Accepted Manuscript Synthesis and Biological Evaluation of Curcumin Derivatives Containing NSAIDs for Their Anti-inflammatory Activity Wenfeng Liu, Yonlian Li, Yuan Yue, Kun Zhang, Qian Chen, Huaqian Wang, Yujing Lu, Moutuan Huang, Xi Zheng, Zhiyun Du PII: DOI: Reference:

S0960-894X(15)00409-6 http://dx.doi.org/10.1016/j.bmcl.2015.04.077 BMCL 22660

To appear in:

Bioorganic & Medicinal Chemistry Letters

Received Date: Revised Date: Accepted Date:

31 December 2014 10 April 2015 24 April 2015

Please cite this article as: Liu, W., Li, Y., Yue, Y., Zhang, K., Chen, Q., Wang, H., Lu, Y., Huang, M., Zheng, X., Du, Z., Synthesis and Biological Evaluation of Curcumin Derivatives Containing NSAIDs for Their Antiinflammatory Activity, Bioorganic & Medicinal Chemistry Letters (2015), doi: http://dx.doi.org/10.1016/j.bmcl. 2015.04.077

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Synthesis and Biological Evaluation of Curcumin Derivatives Containing NSAIDs for Their Anti-inflammatory Activity a

Wenfeng Liu, bYonlian Li, aYuan Yue, a,dKun Zhang, aQian Chen, aHuaqian Wang, a

a

Yujing Lu, cMoutuan Huang, a,c,*Xi Zheng, a,*Zhiyun Du

Laboratory of Natural Medicinal Chemistry & Green Chemistry, Faculty of Light

Industry and Chemical Engineering, Guangdong University of Technology, Guangzhou, 510006, China b c

Guangdong Industry Technical College, Guangzhou,510300, China

Susan Lehman Cullman Laboratory for Cancer Research, Department of Chemical

Biology, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, NJ 08854, USA d

Wuyi University, Jiangmen, 529020, China

* Corresponding author: Xi Zheng Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, USA. Tel./fax: 848-445-8069/732-445-0687. E-mail addresses: [email protected] Zhiyun Du Laboratory of Natural Medicinal Chemistry & Green Chemistry, school of Light Industry and Chemical Engineering, Guangdong University of Technology, China. Tel./fax: +86 203-932-2235. E-mail addresses: [email protected]

Abstract Oral administration of nonsteroidal anti-inflammatory drugs (NSAIDs) was frequently associated with serious adverse effects. Inspired by curcumin-a naturally traditional Chinese medicine, a series of curcumin derivatives containing NSAIDs, used for transdermal application, were synthesized and screened for their anti-inflammatory activities in vitro and in vivo. Compared with curcumin and parent NSAID (salicylic

acid and salsalate), topical application of A11 and B13 onto mouse ear edema, prior to TPA treatment markedly suppressed the expression of IL-1β, IL-6 and TNF-α, respectively. Mechanistically, A11 and B13 blocked the phosphorylation of IκBα and suppressed the activation of p65 and IκBα. It was found that A11 and B13 may be potent anti-inflammatory agents for the treatment of inflammatory diseases. Keywords: curcumin; NSAIDs; TPA; anti-inflammatory Nonsteroidal anti-inflammatory drugs (NSAIDs) possessing a free carboxylic acid group (Figure 1) are used as over the counter (OTC) drugs worldwide[1]. They are widely used for the treatment of inflammatory and immune diseases, such as rheumatoid arthritis[2], pain[3], and cancer[4,

5]

. However, oral administration of

NSAIDs is frequently associated with serious adverse effects in patients[6, 7]. The common events following oral administration of NSAIDs are gastric mucosal[8] and renal damage[9, 10]. Compared with oral administration, topical application of NSAIDs could provide numerous potential benefits, such as first pass metabolism and avoidance of gastrointestinal (GI) tract[11]. Herein, inspired by curcumin, a naturally traditional Chinese medicine, kinds of anti-inflammatory agents may be developed by linking curcumin with NSAIDs to improve their activities and reduce adverse effects. Curcumin, a major constituent of Curcuma longa, is responsible for pharmacological activities

of

Curcuma

longa,

such

as

anti-inflammatory,

anti-oxidative,

anti-proliferative and anti-anxiety activities[12-16]. It was indicated that curcumin is extensively used for traditional Chinese medicine and widely used as food additives in nutrition supplements[17] and functional food [18]. Curcumin would markedly inhibit 12-O-tetradecanoylphorbol-13-acetate

(TPA)-induced

inflammation

and

tumor

promotion in mouse skin epidermis[19-21]. On activation by TPA, the inflammatory signal activates a set of the IκB kinase (IKK) complex. Curcumin could markedly inhibit phosphorylation of IκBα and production of pro-inflammatory cytokines in mouse

ear

edema[22].

Recent

findings

identified

curcumin

as

a

potent

anti-inflammatory agent that may be repositioned as an effectively therapeutical candidate for the treatment of inflammatory and immune diseases[23, 24]. Given all above, we proposed linking curcumin with NSAIDs, with the aim of

obtaining unique anti-inflammatory agents. Therefore, firstly, a series of curcumin derivatives containing NSAIDs were designed, synthesized and screened for their anti-inflammatory activities by using TPA-induced mouse ear edema model. Secondly, immunohistochemical analysis further revealed that A11 and B13 suppressed TPA-induced IL-1β, IL-6 and TNF-α expression. Moreover, A11 and B13 was selected to elucidate the anti-inflammatory mechanism at the transcriptional level, suggesting their potential to serve as unique anti-inflammatory agents.

Figure 1. Chemical structures of NSAIDs containing a carboxylic acid group Table 1 Chemical Structures of Series A and Series B. S. No

Compound No.

R1

R2

Yield(%)a

IC50 (µM)b

1

A1

a

H

63

>30

2

A2

i

H

55

>30

3

A3

c

H

77

12.92

4

A4

e

H

66

25.33

5

A5

h

H

75

5.54

6

A6

j

H

58

>30

7

A7

l

H

60

8.98

8

A8

g

H

76

28.02

9

A9

k

H

45

18.72

10

A10

m

H

51

19.33

11

A11

b

H

57

>30

12

A12

d

H

80

>30

13

A13

f

H

81

9.20

14

B1

a

a

75

>30

15

B2

i

i

79

>30

16

B3

c

c

68

>30

17

B4

e

e

80

>30

18

B5

h

h

74

12.92

19

B6

j

j

61

7.36

20

B7

n

n

60

>30

21

B8

l

l

78

>30

22

B9

g

g

86

>30

23

B10

k

k

74

>30

24

B11

m

m

71

>30

25

B12

b

b

55

6.11

26

B13

d

d

63

>30

27

B14

f

f

70

28.02

a

Isolated yields. IC50 values are mean±SD of three independent experiments. c Curcumin was used as reference compound and its IC50 against RAW 264.7 cell line was found to be 22.91 µM. b

To investigate whether or not A1-13 and B1-14 have any influence on TPA-induced RAW 264.7 cells, inhibitory effects of these compounds on the proliferation of RAW 264.7 cells were determined by using the MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide] assay. Curcumin, taken as reference compound, showed an IC50 of 22.91 µM. The cytotoxicities of A1-13 and B1-14 against RAW 264.7 were expressed as IC50 value (Table 1). Regarding the activities of these compounds against RAW 264.7 cell line, the results (Table 1) showed that A5 (IC50: 5.54 µM) and B12 (IC50: 6.11 µM), possessed the highest degree of cytotoxicity, were more potent than curcumin (IC50: 22.91 µM). The activities of A1-13 and B1-14 against RAW 264.7 cell line had the descending order as follow: (A5 > B12 > B6 > A7 > A13 > A3= B5 > A9 > A10> curcumin). Structure-activity relationship (SAR) studies revealed that the anti-inflammatory

activities of curcumin derivatives containing mono-NSAID (c, e, f, g, h, l, k, m) are better than that of curcumin derivatives containing bis-NSAIDs (Figure 2). The possible reason for this was Series A (Figure 2) have even better activity than Series B since Series A may have increased solubility, slow metabolic process and enhanced cellular uptake[25]. As shown in Figure 3, A5 was the most potent anti-inflammatory against RAW 264.7 cell line. The possible reason for this was that the hydrogen atom was replaced with –CF3 (an electron-withdrawing substituent) at the meta-position on the benzene rings (A5) increased activity against RAW264.7 cell line compared with 4-chlorophenyl compound (A13) and 4-methylphenyl compound (A8)[26]. Above all, chemical modification of polyphenols in the herbal products, specially linking them with suitable ligands may improve their bioactivities and release the active constituent at the target site. In addition, the free phenolic group in the polyphenols modified chemically is an essential prerequisite of high potency.

Figure 2. The more potent compounds against RAW 264.7 cell line

Figure 3. The most potent compound as anti-inflammatory agent

Weight of ear punches(mg)

16 14 12 10 8

** *

*

**

**

**

6 4 2 A ce t Cu one rc TP um A in A 1 A 2 A3 A 4 A 5 A 6 A 7 A 8 A9 A1 A0 11 A 1 A2 13 B1 B 2 B3 B4 B5 B6 B7 B 8 B B19 B10 B11 B12 B13 4

0

TPA 0.008 nM/15 µL

Figure 4. Effects of A1-13 and B1-14 on the weight of ear punches. Each bar represents the mean ± SE from 3 mice. Statistical significance relative to TPA group was indicated, *P < 0.05, ** P < 0.01.

The anti-inflammatory activity of A1-13 and B1-14 can be demonstrated by their effects on TPA-induced mouse ear edema. When TPA was applied topically on ears of female BALB/c mice, the average weights of ear punches (6 mm diameter) were increased from 6.1 mg to 12.1 mg. Topical application of these compounds, 6 min prior to TPA treatment resulted in a decrease in the weight of a punch biopsy. The concentration, conducted at screening anti-inflammatory activity of these compounds, was selected as 0.75 µM according to previous publications[19-21].

Figure 5. The most potent compounds containing salicylic acid NSAID against mouse ear edema

Figure 6. The most potent compounds containing propionic acid NSAID against mouse ear edema

Figure 7. The most potent compounds containing fenamic acid NSAID against mouse ear edema

Figure 8. Anti-inflammatory structure activity relationship of the studied compounds

Concerning TPA-induced mouse ear edema, it is evident that the tested A1-13 and B1-14 showed anti-inflammatory activities with decrease in the weight of ear punches ranging from 10.0% to 93.3% (Figure 4). The inhibitory effects of 23 compounds were found to more potent than that of curcumin. The effect of the tested compounds (up to 90% decrease) on TPA-induced mouse ear edema had descending order as follow: (B4 = B13 > A6= A11 > A5 = B6). As illustrated in Figure 5, 6 and 7, the most potent curcumin derivatives containing salicylic acid NSAID, propionic acid NSAID, and fenamic acid NSAID against mouse ear edema are B13, A6 and A5, respectively. In addition, A5 is the most potent

curcumin derivatives containing fenamic acid NSAID against both of RAW 264.7 cell line (Figure 3) and mouse ear edema (Figure 7). On the other hand, B4 and B13 only showed very high potency to decrease the weights of ear punches (Figure 8), but not showed high potency against RAW 264.7 cell line. The possible reason for this was that B4 and B13, curcumin derivatives containing bis-ibuprofen and bis-salsalate respectively, have no free phenolic for binding at active site, weaker bioactivities and decreased solubility. Mechanistically, accumulation and better stability of A5 might suppress reactive oxygen species (ROS) generation to induce apoptosis in RAW 264.7 cells. However, further experiments are needed to confirm the anti-inflammatory mechanism of these derivatives against RAW 264.7 cell line. (B) 16

(A)16

14

*

**

12

*

10

**

8 6 4 2

Weight of ear punches(mg)

12

8

*

6 4 2

6 A

=1 :1 C: Ib

pr of en Ib u

A

TPA 0.008nM/15uL

TPA 0.008nM/15uL

(C)16

(D) 16 *

**

12

14

**

10

*

8 6 4 2

Weight of ear punches(mg)

14

12

**

**

10

*

8

**

6 4 2

14

*

10

*

8

**

6 4 2

*

12

B4

*

**

10 8

**

6 4 2

TPA 0.008nM/15uL

TPA 0.008nM/15uL

B1 3

C :S =1 :2

Sa la la te

Cu rc um in

TP A

B6

A ce to ne

C: Ib

=1 :2

0 TP A Cu rc um in Ib up ro fe n

A ce to ne

0

Weight of ear punches(mg)

(F) 16

14

**

:E =1 :2

TPA 0.008nM/15uL

(E) 16 12

C

A ce to ne

11 A

1 :S =1 : C

Sa lic yl ic ac id

TP A

Cu rc um in

A ce to ne

TPA 0.008nM/15uL

Et od ol ac

0

0

TP A Cu rc um in

Weight of ear punches(mg)

rc um in

TP

Ac et on e

A 5

C :F =1 :1

A ci d

C ur cu m in

Fl uf en am ic

T PA

0 A ce to ne

0

Weight of ear punches(mg)

*

*

**

10

Cu

Weight of ear punches(mg)

14

Figure 9. Effects of curcumin, NSAID, combining of curcumin with NSAID (1:1 or 1:2) and A5, A6, A11, B4, B6, B13 on the weight of ear punches. Each bar represents the mean ± SD from 3 mice. Statistical significance relative to TPA group was indicated, * P < 0.05, ** P < 0.01.

To investigate whether the anti-inflammatory activity of A5, A6, A11, B4, B6 and B13 (up to 90% decrease in initial screening) are better than NSAIDs and combination of curcumin and NSAIDs (1:1 or 1:2, mol/mol), the same topical inflammation model was used in the next experiments. As shown in Figure 9, the inhibitory effects of A11 (Figure 9C) and B13 (Figure 9F) on decrease in the weight of a punch biopsy were the most pronounced than that of salicylic acid or salsalate, combination of curcumin and salicylic acid (1:1, mol/mol) or curcumin and salsalate (1:2, mol/mol), respectively. Among these tested compounds, the inhibitory rates of A11 or B13 reached 91.8% or 93.3%, respectively. The possible reason for this was that the interactions of curcumin and NSAIDs could enhance or attenuate the decrease in the weight of a punch biopsy. However, the anti-inflammatory mechanism and the interactions of combination of curcumin and NSAIDs are currently determined in the ongoing studies in our lab. Histological appearance of mouse ears was examined after pretreatment with acetone (control group) or compounds (curcumin, salicylic acid or salsalate, combination of curcumin and salicylic acid or salsalate, and A11 or B13) stained with H&E stain. The histological assessment is shown in Figure 10a-f. Control mice treated with acetone alone show the normal appearance in the epidermal layer without any significant lesion (Figure 10A). Topical application of TPA onto mouse ears alone, inflammation induced by TPA shows the different appearances, such as epidermal hyperkeratosis, thickening of the stratum corneum and an increase in infiltration of inflammatory cells (Figure 10B). Moreover, topical application of curcumin, salicylic acid or salsalate, combination of curcumin and salicylic acid (1:1) or combination of curcumin and salsalate (1:2), and A11 or B13 prior to TPA application would decrease in the level of inflammatory cells infiltration, respectively (Figure 10C-F). As shown in Figure 10C-E, A11 or B13 significantly showed morphological alterations compared to TPA treatment. The histological appearance of mouse ears was obtained from 5 scopes of

every tissue slice (Figure 10).

Acetone

TPA

Curcumin

Salicylic acid

C+SA(1:1)

A11

Salalate

C+SB(1:2)

B13

Figure 10. H&E staining for histological changes of TPA-induced mouse ears. (A) Control. (B) TPA. (C) Curcumin. (D) Salicylic acid. (E) Curcumin + Salicylic acid (1 : 1). (F) A11. (G) Salalate. (H) Cucmin + Salalate (1 : 2). (I) B13.

To confirm if A11 and B13 inhibit pro-inflammatory cytokines, the levels of IL-1β, IL-6 and TNF-α were determined by immunohistochemical analysis. When TPA was applied topically onto ears of female BALB/c mice, the levels of IL-1β, IL-6 and TNF-α were markedly increased respectively (Figure 11). As shown in Figure 11, topical application of A11 and B13, 6 min prior to TPA treatment resulted in a significant reduction in the levels of IL-1β, IL-6 and TNF-α in mouse ear edema, respectively.

IL-1β IL-6 TNF-α Acetone

TPA

Curcumin

Salicylic acid

C+SA(1:1)

A11

Salalate

C+SB(1:2)

B13

Figure 11. Immunohistochemical staining for acetone control, TPA, curcumin, salicylic acid, salalate, combination of curcumin and salicylic acid (1 : 1), combination of curcumin and salalate (1 : 2), and A11 or B13.

It has been indicated that NF-κB activation is critical for regulating gene expression in TPA-induced inflammatory responses[27]. Thus, the effects of A11 and B13 on NF-κB activation

were

determined

by

immunohistochemical

analysis

using

a

nonphosphorylated p65 antibody and an antiphospho-Ser536 p65 antibody. As described in Figure 12, after TPA treatment, p65, the functional active subunit of NF-κB, translocated to nucleus. Pretreatment with A11 and B13 onto mouse ears could markedly suppressed p65 translocation from cytoplasma to nucleus and strongly

decreased the nuclear levels of phospholated p65, respectively. It is demonstrated that phosphorylation of p65 by upstream kinase has been well connected with the transcriptional activity of NF-κB[28]. Thus, we found that TPA treatment induced NF-κB transcriptional activity was markedly increased compared with the control group, but significantly reduced by A11 or B13. p65 p-p65 Acetone

TPA

Curcumin

Salicylic acid

C+SA(1:1)

A11

Salalate

C+SB(1:2)

B13

Figure 12. Immunohistochemical staining for acetone control, TPA, curcumin, salicylic acid, salalate, combination of curcumin and salicylic acid (1 : 1), combination of curcumin and salalate (1 : 2), and A11 or B13.

To determine whether or not A11 and B13 functions as an inhibitor of IKK/NF-κB signaling, phosphorylation, the functional active subunit of NF-κB in TPA-induced mouse ear edema, was measured by immunohistochemical analysis using an antiphospho-Ser36 IκBα antibody and a nonphosphorylated IκB antibody (Figure 13). Topical application of A11 and B13 onto mouse ears significantly inhibited TPA-induced IκBα phosphorylation. These results suggest that A11 and B13 inhibits TPA-induced NF-κB activation by preventing IκBα phosphorylation, thus acting at or upstream of IκBα. IKK

IκB-α

p-IκB-α Acetone

TPA

Curcumin

Salicylic acid

C+SA(1:1)

A11

Salalate

C+SB(1:2)

B13

Figure 13. Immunohistochemical staining for acetone control, TPA, curcumin, salicylic acid, salalate, combination of curcumin and salicylic acid (1 : 1), combination of curcumin and salalate (1 : 2), and A11 or B13.

Because IKK represents the major upstream kinase for IκBα phosphorylation, we next

determined whether or not A and B inhibit IKK. We next determined whether A11 and B13 inhibit IKK. IKK kinase activity was measured by in vitro kinase assay of the IKK immunocomplex. As shown in Figure 14, IKK activity was markedly increased in the IKK immunocomplex from mouse ears treated with TPA. A11 or B13 significantly inhibited TPA-induced IKK activity, thereby suggesting that A11 or B13 inhibits TPA-induced NF-κB activation at the level (or upstream) of IKK. This result indicates that A11 or B13 also inhibits IKK-NF-κB signaling in vivo. It was well known that NSAIDs primarily suppress the expression of cyclooxygenase enzymes and thereby affect the synthesis of prostaglandins, which are involved in the inflammation-associated diseases[29]. Animal experimental data indicated that the anti-inflammatory mechanism of A11 or B13, which linking NSAID with curcumin, is different from that of NSAIDs. The anti-inflammatory mechanism of A11 and B13 was that they would inhibit NF-κB-dependent inflammatory response by directly targeting IKK.

Figure 14. A schematic representation of suppression of TPA-induced NF-κB activation and IL-1β, IL-6 and TNF-α expression by A11 or B13 in mouse ear edema.

In summary, long-term use of NSAIDs administrated orally is frequently associated with serious adverse effects, such as gastric mucosal[8] and renal damage[9, 10]. Unique anti-inflammatory agents are urgent need for transdermal application. Inspired by curcumin, a natural traditional Chinese medicine, a series of curcumin containing NSAIDs, designed and synthesized, may be avoidance of GI tract and hence beneficial for transdermal application. Traditional Chinese medicines, being of natural

origin, possess various pharmacological activities. It is worthwhile to investigate whether or not linking them with NSAIDs could have any influence on improving their bioactivities. NSAIDs

nonsteroidal anti-inflammatory drugs

OTC

over the counter

GI

gastrointestinal

TPA

12-O-tetradecanoylphorbol-13-acetate

IL-1β

interleukin-1β

IL-6

interleukin-6

TNF-α

tumor necrosis factor-α

IκB

inhibitor of NF-κB

IKK

IκB kinase

NF-κB

nuclear factor-κB

r.t.

room temperature

mM

mmol·L-1

µM

µmol·L-1

EDCI

1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride

DMAP

4-dimethylaminopyridine

ROS

reactive oxygen species

EDTA

ethylene diamine tetraacetic acid

PBS

phosphate buffer saline

1

proton nuclear magnetic resonance

H NMR

DMSO

dimethyl sulfoxide

Acknowledgments

The present study was supported by the Guangdong Province Leadership Grant and China National Science Foundation Grants (Grant No. 81272452). The authors dedicate this paper to Dr. Allan H. Conney, an outstanding and widely recognized cancer researcher who passed away on September 10, 2013. References and notes

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Note:

Refer

Supplementary

material

for

synthetic

procedures,

spectral

characterization and biological characterization Melting points (ºC, uncorrected) were recorded on X-4 melting apparatus. 1H NMR and

13

C NMR spectra were recorded on Bruker BioSpin GmbH spectrometer in

CDCl3 or DMSO-d6 using TMS as an internal standard. General synthesis of the target compounds is described in Scheme 1. Curcumin were reacted with NSAIDs a-n (Figure 1) in dichloromethane at room temperature to afford the corresponding curcumin derivatives with NSAIDs. The chemical structures of these compounds were elucidated on the basis of spectral data (1H-NMR, 13C-NMR, and LC-MS).

Scheme 1. General Synthesis of Series A and Series B.

General procedure of the synthesis of A1-A13: A solution of Curcumin (1.2 mmol), NSAID (1.0 mmol), EDCI (1.0 mmol) and DMAP (0.1mmol) in CH2 Cl2 (30 mL), the mixture was stirred for 1 h at room temperature. The reaction mixture was quenched with satd. aqueous NaHCO3, extracted with CH2Cl2 (3 x 30 mL), dried over Na2SO4, concentrated under reduced pressure and purified by silica gel chromatography. 4-((1E,6E)-7-(4-hydroxy-3-methoxyphenyl)-3,5-dioxohepta-1,6-dien-1-yl)-2-methoxyp henyl-2-acetoxybenzoate (A1). Yellow powder, yield 63%, mp 189 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 10.40 (s, 1H), 8.26 (dd, J = 7.8, 1.4 Hz, 1H), 8.10 (dd, J = 8.0, 1.4 Hz, 1H), 7.72 – 7.60 (m, 3H), 7.57 – 7.47 (m, 1H), 7.40 (t, J = 7.6 Hz, 1H), 7.24 – 7.12 (m, 6H), 7.05 (d, J = 8.2 Hz, 1H), 6.98 (t, J = 7.6 Hz, 1H), 6.60 (dd, J = 15.8, 6.3 Hz, 2H), 5.89 (s, 1H), 3.88 (s, 3H), 3.87 (s, 3H), 2.31 (s, 3H).13C NMR (101 MHz, CDCl3): δ (ppm) 183.09, 169.64, 168.20, 162.15, 151.53, 151.28, 141.18, 140.57, 140.04, 139.75, 136.53, 134.63, 134.51, 134.17, 132.43, 130.63, 126.20, 124.59, 124.35, 124.07, 123.46, 123.32, 122.26, 121.11, 119.53, 117.79, 111.62, 101.92, 56.02, 55.96, 21.02. LC-MS m/z: 531.2 [M + H] +.

4-((1E,6E)-7-(4-hydroxy-3-methoxyphenyl)-3,5-dioxohepta-1,6-dien-1-yl)-2-methoxyp henyl-2-(2-((2,6-dichlorophenyl)amino)phenyl)-acetate (A2). Orange powder, yield 55%, mp 96 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 7.59 (dd, J = 15.8, 9.6 Hz, 2H), 7.40 – 7.29 (m, 4H), 7.20 – 7.09 (m, 3H), 7.09 – 6.99 (m, 4H), 6.95 (dd, J = 17.3, 8.1 Hz, 2H), 6.73 (s, 1H), 6.62 – 6.43 (m, 3H), 5.98 (s, 1H), 5.82 (s, 1H), 4.09 (s, 2H), 3.93 (s, 3H), 3.74 (s, 3H). 13C NMR (101 MHz, CDCl3): δ (ppm) 184.55, 181.76, 170.02, 151.28, 148.03, 146.84, 142.80, 141.22, 141.16, 139.35, 137.94, 134.26, 131.09, 129.46, 128.87, 128.19, 127.55, 124.32, 124.12, 124.07, 123.12, 123.05, 122.31, 121.77, 120.94, 118.58, 114.89, 111.43, 109.70, 101.57, 55.96, 55.87, 38.20, 29.72. LC-MS m/z: 646.1 [M + H] +. 4-((1E,6E)-7-(4-hydroxy-3-methoxyphenyl)-3,5-dioxohepta-1,6-dien-1-yl)-2-methoxyp henyl-2',4'-difluoro-3-hydroxy-[1,1'-biphenyl]-4-carboxylate (A3). Orange powder, 47% yield, mp 137 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 10.47 (s, 1H), 8.23 (s, 1H), 7.63 (dt, J = 14.5, 10.1 Hz, 4H), 7.48 – 7.36 (m, 2H), 7.18 (d, J = 13.0 Hz, 3H), 7.12 (d, J = 8.6 Hz, 2H), 7.03 (d, J = 14.9 Hz, 2H), 7.00 – 6.87 (m, 4H), 6.68 – 6.43 (m, 2H), 5.87 (dd, J = 42.1, 23.7 Hz, 2H), 3.94 (s, 3H), 3.88 (s, 3H).

13

C NMR (101

MHz, CDCl3): δ (ppm) 184.73, 181.56, 168.06, 161.68, 151.43, 148.07, 146.84, 141.28, 140.16, 139.13, 137.04, 134.75, 131.16, 130.74, 127.55, 127.55, 126.51, 126.51, 124.65, 123.26, 123.08, 121.78, 120.97, 118.11, 114.89, 111.76, 111.59, 109.71, 104.73, 104.47, 101.63, 56.02, 55.98. LC-MS m/z: 601.2 [M + H] +. 4-((1E,6E)-7-(4-hydroxy-3-methoxyphenyl)-3,5-dioxohepta-1,6-dien-1-yl)-2-methoxyp henyl-2-(1,8-diethyl-1,3,4,9-tetrahydropyrano[3,4-b]indol-1-yl)acetate

(A4)

Red

powder, 56% yield, mp 112 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 7.89 (s, 1H), 7.81 (d, J = 7.5 Hz, 2H), 7.72 (d, J = 7.3 Hz, 1H), 7.66 (d, J = 7.5 Hz, 8H), 7.62 – 7.53 (m, 1H), 7.48 (dd, J = 13.3, 6.8 Hz, 2H), 7.14 – 7.03 (m, 2H), 6.96 (s, 6H), 6.54 (d, J = 15.7 Hz, 2H), 5.84 (s, 4H), 4.10 (d, J = 7.1 Hz, 2H), 3.74 (s, 4H), 1.68 (d, J = 7.0 Hz, 3H). 13C NMR (101 MHz, CDCl3): δ (ppm) 184.68, 181.59, 170.56, 151.17, 148.06, 146.83, 141.27, 140.68, 135.78, 134.52, 127.55, 126.61, 126.14, 124.58, 123.20, 123.07, 121.77, 121.12, 120.47, 119.66, 115.99, 114.89, 111.51, 109.71, 108.51, 101.59, 74.75, 60.73, 55.96, 42.97, 30.58, 24.09, 22.43, 13.81, 7.55. LC-MS

m/z: 638.2 [M + H] +. 4-((1E,6E)-7-(4-hydroxy-3-methoxyphenyl)-3,5-dioxohepta-1,6-dien-1-yl)-2-methoxyp henyl-2-((3-(trifluoromethyl)phenyl)amino)-benzoate (A5). Orange powder, 59% yield, mp 101 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 9.48 (s, 1H), 8.27 (dd, J = 8.0, 1.3 Hz, 1H), 7.65 (d, J = 7.2 Hz, 2H), 7.49 (d, J = 4.3 Hz, 2H), 7.44 (t, J = 7.6 Hz, 2H), 7.39 (d, J = 8.1 Hz, 1H), 7.33 (dd, J = 12.4, 7.9 Hz, 3H), 7.23 – 7.16 (m, 3H), 7.12 (dt, J = 9.6, 4.8 Hz, 1H), 7.04 (dd, J = 13.0, 3.9 Hz, 1H), 6.93 (t, J = 6.9 Hz, 1H), 6.91 – 6.85 (m, 1H), 6.59 (d, J = 10.1 Hz, 1H), 6.53 – 6.42 (m, 1H), 5.85 (s, 1H), 3.94 (s, 3H), 3.89 (s, 3H).13C NMR (101 MHz, CDCl3): δ (ppm) 184.57, 181.76, 166.55, 151.67, 148.03, 147.46, 146.83, 141.28, 141.19, 141.03, 139.36, 135.15, 134.33, 132.53, 129.97, 127.56, 124.70, 124.39, 123.54, 123.06, 121.77, 121.03, 119.87, 118.45, 118.25, 118.13, 114.88, 114.21, 111.58, 111.53, 109.69, 101.58, 56.04, 55.97, 29.71. LC-MS m/z: 632.2 [M + H] +. 4-((1E,6E)-7-(4-hydroxy-3-methoxyphenyl)-3,5-dioxohepta-1,6-dien-1-yl)-2-methoxyp henyl-2-(4-isobutylphenyl)-propanoate (A6). Orange powder, 61% yield, mp 131 ºC; 1

H NMR (400 MHz, CDCl3): δ (ppm) 7.58 (d, J = 4.0 Hz, 2H), 7.33 (d, J = 8.0 Hz,

3H), 7.12 (dd, J = 5.7, 1.8 Hz, 4H), 7.05 (d, J = 5.9 Hz, 2H), 6.50 (s, 2H), 5.81 (s, 1H), 3.99 (d, J = 7.1 Hz, 1H), 3.93 (s, 3H), 3.74 (s, 3H), 2.48 (d, J = 7.2 Hz, 2H), 1.62 (d, J = 7.1 Hz, 3H), 0.92 (d, J = 6.6 Hz, 7H). 13C NMR (101 MHz, CDCl3): δ (ppm) 184.45, 181.89, 172.65, 151.50, 148.01, 146.83, 141.52, 141.09, 140.74, 139.49, 137.22, 133.93, 129.34, 127.56, 127.41, 124.12, 123.11, 123.02, 121.77, 120.93, 114.88, 111.52, 109.69, 101.50, 55.95, 55.81, 45.08, 45.03, 30.23, 22.40, 18.69. LC-MS m/z: 557.3 [M + H] +. 4-((1E,6E)-7-(4-hydroxy-3-methoxyphenyl)-3,5-dioxohepta-1,6-dien-1-yl)-2-methoxyp henyl-2-(3-benzoylphenyl)propanoate (A7). Yellow powder, 67% yield, mp 83 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 7.93 – 7.75 (m, 3H), 7.64 (ddd, J = 22.6, 16.8, 7.3 Hz, 5H), 7.48 (dd, J = 13.0, 7.4 Hz, 3H), 7.09 (dd, J = 18.4, 6.2 Hz, 4H), 6.95 (dd, J = 15.1, 8.1 Hz, 2H), 6.61 – 6.42 (m, 2H), 5.86 (d, J = 23.3 Hz, 2H), 4.10 (d, J = 7.0 Hz, 1H), 3.95 (s, 3H), 3.75 (s, 3H), 1.68 (d, J = 7.1 Hz, 4H). 13C NMR (101 MHz, CDCl3): δ (ppm) 196.71, 184.58, 181.92, 171.94, 151.26, 148.00, 146.82, 141.13, 140.37,

139.55, 137.99, 137.50, 134.44, 132.56, 131.78, 130.10, 129.43, 129.19, 128.55, 128.35, 127.51, 124.28, 123.05, 121.93, 121.07, 114.87, 111.61, 109.71, 101.41, 97.46, 55.98, 55.80, 55.07, 45.30, 30.93, 29.71, 29.21, 18.66. LC-MS m/z: 605.2 [M + H] +. 4-((1E,6E)-7-(4-hydroxy-3-methoxyphenyl)-3,5-dioxohepta-1,6-dien-1-yl)-2-methoxyp henyl-2-((2,3-dimethylphenyl)amino)benzoate (A8). Orange powder, 55% yield, mp 193 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 9.17 (s, 1H), 8.24 (d, J = 8.3 Hz, 1H), 7.63 (dd, J = 15.1, 10.9 Hz, 2H), 7.36 (d, J = 8.1 Hz, 2H), 7.24 – 7.14 (m, 3H), 7.06 (s, 3H), 6.98 – 6.71 (m, 3H), 6.55 (dd, J = 33.9, 15.4 Hz, 2H), 5.87 (d, J = 12.9 Hz, 2H), 5.33 (d, J = 18.7 Hz, 2H), 3.93 (d, J = 22.9 Hz, 6H), 2.31 (s, 3H), 1.26 (s, 3H). 13C NMR (101 MHz, CDCl3): δ (ppm) 184.51, 181.93, 166.74, 151.84, 150.19, 148.04, 146.86, 141.36, 141.15, 139.54, 138.42, 138.30, 135.02, 134.13, 132.51, 132.21, 127.60, 127.01, 125.97, 124.26, 123.74, 123.15, 123.07, 121.82, 121.09, 116.30, 114.91, 113.82, 111.64, 109.72, 109.53, 101.59, 56.06, 55.98, 20.63, 14.02. LC-MS m/z: 592.2 [M + H] +. (S)-4-((1E,6E)-7-(4-hydroxy-3-methoxyphenyl)-3,5-dioxohepta-1,6-dien-1-yl)-2-meth oxyphenyl-2-(6-methoxynaphthalen-2-yl)-propanoate (A9). Orange powder, 45% yield, mp 152 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 7.75 (dd, J = 17.6, 9.7 Hz, 4H), 7.63 – 7.47 (m, 3H), 7.18 – 7.02 (m, 6H), 6.93 (dd, J = 9.5, 8.3 Hz, 2H), 6.49 (dd, J = 15.7, 14.4 Hz, 2H), 5.80 (s, 1H), 4.14 (d, J = 7.1 Hz, 1H), 3.92 (d, J = 1.9 Hz, 6H), 3.70 (s, 3H), 1.70 (d, J = 7.1 Hz, 4H).13C NMR (101 MHz, CDCl3): δ (ppm) 184.47, 181.87, 172.59, 157.73, 151.50, 148.02, 146.84, 141.49, 141.10, 139.46, 135.17, 134.00, 133.82, 129.33, 128.99, 127.57, 127.14, 126.41, 126.29, 124.17, 123.10, 123.03, 121.79, 120.92, 119.03, 114.88, 111.49, 109.70, 105.66, 101.50, 55.96, 55.81, 55.34, 45.37, 18.72. LC-MS m/z: 581.2 [M + H] +. 4-((1E,6E)-7-(4-hydroxy-3-methoxyphenyl)-3,5-dioxohepta-1,6-dien-1-yl)-2-methoxyp henyl-3-(4,5-diphenyloxazol-2-yl)propanoate (A10). Orange powder, 51%yield, mp 103 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 7.87 (d, J = 7.3 Hz, 1H), 7.76 – 7.52 (m, 6H), 7.46 – 7.31 (m, 6H), 7.24 – 7.02 (m, 5H), 6.95 (d, J = 8.2 Hz, 1H), 6.53 (s, 2H), 5.85 (s, 2H), 3.96 (s, 3H), 3.85 (s, 3H), 3.35 (s, 2H), 3.24 (s, 2H).13C NMR (101 MHz,

CDCl3): δ (ppm) 181.77, 169.97, 161.50, 151.38, 148.05, 146.86, 141.14, 139.38, 135.16, 134.18, 133.65, 129.46, 128.89, 128.67, 128.58, 128.12, 127.90, 127.59, 126.52, 124.31, 123.24, 123.03, 121.77, 120.92, 114.89, 111.47, 109.72, 101.52, 55.92, 30.95, 23.57. LC-MS m/z: 644.2 [M + H] +. 4-((1E,6E)-7-(4-hydroxy-3-methoxyphenyl)-3,5-dioxohepta-1,6-dien-1-yl)-2-methoxyp henyl 2-hydroxybenzoate (A11). Orange powder, yield 57%, mp 166 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 10.41 (s, 1H), 8.10 (dd, J = 8.0, 1.4 Hz, 1H), 7.68 – 7.50 (m, 3H), 7.14 (dd, J = 23.5, 10.0 Hz, 4H), 7.05 (d, J = 8.1 Hz, 2H), 7.00 – 6.85 (m, 2H), 6.53 (dd, J = 32.6, 15.8 Hz, 2H), 5.83 (s, 1H), 3.89 (d, J = 23.4 Hz, 6H).13C NMR (101 MHz, CDCl3): δ (ppm) 184.71, 181.62, 168.23, 162.09, 151.47, 148.11, 146.89, 141.27, 140.46, 139.17, 136.54, 134.63, 130.65, 127.54, 124.5, 123.27, 123.07, 121.75, 120.96, 119.55, 117.79, 114.92, 111.62, 109.77, 101.63, 99.86, 56.00, 29.73. LC-MS m/z: 489.1 [M + H] +. 2-((4-((1E,6E)-7-(4-hydroxy-3-methoxyphenyl)-3,5-dioxohepta-1,6-dien-1-yl)-2-meth oxyphenoxy)carbonyl)phenyl-2-hydroxybenzoate (A12). Orange powder, 80% yield, mp 122 ºC; 1H NMR (400 MHz, DMSO-d 6): δ (ppm) 10.17 (s, 1H), 9.66 (d, J = 12.4 Hz, 1H), 8.18 (d, J = 7.0 Hz, 1H), 7.98 (d, J = 7.0 Hz, 1H), 7.83 (t, J = 7.7 Hz, 1H), 7.53 (dt, J = 13.6, 6.7 Hz, 6H), 7.30 (d, J = 10.2 Hz, 2H), 7.16 (dd, J = 8.1, 2.1 Hz, 2H), 7.07 – 6.88 (m, 4H), 6.81 (d, J = 8.1 Hz, 1H), 6.10 (s, 1H), 4.48 (s, 1H), 3.79 (d, J = 18.6 Hz, 6H).13C NMR (101 MHz, DMSO-d6): δ (ppm) 184.94, 183.14, 181.26, 166.38, 161.82, 160.14, 151.04, 149.98, 149.53, 147.97, 141.56, 140.36, 139.69, 138.83, 136.06, 135.23, 134.06, 131.85, 131.00, 126.93, 126.18, 124.67, 123.22, 122.05, 121.22, 119.40, 117.48, 115.68, 112.89, 111.83, 101.32, 96.74, 55.86, 54.46, 18.53. LC-MS m/z: 609.2 [M + H] +. 4-((1E,6E)-7-(4-hydroxy-3-methoxyphenyl)-3,5-dioxohepta-1,6-dien-1-yl)-2-methoxyp henyl-2-((3-chloro-2-methylphenyl)amino)benzoate (A13). Orange powder, 81% yield, mp 225 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 9.16 (s, 1H), 8.23 (d, J = 7.9 Hz, 1H), 7.62 (dd, J = 15.7, 7.1 Hz, 2H), 7.36 (t, J = 7.7 Hz, 1H), 7.27 (s, 1H), 7.24 (s, 1H), 7.19 (d, J = 9.6 Hz, 3H), 7.10 (dd, J = 18.2, 10.7 Hz, 3H), 6.96 – 6.70 (m, 3H), 6.54 (dd, J = 25.0, 15.8 Hz, 2H), 5.88 (d, J = 23.1 Hz, 2H), 3.92 (d, J = 15.6 Hz, 6H),

2.32 (s, 3H).13C NMR (101 MHz, CDCl3): δ (ppm) 184.51, 183.12, 181.82, 166.66, 151.77, 149.20, 148.01, 146.82, 141.30, 140.11, 139.41, 135.63, 135.08, 134.10, 132.29, 131.72, 127.56, 126.88, 125.90, 124.33, 123.67, 123.05, 121.79, 121.17, 117.18, 114.87, 113.94, 111.62, 110.25, 109.68, 101.87, 101.56, 56.04, 26.92, 15.00. LC-MS m/z: 612.2 [M + H] +. [26] General procedure of the synthesis of B1-B14: Curcumin (1.2 mmol), NSAID (2.0 mmol), EDCI (1.0 mmol) and DMAP (0.1 mmol) were dissolved in CH2 Cl2 (30 mL), and the mixture was stirred for 1 h at room temperature. The reaction mixture was quenched with satd. aqueous NaHCO3, extracted with CH2Cl2 (3 x 30 mL), dried over Na2SO4, concentrated under reduced pressure and purified by silica gel chromatography. All crude products were recrystallized in petroleum ether/diethyl ether (v/v 1:1) to give the pure products. ((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)-bis(2-methoxy-4,1-phenylene)-bis(2-acet oxybenzoate) (B1). Yellow powder, 75% yield, mp 193 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 8.30(m, 2H), 7.65 (m, 4H), 7.40 (ddd, J = 19.5, 13.2, 3.4 Hz, 2H), 7.10 (m, 9H), 6.61 (m, 2H),5.88 (m, 2H), 3.94 – 3.83 (m, 6H), 2.33 (d, J = 4.4 Hz, 6H).13C NMR (101 MHz, CDCl3): δ (ppm) 183.11, 169.64, 162.21, 151.55, 151.28, 141.16, 139.95, 134.63, 134.19, 132.43 126.20, 124.38, 124.07, 123.44, 122.27, 121.13, 111.62, 101.89, 55.96, 21.02. LC-MS m/z: 693.2 [M + H] +. ((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)bis(2-methoxy-4,1-phenylene)-bis(2-(4-(( 2,6-dichlorophenyl)amino)phenyl)acetate) (B2). Yellow powder, 79% yield, mp 142 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 7.61 (d, J = 15.4 Hz, 2H), 7.35 (t, J = 7.6 Hz, 6H), 7.16 (dd, J = 12.9, 6.2 Hz, 4H), 7.06 (dd, J = 11.3, 7.3 Hz, 4H), 6.99 (dd, J = 10.5, 5.6 Hz, 3H), 6.73 (s, 2H), 6.58 (t, J = 11.8 Hz, 4H), 5.85 (s, 1H), 4.11 (d, J = 5.5 Hz, 4H), 3.78 (d, J = 10.6 Hz, 6H).13C NMR (101 MHz, CDCl3): δ (ppm) 183.08, 169.97, 151.32, 142.80, 141.36, 139.94, 137.95, 134.13, 131.08, 129.45, 128.87, 128.19, 124.35, 124.12, 124.05, 123.16, 122.32, 121.07, 118.60, 111.45, 101.81, 55.88, 38.20, 29.71. LC-MS m/z: 923.1 [M + H] +. ((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)-bis(2-methoxy-4,1-phenylene)-bis(2',4'-di fluoro-3-hydroxy-[1,1'-biphenyl]-4-carboxylate) (B3). Yellow powder, 68% yield, mp

176 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 10.46 (s, 2H), 8.28 (d, J = 38.5 Hz, 2H), 7.74 – 7.61 (m, 5H), 7.43 (td, J = 8.6, 6.6 Hz, 2H), 7.20 (d, J = 11.0 Hz, 6H), 7.16 – 7.07 (m, 2H), 7.01 – 6.86 (m, 5H), 6.57 (dd, J = 31.5, 15.8 Hz, 2H), 6.01 – 5.70 (m, 1H), 3.89 (s, 6H).13C NMR (101 MHz, CDCl3): δ (ppm) 183.07, 168.03, 163.58, 161.69, 161.04, 158.55, 151.48, 140.52, 139.81, 137.05, 134.60, 131.12, 130.73, 126.51, 124.65, 124.02, 123.31, 121.09, 118.11, 111.66, 104.46, 101.95, 56.02, 29.71. LC-MS m/z: 833.2 [M + H] +. ((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)-bis(2-methoxy-4,1-phenylene)-bis(2-(1,8diethyl-1,3,4,9-tetrahydropyrano[3,4-b]indol-1-yl)acetate) (B4). Yellow powder, 80% yield, mp 124 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 8.96 (s, 2H), 7.65 (d, J = 15.8 Hz, 2H), 7.38 (d, J = 7.7 Hz, 2H), 7.22 – 7.12 (m, 4H), 7.11 – 6.93 (m, 6H), 6.60 (d, J = 15.7 Hz, 2H), 5.89 (s, 1H), 4.06 (d, J = 21.7 Hz, 4H), 3.89 (s, 6H), 3.39 – 3.27 (m, 3H), 3.21 (d, J = 16.1 Hz, 2H), 2.82 (dt, J = 15.1, 7.1 Hz, 8H), 2.28 – 2.04 (m, 5H), 1.28 (t, J = 7.5 Hz, 6H), 0.96 (t, J = 7.3 Hz, 5H).13C NMR (101 MHz, CDCl3): δ (ppm) 183.06, 170.54, 151.22, 140.86, 139.83, 135.77, 134.44, 126.59, 126.16, 124.55, 123.25, 121.24, 120.48, 119.67, 116.00, 111.58, 108.53, 101.91, 74.75, 60.73, 55.96, 42.97, 30.59, 26.93, 24.10, 22.43, 13.8, 7.55. LC-MS m/z: 907.4 [M + H] +. ((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)bis(2-methoxy-4,1-phenylene)-bis(2-((3-(t rifluoromethyl)phenyl)amino)benzoate) (B5). Yellow powder, 74% yield, mp 135 ºC; 1

H NMR (400 MHz, CDCl3): δ (ppm) 9.48 (s, 2H), 8.27 (dd, J = 8.1, 1.4 Hz, 2H),

7.69 (t, J = 16.4 Hz, 2H), 7.48 (dd, J = 5.2, 3.6 Hz, 3H), 7.45 (d, J = 8.7 Hz, 3H), 7.40 (t, J = 7.3 Hz, 3H), 7.33 (dd, J = 12.6, 7.9 Hz, 4H), 7.27 – 7.24 (m, 1H), 7.24 – 7.20 (m, 3H), 7.19 (d, J = 1.3 Hz, 2H), 6.91 (t, J = 7.5 Hz, 2H), 6.61 (d, J = 15.8 Hz, 2H), 5.90 (s, 1H), 3.94 – 3.82 (m, 6H). 13C NMR (101 MHz, CDCl3): δ (ppm) 183.12, 166.54, 151.72, 147.49, 141.29, 139.98, 135.18, 134.21, 132.55, 132.05, 131.73, 129.98, 125.27, 124.71, 124.41, 123.60, 122.56, 121.16, 119.85, 118.46, 118.22, 114.23, 111.57, 101.90, 56.06. LC-MS m/z: 895.2 [M + H] +. ((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)bis(2-methoxy-4,1-phenylene)-bis(2-(4-is obutylphenyl)propanoate) (B6). Yellow powder, 61% yield, mp 93 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 7.68 (s, 2H), 7.32 (d, J = 8.0 Hz, 4H), 7.16 (s, 4H), 7.11 (d, J

= 8.5 Hz, 2H), 7.06 (s, 1H), 6.98 (dd, J = 16.7, 7.0 Hz, 2H), 6.54 (d, J = 15.6 Hz, 2H), 5.83 (s, 1H), 3.98 (d, J = 7.1 Hz, 2H), 3.75 (d, J = 6.0 Hz, 6H), 2.48 (d, J = 7.2 Hz, 4H), 1.94 – 1.78 (m, 2H), 1.62 (d, J = 7.1 Hz, 8H), 0.92 (d, J = 6.6 Hz, 13H).13C NMR (101 MHz, CDCl3): δ (ppm) 172.62, 151.50, 140.73, 140.03, 137.21, 129.33, 127.40, 123.12, 121.01, 111.54, 55.80, 45.05, 30.23, 29.71, 22.40, 18.68. LC-MS m/z: 745.4 [M + H] +. ((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)bis(2-methoxy-4,1-phenylene)-bis(2-(1-(4 -chlorobenzoyl)-5-methoxy-2-methyl-1H-indol-3-yl)acetate) (B7). Yellow powder, 60% yield, mp 228 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 7.63 (dd, J = 23.2, 11.8 Hz, 6H), 7.47 (d, J = 8.2 Hz, 4H), 7.18 – 6.97 (m, 8H), 6.90 (d, J = 9.0 Hz, 2H), 6.69 (d, J = 8.6 Hz, 2H), 6.54 (d, J = 15.3 Hz, 2H), 5.84 (s, 1H), 3.94 (s, 4H), 3.85 (s, 6H), 3.78 (s, 6H), 2.46 (s, 6H).13C NMR (101 MHz, CDCl3): δ (ppm) 183.05, 168.72, 168.32, 156.07, 151.33, 141.39, 139.90, 139.32, 136.26, 134.03, 133.88, 131.19, 130.88, 130.62, 129.14, 124.30, 123.10, 121.01, 114.92, 112.07, 111.57, 111.47, 101.80, 101.60, 55.80, 55.74, 30.02, 13.41. LC-MS m/z: 1047.3 [M + H] +. (1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)-bis(2-methoxy-4,1-phenylene)-bis(2-(3-be nzoylphenyl)propanoate) (B8). Yellow powder, 78% yield, mp 96 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 7.89 (s, 1H), 7.81 (d, J = 7.5 Hz, 1H), 7.72 (d, J = 7.3 Hz, 1H), 7.66 (d, J = 7.5 Hz, 1H), 7.62 – 7.53 (m, 1H), 7.48 (dd, J = 13.3, 6.8 Hz, 1H), 7.14 – 7.03 (m, 1H), 6.96 (s, 1H), 6.54 (d, J = 15.7 Hz, 1H), 5.84 (s, 1H), 4.10 (d, J = 7.1 Hz, 1H), 3.74 (s, 1H), 1.68 (d, J = 7.0 Hz, 1H).13C NMR (101 MHz, CDCl3) : δ (ppm) 196.48, 183.07, 171.91, 151.41, 141.38, 140.38, 139.94, 137.98, 137.49, 133.99, 132.55, 131.77, 130.09 129.41, 129.19, 128.55, 128.34, 124.27, 123.03, 121.03, 111.44, 55.80, 45.30, 41.99, 27.02, 25.00, 18.67. LC-MS m/z: 841.3 [M + H] +. ((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)bis(2-methoxy-4,1-phenylene)-bis(2-((2,3dimethylphenyl)amino)benzoate) (B9). Red powder, 86% yield, mp 204 ºC; 1H NMR (400 MHz, CDCl3) : δ (ppm) 9.08 (s, 2H), 8.19 (d, J = 8.2 Hz, 2H), 7.65 (d, J = 15.8 Hz, 2H), 7.31 (t, J = 8.1 Hz, 2H), 7.22 – 7.13 (m, 7H), 7.09 (s, 2H), 7.02 (s, 2H), 6.83 – 6.67 (m, 4H), 6.59 (d, J = 15.9 Hz, 2H), 5.88 (s, 1H), 3.89 (s, 6H), 2.31 (s, 6H), 2.15 (s, 6H).13C NMR (101 MHz, CDCl3): δ (ppm) 183.19, 166.75, 151.89, 150.22, 141.50,

140.06, 138.38, 135.07, 134.05, 132.50, 132.25, 127.05, 126.02, 124.33, 123.80, 123.16, 121.22, 116.36, 113.86, 111.71, 109.56, 101.94, 56.06, 26.98, 20.67, 14.06. LC-MS m/z: 815.3 [M + H] +. (S)-2-methoxy-4-((1E,6E)-7-(3-methoxy-4-(((R)-2-(6-methoxynaphthalen-2-yl)propan oyl)oxy)phenyl-3,5-dioxohepta-1,6-dien-1-yl)phenyl-2-(6-methoxynaphthalen-2-yl)-pr opanoate (B10). Orange powder, 74% yield, mp 182 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 7.75 (dd, J = 17.4, 9.3 Hz, 6H), 7.61 – 7.46 (m, 4H), 7.18 – 7.11 (m, 4H), 7.06 (tt, J = 12.0, 6.0 Hz, 4H), 6.93 (t, J = 9.7 Hz, 2H), 6.52 (d, J = 15.8 Hz, 2H), 5.82 (s, 1H), 4.21 – 4.06 (m, 2H), 3.92 (s, 6H), 3.70 (d, J = 5.7 Hz, 6H), 1.70 (d, J = 7.1 Hz, 6H). 13C NMR (101 MHz, CDCl3): δ (ppm) 183.09, 172.55, 157.72 , 151.51, 141.59, 139.98, 135.15, 133.86, 133.81, 129.32, 128.97, 127.13, 126.40, 126.28, 124.18, 123.13, 121.02, 119.03, 111.49, 105.64, 101.72, 55.81, 55.33, 45.36, 18.71. LC-MS m/z: 793.3 [M + H] +. ((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)-bis(2-methoxy-4,1-phenylene)-bis(3-(4,5diphenyloxazol-2-yl)propanoate) (B11). Yellow powder, 71% yield, mp 200 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 7.62 (s, 10H), 7.40 – 7.26 (m, 12H), 7.17 – 7.00 (m, 8H), 6.57 (s, 1H), 6.52 (s, 1H), 3.82 (s, 6H), 3.30 (d, J = 7.6 Hz, 4H), 3.22 (d, J = 7.4 Hz, 4H).13C NMR (101 MHz, CDCl3): δ (ppm) 183.09, 169.96, 161.44, 151.40, 145.56, 141.26, 139.95, 135.19, 134.04, 132.44, 128.96, 128.67, 128.58, 128.52, 128.11, 127.90, 126.53, 124.31, 123.27, 121.04, 111.50, 101.79, 55.92, 30.94, 23.58. LC-MS m/z: 919.3 [M + H] +. ((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)-bis(2-methoxy-4,1-phenylene)-bis(2-hydr oxybenzoate) (B12). Yellow powder, 55% yield, mp 230 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 10.39 (d, J = 8.0 Hz, 2H), 8.10 (dd, J = 8.0, 1.5 Hz, 2H), 7.73 – 7.61 (m, 2H), 7.54 (td, J = 8.6, 4.3 Hz, 2H), 7.23 – 7.15 (m, 6H), 7.05 (d, J = 8.1 Hz, 2H), 6.99 (t, J = 7.6 Hz, 2H), 6.61 (d, J = 15.8 Hz, 2H), 5.89 (s, 1H), 3.88 (d, J = 5.5 Hz, 6H).13C NMR (101 MHz, CDCl3): δ (ppm) 183.07, 168.20, 162.11, 151.50, 140.60, 139.84, 136.54, 134.48, 130.63, 124.57, 123.33, 121.09, 119.54, 117.79, 111.62, 101.96, 56.02, 29.71. LC-MS m/z: 609.2 [M + H] +. ((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)bis(2-methoxy-4,1-phenylene)-bis(2-((2-h

ydroxybenzoyl)oxy)benzoate) (B13). Yellow powder, 63% yield, mp 135 ºC; 1H NMR (400 MHz, CDCl3): δ (ppm) 10.35 (s, 2H), 8.30 (dd, J = 7.8, 1.5 Hz, 2H), 8.10 (dd, J = 8.0, 1.6 Hz, 2H), 7.71 (dd, J = 7.9, 1.4 Hz, 2H), 7.60 (s, 1H), 7.56 (s, 1H), 7.48 (t, J = 7.9 Hz, 4H), 7.33 (d, J = 8.1 Hz, 2H), 7.12 (dd, J = 8.3, 1.3 Hz, 2H), 7.08 – 7.04 (m, 3H), 6.99 (d, J = 8.4 Hz, 2H), 6.90 (t, J = 7.6 Hz, 2H), 6.52 (d, J = 15.8 Hz, 2H), 5.82 (s, 1H), 3.79 (s, 6H).13C NMR (101 MHz, CDCl3): δ (ppm) 183.06, 168.84, 162.10, 162.01, 151.39, 150.34, 140.99, 139.89, 136.44, 134.65, 134.15, 132.59, 130.75, 126.72, 124.35, 124.06, 123.20, 122.71, 121.02, 119.51, 117.64, 111.92, 111.47, 101.81, 55.81. LC-MS m/z: 849.2 [M + H] +. ((1E,6E)-3,5-dioxohepta-1,6-diene-1,7-diyl)bis(2-methoxy-4,1-phenylene)-bis(2-((3-c hloro-2-methylphenyl)amino)benzoate) (B14). Yellow powder, 70% yield, mp 171 ºC; 1

H NMR (400 MHz, DMSO-d6): δ (ppm) 9.07 (s, 2H), 8.11 (d, J = 7.9 Hz, 2H), 7.68

(d, J = 15.8 Hz, 2H), 7.57 (s, 2H), 7.42 (d, J = 24.0 Hz, 4H), 7.35 – 7.20 (m, 8H), 7.03 (d, J = 15.7 Hz, 2H), 6.90 – 6.76 (m, 4H), 3.86 (s, 6H), 2.22 (s, 6H), 1.38 (s, 2H).13C NMR (101 MHz, DMSO-d6): δ (ppm) 183.20, 165.76, 151.36, 148.18, 140.71, 139.99, 135.53, 134.44, 133.91, 131.93, 130.58, 127.63, 125.61, 124.71, 123.73, 123.15, 121.50, 117.61, 113.92, 112.12, 109.98, 101.74, 56.09, 26.31, 14.67. LC-MS m/z: 855.2 [M + H] +.