New Acrimarines from Citrus Plants

HETEROCYCLES
1992.0

Abstract

Five new acrimarines (acridonecoumarin dimers) were isolated from some Citrus plants (Rutaceae) and their structures were elucidated by spectroscopic method. Recently, we reported the isolation and structure el~cidation of acrimarine-A--H, dimeric compounds in which an acridone unit has been coupled to a coumarin. Further examination of the roots of "Yalaha" [several hybrid seedlings resulting from a cross of Duncan grapefruit (Citrus paradisi Macf.) x Dancy tangerine (L Lanlerine Hort. ex Tanaka)] and "Funadoko" (L funadoko Hort. ex Tanaka) afforded five new compounds, named acrimarine-l (I), -J (4), K (5), -L (6) and -M (7) and in this paper we wish to report the isolation and structure elucidation of these new compounds. Acrimarine-1 (1) was obtained as yellow oil, [α]D +27.8°(~~~13). The high resolution ms showed molecular formula C34H31NO7, which was considered to give rise to two characteristic ions from the halves of the molecule at m/z 323 and 242 corresponding to formulae C19H17NO4 and C15H14O3 respectively, indicating the presence of acridone and coumarin structures. The ir (3400, 1720, 1620, 1560 cm⁻¹) and uv [205, 223 (sh), 270 (sh), 296, 328, 348 (sh) nm] spectra suggested the presence of 1-hydroxy-9-acridone moiety. The ¹H nmr spectrum showed the presence of hydrogen bonded hydroxyl group (δ 14.63), ABC type aromatic protons [δ 7.94 (1H, dd, J=7.9, 1.8 Hz), 7.15 (1H, t, J=7.9 Hz), 7.09 (1H, dd, J=7.9, 1.8 Hz)], 1.1-dimethylpyran ring [δ 6.61 (1H, d, J=9.8 Hz), 5.48 (1H, d, J=9.8 Hz), 1.14, 1.46 (each 3H, s)], characteristic H-4', H-3', H-5' and H-8' of coumarin nucleus [δ 7.67, 6.22 (each 1H, d, J=9.2 Hz), 7.62, 6.69 (each 1H, s)], and a prenyl group connected with two aryl moieties [δ 5.94 (1H, d, J=9.8 Hz), 5.74 (1H, d, J=9.8 Hz), 1.80 (3H, s), 1.75 (3H, s)]. Two singlets at δ 3.75 and 3.72 in the ¹H nmr and two quartets at δ 55.9 and 48.5 in the ¹³C nmr spectra showed the presence of each one methoxy and α-methyl group. In nOe experiments, irradiation of the methoxy signal at δ 3.75 gave 20% increments of the signal at δ 6.69 (H-8'). On the other hand, when the α-methyl signal at δ 3.72 was irradiated, 13% increments were observed on the signal at δ 6.61 (H-11). From these results, the location of methoxy group was assigned to C-7' and the angular orientation of the dimethylpyran ring was confirmed. The similarities of signal patterns of ¹H nmr spectrum suggested the presence of 5-hydroxynoracronycine (z) and suherosin (3) moieties in the structure. The location of the suherosin unit was elucidated by ¹H detected heteronuclear multiple bond connectivity (HMBC) spectrum. The H-bonded proton signal at δ 14.63 showed long-range correlations with the carbon signals at δ 162.0 (C-1), 112.6 (C-2), and 107.1 (C-9a). Further, the proton signal at δ 5.74 (H-1'') was correlated with carbon signals at δ 162.0 (C-1), 112.6 (C-2), 159.0 (C-3), 132.9 (C-3''), 128.6 (C-5'), and 130.6 (C-6'). Other CH long range correlations observed are shown by arrows in Figure 1. Based on these spectral data, the structure of acrimarine-1 was concluded to 1. Acrimarine-J (4) was obtained as yellow oil, [α]D *0°(CHCl₃). The mass spectrum showed the molecular ion at m/z 595 which corresponded to C35H33NO8 a difference of CH2O compared with 1. The ¹H nmr features were similar to those of 1, except for appearances of one more methoxy (δ 3.90) signal and change of ABC type aromatic protons to AB type [δ 7.96, 6.97 (each 1H, d, J=8.5 Hz)]. The signals at δ 3.90, 3.84, 3.75 in the ¹H nmr and δ 60.5, 56.5, 48.9 in the ¹³C nmr spectra indicated the presence of two methoxy and one α-methyl group. The location of methoxy groups and the situation of 1.1-dimethylpyran ring were determined by nOe experiments. Irradiation of the signal at δ 3.75 and 3.84 showed 14% and 20% increments of the signals at δ 6.69 (H-11) and 6.83 (H-8'), respectively. When the signal at δ 3.90 was irradiated, no increments were observed on any signals. These results indicated the locations of methoxy groups at C-5 and C-7', phenolic hydroxyl group at C-6, and the angular situation of 1.1-dimethylpyran ring. On the basis of these results, structure (4) was assigned to acrimarine-J. Acrimarine-K (5) was isolated as yellow oil, [α]D *0° (CHCl₃). The molecular formula C30H27NO8 was established by high resolution ms. The ¹H nmr spectrum showed the signals of ortho coupled [δ 8.04, 6.94 (each 1H, d, J=8.8 Hz)], isolated [δ 6.48 (1H, s)] aromatic protons, characteristic signals due to H-4', H-3', H-5' and H-8' of coumarin nucleus [δ 7.88, 6.16 (each 1H, d, J=9.5 Hz), 7.73, 6.82 (each 1H, s)], and signals due to a prenyl group coupled to two aryl groups [δ 1.75 (3H, s), 1.72 (3H, s), 5.83 (1H, d, J=9.5 Hz) and 6.08 (1H, d, J=9.5 Hz)]. The signals at δ 3.98, 3.87, 3.77 in ¹H nmr and δ 40.2, 56.9, 62.2 in ¹³C nmr spectra indicated the presence of two methoxy and one 1-methyl group. The location of methoxy groups was determined by nOe experiments. Irradiation of the signal at δ 3.98 and 3.87 showed 14% and 16% increments of the signal at δ 6.48 (H-4) and 6.82 (H-8'), respectively. On the other hands, when the signal at δ 3.77 was irradiated, no increments were observed at any proton signals. Thus, the location of two methoxy groups was determined to C-5 and C-7'. From the above data, the location of suberosin unit was assigned at C-2 in the acridone skeleton, and the structure of acrimarine-K was determined to 5. The proposed structure was further confirmed by HC long range COSY shown by arrows in Figure 2. Acrimarine-L (6) was isolated as yellow oil, [α]D *0° (CHCl₃), and gave a molecular ion at m/z 529 which corresponded to C30H27NO8 the same as that of acrimarine-K (5). The ¹H nmr spectrum showed a good similarity with that of 5, except for small differences of chemical shifts indicating the presence of suberosin and 1.3.5.6-oxygenated 9-acridone units. The signals at δ 3.97, 3.94 and 3.87 indicated the presence of methoxy groups. In the nOe experiments, irradiation of the signal at δ 3.97 and 3.87 showed 17% and 14% increments of the signal at δ 6.34 (H-2) and 6.92 (H-8'), respectively. On the other hands, when the signal at δ 3.94 was irradiated, no increments were observed. Meanwhile, when the signal at δ 9.14 assumed to NH signal was irradiated, 12% and 9% increments were observed on the signal at δ 5.78 (H-1'') and 5.91 (H-2''), respectively. These results indicated the locations of methoxy groups were at C-3, C-5 and C-7', and the linking positions of acridone and coumarin rings were at C-4 and C-6', respectively. The above data led us to assign structure 6 to acrimarine-L. Acrimarine-M (7) was isolated as yellow oil, [α]D *0° (CHCl₃). The molecular formula, C29H25NO6, was confirmed by high resolution ms. The ¹H nmr spectrum showed, besides signals of suherosin unit, ABCD type [δ 8.38 (1H, d, J=8.1 Hz), 7.76 (1H, t, J=8.1 Hz), 7.73 (1H, d, J=8.1 Hz), 7.30 (1H, t, J=8.1 Hz)] and a lone [δ 6.54 (1H, s)] aromatic proton signal and two methyl singlets (δ 3.82 and 3.86). The location of two methyl signals at δ 3.86 and 3.82 was assigned by nOe experiments. When the signal at δ 3.82 was irradiated, each 14% and 10% increments were observed on the signals at δ 7.73 (H-5) and 6.54 (H-4), respectively. On the other hands, irradiation of the signal at δ 3.86 showed 11% increments of the signal at δ 6.82 (H-8'). The above data showed the presence of a-methyl group, the location of methoxy group at C-7', and the coupling position of the acridone and the coumarin nuclei was at C-2 and C-6'. From these results, the structure 7 was assigned to acrimarine-M. The absolute configurations of these new acrimarines remain to be determined.

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