邵泰明，宋小平 ，韓長(zhǎng)日，陳光英，陳文豪，戴春燕，宋鑫明

海南師范大學(xué)教育部熱帶藥用植物化學(xué)省部共建重點(diǎn)實(shí)驗(yàn)室;海南師范大學(xué)化學(xué)化工學(xué)院，海口571158

Introduction

The genus of Ficus comprises about 1000 species in the world，among which about 120 are distributed in China［1］. F. auriculata distributed in Ledong，Dongfang，Baoting，Lingao of Hainan province China［2］. It has been used as Chinese folk medicine for the treatment of hyperactivity cough，nocturnal emission［3］. The leaves of F.auriculata have antioxidant，antiinflammatory，antidiabetic and hepatoprotective activity，and fruit has a strong antibacterial activity［4-6］.In order to investigate the constituents of this plant，eleven compounds were isolated from the 95% ethanol extracts and identified as genistein(1)，wighteone(2)，5，7，4'，-trihydroxy-6-(2-hydroxy-3-methyl-3-butenyl)isoflavone(3)，5，7，4'-trihydroxy-3'-(2-hydroxy-3-methyl-3-butenyl )isoflavone(4)，alpinumisoflavone(5)，derrone(6)，(3R)-5-formylmellein(7)，3β-(1-Hydroxyethyl)-7-hydroxy-1-isobenzofuranone(8)，trans-p-phydroxycinnamic acid ethyl ester(9)，β-sitosteral(10)，and β-daucosterin(11).Compounds 4，7，8，9 were reported from the genus Ficus for the first time.Compounds 1-10 were isolated from this plant material for the first time.

Experimental

Apparatus and reagents

Melting points were determined on a WRX-4 micromelting-point apparatus(uncorrected).NMR spectra were measured on a Bruker AV-400 instrument with TMS as the internal standard. Silica gel used for column chromatography(CC)was supplied by Qingdao Marine Chemical Factory(Qingdao，China). Sephadex LH-20 was used(Pharmacia，Sweden). TLC and preparative TLC were purchased from Yantai Chemical Industry Institute(Yantai，China).All solvents were purchased from XiLong Chemical Reagent Factory(Shan-Tou，China). The optical density was measured by an enzyme-labeled detector(Elx800，BioTek Instruments，Inc).

Plant materials

The sterm of F. auriculata were collected in Jian feng ling National Forest Park，Hainan Province China，in 2010，and authenticated by associate Prof. Zhong Qiong-Xin，College of Life Science，Hainan Normal University.A voucher specimen was deposited in the Key Laboratory of Tropical Medicinal Plant Chemistry of Ministry of Education，Hainan Normal University.

Extraction and isolation

The dried sterm of F. auriculata(13.5 kg)were powdered and extracted with 95% aqueous ethanol(v/v)three times at room temperature(3 ×5 d).The extract was suspended in 2.0 L of water and then partitioned with petroleum ether，CHCl3，EtOAc successively. The petroleum ether soluble fraction(240 g)was chromatographed on a silica gel column eluted with petroleum ether/EtOAc(100∶1 to 1∶100，v/v)to afford major fractions 1-20.Fraction 4(3.45g)purified by recrystallization to yield compounds 10(750. 3mg);Fraction 6(5.40 g)was subjected to CC(silica gel，petroleum ether/EtOAc，10 ∶1 to 0 ∶1)and purified by CC(Sephadex LH-20，CHCl3/MeOH，2 ∶3)to provide compounds 5(45.4mg)，6(23.5mg)，and 7(13.0 mg);Fraction 7(2.40 g)was further chromatographed over Sephadex LH-20(CHCl3/MeOH，2 ∶3)，followed by PTLC to give compounds 8(13.3 mg)，9(8.7 mg);Fraction 10(4.70 g)was further chromatographed over Sephadex LH-20(CHCl3/MeOH，2 ∶3)，followed by PTLC to give compounds 1(20.1 mg)，2(13.5 mg)，3(24.4 mg)and 4(13.2 mg).The ethyl acetate fraction(23g)was subjected to silica gel CC(CHCl3/MeOH 100∶1→2∶1)to yield 11(14.4 mg).

Identification

Fig.1 Structures of compounds 1-9

Compound 1White powder，C15H10O5，1H NMR(Acetone，400 MHz)，δ:13.04(1H，s，OH-5)，8.18(1H，s，H-2)，7.46(2H，d，J = 7.6 Hz，H-2'，6')，6.91(2H，d，J =7.6 Hz，H-3'，5')，6.42(1H，s，H-8)，6.29 (1H，s，H-6);13C NMR (Acetone，100 MHz)，δ:182.6(C-4)，166.1(C-7)，164.9(C-5)，160.0(C-8a)，159.5(C-4')，155.3(C-2)，132.2(C-2'，6')，125.0(C-3)，124.0(C-1')，117.0(C-3'，5')，107.1(C-4a)，100.8(C-6)，95.5(C-8). It was identified as genistein by comparison of the physical and spectral data with the reported data［7］.

Compound 2 Yellow powder，C20H18O5，1H NMR(MeOD，400 MHz);δ:13.01(1H，s，OH-5)，8.01(1H，s，H-2)，7.36(2H，d，J = 8.0 Hz，H-2'，6')，6.84(2H，d，J =8.0 Hz，H-3'，5')，6.37(1H，s，H-8)，5.23(1H，t，J=7.2 Hz，H-2″)，3.31(2H，d，overlapped，H-2″)，1.78(3H，s，H-4″)，1.66(3H，s，H-5″);13C NMR(MeOD，100 MHz)δ:182.3(C-4)，163.7(C-7)，160.5(C-5)，158.8(C-4')，157.6(C-8a)，154.6(C-2)，132.1(C-3″)，131.5(C-2'，6')，124.6(C-3)，123.5(C-1')，123.4(C-2″)，116.3(C-3'，5')，113.1(C-6)，106.1(C-4a)，93.9(C-8)，26.0(C-5″)，22.3(C-1″)，18.0(C-4″). It was identified as wighteone by comparison of the physical and spectral data with the reported data［8］.

Compound 3 Yellow powder，C20H18O6，1H NMR(MeOD，400 MHz)，δ:13.16(1H，s，OH-5)，8.03(1H，s，H-2)，7.36(2H，d，J = 8.0 Hz，H-2'，6')，6.84(2H，d，J =8.0 Hz，H-3'，5')，6.38(1H，s，H-8)，4.76，4.70(1H each，s，=CH2)，4.41(1H，t，J =6.4 Hz，H-2″)，3.01(1H，dd，J = 6.4 Hz，J = 13.6 Hz，Ha-1″)，2.90(1H，dd，J =7.2 Hz，J =13.6 Hz，Hb-1″)，1.83(3H，s，H-4″);13C NMR(MeOD，100 MHz)，δ:182.3(C-4)，164.6(C-7)，161.3(C-5)，158.8(C-4')，158.0(C-8a)，154.7(C-2)，148.7(C-3″)，131.5(C-2'，6')，124.7(C-3)，123.5(C-1')，116.3(C-3'，5')，111.1 (= CH2)，110.5 (C-6)，106.1(C-4a)，94.4(C-8)，76.3(C-2″)29.8(C-1″)，17.8(C-4″). It was identified as 5，7，4'，2-hydroxy-3-methyl-3-butenyl isoflavone by comparison of the physical and spectral data with the reported data［9］.

Compound 4White powder，C20H18O6，1H NMR(Acetone，400 MHz)，δ:13.06(1H，s，OH-5)，8.15(1H，s，H-2)，7.37(1H，s，H-2')，7.34(1H，d，J =8.0 Hz，H-6')，6.87(1H，d，J =8.0 Hz，H-5')，6.41(1H，s，H-8)，6.28(1H，s，H-6)，4.98，4.78(1H each，s，=CH2)，4.42(1H，t，J =6.0 Hz，H-2″)，2.91(2H，m，overlapped，H-1″)，1.81(3H，s，H-4″);13C NMR(Acetone，100 MHz)，δ:182.6(C-4)，166.2(C-7)，164.9(C-5)，160.0(C-8a)，158.1(C-4')，155.3(C-2)，149.5(C-3″)，134.0(C-2')，130.4(C-6')，127.8(C-3')，125.1(C-3)，124.1(C-1')，117.7(C-5')，111.6(= CH2)，107.1(C-4a)，100.8(C-6)，95.5(C-8)，78.0(C-2″)，39.8(C-1″)，19.3(C-4″). It was identified as 5，7，4'-trihydroxy-3'-(2-hydroxy-3-methyl-3-butenyl)isoflavone by comparison of the physical and spectral data with the reported data［10］.

Compound 5Yellow crystal，C20H16O5，1H NMR(CDCl3，400 MHz)δ:13.08(1H，s，OH-5)，7.81(1H，s，H-2)，7.33(2H，d，J = 8.4 Hz，H-2'，6')，6.83(2H，d，J =8.4 Hz，H-3'，5')，6.72(1H，d，J =10.4 Hz，H-4″)，6.34(1H，s，H-8)，5.62(1H，d，J =10.4 Hz，H-3″)，1.47(6H，s，CH3-5″，6″);13C NMR(CDCl3，100 MHz)δ:181.1(C-4)，159.7(C-7)，157.4(C-5)，156.9(C-8a)，156.1(C-4')，152.7(C-2)，130.4(C-2'，6')，128.3(C-3″)，123.7(C-3)，122.9(C-1')，115.7(C-3'，5')，115.5(C-4″)，106.1(C-4a)，105.7(C-6)，95.0(C-8)，78.2(C-2″)，28.4(C-5″，6″). It was identified as alpinumisoflavone by comparison of the physical and spectral data with the reported data［11］.

Compound 6Yellow crystal，C20H16O5，1H NMR(CDCl3，400 MHz)δ:12.92(1H，s，OH-5)，7.89(1H，s，H-2)，7.40(2H，d，J = 7.6 Hz，H-2'，6')，6.91(2H，d，J =7.6 Hz，H-3'，5')，6.68(1H，d，J =10.0 Hz，H-4″)，6.29(1H，s，H-6)，5.59(1H，d，J =10.0 Hz，H-3″)，1.47(6H，s，CH3-5″，6″);13C NMR(CDCl3，100 MHz)δ:180.9(C-4)，162.3(C-5)，159.6(C-7)，155.8(C-4')，152.3(C-2)，152.2(C-8a)，130.4(C-2'，6')，127.5(C-3″)，123.6(C-3)，123.1(C-1')，115.5(C-3'，5')，114.6(C-4″)，106.0(C-4a)，101.1(C-8)，100.4(C-6)，78.1(C-2″)，28.2(C-5″，6″). It was identified as derrone by comparison of the physical and spectral data with the reported data［12］.

Compound 7 Pale yellow oil C11H10O4，1H NMR(DMSO，400 MHz)，δ:11.81(1H，br s，OH-8)，10.07(1H，s，-CHO)，8.05(1H，d，J =8.8 Hz，H-6)，7.07(1H，d，J =8.8 Hz，H-7)，4.82(1H，m，H-3)，3.87(1H，dd，J =17.6 Hz，2.8 Hz，Ha-4)，3.06 (1H，dd，J=15.2 Hz，12.0 Hz，Hb-4)，1.46(3H，d，J=6.4 Hz，-CH3);13C NMR(DMSO，100 MHz)，δ:190.7(-CHO)，168.8(C-1)，165.1(C-8)，144.2(C-4a)，138.6(C-6)，124.3(C-5)，116.1(C-7)，108.9(C-8a)，75.2(C-3)，29.7(C-4)，20.2(-CH3). It was identified as(3R)-5-formylmellein by comparison of the physical and spectral data with the reported data［13］.

Compound 8Yellow oil C10H10O4，1H NMR(CDCl3，400 MHz)，δ:11.0(1H，s，OH-7)，7.55(1H，t，J=8.0 Hz，H-5)，7.02(2H，t，J =8.0 Hz，H-4，H-6)，4.62(1H，m，H-3)，4.61(1H，m，H-1')，1.53(3H，d，J=5.6 Hz，H-2');13C NMR(CDCl3，100 MHz)，δ:168.4(C-1)，162.0(C-7)，141.1(C-9)，136.9(C-5)，117.9(C-6)，116.1(C-4)，106.6(C-8)，79.9(C-1')，69.2(C-3)，17.9(C-2'). It was identified as 3β-(1β-hydroxyethyl)-7-hydroxy-1-isobenzofuranone by comparison of the physical and spectral data with the reported data［14］.

Compound 9White crystal C11H12O3，1H NMR(CDCl3，400 MHz)，δ:7.63(1H，d，J =16.0 Hz，H-7)，7.42(2H，d，J =8.0 Hz，H-2，H-6)，6.84(2H，d，J=8.0 Hz，H-3，H-5)，6.30(1H，d，J =16.0 Hz，H-8)，5.50(1H，br s，OH-3)，4.25(2H，q，J =7.2 Hz，H-1')，1.33(3H，t，J = 7.2 Hz，H-2');13C NMR(CDCl3，100 MHz)，δ:167.5(C-9)，157.6(C-4)，144.3(C-7)，129.9(C-2，C-6)，127.3(C-1)，115.8(C-3，C-5)，115.7(C-8)，60.4(C-1')，14.3(C-2'). It was identified as trans-p-hydroxycinnamic acid ethyl ester by comparison of the physical and spectral data with the reported data［15］.

Compound 10 White needles crystal;It was characterized by comparing it with authentic sample on TLC. It was identified as β-sitosterol［16］.

Compound 11White amorphous powder;It was characterized by comparing it with authentic sample on TLC. It was identified as β-daucosterol［17］.

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