New diketopiperazine derivatives with cytotoxicity from Nocardiopsis sp YIM M13066
New diketopiperazine derivatives with cytotoxicity from Nocardiopsis sp YIM M13066
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来自诺卡氏菌 YIM M13066 的具有细胞毒性的新型二酮哌嗪衍生物
DOI：
10.1038/ja.2017.46
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发表时间：
2017-06-01
影响因子：
3.3
通讯作者：
Shen, Yuemao
中科院分区：
文献类型：
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作者：
Sun, Mingwei;Chen, Xiaotong;Shen, Yuemao
含二酮哌嗪（DKPs）的天然产物代表了一大类次生代谢产物，主要由细菌、真菌、海洋无脊椎动物和高等生物产生。 DKP 是通过色氨酸、脯氨酸、组氨酸和苯丙氨酸等两种氨基酸缩合而生物合成的。 1–3 对 DKP 的兴趣是由于它们在抗菌、抗真菌、抗病毒和免疫抑制等各种药理学测定中的活性。 4-7 在最近的研究中，诺卡嗪是新定义的抗菌和细胞毒性环状二肽家族，由诺卡氏菌 (Nocardiopsis dassonvillei) 和白诺卡氏菌 (N. alba) 产生。 8, 9 我们对深海沉积物菌株 YIM M13066（被鉴定为诺卡氏菌）进行的调查发现，它能产生 6 种 DKP，包括两种新的 DKP：诺卡嗪 F (1) 和 G (2)，以及四种已知的 DKP (3-6)。在这项研究中，来自诺卡氏菌的化合物 1-6（图 1）的发酵、分离、结构阐明和生物活性。描述了 YIM M13066。菌株 YIM M13066 首先在培养皿中培养，其中含有约20ml MP琼脂培养基培养7天以获得种子培养物。将种子通过平板划线法接种到MP琼脂培养基(20μl)上，并在28℃下培养10天。发酵后的培养物用EtOAc-MeOH(85:15，体积比，20μl)在室温下提取，得到粗提物。将粗提取物用等体积的EtOAc(0.5l)和H 2 O分配。将EtOAc部分真空浓缩，得到EtOAc萃取物(7.8g)。提取物经Sephadex LH-20（25-100μm；GE Healthcare，瑞典；柱尺寸：200×20mm）柱层析，用MeOH洗脱，并在TLC检测的基础上合并，得到Fr。 1-10。神父。通过MPLC(310×2.6mm)在RP-18硅胶(80g)上分离7(1.1g)，得到Fr.7。 7a-7h。通过从Fr中结晶得到化合物5(30.0mg)和6(40mg)。 7d 和神父。分别为7e。神父。 7f 通过 HPLC（SunFire Prep C18 OBD，19×150mm，15mlmin−1，UV 365nm）纯化，用 70% 乙腈洗脱，得到 4（tR 11.7 分钟，6.8 mg）和 3（tR 13.5 分钟，7.1 mg）。神父。通过MPLC在RP-18硅胶(80g)上分离10(108.0mg)，得到Fr.10(108.0mg)。 10a-10j。神父。 10e 通过 HPLC（SunFire Prep C18 OBD，19×150mm，15mlmin−1，UV365nm）纯化，用 40% 乙腈洗脱，分别得到 1（tR 7.1 分钟，29.2 mg）和 2（tR 9.2 分钟，20.1 mg）。 Nocazine F (1) 为黄色无定形粉末，根据 HRESI-MS 测得其分子式为 C20H18N2O4，m/z= 351.1297 [M+ H]+（计算值 351.1339）。其 1H 和 13C NMR（在 DMSO-d6 中，表 1）对应于 1 个 NH 基团（δ 9.86 (s)）、2 个 OCH3（δH/δC 3.98/54.8 和 3.79/55.7）、10 个烯属 CH、7 个季碳原子和 1 个酰胺羰基（δ 160.2）。通过 δ 8.02 (d, J= 8.7 Hz)、6.83 (d, J= 8.7 Hz) 和 δ 7.50 (d, J= 8.7 Hz)、6.99 (d, J= 8.8 Hz) 处的两组耦合 1H 信号确定是否存在两个 1, 4-二取代苯环。从 18-OH (δ 9.91) 到 C-17/19 (δ 115.9) 和 C-18 (δ 158.8)，以及从 11-OCH3 (δ 3.79) 到 C-11 (δ 159.5) 的 HMBC 相关性表明，两个 1, 4-二取代苯环分别与羟基和甲氧基连接。 HMBC 相关性从 5-OCH3 (δ 3.98) 到 C-5 (δ 154.6)、从 NH-1 (δ 9.86) 到 C-5 和 C-3、从 H-7 (δ 6.46) 到 C-5 和 C-9/13 (δ 131.2)、从 H-14 (δ 7.03) 到 C-2 (δ 160.2) 和C-16/20 (δ 133.8) 表示由 p-羟基-Phe (Tyr) 和 p-MeO-Phe 部分形成的特定 DKP 单元，类似于 nocazine A。 8 Δ6 和 Δ3 (14) 双键的 Z 构型由 5-OCH3 (δ 3.98) 和 H-20 (δ 8 ...
The diketopiperazines (DKPs)-containing natural products represent a large class of secondary metabolites mainly produced by bacteria, fungi, marine invertebrates and higher organisms. DKPs were biosynthesized by condensation of two amino acids including tryptophan, proline, histidine and phenylalanine. 1–3 Interest in DKPs is due to their activities in various pharmacological assays ranging from antibacterial, antifungal, antiviral to immunosuppressive. 4–7 In recent studies, the nocazines are a newly defined family of antibacterial and cytotoxic cyclic dipeptides produced by Nocardiopsis dassonvillei and N. alba. 8, 9 Our investigation with a deep-sea sediment strain, YIM M13066, identified as a Nocardiopsis sp., was found to produce six DKPs, including two new DKPs, nocazines F (1) and G (2), and four known DKPs (3–6). In this study, the fermentation, isolation, and structural elucidation and bioactivity of compounds 1–6 (Figure 1) from Nocardiopsis sp. YIM M13066 were described. The strain YIM M13066 was first cultured in Petri dishes with ca. 20 ml MP agar medium for 7 days to obtain seed cultures. The seeds were inoculated on MP agar media (20 l) by streaking plate method and cultured for 10 days at 28 C. The fermented culture was extracted with EtOAc–MeOH (85: 15, volume per volume, 20l) at room temperature to obtain a crude extract. The crude extract was partitioned with equal volume EtOAc (0.5 l) and H2O. The EtOAc portion was concentrated under vacuum to afford the EtOAc extract (7.8 g). The extract was subjected to column chromatography over Sephadex LH-20 (25–100μm; GE Healthcare, Sweden; column dimensions: 200× 20mm) eluted with MeOH and pooled on the basis of TLC detection to obtain Fr. 1–10. Fr. 7 (1.1 g) was separated by MPLC (310× 2.6 mm) over RP-18 silica gel (80g) to afford Fr. 7a–7h. Compounds 5 (30.0 mg) and 6 (40mg) were obtained by crystallization from Fr. 7d and Fr. 7e, respectively. Fr. 7f was purified by HPLC (SunFire Prep C18 OBD, 19× 150mm, 15mlmin− 1, UV 365nm) eluted with 70% acetonitrile to afford 4 (tR 11.7 min, 6.8 mg) and 3 (tR 13.5 min, 7.1 mg). Fr. 10 (108.0 mg) was separated by MPLC over RP-18 silica gel (80g) to afford Fr. 10a–10j. Fr. 10e was purified by HPLC (SunFire Prep C18 OBD, 19× 150mm, 15mlmin− 1, UV365nm) eluted with 40% acetonitrile to yield 1 (tR 7.1 min, 29.2 mg) and 2 (tR 9.2 min, 20.1 mg), respectively. Nocazine F (1) was obtained as a yellow amorphous powder with the molecular formula C20H18N2O4 according to its HRESI-MS at m/z= 351.1297 [M+ H]+(calcd 351.1339). Its 1H and 13C NMR (in DMSO-d6, Table 1) corresponded to 1 NH group (δ 9.86 (s)), 2 OCH3 (δH/δC 3.98/54.8 and 3.79/55.7), 10 olefinic CH, 7 quaternary C-atoms and 1 amidocarbonyl (δ 160.2). The presence of two 1, 4-disubstituted benzene rings was determined by two sets of coupled 1H signals at δ 8.02 (d, J= 8.7 Hz), 6.83 (d, J= 8.7 Hz) and δ 7.50 (d, J= 8.7 Hz), 6.99 (d, J= 8.8 Hz). HMBC correlations from 18-OH (δ 9.91) to C-17/19 (δ 115.9) and C-18 (δ 158.8), and from 11-OCH3 (δ 3.79) to C-11 (δ 159.5) revealed that the two 1, 4-disubstituted benzene rings were connected with hydroxyl and methoxyl, respectively. HMBC correlations from 5-OCH3 (δ 3.98) to C-5 (δ 154.6), from NH-1 (δ 9.86) to C-5 and C-3, from H-7 (δ 6.46) to C-5 and C-9/13 (δ 131.2), and from H-14 (δ 7.03) to C-2 (δ 160.2) and C-16/20 (δ 133.8) indicated a specific DKP unit formed from a p-hydroxyl-Phe (Tyr) and a p-MeO-Phe moieties, similar to nocazine A. 8 The Z configurations of both Δ6 and Δ3 (14) double bonds were determined by the NOE correlations between 5-OCH3 (δ 3.98) and H-20 (δ 8 …