Phanerosides A–X, Phenylpropanoid Esters of Sucrose from the Rattans of Phanera championii Benth

Abstract

Twenty-four new phenylpropanoid esters of sucrose, phanerosides A–X (1–24), were isolated from an EtOH extract of the rattans of Phanera championii Benth. (Fabaceae). Their structures were elucidated on the basis of comprehensive spectroscopic data analysis. A wide range of structural analogues were presented due to the different numbers and positions of acetyl substituents and the structures of phenylpropanoid moieties. Phenylpropanoid esters of sucrose were isolated from the Fabaceae family for the first time. Biologically, the inhibitory effects of compounds 6 and 21 on NO production in LPS-induced BV-2 microglial cells were better than that of the positive control, with IC₅₀ values of 6.7 and 5.2 μM, respectively. The antioxidant activity assay showed that compounds 5, 15, 17, and 24 displayed moderate DPPH radical scavenging activity, with IC₅₀ values ranging from 34.9 to 43.9 μM.

1. Introduction

Phenylpropanoid esters of sucrose are characterized by the esterification of different hydroxy groups of sucrose with phenylpropanoid moieties [1,2]. These compounds have been isolated mainly from the families Arecaceae, Brassicaceae, Liliaceae, Polygonaceae, Rosaceae, Rutaceae, Smilacaceae, and Sparganiaceae [1,3]. The structural diversity of phenylpropanoid esters of sucrose generates extensive pharmacological antitumor [4,5,6], anti-inflammatory [7,8], antioxidant [9], an anti-HIV [10] activities, which are extremely thrilling to those engaged in medicinal chemistry [11,12].

Fabaceae is the third largest plant family, comprising approximately 800 genera and 20,000 species worldwide [13,14]. As the largest of the several genera in the Fabaceae family, Phanera results from the reorganization of Bauhinia sensu lato. The genus Phanera encompasses about 90–100 species, mainly distributed in tropical Asia and Australasia [15]. As one of them, Phanera championii Benth. is widely distributed in Guangxi Province and was first recorded in ‘Nanning Drug Chi’ [16]. It is a well-known folk medicine with a rich history of being used as medication to treat rheumatoid arthritis and epigastric pain [17,18]. Phytochemical investigations of this plant have primarily resulted in the isolation of flavonoids, nitrile glucoside, dibenzofurans, and diterpenoid [16,19,20,21]. In our ongoing research in pursuit of novel and biologically active metabolites from ethnic medicines in Guangxi, 24 new phenylpropanoid esters of sucrose (1–24) were isolated from the rattans of P. championii. Herein, the isolation, structural elucidation, anti-inflammatory, and antioxidant activities in vitro of all the isolated compounds are described.

2. Results and Discussion

Structural Elucidation

Compound 1 (Figure 1) was isolated as a white amorphous powder with a molecular formula of C₃₀H₃₄O₁₅, as determined using HRESIMS (m/z 657.1798 [M + Na]⁺, calcd for 657.1790) and ¹³C NMR data. The ¹H NMR spectrum (

Table 1. ¹H NMR data of compounds 1–5 in δ ppm (J coupling in Hertz).

Position	1 a	2 b	3 a	4 a	5 a
1	3.51, d (12.0) 3.31, d (12.0)	3.51, d (12.0) 3.31, overlapped	3.52, d (12.0) 3.32, d (12.0)	3.51, d (12.0) 3.30, d (12.0)	3.51, d (11.6) 3.31, overlapped
3	4.22, d (8.8)	4.22, d (9.0)	4.22, d (8.8)	4.22, d (8.4)	4.22, d (8.8)
4	4.04, t (8.8)	4.04, t (9.0)	4.04, t (8.8)	4.07, dd (9.2, 8.4)	4.07, t (8.8)
5	3.79, m	3.78, m	3.79, m	3.79, m	3.79, m
6	3.88, dd (12.0, 7.2) 3.77, m	3.88, dd (12.0, 7.2) 3.77, dd (12.0, 2.4)	3.88, m 3.77, m	3.88, m 3.77, m	3.89, overlapped 3.78, m
1′	5.60, d (3.6)	5.60, d (3.6)	5.60, d (4.0)	5.60, d (3.6)	5.61, d (3.6)
2′	4.73, dd (10.0, 3.6)	4.73, dd (9.6, 3.6)	4.73, dd (10.0, 4.0)	4.73, dd (9.6, 3.6)	4.73, dd (9.6, 3.6)
3′	4.01, dd (10.0, 9.2)	4.01, t (9.6)	4.01, t (10.0)	4.01, t (9.6)	4.01, t (9.6)
4′	3.45, t (9.2)	3.46, t (9.6)	3.46, t (10.0)	3.45, t (9.6)	3.45, t (9.6)
5′	4.19, ddd (9.2, 6.0, 2.0)	4.20, m	4.18, dd (10.0, 6.0, 2.0)	4.20, m	4.20, m
6′	4.54, dd (12.0, 2.0) 4.31, dd (12.0, 6.0)	4.56, br d (12.0) 4.34, dd (12.0, 6.6)	4.54, dd (12.0, 2.0) 4.31, dd (12.0, 6.0)	4.54, d (12.0) 4.31, dd (12.0, 6.4)	4.54, br d (12.0) 4.31, dd (12.0, 6.4)
2″	7.49, d (8.8)	7.49, d (8.4)	7.21, d (2.0)	7.48, d (8.4)	7.21, br s
3″	6.82, d (8.8)	6.81, d (8.4)		6.81, d (8.4)
5″	6.82, d (8.8)	6.81, d (8.4)	6.82, d (8.0)	6.81, d (8.4)	6.82, d (8.4)
6″	7.49, d (8.8)	7.49, d (8.4)	7.11, dd (8.0, 2.0)	7.48, d (8.4)	7.11, br d (8.4)
7″	7.72, d (16.0)	7.72, d (16.2)	7.72, d (16.0)	7.72, d (16.0)	7.72, d (16.0)
8″	6.39, d (16.0)	6.39, d (16.2)	6.42, d (16.0)	6.39, d (16.0)	6.42, d (16.0)
2‴	7.51, d (8.8)	7.65, m	7.50, d (8.4)	7.25, br s	7.25, br s
3‴	6.81, d (8.8)	7.41, overlapped	6.81, d (8.4)
4‴		7.41, overlapped
5‴	6.81, d (8.8)	7.41, overlapped	6.81, d (8.4)	6.81, d (8.4)	6.81, d (8.4)
6‴	7.51, d (8.8)	7.65, m	7.50, d (8.4)	7.10, br d (8.4)	7.11, br d (8.4)
7‴	7.67, d (16.0)	7.75, d (16.2)	7.67, d (16.0)	7.65, d (16.0)	7.66, d (16.0)
8‴	6.43, d (16.0)	6.64, d (16.2)	6.43, d (16.0)	6.46, d (16.0)	6.46, d (16.0)
3″-OMe			3.90, s		3.90, s
3‴-OMe				3.89, s	3.90, s

^a In MeOH-d₄, ¹H NMR at 400 MHz. ^b In MeOH-d₄, ¹H NMR at 600 MHz. s = singlet; d = doublet; t = triplet; m = multiplet; br = broad.

) revealed two groups of AA′BB′ aromatic signals [δ_H 7.49 (2H, d, J = 8.8 Hz, H-2″/6″), 6.82 (2H, d, J = 8.8 Hz, H-3″/5″) and 7.51 (2H, d, J = 8.8 Hz, H-2‴/6‴), 6.81 (2H, d, J = 8.8 Hz, H-3‴/5‴)] and two sets of trans-olefinic protons [δ_H 7.72 (1H, d, J = 16.0 Hz, H-7″), 6.39 (1H, d, J = 16.0 Hz, H-8″) and 7.67 (1H, d, J = 16.0 Hz, H-7‴), 6.43 (1H, d, J = 16.0 Hz, H-8‴]. In addition, a characteristic doublet with a small coupling constant (J = 3.6 Hz) at δ_H 5.60 was also shown in the ¹H NMR spectrum, which together with 12 oxygen-bearing carbon signals (including two anomers at δ_C 105.7 and 90.6) in the ¹³C NMR data (

Table 2. ¹³C NMR data of compounds 1–5 in δ ppm.

Position	1 a	2 b	3 a	4 a	5 a
1	62.9	62.9	62.9	62.9	63.0
2	105.7	105.7	105.7	105.6	105.7
3	77.0	77.0	77.0	77.0	77.0
4	75.6	75.6	75.6	75.7	75.7
5	84.0	84.0	84.0	84.0	84.0
6	64.2	64.2	64.2	64.2	64.2
1′	90.6	90.6	90.6	90.5	90.5
2′	74.4	74.4	74.4	74.4	74.4
3′	71.9	71.9	71.9	72.0	72.0
4′	72.0	72.0	72.0	72.0	72.0
5′	71.9	71.9	71.9	71.9	71.9
6′	65.0	65.2	65.0	65.0	65.1
1″	127.1	127.0	127.6	127.0	127.7
2″	131.3	131.3	111.8	131.3	111.8
3″	116.9	116.9	149.4	116.9	149.4
4″	161.6	161.7	151.0	161.6	151.0
5″	116.9	116.9	116.5	116.9	116.5
6″	131.3	131.3	124.3	131.3	124.3
7″	147.5	147.5	147.8	147.5	147.8
8″	114.7	114.7	114.9	114.7	115.0
9″	168.8	168.8	168.8	168.8	168.8
1‴	127.0	135.8	127.1	127.7	127.6
2‴	131.3	129.4	131.3	111.6	111.6
3‴	116.8	130.0	116.8	149.4	149.4
4‴	161.4	131.6	161.4	150.7	150.8
5‴	116.8	130.0	116.8	116.4	116.4
6‴	131.3	129.4	131.3	124.3	124.3
7‴	146.9	146.7	146.9	147.1	147.1
8‴	114.9	118.7	115.0	115.2	115.2
9‴	169.2	168.5	169.2	169.1	169.1
3″-OMe			56.4		56.5
3‴-OMe				56.5	56.5

^a In MeOH-d₄, ¹³C NMR at 100 MHz. ^b In MeOH-d₄, ¹³C NMR at 150 MHz.

) supposed the presence of a disaccharide moiety. Furthermore, detailed analysis of the 2D NMR correlations (Figure 2) and chiral HPLC analysis of monosaccharides after acid hydrolysis of 1 revealed that the two sugars were β-D-fructose and α-D-glucose units connected via C-2→C-1′ to construct a D-sucrose moiety. The ¹³C NMR spectrum showed 30 carbon signals, apart from the 12 carbon signals occupied by the D-sucrose moiety; the other 18 ones were classified as 2 carbonyl carbons (δ_C 169.2, 168.8) and 16 olefinic or aromatic carbons with the assistance of the HSQC data. Two discrete spin systems, H-7″/H-8″ and H-7‴/H-8‴, in the ¹H-¹H COSY spectrum, and the correlations from H-7″ to C-1″/C-2″/C-6″/C-9″ and H-7‴ to C-1‴/C-2‴/C-6‴/C-9‴ in the HMBC spectrum (Figure 2), established two trans-p-coumaroyl moieties. Subsequently, the key HMBC correlations from H-2′ to C-9″ as well as from H₂-6′ to C-9‴ suggested that the two trans-p-coumaroyl moieties were located at C-2′ and C-6′. Thus, the structure of 1 was identified and named phaneroside A.

Compounds 2–24 were determined to be structurally related to 1 according to their extremely similar physicochemical properties and NMR data. The latter all showed the characteristics of a core D-sucrose unit, while the variation of substituents and their positions obtained different structures. The structural elucidation of 2–24 was as follows.

The molecular formula of compound 2 was assigned as C₃₀H₃₄O₁₄ based on its HRESIMS (m/z 641.1847 [M + Na]⁺, calcd for 641.1841) and ¹³C NMR data, with 16 mass units less than 1. The analogous ¹H NMR data (Table 1) of 1 and 2 indicated that they were structural analogues, excepting of the presence of a monosubstituted aromatic moiety [δ_H 7.65 (1H, m, H-2‴/6‴), 7.41 (3H, overlapped, H-3‴/4‴/5‴)] and the absence of an AA′BB′ aromatic unit in 2. The long-range correlations from phenyl protons to olefinic carbons and correlations from olefinic protons to ester carbonyl carbons in the HMBC spectrum indicated that a trans-p-coumaroyl group and a trans-cinnamoyl group were presented (Figure 3). The HMBC correlations from H-2′ (δ_H 4.73) to C-9″ and H₂-6′ (δ_H 4.56, 4.34) to C-9‴ confirmed the structure of 2 as shown, and this compound was named phaneroside B.

Compounds 3 and 4 had the same molecular formula, C₃₁H₃₆O₁₆, as determined using the HRESIMS peaks at m/z 687.1895 and 687.1896 ([M + Na]⁺, calcd for 687.1896), respectively, indicating 30 mass units more than 1, ascribed to a methoxy group. The ¹H NMR data (Table 1) of 3 and 4 were similar to those of 1, except for the presence of an ABX aromatic moiety [δ_H 7.21 (1H, d, J = 2.0 Hz, H-2″), 7.11 (1H, dd, J = 8.0, 2.0 Hz, H-6″), 6.82 (1H, d, J = 8.0 Hz, H-5″) in 3 and 7.25 (1H, br s, H-2‴), 7.10 (1H, br d, J = 8.4 Hz, H-6‴), 6.81 (1H, d, J = 8.4 Hz, H-5‴) in 4] and a methoxy [δ_H 3.90 (3H, s) in 3 and 3.89 (3H, s) in 4] and the absence of an AA′BB′ aromatic unit. The HMBC correlations from H-5″/3″-OMe to C-3″ in 3 and from H-5‴/3‴-OMe to C-3‴ in 4, together with the ¹H-¹H COSY and HMBC correlations as mentioned previously, suggested that there were a trans-feruloyl group and a trans-p-coumaroyl group in both 3 and 4 (Figure 3). The complete structures of 3 and 4 were further established by the HMBC correlations from the protons of the sucrose unit to carbonyl carbons. Therefore, compounds 3 and 4 were determined to be phanerosides C and D, respectively.

Compound 5 gave the molecular formula of C₃₂H₃₈O₁₇, as determined by an HRESIMS ion at m/z 717.1990 ([M + Na]⁺, calcd for 717.2001). The NMR data (Table 1 and Table 2) indicated the presence of two trans-feruloyl moieties and a D-sucrose unit. The key HMBC cross-peaks from H-2′ (δ_H 4.73) to C-9″ and H₂-6′ (δ_H 4.54, 4.31) to C-9‴ suggested that the two trans-feruloyl moieties were linked to C-2′ and C-6′ of glucopyranose unit (Figure 3). Thus, the structure of compound 5 was identified and named phaneroside E.

Compounds 6–8 shared the same molecular formula of C₃₂H₃₆O₁₆, as obtained from their respective HRESIMS data. Their molecular masses were 42 mass units more than that of 1, which combined with the 1D NMR data of 6–8 (

Table 3. ¹H NMR data (400 MHz) of compounds 6–11 in MeOH-d₄.

Position	6	7	8	9	10	11
1	4.11, d (11.6) 4.06, d (11.6)	3.54, d (12.0) 3.35, d (12.0)	3.57, d (12.0) 3.40, d (12.0)	4.11, d (12.0) 4.06, d (12.0)	3.54, d (12.0) 3.34, d (12.0)	3.57, br d (12.0) 3.40, br d (12.0)
3	4.08, d (8.8)	5.43, d (8.4)	4.47, d (8.4)	4.08, d (6.8)	5.44, d (8.4)	4.47, d (8.4)
4	4.04, t (8.8)	4.35, t (8.4)	5.30, t (8.4)	4.07, t (6.8)	4.39, t (8.4)	5.28, dd (8.4, 7.6)
5	3.79, m	3.90, m	3.91, m	3.79, m	3.91, m	3.91, overlapped
6	3.88, dd (11.6, 7.2) 3.75, m	3.84, dd (12.0, 6.4) 3.78, dd (12.0, 3.2)	3.92, m 3.77, dd (11.2, 2.4)	3.89, overlapped 3.76, m	3.85, dd (11.6, 6.8) 3.78, dd (11.6, 2.8)	3.93, m 3.80, dd (11.2, 2.8)
1-OAc	2.01, s			2.01, s
3-OAc		2.18, s			2.18, s
4-OAc			2.05, s			2.04, s
1′	5.65, d (3.6)	5.63, d (3.6)	5.65, d (3.6)	5.66, d (3.6)	5.63, d (3.6)	5.65, d (3.6)
2′	4.79, dd (10.4, 3.6)	4.72, dd (10.0, 3.6)	4.76, dd (10.0, 3.6)	4.80, dd (10.0, 3.6)	4.73, dd (10.4, 3.6)	4.76, dd (10.0, 3.6)
3′	4.00, dd (10.4, 9.2)	3.88, t (10.0)	4.01, dd (10.0, 9.6)	4.00, dd (10.0, 8.8)	3.88, overlapped	4.01, t (10.0)
4′	3.46, dd (10.0, 9.2)	3.47, t (10.0)	3.43, t (9.6)	3.45, dd (10.0, 8.8)	3.45, dd (10.0, 8.8)	3.44, t (10.0)
5′	4.20, ddd (10.0, 6.0, 2.0)	4.16, ddd (10.0, 6.0, 2.0)	4.23, m	4.21, ddd (10.0, 6.8, 2.0)	4.19, ddd (10.0, 6.8, 2.0)	4.24, ddd (10.0, 6.4, 1.6)
6′	4.54, dd (12.0, 2.0) 4.31, dd (12.0, 6.0)	4.56, dd (12.0, 2.0) 4.32, dd (12.0, 6.0)	4.57, dd (12.0, 1.6) 4.31, dd (12.0, 6.4)	4.54, dd (12.0, 2.0) 4.30, dd (12.0, 6.8)	4.56, dd (12.0, 2.0) 4.31, dd (12.0, 6.8)	4.57, dd (12.0, 1.6) 4.32, dd (12.0, 6.4)
2″	7.49, d (8.8)	7.48, d (8.8)	7.49, d (8.4)	7.49, d (8.8)	7.48, d (8.8)	7.49, d (8.8)
3″	6.80, d (8.8)	6.82, d (8.8)	6.81, d (8.4)	6.81, d (8.8)	6.81, d (8.8)	6.81, d (8.8)
5″	6.80, d (8.8)	6.82, d (8.8)	6.81, d (8.4)	6.81, d (8.8)	6.81, d (8.8)	6.81, d (8.8)
6″	7.49, d (8.8)	7.48, d (8.8)	7.49, d (8.4)	7.49, d (8.8)	7.48, d (8.8)	7.49, d (8.8)
7″	7.71, d (16.0)	7.70, d (16.0)	7.70, d (16.0)	7.71, d (16.0)	7.69, d (16.0)	7.70, d (16.0)
8″	6.39, d (16.0)	6.37, d (16.0)	6.40, d (16.0)	6.41, d (16.0)	6.37, d (16.0)	6.41, d (16.0)
2‴	7.50, d (8.8)	7.50, d (8.8)	7.51, d (8.4)	7.26, d (2.0)	7.25, d (2.0)	7.25, d (2.0)
3‴	6.80, d (8.8)	6.80, d (8.8)	6.80, d (8.4)
5‴	6.80, d (8.8)	6.80, d (8.8)	6.80, d (8.4)	6.81, d (8.0)	6.81, d (8.0)	6.81, d (8.4)
6‴	7.50, d (8.8)	7.50, d (8.8)	7.51, d (8.4)	7.11, dd (8.0, 2.0)	7.09, dd (8.0, 2.0)	7.11, dd (8.4, 2.0)
7‴	7.66, d (16.0)	7.66, d (16.0)	7.65, d (16.0)	7.65, d (15.6)	7.65, d (16.0)	7.65, d (16.0)
8‴	6.43, d (16.0)	6.43, d (16.0)	6.49, d (16.0)	6.47, d (15.6)	6.47, d (16.0)	6.52, d (16.0)
3″-OMe						3.90, s
3‴-OMe				3.90, s	3.89, s

s = singlet; d = doublet; t = triplet; m = multiplet; br = broad.

and Table 5) suggested that an acetyl group [δ_H 2.01 (3H, s), δ_C 172.0, 20.6 in 6; δ_H 2.18 (3H, s), δ_C 172.2, 20.8 in 7; δ_H 2.05 (3H, s), δ_C 172.4, 20.8 in 8] existed in each compound. The key HMBC correlations from H₂-1 (δ_H 4.11, 4.06) to carbonyl (δ_C 172.0) in 6, from H-3 (δ_H 5.43) to carbonyl (δ_C 172.2) in 7, and from H-4 (δ_H 5.30) to carbonyl (δ_C 172.4) in 8 indicated that the acetyl group was attached to C-1, C-3, and C-4 of the fructofuranose unit in compounds 6, 7, and 8, respectively (Figure 4). In addition, the chemical shifts of the protons linked to the acetyl group in 6–8 were obviously shifted downfield compared to those of 1, which also supported the above description. Thus, the structures of 6, 7, and 8 were established and named phanerosides F, G, and H, respectively.

Compounds 9–11 were assigned the same molecular formula, C₃₃H₃₈O₁₇, according to their HRESIMS and ¹³C NMR data. Compounds 9–11 were determined to be acetylated derivatives of 4, as their molecular masses were 42 mass units more than that of 4. Their ¹H and ¹³C NMR data (Table 3 and Table 5) showed the characteristics of the acetyl group [δ_H 2.01 (3H, s), δ_C 172.0, 20.6 in 9; δ_H 2.18 (3H, s), δ_C 172.2, 20.8 in 10; δ_H 2.04 (3H, s), δ_C 172.4, 20.8 in 11]. Furthermore, the key HMBC correlations from H₂-1 (δ_H 4.11, 4.06) to carbonyl (δ_C 172.0) in 9, from H-3 (δ_H 5.44) to carbonyl (δ_C 172.2) in 10, and from H-4 (δ_H 5.28) to carbonyl (δ_C 172.4) in 11 located the acetyl group at C-1, C-3, and C-4 for compounds 9, 10, and 11, respectively. Thus, the structures of compounds 9–11 (phanerosides I–K) were defined as shown.

Compounds 12–14 possessed identical molecular formula, C₃₃H₃₈O₁₇, as determined by their respective HRESIMS ion peaks at m/z 729.2018, 729.2010, and 729.1990 ([M + Na]⁺, calcd for 729.2001), which were 42 mass units more than that of 3. Detailed analysis of their 1D NMR data (

Table 4. ¹H NMR data of compounds 12–19.

Position	12 a	13 a	14 a	15 a	16 b	17 a	18 b	19 b
1	4.14, d (12.4) 4.03, d (12.4)	3.55, d (11.6) 3.36, d (11.6)	3.59, d (12.0) 3.42, d (12.0)	4.14, d (11.6) 4.02, d (11.6)	3.54, d (12.0) 3.34, d (12.0)	3.59, d (12.0) 3.42, d (12.0)	3.55, d (12.0) 3.39, d (12.0)	3.58, d (12.0) 3.42, d (12.0)
3	4.09, d (8.4)	5.43, d (8.4)	4.47, d (8.0)	4.09, d (8.4)	5.44, d (8.4)	4.48, d (8.8)	4.47, d (8.4)	4.46, d (8.4)
4	4.04, t (8.4)	4.35, t (8.4)	5.31, t (8.0)	4.08, t (8.4)	4.39, t (8.4)	5.29, dd (8.8, 7.6)	5.26, t (8.4)	5.29, t (8.4)
5	3.79, m	3.90, overlapped	3.91, overlapped	3.80, m	3.91, overlapped	3.91, overlapped	3.90, overlapped	3.90, overlapped
6	3.88, m 3.75, dd (11.6, 2.8)	3.84, dd (12.0, 6.4) 3.78, dd (12.0, 3.2)	3.92, overlapped 3.78, m	3.89, overlapped 3.76, dd (11.6, 2.8)	3.85, dd (12.0, 6.6) 3.79, dd (12.0, 2.4)	3.92, overlapped 3.81, dd (11.2, 2.4)	3.92, overlapped 3.78, dd (10.8, 2.4)	3.90, overlapped 3.78, m
1-OAc	2.02, s			2.02, s
3-OAc		2.18, s			2.18, s
4-OAc			2.04, s			2.03, s	2.08, s	2.05, s
1′	5.64, d (3.6)	5.63, d (4.0)	5.67, d (3.6)	5.65, d (4.0)	5.64, d (3.6)	5.66, d (4.0)	5.62, d (3.6)	5.66, d (3.6)
2′	4.82, dd (10.4, 3.6)	4.72, dd (10.4, 4.0)	4.76, dd (10.0, 3.6)	4.82, dd (10.0, 4.0)	4.72, dd (10.2, 3.6)	4.76, dd (10.0, 4.0)	4.75, dd (10.2, 3.6)	4.73, dd (10.2, 3.6)
3′	4.00, dd (10.4, 9.2)	3.88, overlapped	4.01, t (10.0)	4.00, dd (10.0, 8.8)	3.87, t (10.2)	4.01, dd (10.0, 8.8)	3.98, dd (10.2, 9.6)	3.99, dd (10.2, 9.0)
4′	3.46, dd (10.0, 9.2)	3.47, t (10.0)	3.43, t (10.0)	3.46, dd (10.0, 8.8)	3.45, t (10.2)	3.44, dd (10.0, 8.8)	3.43, t (9.6)	3.43, dd (10.2, 9.0)
5′	4.20, ddd (10.0, 6.4, 2.0)	4.16, ddd (10.0, 6.0, 2.0)	4.23, dd (10.0, 6.4)	4.22, ddd (10.0, 6.4, 2.0)	4.19, m	4.23, ddd (10.0, 6.4, 1.6)	4.23, ddd (9.6, 6.6, 1.8)	4.19, ddd (10.2, 5.4, 1.8)
6′	4.54, dd (12.0, 2.0) 4.31, dd (12.0, 6.4)	4.56, dd (12.0, 2.0) 4.32, dd (12.0, 6.0)	4.57, br d (12.0) 4.31, dd (12.0, 6.4)	4.54, dd (12.0, 2.0) 4.30, dd (12.0, 6.4)	4.56, br d (12.0) 4.31, dd (12.0, 6.6)	4.57, dd (12.0, 1.6) 4.32, dd (12.0, 6.4)	4.56, dd (12.0, 1.8) 4.31, dd (12.0, 6.6)	4.53, dd (12.0, 1.8) 4.30, dd (12.0, 5.4)
2″	7.26, d (2.0)	7.21, d (2.0)	7.23, d (2.0)	7.26, d (2.4)	7.21, br s	7.23, d (2.0)	7.93, d (2.4)	7.23, d (1.8)
5″	6.81, d (8.4)	6.82, d (8.0)	6.82, d (8.0)	6.81, d (8.0)	6.82, d (7.8)	6.81, d (8.0)	6.77, d (8.4)	6.81, d (8.4)
6″	7.09, dd (8.4, 2.0)	7.10, dd (8.0, 2.0)	7.09, dd (8.0, 2.0)	7.09, dd (8.0, 2.4)	7.10, overlapped	7.09, dd (8.0, 2.0)	7.19, dd (8.4, 2.4)	7.09, dd (8.4, 1.8)
7″	7.70, d (15.6)	7.69, d (15.6)	7.69, d (16.0)	7.70, d (16.0)	7.69, d (16.2)	7.69, d (16.0)	6.92, d (12.6)	7.68, d (15.6)
8″	6.45, d (15.6)	6.39, d (15.6)	6.44, d (16.0)	6.45, d (16.0)	6.40, d (16.2)	6.44, d (16.0)	5.88, d (12.6)	6.44, d (15.6)
2‴	7.51, d (8.4)	7.50, d (8.8)	7.51, d (8.0)	7.26, d (2.4)	7.26, br s	7.24, d (2.0)	7.25, d (2.4)	7.87, d (1.8)
3‴	6.81, d (8.4)	6.80, d (8.8)	6.80, d (8.0)
5‴	6.81, d (8.4)	6.80, d (8.8)	6.80, d (8.0)	6.81, d (8.0)	6.80, d (7.8)	6.81, d (8.0)	6.81, d (8.4)	6.77, d (8.4)
6‴	7.51, d (8.4)	7.50, d (8.8)	7.51, d (8.0)	7.11, dd (8.0, 2.4)	7.10, overlapped	7.11, dd (8.0, 2.0)	7.11, dd (8.4, 2.4)	7.15, dd (8.4, 1.8)
7‴	7.66, d (15.6)	7.66, d (15.6)	7.65, d (16.0)	7.65, d (16.0)	7.65, d (16.2)	7.65, d (16.0)	7.65, d (16.2)	6.88, d (13.2)
8‴	6.43, d (15.6)	6.43, d (15.6)	6.49, d (16.0)	6.47, d (16.0)	6.47, d (16.2)	6.52, d (16.0)	6.53, d (16.2)	5.90, d (13.2)
3″-OMe	3.92, s	3.90, s	3.92, s	3.92, s	3.90, s	3.92, s	3.88, s	3.92, s
3‴-OMe				3.90, s	3.89, s	3.90, s	3.90, s	3.89, s

^a In MeOH-d₄, ¹H NMR at 400 MHz. ^b In MeOH-d₄, ¹H NMR at 600 MHz. s = singlet; d = doublet; t = triplet; m = multiplet; br = broad.

and

Table 5. ¹³C NMR data of compounds 12–19.

Position	6 a	7 a	8 a	9 a	10 a	11 a	12 a	13 a	14 a	15 a	16 b	17 a	18 b	19 b
1	64.8	64.4	62.5	64.8	64.5	62.5	64.8	64.5	62.5	64.8	64.5	62.5	62.5	62.3
2	103.9	105.0	106.0	103.8	104.9	106.0	103.9	105.0	106.1	103.8	104.9	106.1	105.9	106.2
3	78.5	78.6	75.2	78.5	78.6	75.2	78.4	78.7	75.2	78.4	78.6	75.2	75.2	75.2
4	75.3	73.8	78.0	75.4	73.8	78.1	75.4	73.8	78.0	75.4	73.8	78.2	78.2	77.9
5	84.1	84.2	82.2	84.1	84.2	82.3	84.2	84.2	82.3	84.2	84.2	82.3	82.3	82.3
6	64.1	63.9	64.8	64.2	64.0	64.8	64.2	63.9	64.9	64.3	64.0	64.9	64.8	64.8
1-OAc	172.0 20.6			172.0 20.6			172.0 20.6			172.0 20.6
3-OAc		172.2 20.8			172.2 20.8			172.2 20.8			172.2 20.8
4-OAc			172.4 20.8			172.4 20.8			172.3 20.8			172.4 20.8	172.4 20.9	172.3 20.8
1′	90.9	90.5	91.1	90.9	90.4	91.1	91.0	90.6	91.2	90.9	90.4	91.2	91.0	91.3
2′	74.1	74.3	74.2	74.1	74.3	74.3	74.1	74.3	74.2	74.1	74.3	74.2	73.9	74.2
3′	72.1	72.3	72.0	72.2	72.3	72.0	72.2	72.3	72.0	72.2	72.3	72.1	71.9	72.0
4′	72.0	71.8	72.0	72.0	71.9	72.1	72.1	72.0	72.1	72.0	71.9	72.0	72.1	71.9
5′	72.0	72.0	72.0	72.0	72.0	72.0	72.0	71.8	72.0	72.0	72.0	72.0	72.0	71.8
6′	65.0	64.9	65.1	65.1	65.1	65.1	65.0	64.9	65.1	65.1	65.1	65.1	65.1	64.5
1″	127.1	127.1	127.0	127.2	127.0	127.1	127.7	127.5	127.7	127.8	127.5	127.6	127.9	127.5
2″	131.3	131.3	131.3	131.4	131.3	131.4	111.7	111.8	111.7	111.6	111.8	111.8	115.3	111.7
3″	116.9	116.8	116.9	116.8	116.9	116.8	149.4	149.5	149.4	149.4	149.5	149.4	148.3	149.5
4″	161.7	161.6	161.6	161.4	161.6	161.4	150.8	151.5	150.8	150.8	151.1	150.9	149.9	150.9
5″	116.9	116.8	116.9	116.8	116.9	116.8	116.4	116.6	116.4	116.4	116.5	116.4	115.6	116.5
6″	131.3	131.3	131.3	131.4	131.3	131.4	124.5	124.3	124.4	124.5	124.3	124.4	127.4	124.5
7″	147.5	147.5	147.4	147.5	147.5	147.3	147.7	147.8	147.5	147.7	147.8	147.6	147.0	147.5
8″	114.8	114.6	114.9	114.9	114.6	114.9	115.2	114.9	115.2	115.2	114.9	115.2	115.8	115.1
9″	168.8	168.7	168.8	168.8	168.7	168.8	168.7	168.7	168.8	168.7	168.7	168.8	167.5	168.8
1‴	127.1	127.0	127.2	127.7	127.6	127.8	127.2	127.1	127.3	127.7	127.6	127.8	127.7	128.0
2‴	131.4	131.3	131.4	111.6	111.5	111.9	131.3	131.3	131.3	111.7	111.5	111.7	111.8	115.1
3‴	116.9	116.9	116.8	149.4	149.4	149.4	116.8	116.9	116.8	149.4	149.5	149.4	149.4	148.4
4‴	161.6	161.4	161.4	150.7	150.8	150.6	161.4	161.5	161.3	150.7	150.9	150.7	150.8	149.7
5‴	116.9	116.9	116.8	116.4	116.4	116.4	116.8	116.9	116.8	116.4	116.4	116.5	116.4	115.7
6‴	131.4	131.3	131.4	124.4	124.4	124.4	131.3	131.3	131.3	124.4	124.5	124.4	124.4	127.0
7‴	146.9	146.9	146.8	147.1	147.1	147.1	146.9	146.9	146.8	147.1	147.2	147.1	147.1	146.0
8‴	114.9	114.9	115.2	115.3	115.2	115.4	115.0	114.9	115.3	115.3	115.2	115.4	115.4	116.1
9‴	169.2	169.2	169.3	169.1	169.1	169.3	169.2	169.2	169.3	169.1	169.1	169.3	169.3	168.1
3″-OMe						56.5	56.5	56.4	56.5	56.5	56.5	56.5	56.4	56.4
3‴-OMe				56.5	56.5					56.5	56.4	56.5	56.5	56.4

^a In MeOH-d₄, ¹³C NMR at 100 MHz. ^b In MeOH-d₄, ¹³C NMR at 150 MHz.

) indicated that compounds 12–14 were acetylated derivatives of 3, with a difference in the position of the acetyl group. In their HMBC spectra, the correlations from H₂-1 (δ_H 4.14, 4.03) to carbonyl (δ_C 172.0) in 12, from H-3 (δ_H 5.43) to carbonyl (δ_C 172.2) in 13, and from H-4 (δ_H 5.31) to carbonyl (δ_C 172.3) in 14 suggested that the acetyl group was linked to C-1, C-3, and C-4 in 12, 13, and 14, respectively. Therefore, the structures of compounds 12–14 (phanerosides L–N) were established as shown.

Compounds 15–17 were determined to have the same molecular formula, C₃₄H₄₀O₁₈, on the basis of their HRESIMS and ¹³C NMR data, displaying 42 mass units more than that of 5. Compounds 15–17 were suggested to be acetylated derivatives of 5 after an analysis of their 1D NMR data (Table 4 and Table 5). The acetyl group was, respectively, located at C-1, C-3, and C-4 of the fructofuranose unit in 15, 16, and 17, which was confirmed using the key HMBC correlations from the protons of the sugar unit to corresponding carbonyl carbons. Accordingly, the structure of compounds 15–17 (phanerosides O–Q) were determined as shown.

Compounds 18 and 19 had the same molecular formula, C₃₄H₄₀O₁₈, as 17 according to their HRESIMS and ¹³C NMR data. The 1D NMR data (Table 4 and Table 5) closely resembled those of 17, with the exception that one of the two trans-feruloyl groups was replaced by a cis-feruloyl group according to the smaller coupling constants of ³J_7″,8″ (12.6 Hz) in 18 and ³J_7‴,8‴ (13.2 Hz) in 19. The cis-feruloyl group was linked to C-2′ and C-6′ of the glucopyranose unit in 18 and 19, respectively, which was proved by the key HMBC cross-peaks from H-2′/H-7″ to C-9″ in 18 and from H-6′/H-7‴ to C-9‴ in 19 (Figure 5). Thus, the structures of compounds 18 and 19 were identified and named phanerosides R and S, respectively.

Compounds 20 and 21 were suggested to have the same molecular formula, C₃₄H₃₈O₁₇, owing to their coincident positive HRESIMS ion at m/z 741.2001 [M + Na]⁺ (calcd for 741.2001), indicating 42 mass units more than that of 7. Comparison of their 1D NMR data (

Table 6. ¹H NMR data of compounds 20–24.

Position	20 a	21 a	22 b	23 a	24 a
1	4.17, d (12.0) 4.00, d (12.0)	3.57, d (12.0) 3.45, d (12.0)	3.58, d (12.0) 3.47, d (12.0)	4.16, d (11.6) 3.99, d (11.6)	3.59, d (12.0) 3.48, d (12.0)
3	5.29, d (8.0)	5.65, d (7.6)	5.65, d (7.8)	5.30, d (8.4)	5.65, d (7.6)
4	4.34, t (8.0)	5.48, t (7.6)	5.50, t (7.8)	4.38, t (8.4)	5.49, t (7.6)
5	3.89, m	4.07, m	4.08, m	3.90, overlapped	4.08, td (7.6, 4.4)
6	3.84, dd (11.6, 6.8) 3.77, dd, (11.6, 2.8)	3.87, dd (12.0, 6.8) 3.77, dd (12.0, 4.4)	3.87, dd (12.0, 7.2) 3.78, dd (12.0, 4.2)	3.85, dd (12.0, 6.8) 3.77, dd (12.0, 2.8)	3.87, overlapped 3.80, dd (12.0, 4.4)
1-OAc	2.02, s			2.02, s
3-OAc	2.17, s	2.11, s	1.98, s	2.18, s	2.11, s
4-OAc		1.99, s	2.10, s		1.97, s
1′	5.66, d (3.6)	5.62, d (3.6)	5.62, d (3.6)	5.66, d (3.6)	5.62, d (3.6)
2′	4.78, dd (10.0, 3.6)	4.76, dd (10.0, 3.6)	4.77, dd (10.2, 3.6)	4.78, dd (10.0, 3.6)	4.77, dd (10.0, 3.6)
3′	3.88, dd (10.0, 9.2)	3.91, dd (10.0, 9.2)	3.91, m	3.88, m	3.91, overlapped
4′	3.47, t (9.2)	3.47, t (9.2)	3.46, t (10.2)	3.46, dd (10.0, 9.2)	3.47, m
5′	4.16, m	4.19, ddd (10.0, 6.0, 2.0)	4.18, ddd (10.2, 6.0, 1.8)	4.18, m	4.19, ddd (10.0, 6.4, 2.0)
6′	4.57, dd (12.0, 2.0) 4.32, dd (12.0, 6.0)	4.60, dd (12.0, 2.0) 4.33, dd (12.0, 6.0)	4.59, dd (12.0, 1.8) 4.32, dd (12.0, 6.0)	4.57, dd (12.0, 2.0) 4.30, dd (12.0, 6.8)	4.60, dd (12.0, 2.0) 4.33, dd (12.0, 6.4)
2″	7.49, d (8.8)	7.49, d (8.8)	7.25, br s	7.49, d (8.8)	7.24, d (2.0)
3″	6.81, d (8.8)	6.81, d (8.8)		6.81, d (8.8)
5″	6.81, d (8.8)	6.81, d (8.8)	6.81, d (8.4)	6.81, d (8.8)	6.82, d (8.0)
6″	7.49, d (8.8)	7.49, d (8.8)	7.10, dd (8.4, 1.8)	7.49, d (8.8)	7.10, dd (8.0, 2.0)
7″	7.68, d (15.6)	7.69, d (16.0)	7.69, d (16.2)	7.68, d (16.0)	7.69, d (16.0)
8″	6.38, d (15.6)	6.41, d (16.0)	6.45, d (16.2)	6.38, d (16.0)	6.45, d (16.0)
2‴	7.50, d (8.8)	7.50, d (8.8)	7.49, d (8.4)	7.26, d (2.0)	7.24, d (2.0)
3‴	6.81, d (8.8)	6.81, d (8.8)	6.80, d (8.4)
5‴	6.81, d (8.8)	6.81, d (8.8)	6.80, d (8.4)	6.81, d (8.0)	6.82, d (8.0)
6‴	7.50, d (8.8)	7.50, d (8.8)	7.49, d (8.4)	7.09, dd (8.0, 2.0)	7.10, dd (8.0, 2.0)
7‴	7.66, d (16.0)	7.65, d (16.0)	7.65, d (16.2)	7.65, d (16.0)	7.65, d (16.0)
8‴	6.43, d (16.0)	6.45, d (16.0)	6.44, d (16.2)	6.47, d (16.0)	6.49, d (16.0)
3″-OMe			3.91, s		3.90, s
3‴-OMe				3.89, s	3.91, s

^a In MeOH-d₄, ¹H NMR at 400 MHz. ^b In MeOH-d₄, ¹H NMR at 600 MHz. s = singlet; d = doublet; t = triplet; m = multiplet; br = broad.

and

Table 7. ¹³C NMR data of compounds 20–24.

Position	20 a	21 a	22 b	23 a	24 a
1	66.1	63.6	63.4	66.2	63.5
2	103.3	106.1	106.2	103.2	106.2
3	79.5	76.6	76.6	79.5	76.7
4	73.5	76.5	76.5	73.5	76.6
5	84.2	82.7	82.7	84.3	82.7
6	63.8	64.1	64.2	63.9	64.3
1-OAc	172.0 20.6			171.9 20.6
3-OAc	172.1 20.7	171.8 20.7	172.0 20.7	172.1 20.7	171.7 20.7
4-OAc		172.0 20.7	171.8 20.7		172.0 20.7
1′	91.0	91.3	91.4	90.9	91.4
2′	74.0	74.1	74.1	74.0	74.1
3′	72.3	72.2	72.2	72.4	72.2
4′	71.8	71.8	71.9	71.9	71.9
5′	72.2	72.2	72.2	72.2	72.2
6′	64.9	64.9	64.9	65.1	64.9
1″	127.1	127.2	127.4	127.1	127.6
2″	131.3	131.3	111.7	131.4	111.7
3″	116.8	116.8	149.5	116.8	149.4
4″	161.5	161.6	151.2	161.5	150.7
5″	116.8	116.8	116.5	116.8	116.4
6″	131.3	131.3	124.6	131.4	124.3
7″	147.5	147.5	147.8	147.5	147.8
8″	114.7	114.8	115.0	114.8	115.1
9″	168.7	168.7	168.7	168.7	168.7
1‴	127.1	127.0	127.1	127.7	127.8
2‴	131.4	131.4	131.3	111.5	111.8
3‴	116.8	116.9	116.9	149.4	149.4
4‴	161.4	161.4	161.6	150.7	150.8
5‴	116.8	116.9	116.9	116.4	116.4
6‴	131.4	131.4	131.3	124.5	124.5
7‴	146.9	146.8	146.9	147.1	147.1
8‴	114.9	115.1	115.0	115.3	115.4
9‴	169.2	169.2	169.2	169.1	169.2
3″-OMe			56.4		56.5
3‴-OMe				56.5	56.5

^a In MeOH-d₄, ¹³C NMR at 100 MHz. ^b In MeOH-d₄, ¹³C NMR at 150 MHz.

) with those of 7 disclosed that one more acetyl group was presented in both 20 and 21. The key HMBC correlations from H₂-1 (δ_H 4.17, 4.00) to carbonyl (δ_C 172.0) and H-3 (δ_H 5.29) to carbonyl (δ_C 172.1) in 20, and from H-3 (δ_H 5.65) to carbonyl (δ_C 171.8) and H-4 (δ_H 5.48) to carbonyl (δ_C 172.0) in 21, indicated that the two acetyl groups were located at C-1 and C-3 in 20 and C-3 and C-4 in 21 (Figure 6). Thus, the structures of compounds 20 and 21 were characterized and named phanerosides T and U, respectively.

Compounds 22 and 23 showed the same molecular formula, C₃₅H₄₀O₁₈, as determined by their ¹³C NMR data and respective HRESIMS ion peaks at m/z 771.2093 and 771.2112 ([M + Na]⁺, calcd for 771.2107). Analysis of the NMR data (Figures S194–S199 and S203–S208) of 22 and 23 proclaimed that the sugar moieties in both compounds were acylated by a trans-ferulic acid, a trans-p-coumaric acid, and two acetic acids. The trans-feruloyl and trans-p-coumaroyl units were, respectively, located at C-2′ and C-6′ in 22 based on the key HMBC correlations from H-2′ to C-9″ and H₂-6′ to C-9‴, while the locations of these two substituents in 23 were the opposite. The key HMBC correlations from H-3 (δ_H 5.65) to carbonyl (δ_C 172.0) and H-4 (δ_H 5.50) to carbonyl (δ_C 171.8) in 22, and from H₂-1 (δ_H 4.16, 3.99) to carbonyl (δ_C 171.9) and H-3 (δ_H 5.30) to carbonyl (δ_C 172.1) in 23, indicated that the two acetyl groups were placed at C-3 and C-4 in 22 and C-1 and C-3 in 23. Accordingly, the structures of compounds 22 and 23 (phanerosides V and W) were defined as shown.

The molecular formula of compound 24 was assigned as C₃₆H₄₂O₁₉ based on the HRESIMS (m/z 801.2226 [M + Na]⁺, calcd for 801.2213) and ¹³C NMR data. The NMR data (Figures S212–S217) indicated that it possessed two trans-feruloyl moieties, D-sucrose, and two acetyl groups. The two trans-feruloyl moieties were linked to C-2′ and C-6′ of the glucopyranose ring, and two acetyl groups were attached to C-3 and C-4 of the fructofuranose ring, according to the key HMBC correlations as shown in Figure 6. Thus, the structure of compound 24 was identified and named phaneroside X.

• In Vitro Anti-inflammatory Effects of Compounds 1–24

Nitric oxide (NO) is one of the major inflammatory mediators, and phenylpropanoid esters of sucrose have been previously reported to possess potent anti-inflammatory activity [3,7,22]. Therefore, all the isolates were evaluated in vitro for their anti-inflammatory potential via the Griess reaction in LPS-induced BV-2 microglial cells (Figure 7) [23]. Especially, compounds 6 and 21 exhibited potent inhibitory activities on NO production, with IC₅₀ values of 6.7 ± 1.7 and 5.2 ± 3.5 μM, which were better than the positive control, L-NMMA (IC₅₀ = 7.0 ± 2.7 μM). Compounds 10, 14, and 19 showed moderate inhibitory effects on NO production, with IC₅₀ values of 72.7, 46.0, and 57.7 μM, respectively. These results suggested that the anti-inflammatory activities of these compounds were not determined by a single variable, while the type, number, and position of the substituents may all affect their inhibitory activities.

b.

Antioxidant Effects of Compounds 1–24

Many isolated phenylpropanoid esters of sucrose are thought to act as potential antioxidants [3]. Consequently, their antioxidant activities were also tested using the DPPH radical scavenging assay [24]. Compounds 5, 15, 17, and 24 exhibited moderate inhibitory effects with EC₅₀ values of 43.9 ± 0.2, 43.8 ± 0.1, 34.9 ± 0.1, 39.4 ± 0.3 μM, respectively. As it stands, the compounds whose C-2′ and C-6′ of the glucopyranose ring were both substituted by trans-feruloyl groups showed a more positive impact on their antioxidant effects.

3. Materials and Methods

General Experimental Procedures

Optical rotations were obtained on a JASCO P-2000 polarimeter. UV absorption spectra were determined on a PerkinElmer 650 spectrophotometer. NMR spectra were acquired on a 400 or 600 MHz Bruker AVANCE apparatus. Chemical shifts are expressed in δ (ppm) and referenced to the solvent residual peak. HRESIMS data were obtained on an Agilent 6545 Q-TOF LC-MS spectrometer. The other instruments and materials serving for the isolation and purification of compounds were coincident with previous papers [25,26].

• Plant Material

The rattans of P. championii Benth. were collected in November 2020 in Guilin, Guangxi Province, People’s Republic of China (GPS: 24°47′32.7″ N 110°27′36.8″ E). The specimen (No. PC-202011) was authenticated by Professor Shao-Qing Tang (College of Life Science, Guangxi Normal University) and deposited at the State Key Laboratory for Chemistry and Molecular Engineering of Medicinal Resources, Guangxi Normal University.

b.

Extraction and Isolation

The dried rattans of P. championii (21.0 kg) were soaked for 12 h in 95% aqueous EtOH (100 L) at room temperature, and then extracted three times with 95% aqueous EtOH (3 × 100 L) via refluxing. The filtrate was concentrated under reduced pressure to afford 4.5 kg of crude extract, which was suspended in H₂O and successively partitioned with EtOAc and n-BuOH. The EtOAc partition (1.8 kg) was separated via silica gel (200–300 mesh) column chromatography (CC), eluting with a gradient of CH₂Cl₂/MeOH (from 1:0 to 1:1) to give 11 fractions (Frs.1–11).

Fr.6 (20.7 g) was separated via C₁₈ reversed-phase (RP) CC, eluting with a gradient of MeOH-H₂O (30:70 to 70:30) to give 11 subfractions (Frs.6.1–6.11).

Fr.6.2 (656.8 mg) was subjected to Sephadex LH-20 CC (MeOH) to yield six subfractions (Frs.6.2.1–6.2.6). Fr.6.2.5 (183.9 mg) was applied to silica gel (200–300 mesh) CC, eluting with CH₂Cl₂/MeOH (50:1 to 5:1) to obtain nine subfractions (Frs.6.2.5.1–6.2.5.9). Fr.6.2.5.8 (14.3 mg) was further purified using semipreparative RP-HPLC (CH₃CN/H₂O, 22:78, 8.0 mL/min) to afford compound 5 (10.0 mg, t_R 37.1 min). Compounds 3 (12.0 mg, t_R 38.5 min) and 4 (35.0 mg, t_R 42.4 min) were obtained using semipreparative RP-HPLC (CH₃CN/H₂O, 22:78, 8.0 mL/min) from Fr.6.2.5.9 (91.5 mg).

Fr.6.3 (475.2 mg) was separated via a Sephadex LH-20 column, eluting with MeOH to yield six subfractions (Frs.6.3.1–6.3.6). Fr.6.3.5 (223.1 mg) was fractionated using silica gel (200–300 mesh) with gradient elution (CH₂Cl₂/MeOH, 50:1 to 5:1) to provide seven subfractions (Frs.6.3.5.1–6.3.5.7). Purification of Fr.6.3.5.3 (25.4 mg) using semipreparative RP-HPLC (CH₃CN/H₂O, 24:76, 8.0 mL/min) to yield compounds 18 (4.0 mg, t_R 54.9 min), 17 (12.0 mg, t_R 64.4 min), and 19 (3.0 mg, t_R 77.7 min). Fr.6.3.5.4 (93.5 mg) was purified using semipreparative RP-HPLC (CH₃CN/H₂O, 24:76, 8.0 mL/min) to obtain compounds 14 (15.0 mg, t_R 48.1 min), 16 (3.0 mg, t_R 49.2 min), and 11 (25.0 mg, t_R 52.0 min). Fr.6.3.5.5 (32.6 mg) was further purified using semipreparative RP-HPLC (CH₃CN/H₂O, 24:76, 8.0 mL/min) to afford compounds 13 (8.0 mg, t_R 40.2 min) and 10 (18.5 mg, t_R 44.4 min).

Fr.6.5 (532.6 mg) was separated via Sephadex LH-20 CC (MeOH) and then silica gel (200–300 mesh) CC (CH₂Cl₂/MeOH, 50:1 to 8:1) to provide nine subfractions (Frs.6.5.1–6.5.9). Fr.6.5.3 (15.0 mg) was further purified using semipreparative RP-HPLC (CH₃CN/H₂O, 25:75, 8.0 mL/min) to produce compound 24 (6.0 mg, t_R 77.7 min). Fr.6.5.4 (59.0 mg) was chromatographed on a Sephadex LH-20 column using MeOH as a solvent and then purified using semipreparative RP-HPLC (CH₃CN/H₂O, 27:73, 8.0 mL/min) to give compound 22 (3.5 mg, t_R 89.4 min). Fr.6.5.5 (35.4 mg) and Fr.6.5.9 (11.9 mg) were further purified using semipreparative RP-HPLC (CH₃CN/H₂O, 27:73, 8.0 mL/min) to produce compounds 23 (11.0 mg, t_R 50.0 min) and 2 (3.0 mg, t_R 58.1 min), respectively. Further purification of Fr.6.5.6 (26.5 mg) using semipreparative RP-HPLC (CH₃CN/H₂O, 26:74, 8.0 mL/min) yielded compound 15 (7.0 mg, t_R 42.9 min). Purification of Fr.6.5.7 (26.5 mg) using semipreparative RP-HPLC (CH₃CN/H₂O, 25:75, 8.0 mL/min) gave compounds 12 (6.0 mg, t_R 42.3 min) and 9 (11.0 mg, t_R 45.0 min).

Fr.7 (14.0 g) was chromatographed over an MCI-gel column, eluting with MeOH/H₂O (20:80 to 85:15) to give 12 subfractions (Frs.7.1–7.12).

Fr.7.4 (527.6 mg) was fractionated using a silica gel (200–300 mesh) column (CH₂Cl₂/MeOH, 50:1 to 5:1) to obtain seven subfractions (Frs.7.4.1–7.4.7). Fr.7.4.6 (252.4 mg) was purified using semipreparative RP-HPLC (CH₃CN/H₂O, 20:80, 8.0 mL/min) to give compound 1 (80.0 mg, t_R 49.1 min).

Fr.7.6 (740.5 mg) was partitioned into six subfractions (Frs.7.6.1–7.6.6) using a Sephadex LH-20 column (MeOH). Fr.7.6.4 (449.3 mg) was separated via silica gel (200–300 mesh) CC to provide eight subfractions (Frs.7.6.4.1–7.6.4.8). Fr.7.6.4.4 (77.3 mg) was further purified using semipreparative RP-HPLC (CH₃CN/H₂O, 24:76, 8.0 mL/min) to yield compound 8 (25.0 mg, t_R 39.8 min). Fr.7.6.4.6 (80.6 mg) was purified using semipreparative RP-HPLC (CH₃CN/H₂O, 24:76, 8.0 mL/min) to obtain compounds 7 (30.0 mg, t_R 44.1 min) and 6 (8.0 mg, t_R 56.2 min).

Fr.7.7 (833.7 mg) was separated using Sephadex LH-20 (MeOH) to obtain eight subfractions (Frs.7.7.1–7.7.8). Fr.7.7.2 (258.5 mg) was then fractionated via silica gel (200–300 mesh) CC to give six subfractions (Frs.7.7.2.1–7.7.2.6). Fr.7.7.2.2 (74.3 mg) and Fr.7.7.2.3 (69.8 mg) were purified using semipreparative RP-HPLC (CH₃CN/H₂O, 27:73, 8.0 mL/min) to yield compounds 21 (25.0 mg, t_R 54.4 min) and 20 (35.0 mg, t_R 50.2 min), respectively.

c.

Physicochemical Properties and Spectroscopic Data of Compounds 1–24

Phaneroside A (1): white amorphous powder; [α]${}_D^{20}$ + 33 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 211 (3.87), 228 (3.92), 314 (4.27) nm; IR (KBr) ν_max 3348, 1696, 1605, 1516, 1171, 1054 cm⁻¹; (+) HRESIMS m/z 657.1798 [M + Na]⁺ (calcd for C₃₀H₃₄O₁₅Na, 657.1790); ¹H and ¹³C NMR data, see Table 1 and Table 2. All significant data are presented in Supplementary Materials (Figures S1–S9).

Phaneroside B (2): white amorphous powder; [α]${}_D^{20}$ + 36 (c 0.08, MeOH); UV (MeOH) λ_max (log ε) 217 (3.95), 320 (4.25) nm; IR (KBr) ν_max 3432, 1696, 1605, 1516, 1271, 1169, 1050 cm⁻¹; (+) HRESIMS m/z 641.1847 [M + Na]⁺ (calcd for C₃₀H₃₄O₁₄Na, 641.1841); ¹H and ¹³C NMR data, see Table 1 and Table 2. All significant data are presented in Supplementary Materials (Figures S11–S19).

Phaneroside C (3): white amorphous powder; [α]${}_D^{20}$ + 22 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 219 (3.82), 320 (4.10) nm; IR (KBr) ν_max 3417, 1691, 1631, 1605, 1516, 1170, 1052 cm⁻¹; (+) HRESIMS m/z 687.1895 [M + Na]⁺, (calcd for C₃₁H₃₆O₁₆Na, 687.1896); ¹H and ¹³C NMR data, see Table 1 and Table 2. All significant data are presented in Supplementary Materials (Figures S20–S28).

Phaneroside D (4): white amorphous powder; [α]${}_D^{20}$ + 19 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 231 (3.94), 320 (4.25) nm; IR (KBr) ν_max 3348, 1694, 1632, 1604, 1516, 1270, 1170 cm⁻¹; (+) HRESIMS m/z 687.1896 [M + Na]⁺ (calcd for C₃₁H₃₆O₁₆Na, 687.1896); ¹H and ¹³C NMR data, see Table 1 and Table 2. All significant data are presented in Supplementary Materials (Figures S29–S37).

Phaneroside E (5): white amorphous powder; [α]${}_D^{20}$ + 36 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 217 (4.07), 237 (4.01), 328 (4.28) nm; IR (KBr) ν_max 3418, 1694, 1606, 1517, 1246, 1170, 1053 cm⁻¹; (+) HRESIMS m/z 717.1990 [M + Na]⁺ (calcd for C₃₂H₃₈O₁₇Na, 717.2001); ¹H and ¹³C NMR data, see Table 1 and Table 2. All significant data are presented in Supplementary Materials (Figures S38–S46).

Phaneroside F (6): white amorphous powder; [α]${}_D^{20}$ + 31 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 211 (3.86), 228 (3.91), 314 (4.26) nm; IR (KBr) ν_max 3431, 1693, 1605, 1516, 1171, 1051 cm⁻¹; (+) HRESIMS m/z 699.1915 [M + Na]⁺ (calcd for C₃₂H₃₆O₁₆Na, 699.1896); ¹H and ¹³C NMR data, see Table 3 and Table 5. All significant data are presented in Supplementary Materials (Figures S47–S55).

Phaneroside G (7): white amorphous powder; [α]${}_D^{20}$ + 36 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 211 (3.88), 228 (3.93), 315 (4.28) nm; IR (KBr) ν_max 3421, 1694, 1606, 1516, 1259, 1171, 1059 cm⁻¹; (+) HRESIMS m/z 699.1898 [M + Na]⁺ (calcd for C₃₂H₃₆O₁₆Na, 699.1896); ¹H and ¹³C NMR data, see Table 3 and Table 5. All significant data are presented in Supplementary Materials (Figures S56–S64).

Phaneroside H (8): white amorphous powder; [α]${}_D^{20}$ + 33 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 211 (3.93), 228 (3.98), 314 (4.32) nm; IR (KBr) ν_max 3326, 1696, 1605, 1516, 1171, 1063 cm⁻¹; (+) HRESIMS m/z 699.1899 [M + Na]⁺ (calcd for C₃₂H₃₆O₁₆Na, 699.1896); ¹H and ¹³C NMR data, see Table 3 and Table 5. All significant data are presented in Supplementary Materials (Figures S65–S73).

Phaneroside I (9): white amorphous powder; [α]${}_D^{20}$ + 23 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 217 (4.00), 319 (4.29) nm; IR (KBr) ν_max 3427, 1695, 1632, 1605, 1516, 1270, 1170, 1051 cm⁻¹; (+) HRESIMS m/z 729.2015 [M + Na]⁺ (calcd for C₃₃H₃₈O₁₇Na, 729.2001); ¹H and ¹³C NMR data, see Table 3 and Table 5. All significant data are presented in Supplementary Materials (Figures S74–S82).

Phaneroside J (10): white amorphous powder; [α]${}_D^{20}$ + 23 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 216 (3.98), 320 (4.28) nm; IR (KBr) ν_max 3436, 1691, 1632, 1605, 1516, 1268, 1170 cm⁻¹; (+) HRESIMS m/z 729.2012 [M + Na]⁺ (calcd for C₃₃H₃₈O₁₇Na, 729.2001); ¹H and ¹³C NMR data, see Table 3 and Table 5. All significant data are presented in Supplementary Materials (Figures S83–S91).

Phaneroside K (11): white amorphous powder; [α]${}_D^{20}$ + 19 (c 0.05, MeOH); UV (MeOH) λ_max (log ε) 216 (4.01), 319 (4.29) nm; IR (KBr) ν_max 3429, 1695, 1632, 1605, 1516, 1268, 1170, 1053 cm⁻¹; (+) HRESIMS m/z 729.1989 [M + Na]⁺ (calcd for C₃₃H₃₈O₁₇Na, 729.2001); ¹H and ¹³C NMR data, see Table 3 and Table 5. All significant data are presented in Supplementary Materials (Figures S92–S100).

Phaneroside L (12): white amorphous powder; [α]${}_D^{20}$ + 19 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 218 (4.03), 319 (4.30) nm; IR (KBr) ν_max 3436, 1695, 1632, 1605, 1516, 1274, 1171 cm⁻¹; (+) HRESIMS m/z 729.2018 [M + Na]⁺ (calcd for C₃₃H₃₈O₁₇Na, 729.2001); ¹H and ¹³C NMR data, see Table 4 and Table 5. All significant data are presented in Supplementary Materials (Figures S101–S109).

Phaneroside M (13): white amorphous powder; [α]${}_D^{20}$ + 22 (c 0.05, MeOH); UV (MeOH) λ_max (log ε) 217 (4.04), 319 (4.32) nm; IR (KBr) ν_max 3436, 1695, 1632, 1605, 1516, 1268, 1170, 1053 cm⁻¹; (+) HRESIMS m/z 729.2010 [M + Na]⁺ (calcd for C₃₃H₃₈O₁₇Na, 729.2001); ¹H and ¹³C NMR data, see Table 4 and Table 5. All significant data are presented in Supplementary Materials (Figures S110–S118).

Phaneroside N (14): white amorphous powder; [α]${}_D^{20}$ + 28 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 232 (4.03), 319 (4.32) nm; IR (KBr) ν_max 3432, 1692, 1606, 1518, 1172, 1051 cm⁻¹; (+) HRESIMS m/z 729.1990 [M + Na]⁺ (calcd for C₃₃H₃₈O₁₇Na, 729.2001); ¹H and ¹³C NMR data, see Table 4 and Table 5. All significant data are presented in Supplementary Materials (Figures S119–S127).

Phaneroside O (15): white amorphous powder; [α]${}_D^{20}$ + 22 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 217 (4.13), 237 (4.06), 327 (4.34) nm; IR (KBr) ν_max 3429, 1695, 1632, 1602, 1516, 1273, 1163, 1051 cm⁻¹; (+) HRESIMS m/z 759.2124 [M + Na]⁺ (calcd for C₃₄H₄₀O₁₈Na, 759.2107); ¹H and ¹³C NMR data, see Table 4 and Table 5. All significant data are presented in Supplementary Materials (Figures S128–S136).

Phaneroside P (16): white amorphous powder; [α]${}_D^{20}$ + 27 (c 0.05, MeOH); UV (MeOH) λ_max (log ε) 217 (4.22), 236 (4.15), 325 (4.37) nm; IR (KBr) ν_max 3432, 1690, 1605, 1517, 1169, 1056 cm⁻¹; (+) HRESIMS m/z 759.2095 [M + Na]⁺ (calcd for C₃₄H₄₀O₁₈Na, 759.2107); ¹H and ¹³C NMR data, see Table 4 and Table 5. All significant data are presented in Supplementary Materials (Figures S137–S145).

Phaneroside Q (17): white amorphous powder; [α]${}_D^{20}$ + 28 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 217 (4.07), 237 (4.01), 327 (4.29) nm; IR (KBr) ν_max 3426, 1697, 1632, 1598, 1516, 1272, 1161 cm⁻¹; (+) HRESIMS m/z 759.2097 [M + Na]⁺ (calcd for C₃₄H₄₀O₁₈Na, 759.2107); ¹H and ¹³C NMR data, see Table 4 and Table 5. All significant data are presented in Supplementary Materials (Figures S146–S154).

Phaneroside R (18): white amorphous powder; [α]${}_D^{20}$ + 27 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 217 (4.13), 234 (4.04), 324 (4.20) nm; IR (KBr) ν_max 3428, 1696, 1605, 1517, 1261, 1170, 1054 cm⁻¹; (+) HRESIMS m/z 759.2096 [M + Na]⁺ (calcd for C₃₄H₄₀O₁₈Na, 759.2107); ¹H and ¹³C NMR data, see Table 4 and Table 5. All significant data are presented in Supplementary Materials (Figures S155–S163).

Phaneroside S (19): white amorphous powder; [α]${}_D^{20}$ + 23 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 217 (4.17), 235 (4.07), 325 (4.26) nm; IR (KBr) ν_max 3397, 2921, 2850, 1646, 1516, 1272 cm⁻¹; (+) HRESIMS m/z 759.2097 [M + Na]⁺ (calcd for C₃₄H₄₀O₁₈Na, 759.2107); ¹H and ¹³C NMR data, see Table 4 and Table 5. All significant data are presented in Supplementary Materials (Figures S164–S172).

Phaneroside T (20): white amorphous powder; [α]${}_D^{20}$ + 25 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 211 (3.92), 229 (3.97), 314 (4.32) nm; IR (KBr) ν_max 3420, 1697, 1606, 1516, 1262, 1171, 1056 cm⁻¹; (+) HRESIMS m/z 741.2001 [M + Na]⁺ (calcd for C₃₄H₃₈O₁₇Na 741.2001); ¹H and ¹³C NMR data, see Table 6 and Table 7. All significant data are presented in Supplementary Materials (Figures S173–S181).

Phaneroside U (21): white amorphous powder; [α]${}_D^{20}$ + 26 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 211 (3.89), 228 (3.94), 314 (4.28) nm; IR (KBr) ν_max 3432, 1695, 1606, 1516, 1170, 1061 cm⁻¹; (+) HRESIMS m/z 741.2001 [M + Na]⁺ (calcd for C₃₄H₃₈O₁₇Na, 741.2001); ¹H and ¹³C NMR data, see Table 6 and Table 7. All significant data are presented in Supplementary Materials (Figures S182–S190).

Phaneroside V (22): white amorphous powder; [α]${}_D^{20}$ + 26 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 230 (4.02), 316 (4.28) nm; IR (KBr) ν_max 3429, 1697, 1606, 1516, 1269, 1170, 1054 cm⁻¹; (+) HRESIMS m/z 771.2093 [M + Na]⁺ (calcd for C₃₅H₄₀O₁₈Na, 771.2107); ¹H and ¹³C NMR data, see Table 6 and Table 7. All significant data are presented in Supplementary Materials (Figures S191–S199).

Phaneroside W (23): white amorphous powder; [α]${}_D^{20}$ + 23 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 217 (4.04), 319 (4.33) nm; IR (KBr) ν_max 3436, 1696, 1633, 1605, 1516, 1268, 1170, 1053 cm⁻¹; (+) HRESIMS m/z 771.2112 [M + Na]⁺ (calcd for C₃₅H₄₀O₁₈Na, 771.2107); ¹H and ¹³C NMR data, see Table 6 and Table 7. All significant data are presented in Supplementary Materials (Figures S200–S209).

Phaneroside X (24): white amorphous powder; [α]${}_D^{20}$ + 25 (c 0.06, MeOH); UV (MeOH) λ_max (log ε) 217 (4.16), 238 (4.10), 328 (4.37) nm; IR (KBr) ν_max 3436, 1712, 1633, 1601, 1515, 1273, 1177 cm⁻¹; (+) HRESIMS m/z 801.2226 [M + Na]⁺ (calcd for C₃₆H₄₂O₁₉Na, 801.2213); ¹H and ¹³C NMR data, see Table 6 and Table 7. All significant data are presented in Supplementary Materials (Figures S210–S217).

d.

Acid Hydrolysis of Compound 1

Compound 1 (4.0 mg) was added to 5.0 mL of 9% aqueous HCl in a sealed flask, which was refluxed at 80 °C for 5 h. The acidic aqueous mixture was dried, H₂O (2 mL) was added, and the mixture was extracted with EtOAc (3 × 2 mL). The aqueous layer was concentrated to obtain the sugar fraction, which was dissolved with MeOH and analyzed using chiral-phase HPLC equipped with a Daicel Chiralpak AD-H column (250 × 4.6 mm, 5 μm) and an evaporative light-scattering detector (ELSD) using n-hexane:EtOH (82:18) as the mobile phase (0.7 mL/min) [27]. The sugars were confirmed to be D-glucose and D-fructose by comparing their retention times with those of D-glucose (17.4 min), L-glucose (18.2 min), D-fructose (25.6 min), and L-fructose (26.4 min) (Figure S10).

e.

NO Production Measurements and Cell Viability Assays

The inhibitory effects of the isolated compounds on LPS-stimulated NO production were evaluated using the Griess reaction, and the cytotoxicities of compounds on BV-2 microglial cells were evaluated using MTT assays, as described in our previous report [23]. The result is shown in Figure 7.

f.

Antioxidant Activity Assay

The antioxidant activity of the isolated compounds was tested using a DPPH radical scavenging assay as previously described, and vitamin C was used as the positive control [24].

4. Conclusions

In conclusion, 24 new phenylpropanoid esters of sucrose were isolated from the rattans of P. championii. Their structures were determined via extensive spectroscopic methods. The configuration of sugar moiety was determined via chiral-phase HPLC equipped with an evaporative light-scattering detector (ELSD) after acid hydrolysis of compound 1. This is the first report of phenylpropanoid esters of sucrose isolated from the family Fabaceae. Structurally, these compounds revealed a huge structural diversity in terms of the number and position of phenylpropanoid and acetyl substituents. Biologically, all the isolated compounds were evaluated for their anti-inflammatory and antioxidant activities, and several compounds showed potent or moderate effects. Additionally, the structure–activity relationship was briefly discussed. These compounds may serve as potential leads for the development of anti-inflammatory agents.