-(3-(3,4-dihydroxyphenyl)acryloyl)-5-hydroxyphenyl benzoate; (E)-2-(3-(3,4-dihydroxyphenyl)acryloyl)-5-hydroxyphenyl 4-methylbenzoate; (E)-2-(3-(3,4-dihydroxyphenyl)acryloyl)-5-hydroxyphenyl 4-methoxybenzoate, and (E)-2-(3-(3,4-dihydroxyphenyl)acryloyl)-5-hydroxyphenyl 4-chlorobenzoate.

4. The compound or pharmaceutically, food, or cosmetically acceptable salt thereof of claim 1, wherein the compound or pharmaceutically, food, or cosmetically acceptable salt thereof is for treatment, prevention, or amelioration of lymphedema.

5. A pharmaceutical composition for treating or preventing lymphedema, comprising a compound of Formula 1 or Formula 2, or pharmaceutically acceptable salt thereof: ##STR00032## wherein, in this formula, R1 and R2 are each independently one or more selected from the group consisting of —H, —OH, —OR3, —OCOR3, —NO.sub.2, —CN, —SH, —SR3, —COR4, —COOR4, —CONHR4, —CON(R4).sub.2, —NH.sub.2, —NHR4, —N(R4).sub.2, —NHCOR4, —N(R5)COR.sub.4, a C.sub.1-C.sub.6 alkyl group, a C.sub.2-C.sub.6 alkenyl group, a C.sub.2-C.sub.6 alkynyl group, halogen, an allyloxy group, a C.sub.3-C.sub.7 cycloalkyl group, an allyl group, a heteroallyl group having at least one hetero atom and consisting of 3-7 atoms, and a heterocycloalkyl group having at least one hetero atom and consisting of 3-7 atoms, R3 are each independently selected from the group consisting of a C.sub.1-C.sub.6alkyl group, a C.sub.3-C.sub.10 aryl group, ##STR00033## R6 are each independently one or more selected from the group consisting of —H, —OH, —OR7, —NO.sub.2, —CN, a C.sub.1-C.sub.6 alkyl group, a C.sub.1-C.sub.6 alkoxy group, halogen, —NH.sub.2, —NHR7, and —N(R7).sub.2, R7 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, R4 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, R5 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, provided that butein is excluded from the compound of Formula 1; [Formula 2] ##STR00034## wherein, in this formula, R8 and R9 are each independently one or more selected from the group consisting of —H, —OH, —OR10, —NO.sub.2, —CN, —SH, —SR10, —COR11, —COOR11, —CONHR11, —CON(R11).sub.2, —NH.sub.2, —NHR11, —N(R11).sub.2, —NHCOR.sub.11, —N(R12)COR11, a C.sub.1-C.sub.6 alkyl group, a C.sub.2-C.sub.6 alkenyl group, a C.sub.2-C.sub.6 alkynyl group, halogen, an allyloxy group, a C.sub.3-C.sub.7 cycloalkyl group, an allyl group, a heteroallyl group having at least one hetero atom and consisting of 3-7 atoms, and a heterocycloalkyl group having at least one hetero atom and consisting of 3-7 atoms, R10 is each independently selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, a C.sub.3-C.sub.10 aryl group, ##STR00035## R13 is each independently one or more selected from the group consisting of —H, —OH, —NO.sub.2, —CN, a C.sub.1-C.sub.6 alkyl group, a C.sub.1-C.sub.6 alkoxy group, halogen, —NH.sub.2, —NHR14, and —N(R14).sub.2, R14 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, R11 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, and R12 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group.

6. A method of treating or preventing lymphedema comprising administering to a subject a compound of Formula 1 or Formula 2, or a pharmaceutically acceptable salt thereof: ##STR00036## wherein, in this formula, R1 and R2 are each independently one or more selected from the group consisting of —H, —OH, —OR3, —OCOR3, —NO.sub.2, —CN, —SH, —SR3, —COR4, —COOR4, —CONHR4, —CON(R4).sub.2, —NH.sub.2, —NHR4, —N(R4).sub.2, —NHCOR4, —N(R5)COR.sub.4, a C.sub.1-C.sub.6 alkyl group, a C.sub.2-C.sub.6 alkenyl group, a C.sub.2-C.sub.6 alkynyl group, halogen, an allyloxy group, a C.sub.3-C.sub.7 cycloalkyl group, an allyl group, a heteroallyl group having at least one hetero atom and consisting of 3-7 atoms, and a heterocycloalkyl group having at least one hetero atom and consisting of 3-7 atoms, R3 are each independently selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, a C.sub.3-C.sub.10 aryl group, ##STR00037## R6 are each independently one or more selected from the group consisting of —H, —OH, —OR7, —NO.sub.2, —CN, a C.sub.1-C.sub.6 alkyl group, a C.sub.1-C.sub.6 alkoxy group, halogen, —NH.sub.2, —NHR7, and —N(R7).sub.2, R7 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, R4 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, R5 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, provided that butein is excluded from the compound of Formula 1; ##STR00038## wherein, in this formula, R8 and R9 are each independently one or more selected from the group consisting of —H, —OH, —OR10, —NO.sub.2, —CN, —SH, —SR10, —COR11, —COOR11, —CONHR11, —CON(R11).sub.2, —NH.sub.2, —NHR11, —N(R11).sub.2, —NHCOR.sub.11, —N(R12)COR11, a C.sub.1-C.sub.6 alkyl group, a C.sub.2-C.sub.6 alkenyl group, a C.sub.2-C.sub.6 alkynyl group, halogen, an allyloxy group, a C.sub.3-C.sub.7 cycloalkyl group, an allyl group, a heteroallyl group having at least one hetero atom and consisting of 3-7 atoms, and a heterocycloalkyl group having at least one hetero atom and consisting of 3-7 atoms, R10 is each independently selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, a C.sub.3-C.sub.10 aryl group, ##STR00039## R13 is each independently one or more selected from the group consisting of —H, —OH, —NO.sub.2, —CN, a C.sub.1-C.sub.6 alkyl group, a C.sub.1-C.sub.6 alkoxy group, halogen, —NH.sub.2, —NHR14, and —N(R14).sub.2, R14 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, R11 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group, and R12 is selected from the group consisting of a C.sub.1-C.sub.6 alkyl group, an allyl group, and a heteroallyl group.

Description

BRIEF DESCRIPTION OF DRAWINGS

[0092] FIG. 1 is a graph showing the level of LPS-induced TNFα production in mouse peritoneal macrophages.

[0093] FIG. 2A shows images showing the change in the size of lymphedema on the 7.sup.th day after surgery in the normal group, the control group, and the compound-treated group of the disclosure with respect to lymphedema induced in mice, and FIG. 2B is a graph in which the volumes of lymphedema in the respective groups are compared.

[0094] FIG. 3A shows an H&E image of the right hind leg of a mouse taken through immunofluorescence analysis in a normal group, a control group, and a group treated with the compound of the disclosure.

[0095] FIG. 3B is a graph in which mRNA expression of TNFα in a lymphedema tissue of the right hind limb of a mouse can be compared.

[0096] FIG. 4A shows an H&E image of the right hind leg of a mouse taken through immunofluorescence analysis in a normal group, a control group, and a group treated with the compound of the disclosure.

[0097] FIG. 4B shows image in which the distribution of PPARγ, which is a major regulator of lipid production in the epidermal layer of lymphedema tissue, can be identified.

[0098] FIG. 4C shows a western blot image in which the expression of PPARγ and a target protein Fabp4 thereof in lymphedema tissue can be identified.

[0099] FIG. 4D is a graph in which the expression of PPARγ and a target protein Fabp4 thereof in lymphedema tissue can be compared.

DETAILED DESCRIPTION

[0100] Hereinafter, the disclosure will be described in more detail using Examples. However, these examples are for illustrative purposes only, and the scope of the disclosure is not limited by these examples. In addition, those of ordinary skill in the art will be able to make various changes and modifications to the disclosure within the scope that does not impair the spirit of the disclosure. Terms not specifically defined in this specification should be understood to have meanings commonly used in the technical field to which the disclosure pertains.

[0101] All animal tests in the following examples were performed after obtaining approvals by the Institutional Animal Care and Use Committee (IACUC) of Sungkyunkwan University School of Medicine (Approval No.: SKKUIACUC-20150037). All methods were performed according to the approved guidelines. A murine model for acquired lymphedema was surgically induced in the legs of male ICR mice using an established protocol. Briefly, major lymphatic sites including inguinal lymph nodes, popliteal lymph nodes, deep inguinal lymph nodes and femoral lymphatic vessels were removed, and the lower right leg of the mouse was limitedly cauterized under anesthesia.

[0102] Compounds of Formula 1 or 2 were prepared according to Scheme 1 or Scheme 2. For example, compounds 1, 7a-m, 8a-b, and 9 described below were prepared according to scheme 1, and compounds 14a-d was prepared according to scheme 2.

##STR00006##

[0103] Reagents and conditions in Scheme 1 are as follows:

[0104] (i) Preparation A: 60% KOH(aq), ethanol, room temperature, 6 h, 60-72%; Preparation B: 10% CuBr.sub.2, ethyl acetate, 80° C., 8 h, 42-60%;

[0105] (ii) acetic anhydride, reflux, overnight, 93% or benzoyl chloride, TEA, THF, 6 h, 65%; and

[0106] (iii) KOH(s), methanol, reflux, 4 h, 60%.

##STR00007##

[0107] Reagents and conditions in Scheme 2 are as follows:

[0108] (iv) methoxymethyl chloride, K.sub.2CO.sub.3, actone, room temperature, 4 h, 61%;

[0109] (v) methoxymethyl chloride, K.sub.2CO.sub.3, acetone, reflux, 2.5 h, 70%;

[0110] (vi) 60% KOH(aq), ethanol, room temperature, 6 h, 70%;

[0111] (vii) acyl chloride, TEA, DMF, room temperature, 2-3 h, 60%-73%; and

[0112] (viii) 6N HCl (aq), ethanol, room temperature, 4 h, 60%-75%.

[0113] Hereinafter, specific preparation steps for the preparation of the compound according to the disclosure and representative examples corresponding thereto are described together. In the case of compounds having different substituents, these compounds were actually prepared through similar steps, but not all preparation examples are specified in the present specification. Those of ordinary skill in the art will be able to easily prepare compounds of Formula 1 or 2 having different substituents with reference to the following representative examples.

Preparation A

Compound 1: (E)-1-(2,4-dihydroxyphenyl)-3-(3,4-dihydroxyphenyl)prop-2-en-1-one

[0114] ##STR00008##

[0115] Compound 1 refers to butein.

[0116] 3,4-dihydroxybenzaldehyde (compound 6a, 1.0 g, 7.2 mmol) and 1-(2,4-dihydroxyphenyl)ethan-1-one (compound 5a, 1.1 g, 7.2 mmol) were dissolved in ethanol (10 mL). 60% KOH (aq) (1 mL) was added and the mixture was left at room temperature for 6 h. The solvent was removed therefrom under reduced pressure, and the residue was extracted three times with 30 ml of diethyl ether. The diethyl ether layer was washed three times with water (30 ml×3) and dried using anhydrous MgSO.sub.4.

[0117] After removing the solvent, compound 1 was obtained by silica gel column chromatography (silica gel: n-hexane-ethyl acetate=1:1) (yellow solid, 42% yield). .sup.1H NMR, .sup.13C NMR, LRMS, HRMS, and M.P data were measured as follows:

[0118] .sup.1H NMR (MeOD, 400 MHz) b 7.95 (1H, d, J=8.8 Hz), 7.73 (1H, d, J=15.6 Hz), 7.54 (1H, d, J=15.6 Hz), 7.20 (1H, d, J=2.0 Hz), 7.12 (1H, dd, J=8.4, 2.4 Hz), 6.83 (1H, d, J=8.4 Hz), 6.43 (1H, dd, J=8.8, 2.4 Hz), 6.31 (1H, d, J=2.4 Hz); .sup.13C NMR (100 MHz, MeOD) b 193.50, 167.52, 166.37, 149.95, 146.86, 146.11, 133.30, 128.45, 123.64, 118.31, 116.63, 115.84, 114.74, 109.17, 103.85; LRMS (ESI) calcd. for C.sub.15H.sub.13O.sub.5 [M+H].sup.+: 273.08, found: 273.10; HRMS (ESI) calcd. for C.sub.15H.sub.13O.sub.5 [M+H].sup.+: 273.0685, found: 273.0754; M.P=214.7° C.

Preparation B

[0119] 1-(4-hydroxy-2-methylphenyl)ethan-1-one (compound 5b, 150 mg, 1.0 mmol), 2,4-dihydroxybenzaldehyde (compound 6a, 138 mg, 1.0 mmol) and CuBr.sub.2 (22 mg, 0.1 mmol) were dissolved in 5 ml ethyl acetate at room temperature in a pressurized tube, and the reaction mixture was stirred at 80° C. After 12 h, the mixture was cooled to room temperature and filtered through Celite.

[0120] The organic layer was concentrated in a vacuum condition, and compound 7h was obtained by silica gel column chromatography (silica gel:n-hexane-ethylacetate=1:1) (yellow solid, 66% yield).

Compound 7a: (E)-1-(2,4-dihydroxyphenyl)-3-(4-hydroxy-3-methoxyphenyl)prop-2-en-1-one

[0121] ##STR00009##

[0122] Compound 7a was isolated via Preparation A (yellow solid, 46% yield), and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0123] .sup.1H NMR (MeOD, 400 MHz) b 8.06 (1H, d, J=8.8 Hz), 7.69 (1H, d, J=15.2 Hz), 7.55 (1H, d, J=15.2 Hz), 7.27 (1H, d, J=2.0 Hz), 7.12 (1H, dd, J=8.4, 2.0 Hz), 6.75 (1H, d, J=8.0 Hz), 6.31 (1H, dd, J=8.8, 2.4 Hz), 6.19 (1H, d, J=2.4 Hz) 3.84 (3H, s); .sup.13C NMR (100 MHz, MeOD) b 193.4, 167.5, 166.4, 151.7, 148.0, 145.5, 133.4, 129.5, 123.5, 119.2, 115.2, 114.7, 112.5, 109.2, 103.8, 56.56; LRMS (ESI) calcd. for C.sub.16H.sub.15O.sub.5 [M+H].sup.+: 287.09, found: 287.10; HRMS (ESI) calcd. for C.sub.16H.sub.15O.sub.5 [M+H].sup.+: 287.0914, found: 287.0908; M.P=198.1° C.

Compound 7b: (E)-1-(2,4-dihydroxyphenyl)-3-(3-hydroxy-4-methoxyphenyl)prop-2-en-1-one

[0124] ##STR00010##

[0125] Compound 7b was isolated via Preparation A (yellow solid, 48% yield), and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0126] .sup.1H NMR (400 MHz, MeOD) b 7.96 (1H, d, J=8.0 Hz), 7.74 (1H, d, J=15.6 Hz), 7.59 (1H, d, J=15.2 Hz), 7.25 (1H, d, J=2.4 Hz), 7.20 (1H, dd, J=8.4, 2.0 Hz), 6.98 (1H, d, J=8.4 Hz), 6.43 (1H, dd, J=8.8, 2.4 Hz), 6.31 (1H, d, J=2.4 Hz) 3.92 (3H, s); .sup.13C NMR (100 MHz, MeOD) b 193.4, 167.5, 166.4, 151.7, 148.0, 145.5, 133.4, 129.5, 123.5, 119.2, 115.2, 114.7, 112.5, 109.2, 103.8, 56.39; LRMS (ESI) calcd. for C.sub.16H.sub.15O.sub.5 [M+H].sup.+: 287.09, found: 287.10; HRMS (ESI) calcd. for C.sub.16H.sub.15O.sub.5 [M+H].sup.+: 287.0914, found: 287.0908; M.P=206.4° C.

Compound 7c: (E)-1-(2,4-dihydroxyphenyl)-3-(3-fluoro-4-hydroxyphenyl)prop-2-en-1-one

[0127] ##STR00011##

[0128] Compound 7c was isolated via Preparation A (yellow solid, 53% yield), and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0129] .sup.1H NMR (400 MHz, MeOD δ 7.99 (1H, d, J=8.8 Hz), 7.75 (1H, d, J=15.2 Hz), 7.65 (1H, d, J=15.6 Hz), 7.57 (1H, dd, J=12.4, 2.0 Hz), 7.39 (1H, dd, J=8.4, 1.6 Hz), 6.97 (1H, t, J=8.8 Hz), 6.43 (1H, dd, J=8.8, 2.4 Hz), 6.30 (1H, d, J=2.4 Hz); .sup.13C NMR (100 MHz, MeOD) δ 193.2, 167.5, 166.5, 154.2, 151.9, 149.1, 144.1, 133.5, 127.4, 119.9, 118.9, 116.5, 114.6, 109.2, 103.8; LRMS (ESI) calcd. for C.sub.15H.sub.12O.sub.4F [M+H].sup.+: 275.07, found: 275.10; HRMS (ESI) calcd. for C.sub.15H.sub.12O.sub.4F [M+H].sup.+: 275.0714, found: 275.0709; M.P=210.5° C.

Compound 7d: (E)-1-(2,4-dihydroxyphenyl)-3-(4-fluoro-3-hydroxyphenyl)prop-2-en-1-one

[0130] ##STR00012##

[0131] Compound 7d was isolated via Preparation A (yellow solid, 52% yield) and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0132] .sup.1H NMR (400 MHz, MeOD) δ 7.98 (1H, d, J=8.8 Hz), 7.74 (1H, d, J=15.6 Hz), 7.67 (1H, d, J=15.6 Hz), 7.32 (1H, dd, J=8.4, 2.0 Hz), 7.25-7.21 (1H, m), 7.15-7.10 (1H, m), 6.44 (1H, dd, J=8.8, 2.4 Hz), 6.31 (1H, d, J=2.4 Hz); .sup.13C NMR (100 MHz, MeOD) δ 193.2, 167.6, 166.7, 155.7, 146.7, 144.3, 133.5, 133.2, 122.0, 121.4, 118.5, 117.4, 114.6, 109.3, 103.8; LRMS (ESI) calcd. for C.sub.15H.sub.12O.sub.4F [M+H].sup.+: 275.07, found: 275.10; HRMS (ESI) calcd. for C.sub.15H.sub.12O.sub.4F [M+H].sup.+: 275.0714, found: 275.0709; M.P=252.4° C.

Compound 7e: (E)-1-(2,4-dihydroxyphenyl)-3-(4-hydroxyphenyl)prop-2-en-1-one

[0133] ##STR00013##

[0134] Compound 7e was isolated via Preparation A (yellow solid, 43% yield), and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0135] .sup.1H NMR (400 MHz, MeOD) δ 7.98 (1H, d, J=9.2 Hz), 7.80 (1H, d, J=15.2 Hz), 7.64-7.60 (3H, m), 6.86 (2H, d, J=8.0 Hz), 6.43 (1H, dd, J=8.8, 2.4 Hz), 6.31 (1H, d, J=2.4 Hz); .sup.13C NMR (100 MHz, MeOD) δ 193.5, 167.5, 166.3, 161.5, 145.6, 133.3, 131.8, 127.8, 118.5, 116.9, 114.7, 109.1, 103.8; LRMS (ESI) calcd. for C.sub.15H.sub.13O.sub.4 [M+H].sup.+: 257.08, found: 257.10; HRMS (ESI) calcd. for C.sub.15H.sub.13O.sub.4 [M+H].sup.+: 257.0801, found: 257.0801; M.P=205.7° C.

Compound 7f: (E)-1-(2,4-dihydroxyphenyl)-3-(3-hydroxyphenyl)prop-2-en-1-one

[0136] ##STR00014##

[0137] Compound 7f was isolated via Preparation A (yellow solid, 52% yield), and 1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0138] .sup.1H NMR (400 MHz, MeOD) δ 8.00 (2H, d, J=8.8 Hz), 7.66 (1H, d, J=15.6 Hz), 7.51 (1H, d, J=15.2 Hz), 7.19 (1H, d, J=2.4 Hz), 7.11 (1H, dd, J=8.0, 2.0 Hz), 6.91 (2H, d, J=8.8 Hz), 6.83 (1H, d, J=8.0 Hz); .sup.13C NMR (100 MHz, MeOD) δ 191.0, 163.7, 149.8, 146.2, 132.2, 131.2, 128.4, 123.4, 119.5, 116.6, 116.4, 115.6; LRMS (ESI) calcd. for C.sub.15H.sub.13O.sub.4 [M+H].sup.+: 257.08, found: 257.10; HRMS (ESI) calcd. for C.sub.15H.sub.13O.sub.4 [M+H].sup.+: 257.0808, found: 257.0803; M.P=208.0° C.

Compound 7g: (E)-3-(3,4-dihydroxyphenyl)-1-(2-fluoro-4-hydroxyphenyl)prop-2-en-1-one

[0139] ##STR00015##

[0140] Compound 7g was isolated via Preparation A (yellow solid, 53% yield) and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0141] .sup.1H NMR (400 MHz, MeOD) δ 7.75 (1H, t, J=8.8 Hz), 7.62 (1H, dd, J=15.6, 2.0 Hz), 7.28 (1H, dd, J=15.2, 2.4 Hz), 7.14 (1H, d, J=2.0 Hz), 7.19 (1H, d, J=2.4 Hz), 7.04 (1H, dd, J=8.4, 2.0 Hz), 6.82 (1H, d, J=8.0 Hz), 6.72 (1H, dd, J=8.8, 2.4 Hz), 6.61 (1H, dd, J=13.2, 2.4 Hz); .sup.13C NMR (100 MHz, MeOD) δ 189.5, 165.0, 150.0, 146.8, 133.6, 128.2, 123.6, 123.2, 119.6, 116.6, 115.4, 113.1, 104.2, 103.9; LRMS (ESI) calcd. for C.sub.15H.sub.12O.sub.4F[M+H].sup.+: 275.08, found: 275.10; HRMS (ESI) calcd. for C.sub.15H.sub.12O.sub.4F [M+H].sup.+: 275.0714, found: 275.0708; M.P=207.8° C.

Compound 7h: (E)-3-(3,4-dihydroxyphenyl)-1-(4-hydroxy-2-methylphenyl)prop-2-en-1-one

[0142] ##STR00016##

[0143] Compound 7h was isolated via Preparation B (yellow solid, 66% yield) and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0144] .sup.1H NMR (400 MHz, MeOD) δ 7.54 (1H, d, J=9.2 Hz), 7.41 (1H, d, J=15.6 Hz), 7.12 (1H, d, J=2.4 Hz), 7.09 (1H, d, J=16.0 Hz), 7.02 (1H, dd, J=8.0, 2.0 Hz), 6.81 (1H, d, J=8.4 Hz), 6.73-6.70 (2H, m), 2.42 (3H, s); .sup.13C NMR (100 MHz, MeOD) δ 197.0, 161.3, 149.9, 147.3, 146.8, 141.7, 132.5, 131.6, 128.1, 124.0, 123.4, 119.2, 116.5, 115.4, 113.3, 21.26; LRMS (ESI) calcd. for C.sub.16H.sub.15O.sub.4 [M+H].sup.+: 271.10, found: 271.10; HRMS (ESI) calcd. for C.sub.16H.sub.15O.sub.4 [M+H].sup.+: 271.0965, found: 271.0958; M.P=208.2° C.

Compound 7i: (E)-3-(3,4-dihydroxyphenyl)-1-(4-hydroxy-2-methoxyphenyl)prop-2-en-1-one

[0145] ##STR00017##

[0146] Compound 7i was isolated via Preparation B (yellow solid, 46% yield), and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0147] .sup.1H NMR (400 MHz, MeOD) δ 7.59 (1H, d, J=8.8 Hz), 7.51 (1H, d, J=15.6 Hz), 7.38 (1H, d, J=15.2 Hz), 7.12 (1H, d, J=2.0 Hz), 7.00 (1H, dd, J=8.4, 2.0 Hz), 6.80 (1H, d, J=8.0 Hz), 6.53 (1H, d, J=2.0 Hz), 6.47 (1H, dd, J=8.4, 2.0 Hz), 3.91 (3H, s); .sup.13C NMR (100 MHz, MeOD) δ 193.0, 164.5, 162.5, 149.5, 146.8, 144.5, 133.7, 128.6, 125.1, 123.3, 121.7, 116.5, 115.2, 108.9, 100.1, 56.15; LRMS (ESI) calcd. for C.sub.16H.sub.15O.sub.5 [M+H].sup.+: 287.09, found: 287.10; HRMS (ESI) calcd. for C.sub.16H.sub.15O.sub.5 [M+H].sup.+: 287.0914, found: 287.0906; M.P=200.4° C.

Compound 7j: (E)-3-(3,4-dihydroxyphenyl)-1-(4-hydroxy-2-isopropoxyphenyl)prop-2-en-1-one

[0148] ##STR00018##

[0149] Compound 7j was isolated via Preparation A (yellow solid, 53% yield), and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0150] .sup.1H NMR (400 MHz, MeOD) δ 7.58 (1H, d, J=8.8 Hz), 7.47 (2H, d, J=2.8 Hz), 7.11 (1H, d, J=2.0 Hz), 7.00 (1H, dd, J=8.4, 2.4 Hz), 6.81 (1H, d, J=8.0 Hz), 6.51 (1H, d, J=2.4 Hz), 6.45 (1H, dd, J=8.8, 2.4 Hz), 4.69 (1H, hept, 6.0 Hz), 1.40 (6H, d, J=6.0 Hz); .sup.13C NMR (100 MHz, MeOD) δ 193.4, 160.6, 146.8, 143.9, 133.7, 128.7, 125.5, 123.1, 122.9, 116.5, 115.2, 109.1, 102.2, 72.21, 22.43; LRMS (ESI) calcd. for C.sub.18H.sub.19O.sub.5 [M+H].sup.+: 315.12, found: 315.20; HRMS (ESI) calcd. for C.sub.18H.sub.19O.sub.5 [M+H].sup.+: 315.1227, found: 315.1219; M.P=80.4° C.

Compound 7k: (E)-3-(3,4-dihydroxyphenyl)-1-(2,4-dimethoxyphenyl)prop-2-en-1-one

[0151] ##STR00019##

[0152] Compound 7k was isolated via Preparation A (yellow solid, 63% yield) and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0153] .sup.1H NMR (400 MHz, MeOD) δ 7.65 (1H, d, J=8.4 Hz), 7.51 (1H, d, J=15.6 Hz), 7.34 (1H, d, J=15.6 Hz), 7.12 (1H, d, J=2.0 Hz), 7.00 (1H, dd, J=8.4, 2.0 Hz), 6.81 (1H, d, J=8.4 Hz), 6.65-6.61 (2H, m), 3.93 (3H, s), 3.89 (3H, s); .sup.13C NMR (100 MHz, MeOD) δ 193.3, 166.0, 162.0, 149.7, 146.8, 145.0, 133.4, 128.5, 124.9, 123.4, 116.5, 115.2, 106.7, 99.53, 56.20; LRMS (ESI) calcd. for C.sub.17H.sub.17O.sub.5 [M+H].sup.+: 301.10, found: 301.10; HRMS (ESI) calcd. for C.sub.17H.sub.17O.sub.5 [M+H].sup.+: 301.1071, found: 301.1065; M.P=162.9° C.

Compound 7l: (E)-3-(3,4-dihydroxyphenyl)-1-(p-tolyl)prop-2-en-1-one

[0154] ##STR00020##

[0155] Compound 7l was isolated via Preparation A (yellow solid, 66% yield) and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0156] .sup.1H NMR (400 MHz, MeOD) δ 7.96 (2H, d, J=8.0 Hz), 7.68 (1H, d, J=15.6 Hz), 7.50 (1H, d, J=15.2 Hz), 7.36 (2H, d, J=8.0 Hz), 7.20 (1H, d, J=2.0 Hz), 7.11 (1H, dd, J=8.4, 2.4 Hz), 6.83 (1H, d, J=8.4 Hz), 2.44 (3H, s); .sup.13C NMR (100 MHz, MeOD) δ 192.2, 150.0, 147.0, 146.8, 145.0, 137.1, 130.4, 129.6, 128.3, 123.6, 119.6, 116.6, 115.7, 21.64; LRMS (ESI) calcd. for C.sub.16H.sub.15O.sub.3 [M+H].sup.+: 255.10, found: 255.10; HRMS (ESI) calcd. for C.sub.16H.sub.15O.sub.3 [M+H].sup.+: 255.1016, found: 255.1014; M.P=190.9° C.

Compound 7m: (E)-1-(4-chlorophenyl)-3-(3,4-dihydroxyphenyl)prop-2-en-1-one

[0157] ##STR00021##

[0158] Compound 7m was isolated via Preparation A (yellow solid, 63% yield), and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0159] .sup.1H NMR (400 MHz, MeOD) δ 8.06 (2H, d, J=8.8 Hz), 7.71 (1H, d, J=15.6 Hz), 7.56 (2H, d, J=8.8 Hz), 7.49 (1H, d, J=15.6 Hz), 7.21 (1H, d, J=2.0 Hz), 7.14 (1H, dd, J=8.4, 2.4 Hz), 6.84 (1H, d, J=8.4 Hz); .sup.13C NMR (100 MHz, MeOD) δ 191.1, 147.8, 146.9, 140.0, 138.2, 131.1, 129.9, 128.1, 123.8, 119.1, 116.6, 115.8; LRMS (ESI) calcd. for C.sub.15H.sub.12O.sub.3Cl[M+H].sup.+: 275.05, found: 275.10; HRMS (ESI) calcd. for C.sub.15H.sub.12O.sub.3Cl [M+H].sup.+: 275.0469, found: 275.0465; M.P=205.4° C.

Compound 8a: (E)-4-(3-(2,4-diacetoxyphenyl)-3-oxoprop-1-en-1-yl)-1,2-phenylene diacetate

[0160] ##STR00022##

[0161] The solution of (E)-1-(2,4-dihydroxyphenyl)-3-(3,4-dihydroxyphenyl)prop-2-en-1-one (compound 1, 500 mg, 1.8 mmol)) in Ac.sub.2O (8 mL) was heated to 120° C.-130° C. and stirred at this temperature for 5 h. After cooling to room temperature, the resulting mixture was concentrated in a vacuum condition. From the residue, 397 mg of compound 8a was obtained by silica gel column chromatography (silica gel:n-hexane-ethylacetate=1:2) (white solid, 90% yield). .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0162] .sup.1H NMR (400 MHz, MeOD) b 7.86 (2H, d, J=8.4 Hz), 7.63-7.61 (2H, m), 7.58 (1H, d, J=16.0 Hz), 7.35 (1H, d, J=12.4 Hz), 7.31 (1H, d, J=5.2 Hz), 7.22 (1H, dd, J=8.4, 2.4 Hz), 7.10 (1H, d, J=2.4 Hz), 2.33 (3H, s), 2.31 (6H, d, J=3.2 Hz), 2.23 (3H, s); .sup.13C NMR (100 MHz, MeOD) b 188.8, 168.6, 168.0, 153.4, 149.5, 143.7, 142.4, 142.3, 133.1, 131.2, 128.9, 127.5, 125.4, 124.1, 123.3, 119.6, 117.6, 20.76, 20.32; LRMS (ESI) calcd. for C.sub.23H.sub.21O.sub.9 [M+H].sup.+: 441.12, found: 441.10; HRMS (ESI) calcd. for C.sub.23H.sub.21O.sub.9 [M+H].sup.+: 441.1180, found: 441.1171; M.P=131.3° C.

Compound 8b: (E)-4-(3-(2,4-bis(benzoyloxy)phenyl)-3-oxoprop-1-en-1-yl)-1,2-phenylene dibenzoate

[0163] ##STR00023##

[0164] (E)-1-(2,4-dihydroxyphenyl)-3-(3,4-dihydroxyphenyl)prop-2-en-1-one (compound 1, 500 mg, 1.8 mmol) was dissolved in anhydrous pyridine (5 mL), treated with benzoylchloride (2.1 ml, 18.4 mmol), refluxed for 4 h, left at 22° C.-24° C. overnight, and diluted with cold H.sub.2O the next day. The ethyl acetate layer was washed three times with water (30 ml×3) and dried using anhydrous MgSO.sub.4.

[0165] After removing the solvent, 413 mg of compound 8b was obtained by silica gel column chromatography (silica gel: n-hexane-ethyl acetate=1:2) (white solid, 60% yield). .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0166] .sup.1H NMR (400 MHz, MeOD) b 8.24-8.21 (2H, m), 8.16-8.13 (2H, m), 8.05-7.99 (6H, m), 7.65-7.57 (7H, m), 7.51-7.37 (11H, m); .sup.13C NMR (100 MHz, MeOD) b 188.6, 164.2, 163.4, 153.7, 149.7, 143.7, 142.4, 134.2, 133.4, 132.7, 131.1, 129.7, 129.6, 129.4, 129.2, 128.9, 128.4, 128.1, 127.8, 125.7, 124.1, 123.2, 120.1, 118.0; LRMS (ESI) calcd. for C.sub.43H.sub.29O.sub.9 [M+H].sup.+: 689.18, found: 689.20; HRMS (ESI) calcd. for C.sub.43H.sub.29O.sub.9 [M+H].sup.+: 689.1812, found: 689.1794; M.P=36.7° C.

Compound 10: 1-(2-hydroxy-4-(methoxymethoxy)phenyl)ethanone

[0167] 1-(2,4-dihydroxyphenyl)ethanone (compound 5a, 1.5 g, 10 mmol) in 12 mL acetone was added to K.sub.2CO.sub.3 (1.4 g, 10 mmol), and at room temperature, in an Ar atmospheric condition, chloro(methoxy)methane (885 mg, 11 mmol) was added thereto with slow stirring.

[0168] The reaction mixture was stirred for 4 h at room temperature and filtered. The solid was washed with EtOAc (80 mL) and the filtrate was washed with NaH.sub.2PO.sub.4 (sat. 50 mL) and water (80 mL), dried using MgSO.sub.4 and concentrated.

[0169] From the crude product, 1.2 g of compound 10 was obtained by silica gel column chromatography (silica gel: n-hexane-ethyl acetate=1:4) (yellow white solid, 61% yield). .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0170] .sup.1H NMR (400 MHz, DMSO-d6) δ 12.49 (1H, s), 7.85 (1H, d, J=8.8 Hz) 6.60 (1H, dd, J=8.8, 2.4 Hz), 5.53 (1H, d, J=2.4 Hz), 5.27 (2H, s), 3.39 (3H, s), 2.57 (3H, s); .sup.13C NMR (100 MHz, DMSO-d6) b 203.1, 163.5, 133.2, 114.5, 108.0, 102.9, 93.62, 55.90, 26.71; HRMS (ESI) calcd. for C.sub.10H.sub.12O.sub.4 [M+H].sup.+: 197.08, found: 197.10; HRMS (ESI) calcd. for C.sub.10H.sub.12O.sub.4 [M+H].sup.+: 197.0825, found: 197.0815; M.P=60.2° C.

Compound 11: 3,4-bis(methoxymethoxy)benzaldehyde

[0171] Potassium carbonate (4.0 g, 19 mmol) and methoxymethyl chloride (2.3 g, 2.2 mL, 19 mmol) were added to a solution of 3,4-dihydroxybenzaldehyde (compound 6a, 1 g, 7.2 mmol) in acetone (40 mL). The mixture was refluxed for 2.5 hours, and the precipitate was filtered therefrom. The solvent was then removed under reduced pressure.

[0172] The residue was dissolved in CH.sub.2C.sub.12 (30 mL), washed with water (80 mL), dried using MgSO.sub.4 and concentrated.

[0173] From the crude product, 1.1 g of compound 11 was obtained by chromatography on silica (n-hexane-ethyl acetate, 1:4) (white solid, 70% yield). .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0174] .sup.1H NMR (400 MHz, DMSO-d6) δ 9.85 (1H, s), 7.60-7.58 (2H, m), 7.31 (1H, d, J=8.0 Hz), 5.30 (4H, d, J=23.6 Hz), 3.42 (6H, d, J=2.0 Hz); .sup.13C NMR (100 MHz, DMSO-d6) b 191.3, 152.1, 146.9, 130.5, 126.1, 115.6, 115.3, 94.67, 94.34, 55.97, 55.79; HRMS (ESI) calcd. for C.sub.11H.sub.14O.sub.5 [M+H].sup.+: 227.09, found: 227.10; HRMS (ESI) calcd. for C.sub.11H.sub.14O.sub.5 [M+H].sup.+: 227.0935, found: 227.0935; M.P=44.6° C.

Compound 12: (E)-3-(3,4-bis(methoxymethoxy)phenyl)-1-(2-hydroxy-4-(methoxymethoxy)-phenyl)-prop-2-ene-1-one

[0175] 1-(2-hydroxy-4-(methoxymethoxy)phenyl)-ethanone (compound 10, 1.0 g, 5.09 mmol) and 3,4-bis(methoxymethoxy)benzaldehyde (compound 11, 1.2 g, 5.09 mmol) were dissolved in ethanol (10 mL). 60% KOH (aq) (2 mL) was added thereto and the mixture was left at room temperature for 6 h. The solvent was removed therefrom under reduced pressure, and the residue was extracted three times with 30 ml of diethyl ether. The diethyl ether layer was washed three times with water (30 ml×3) and dried using anhydrous MgSO.sub.4.

[0176] After removing the solvent, 1.3 g of compound 12 was obtained by silica gel column chromatography (silica gel: n-hexane-ethyl acetate=1:2) (yellow solid, 63% yield). .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0177] .sup.1H NMR (700 MHz, DMSO-d6) δ 13.36 (1H, s), 8.25 (1H, dd, J=9.1, 3.5 Hz), 7.88 (1H, dd, J=15.4, 3.5 Hz), 7.77 (1H, dd, J=15.4, 3.5 Hz), 7.76-7.68 (1H, m), 7.52-7.50 (1H, m), 7.18 (1H, dd, J=8.4, 4.2 Hz), 6.66-6.64 (1H, m), 6.59-6.58 (1H, m) 5.29 (3H, dd, J=13.3, 4.2 Hz) 3.45 (3H, d, J=4.2 Hz), 3.41 (6H, dd, J=7.7, 4.2 Hz); .sup.13C NMR (175 MHz, DMSO-d6) b 133.0, 129.2, 125.2, 120.1, 117.7, 117.0, 115.2, 108.6, 103.6, 95.53, 95.12, 94.33, 56.51, 56.43, 56.37; HRMS (ESI) calcd. for C.sub.21H.sub.24O.sub.8 [M+H].sup.+: 405.16, found: 405.10; HRMS (ESI) calcd. for C.sub.21H.sub.24O.sub.8 [M+H].sup.+: 405.1582, found: 405.1592; M.P=107.8° C.

Compound 13a: (E)-2-(3-(3,4-bis(methoxymethoxy)phenyl)acryloyl)-5-(methoxymethoxy)phenyl benzoate

[0178] (E)-3-(3,4-bis(methoxymethoxy)phenyl)-1-(2-hydroxy-4-(methoxymethoxy)phenyl)prop-2-en-1-one (Compound 12, 500 mg, 1.0 mmol) was dissolved in methylene chloride (10 mL). TEA (164 μL, 1.18 mmol) and benzoylchloride (137 μL, 1.18 mmol) were added and the mixture was left at room temperature for 6 hours. The solvent was removed under reduced pressure, and the residue was extracted three times with 20 ml of ethyl acetate. The ethyl acetate layer was washed three times with water (20 ml×3) and dried using anhydrous MgSO.sub.4.

[0179] After removing the solvent, 430 mg of compound 13a was obtained by silica gel column chromatography (silica gel: n-hexane-ethyl acetate=1:2) (yellow solid, 86% yield). .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0180] .sup.1H NMR (700 MHz, DMSO-d6) δ 7.97 (2H, d, J=7.7 Hz), 7.92 (1H, d, J=9.1 Hz), 7.43 (1H, d, J=11.9 Hz), 7.35 (1H, d, J=14.7 Hz), 7.32 (1H, d, J=7.7 Hz), 7.25 (1H, d, J=8.4 Hz), 7.12 (1H, d, J=8.4 Hz), 7.08 (2H, d, J=9.1 Hz), 5.33 (2H, s), 5.23 (4H, d, J=19.6 Hz), 3.42 (9H, t, J=8.1 Hz), 2.38 (3H, s).sup.13C NMR (175 MHz, DMSO-de) b 188.8, 164.8, 160.7, 150.9, 149.8, 147.2, 144.8, 143.8, 132.1, 130.3, 129.7, 129.5, 129.0, 126.4, 126.1, 124.4, 124.0, 117.2, 116.9, 114.1, 111.7, 95.49, 95.12, 94.52, 56.43, 56.36, 56.32, 21.63; HRMS (ESI) calcd. for C.sub.29H.sub.30O.sub.9 [M+H].sup.+: 523.20, found: 523.20; HRMS (ESI) calcd. for C.sub.29H.sub.30O.sub.9 [M+H].sup.+: 523.2006, found: 523.2006; M.P=47.2° C.

Compound 14a: (E)-2-(3-(3,4-dihydroxyphenyl)acryloyl)-5-hydroxyphenyl benzoate

[0181] ##STR00024##

[0182] (E)-2-(3-(3,4-bis(methoxymethoxy)phenyl)acryloyl)-5-(methoxymethoxy)phenyl benzoate (compound 13a, 400 mg, 0.787 mmol) was dissolve in methanol (1.5 mL) and 0.25 mL of concentrated hydrochloric acid was added thereto. The mixture was then heated to 45° C. for 2 h. The solvent was removed under reduced pressure to obtain a solid residue, which was dissolved in ethyl acetate (10 mL) and washed with saturated NaHCO.sub.3-saturated solution (3×20 mL). The organic phase was dried using MgSO.sub.4.

[0183] After removing the solvent, 220 mg of compound 14a was obtained by silica gel column chromatography (silica gel: n-hexane-ethyl acetate=1:1) (yellow solid, 74% yield). .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0184] .sup.1H NMR (400 MHz, MeOD) b 8.12 (2H, dd, J=8.4, 1.6 Hz), 7.75 (1H, d, J=8.4 Hz), 7.65-7.60 (1H, m), 7.47 (2H, t, J=8.4 Hz), 7.42 (1H, d, J=15.6 Hz), 7.11 (1H, d, J=16.0 Hz), 6.89-6.85 (2H, m), 6.73-6.70 (2H, m); .sup.13C NMR (100 MHz, MeOD) b 192.1, 166.5, 152.5, 149.8, 146.7, 134.9, 133.1, 131.2, 130.4, 129.7, 1.9, 125.2, 123.4, 123.0, 116.4, 115.5, 114.3, 111.4; LRMS (ESI) calcd. for C.sub.22H.sub.17O.sub.6 [M+H].sup.+: 377.10, found: 377.10; HRMS (ESI) calcd. for C.sub.22H.sub.17O.sub.6 [M+H].sup.+: 377.1020, found: 377.1007; M.P=204.9° C.

Compound 14b: (E)-2-(3-(3,4-dihydroxyphenyl)acryloyl)-5-hydroxyphenyl 4-methylbenzoate

[0185] ##STR00025##

[0186] Compound 14b was isolated (yellow solid, 51% yield) and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0187] .sup.1H NMR (400 MHz, MeOD) b 7.98 (2H, d, J=8.0 Hz), 7.74 (1H, d, J=15.6 Hz), 7.39 (1H, d, J=8.4 Hz), 7.26 (2H, d, J=7.6 Hz), 7.08 (1H, d, J=16.0 Hz), 6.92 (1H, d, J=2.0 Hz), 6.86-6.82 (2H, m), 6.71-6.68 (2H, m), 2.40 (3H, s); .sup.13C NMR (100 MHz, MeOD) b 192.1, 166.6, 163.5, 152.6, 149.8, 146.6, 146.1, 133.1, 131.2, 130.3, 128.0, 127.5, 125.3, 123.4, 123.2, 116.3, 115.4, 114.3, 111.4, 21.72; LRMS (ESI) calcd. for C.sub.23H.sub.19O.sub.6 [M+H].sup.+: 391.12, found: 391.10; HRMS (ESI) calcd. for C.sub.23H.sub.19O.sub.6 [M+H].sup.+: 391.1176, found: 391.1171; M.P=185.7° C.

Compound 14c: (E)-2-(3-(3,4-dihydroxyphenyl)acryloyl)-5-hydroxyphenyl 4-methoxybenzoate

[0188] ##STR00026##

[0189] Compound 14c was isolated (brown solid, 44% yield) and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0190] .sup.1H NMR (400 MHz, MeOD) b 8.05 (2H, d, J=8.8 Hz), 7.73 (1H, d, J=8.4 Hz), 7.39 (1H, d, J=15.6 Hz), 7.09 (1H, d, J=15.6 Hz), 6.95-6.93 (3H, m), 6.86-6.82 (2H, m), 6.71-6.69 (2H, m), 3.85 (3H, s); .sup.13C NMR (100 MHz, MeOD) b 192.1, 166.2, 165.7, 163.5, 152.7, 149.8, 146.6, 146.4, 133.4, 133.1, 128.0, 125.5, 123.4, 123.3, 122.3, 116.3, 115.4, 115.0, 114.3, 111.4, 56.07; LRMS (ESI) calcd. for C.sub.23H.sub.19O.sub.7 [M+H].sup.+: 407.11, found: 407.10; HRMS (ESI) calcd. for C.sub.23H.sub.19O.sub.7 [M+H].sup.+: 407.1125, found: 407.1116; M.P=161.1° C.

Compound 14d: (E)-2-(3-(3,4-dihydroxyphenyl)acryloyl)-5-hydroxyphenyl 4-methylbenzoate

[0191] ##STR00027##

[0192] Compound 14d was isolated (yellow solid, 61% yield) and .sup.1H NMR, .sup.13C NMR, LRMS, HRMS and MP data were measured as follows:

[0193] .sup.1H NMR (400 MHz, MeOD) b 8.08 (2H, d, J=8.4 Hz), 7.76 (1H, d, J=8.8 Hz), 7.47 (2H, d, J=8.8 Hz), 7.40 (1H, d, J=16.0 Hz), 7.09 (1H, d, J=16.0 Hz), 6.96 (1H, d, J=2.4 Hz), 6.88-6.85 (2H, m), 6.73-6.71 (2H, m); .sup.13C NMR (100 MHz, MeOD) b 191.9, 163.6, 152.5, 149.9, 146.7, 146.6, 133.2, 130.0, 129.1, 127.9, 125.0, 123.4, 123.0, 116.4, 115.4, 114.4, 111.4; LRMS (ESI) calcd. for C.sub.22H.sub.16O.sub.6Cl[M+H].sup.+: 411.06, found: 411.10; HRMS (ESI) calcd. for C.sub.22H.sub.16O.sub.6Cl [M+H].sup.+: 411.0630, found: 411.0620; M.P=102.8° C.

EXPERIMENTAL EXAMPLES

[0194] Compound 1 is butein and the therapeutic effect on lymphedema thereof is disclosed in Korean Patent Publication No. 10-1533197 (published on Jul. 2, 2015). However, compound 1 has poor physiological and chemical properties such as water solubility (79 μM, 21 μg/mL) and partition coefficient (log P, 0.42), and also, when treated in vitro at the concentration of 20 μM, IC.sub.50 (43 μM) and efficacy (10% inhibition) were poor. This indicates that compound 1 is not the most suitable active ingredient for the treatment of lymphedema.

[0195] Accordingly, the inventors of the present application have studied to develop, through structural modification, a compound of which physicochemical properties (solubility, log P), and pharmacokinetic parameters, such as maximum concentration (Cmax) of the drug after oral administration, bioavailability (BA), half-life (T.sub.1/2), total concentration (AUC), etc., are improved.

[0196] Anti-Inflammatory Effect

[0197] To isolate peritoneal macrophages, male Balb/c mice were intraperitoneally injected with 2 ml of 3.5% sterile thioglycolate (B D, Sparks, Md., USA) 4 days before being sacrificed. Mice were sacrificed by cervical dislocation, and peritoneal secretory cells were aseptically isolated through peritoneal washing with cold DMEM (HyClone, Logan, Utah, USA) containing 10% FBS (fetal bovine serum; HyClone) and 1% penicillin-streptomycin. After centrifugation, cells were resuspended and counted using a TC20 Cell Counter (Bio-Rad Laboratories, Hercules, Calif., USA). Peritoneal secretory cells were left in a 24-well plate overnight at 37° C., and non-adherent cells were removed. The cells were then stimulated with 100 ng/ml lipopolysaccharide (LPS) for 24 hours. The supernatant was collected during cytokine analysis, and the level of TNFα in the supernatant was measured using BD OptEIA mouse ELISA sets (BD Biosciences, San Diego, Calif., USA) according to the manufacturer's protocol.

[0198] The measurements of the LPS-induced TNFα production level in mouse peritoneal macrophages exposed to the compound having a concentration of 20 μM are shown in FIG. 1. Data are mean±standard deviation of values for three mice, respectively (with respect to LPS, *: P<0.01, **: P<0.001; with respect to compound 1 #: P<0.01, ##: P<0.001). Referring to FIG. 1, it can be seen that all of the compounds 14a to 14d, 8a and 8b treatment groups have significantly better TNFα production inhibitory ability than the positive control butein treatment group.

[0199] In addition, in order to confirm the anti-inflammatory activity of the compound, the IC.sub.50 values in relation to the dose-dependent inhibition level of TNFα production in macrophages are shown in Table 1 below. IC.sub.50 (μM) values were measured twice, respectively, and expressed as mean±standard deviation. As can be seen in Table 1 below, the IC.sub.50 value of the compound 7h treatment group was similar to that of the positive control butein treatment group, and compounds 7i, 7l, 7m, 14a to 14d, and 8a all showed significantly superior IC.sub.50 values compared to butein.

TABLE-US-00001 TABLE 1 IC.sub.50(μM) compound 1 43.3 ± 1.72 compound 7h 48.4 ± 2.35 compound 7i 32.2 ± 2.15 compound 7l 18.6 ± 1.50 compound 7m 17.3 ± 2.11 compound 14a 14.6 ± 1.56 compound 14b 18.9 ± 1.15 compound 14c 20.8 ± 1.62 compound 14d 17.9 ± 1.84 compound 8a 12.8 ± 1.24

[0200] Physicochemical and Pharmacokinetic Parameters

[0201] Compounds were administered to 5-week-old male CD-1 mice through 1 mg/kg tail vein administration (administration volume of 5 mL/kg), 20 mg/kg oral administration, and 100 mg/kg oral administration, respectively. Three mice were used for each independent experiment. Blood samples were taken in an amount of 20 μL to 30 μL via the saphenous vein at 5, 15, 30, 60, 120, 180, 360, and 1440 minutes.

[0202] Table 2 below shows the physicochemical properties of representative compounds confirmed by parallel artificial membrane permeability assay (PAMPA). As can be seen in Table 2 below, it was confirmed that in terms of the solubility and permeability, the compound of the disclosure exhibited better properties compared than Compound 1.

TABLE-US-00002 TABLE 2 pKa logP Permeability Solubility compound 1 7.96/11.65 0.42 −5.35 21 μg/mL compound 7m 9.23/11.58 3.77 −4.33 28 μg/mL compound 14a 3.38/8.63/9.55 3.47 −5.67 136 μg/mL 

[0203] In addition, pharmacokinetic parameters were calculated from non-compartmental analysis using Win Non-lin. Table 3 below shows the results of pharmacokinetic analysis of compounds 1 and 14a.

TABLE-US-00003 TABLE 3 Route of Dosage Cmax AUC∞ Com- adminis- (mg/ T.sub.1/2 Tmax (ng/ (hr*ng/ BA pound tration Kg) (hr) (hr) mL) mL) (%) com- I.V 1 — 0.1 41 6.7 — pound P.O 20 — 0.3 43 21 15 1 P.O 100 — 0.3 601 251 37 com- I.V 1 1.36 0.2 110 163 — pound P.O 20 0.57 0.4 83 141 4 14a P.O 100 0.35 0.4 1600 1224 7.5

[0204] In Table 3, T.sub.1/2 denotes the terminal half-life, Cmax denotes the maximum plasma concentration, AUC-O denotes the area under the plasma concentration-time curve from time zero to infinity, and indicates exposure concentration of whole blood, and BA represents bioavailability expressed as (AUCpo/AUCiv)×100. Compound 1 had too short terminal half-life and measurements thereof are two low, and thus T.sub.1/2 could not be measured. Cmax and AUC∞ are mean±SD values obtained from three male ICR mice. Regarding blood exposure during systemic circulation of the compound, the compound was orally administered at an amount of 100 mg/kg, and then, AUC (area under the curve) was measured, and the obtained measurements were compared.

[0205] Pharmacokinetic analysis through oral administration showed that compared to the case where unmodified compound 1 (AUC 251 h*ng/mL) was used, the case where compound 1 was delivered to the systemic circulation using compound 14a (AUC 1,224 h*ng/m) was significantly more effective.

[0206] Prophylactic Effect on Lymphedema Formation

[0207] Compounds 1, 7m, and 14a were each orally administered to mice for 7 days before surgical induction of lymphedema in mice.

[0208] The normal group did not undergo surgery and did not induce lymphedema. On the other hand, in the control group, the compound 1 treatment group, the compound 7m treatment group, and the compound 14a treatment group, lymphedema was induced through surgery. After surgery, the compounds were administered once every two days.

[0209] As can be seen in FIG. 2A, it was observed that the expansion of lymphedema in the right hind leg of the mouse was significantly suppressed. The volume of lymphedema was measured on the 7.sup.th day after surgery, and the size of the lymphedema was recorded using a caliper ruler by a person who was blinded to the treatment status of the mice.

[0210] As can be seen in FIG. 2B, the volume of lymphedema on the 7.sup.th day after surgery in the compound 1 treatment group, the compound 7m treatment group, and the compound 14a treatment group was reduced by 52%, 53%, and 70%, respectively, (with respect to the control group, *: P<0.005; with respect to compound 1, #: P<0.001).

[0211] Accordingly, it was confirmed that the modification of the pharmacokinetic properties of compound 14a ultimately significantly increased the prophylactic effect of lymphedema.

[0212] Histological and Immunofluorescence Analysis

[0213] Tissue samples were collected on the 7.sup.th day after surgery to induce lymphedema at a location 15 mm-25 mm from the right leg of the mouse. The harvested tissue samples were fixed with 10% formalin solution, and the specimens were cut to a thickness of 5 μm. Sections were deparaffinized in 100% xylene, rehydrated with a series of 100% ethanol, 95% ethanol, 90% ethanol, 80% ethanol, and 70% ethanol, and washed with water. Sections were reacted with hematoxylin solution at room temperature. After washing in PBS, sections were stained with eosin at room temperature. Images were acquired by a Pannoramic viewer microscope (3DHISTECH Ltd., Hungary). Prepared slides were incubated using anti-mouse PPARγ (Santa Cruz Biotechnology, USA) and Alexa Fluor 488 and DAPI (Life Technologies, USA). Samples were analyzed using a Nikon ECLIPSE 80i microscope (Nikon Instruments Inc., Japan).

[0214] FIG. 3A shows H&E images of the right hind legs of mice taken through histological and immunofluorescence analysis in the normal group, the control group (lymphedema-inducing carrier injection group), the compound 1 treated group, and the compound 14a treated group. In FIG. 3A, the scale bar of the upper panel is 500 μm, and the scale bar of the lower panel is 100 μm. As can be seen in FIG. 3A, it can be seen that compound 14a decreased neutrophil infiltration and ameliorated the epidermal layer of lymphedema tissue.

[0215] In addition, the result of analyzing the mRNA expression of TNFα in lymphedema tissue is shown in FIG. 3B. As a result, it was confirmed that the expression level of TNFα mRNA was decreased in the compound 1 treatment group and the compound 14a treatment group (with respect to the control group, *: P<0.05; with respect to compound 1, #: P<0.05).

[0216] Next, the modulating effects of compound 1 and compound 14a on adipogenesis observed in lymphedema were analyzed.

[0217] FIG. 4A shows H&E images of the right hind legs of mice taken through histological and immunofluorescence analysis in the normal group, the control group (lymphedema-inducing carrier injection group), the compound 1 treated group, and the compound 14a treated group. In FIG. 4A, the scale bar of the upper panel is 200 μm, and the scale bar of the lower panel is 100 μm. As can be seen in FIG. 4A, it was confirmed that compound 1 and compound 14a had the prophylactic effect on lymphedema tissue by inhibiting the development of adipose tissue in the epidermal layer.

[0218] In addition, peroxisome activated receptor γ (PPARγ) is a major regulator of lipogenesis, and as shown in FIG. 4B showing the results of immunofluorescence staining, the distribution of PPARγ in the epidermal layer of lymphedema tissue after treatment with compound 1 and compound 14a was suppressed.

[0219] As can be seen in FIG. 4C showing a western blot image, it was confirmed that compound 1 and compound 14a reduced the expression of PPARγ, which is a lipogenic biomarker, and a target protein Fabp4 (fatty acid binding protein 4) thereof in lymphedema tissue. In particular, in FIG. 4D showing the levels of PPARγ and Fabp4 mRNA, it can be seen that compound 1 exhibits superior efficacy compared to the control group, but compound 14a exhibits more significant efficacy even compared to compound 1 (* P<0.05, ** P<0.01, *** P<0.001).

[0220] These two histological tests show that inflammation and lipogenesis affect the regulation of the pathophysiological properties of lymphedema, and compound 14a significantly regulates lymphedema through the anti-inflammatory and anti-lipogenic effects compared to the control group and even to compound 1.