OPB-01
骨架蛋白 cofilin 在肿瘤转移中的机制探讨与干预研究
黄洵，丁健*，耿美玉*
摘要
细胞骨架蛋白及其相关调节因子作为细胞形态的结构基础和细胞运动的动
力基础，决定肿瘤细胞迁移和侵袭能力，其调控异常与肿瘤转移密切相关。本论
文对细胞骨架的重要成员 cofilin 转录后调控进行研究，重点关注了 cofilin
翻译后 O-GlcNAc 糖基化修饰在调控 cofilin 介导的肿瘤细胞迁移侵袭重要机制，
并在此基础上研究干预 cofilin 功能的抗肿瘤转移作用。本研究首次发现位于
cofilin Ser108 位 O-GlcNAc 糖基化修饰对细胞侵袭性伪足结构功能维持发挥关
键作用。进一步研究发现，O-GlcNAc 糖基化修饰缺失导致侵袭性伪足结构不稳
定，降低细胞侵袭能力，抑制肿瘤转移。本研究从内源性 O-GlcNAc 糖基化修饰
对 cofilin 功能调节入手，阐明了 cofilin 糖基化修饰在肿瘤细胞迁移侵袭和肿
瘤转移中的重要作用。
关键词：O-GlcNAc 糖基化修饰；骨架蛋白 cofilin；片状伪足；侵袭性伪足；肿
瘤转移
OPB-02
Base lesions recognition by ABH2 and ABH3
Baoen Chen and Cai-Guang Yang*
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road,
Shanghai 201203, China
Abstracts
Aim: To investigate: i) mechanistic insight into DNA substrate selection by ABH2
and ABH3, and ii) how ABH2 detect and process base lesions in the genome.
Methods: ABH2 and ABH3 proteins were overexpressed in E. coli BL21 (DE3)
(Stratagene) and purified using Ni-NTA chromatography (GE Healthcare). Mutation
was introduced using the QuikChange Site-Directed Mutagenesis Kit (Stratagene).
DNA oligonucleotides were synthesized on an Expedite DNA synthesizer according
to the standard solid-phase protocol. Cross-linking reactions between protein and
dsDNA were performed as previously described. Crystals were grown by hanging
drop vapour diffusion. All X-ray data were integrated and scaled using HKL2000
program suite, and converted to structure factors with the CCP4 program. Results:
Swapping the RED motif in ABH3 to equivalent VFG residues in ABH2 endowed
ABH3 with activity in dsDNA repair as efficient as wild-type ABH2. This result
indicates that the RED motif most likely prevents ABH3 binding and repair of dsDNA.
Consistently, swapped ABH3 cross-linked with dsDNA very well, confirming the
determining roles of RED in the initial DNA strand recognition. Crystal structures of
ABH2 bound to duplex DNA containing central C:G, C:I (inosine) pairs have been
determined, respectively. Together with fluorescence studies, the results suggest that
ABH2 probes the stability of base pairs in searching for base damage. Conclusion: i)
The β-hairpin motif plays a crucial role in dsDNA recognition for ABH3. ii) It is
likely that ABH2 detects and flips out damage base in an active way.
Keywords: oxidative demethylation; ABH2; ABH3; mutagenesis; disulfide
cross-linking
OPB-03
Lithium enhances the generation of iPS
Quan Wang, Xin Xie
Abstracts
Somatic cells can be reprogrammed into induced pluripotent stem cells (iPSCs)
by defined factors. The iPSC technology has profound meaning due to its potential
biomedical applications. Patient-specific iPSCs could be created by reprogramming
using this technology and they could be differentiated into functional cells for
cell-based therapy without immuno-compatibility issues and ethical concerns.
However, iPS cell applications are limited by safety issues due to the genomic
integration of oncogenes and incorporation of viral DNA. Many efforts have been
made to put the iPSCs more safety, such as using less number of ectopic factors,
non-integrating gene delivery methods, but under such conditions the reprogramming
efficiency is extremely low. Small molecules that can enhance the generation of iPSCs
or replace the oncogenic factors will be highly valuable. Here we report Li, a drug
used to treat mood disorders, greatly enhances reprogramming in both mouse
embryonic fibroblast (MEF) and human umbilical vein endothelial cells (HUVEC). It
also facilitates the reprogramming of one factor (Oct4)-hiPSCs with combinations of
other compounds. The effect of Li on promoting reprogramming only partially
depends on its major target GSK3. Unlike other GSK3 inhibitors, Li not only
increases the expression of Nanog, it also enhances the transcriptional activity of
Nanog. We also found Li exerts its effect by promoting epigenetic modifications via
downregulation of LSD1, a H3K4 specific histone demethylase. Knocking down
LSD1 partially mimics Li’s effect in enhancing reprogramming.
OPB-04
Liquid-liquid extraction with small n-hexane to sample volume ratio
increases sensitivity of GC/MS-based assay for measurement of
borneol, isoborneol and their metabolite camphor in rat plasma
Chen CHENG, Feifei DU, Fang XU, Meijuan LI,Yan SUN, Chuan LI*
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China
Abstracts
Aims: Bingpian (Borneolum) is an important and classical traditional Chinese
medicine. Chinese pharmacopoeia included borneolum and borneolum syntheticum,
the former dominantly contains (+)-borneol while the latter includes (±)-borneol and
(±)-isoborneol. Because of the present unreliable analytical methods (low sensitivity
or loss of analytes during the solvent evaporation in sample preparation), the earlier
pharmacokinetic studies of borneol were unsuccessful. The aim of this study was to
develop a small organic solvent volume-based liquid-liquid extraction method for
simultaneous determination of borneol, isoborneol and their metabolite camphor in rat
plasma samples. Methods: Different extracting solvent to plasma sample volume
ratios were applied while the relative measurement responses (peak area) and
extraction efficiency of test compounds were examined. The solubility of all analytes
in plasma water and extracting solvent were measured, as well as calculated using
software. Finally, this method was successfully applied to a pharmacokinetic study of
rats after p.o. administration of natural Bingpian (20 mg/kg), synthesized Bingpian
(20 mg/kg), or Fufang Danshen tablets (20 mg synthesized Bingpian/kg) or i.v.
administration of natural Bingpian or synthesized Bingpian (5 mg/kg). Results:
Analytes with high solubility in extraction solvent but much lower solubility in
plasma water were found to be easily extracted using a quite small volume of
extraction solvent with high extraction efficiency. Plasma/extraction solvent
(n-hexane) volume ratio 1:1 was taken for the isolation of all analytes from real rat
plasma samples and the supernatant was directly sent to GC–MS analysis after
centrifugation. The newly developed bioanalytical assay processed favorable accuracy
and precision with lower limits of quantification of 1.95 nM for borneol, 5.8 nM for
isoborneol, and 6.0 nM for camphor, and was successfully applied to pharmacokinetic
studies in rats. Conclusion: Liquid-liquid extraction with small n-hexane to sample
volume ratio increases sensitivity of GC/MS-based assay for measurement of borneol,
isoborneol and their metabolite camphor in rat plasma. The experimental strategies
illustrated in our report may be adopted for measurement of other drugs subject to
volatility or sensitivity problems.
Keywords: Gas chromatography/mass spectrometry；Liquid-liquid extraction with
small n-hexane to sample volume ratio; Borneol; Isoborneol; Camphor; Bingpian
OPB-05
Absorption and Disposition of Scutellarin in Rats: a Pharmacokinetic
Explanation for the High Exposure of its Isomeric Metabolite
Chunying Gao, Xiaoyan Chen and Dafang Zhong
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China
Abstracts
Scutellarin or scutellarein-7-O-glucuronide (S-7-G) is a flavonoid used in the
treatment of cardiovascular diseases. After oral administration to humans, S-7-G can
hardly be detected, whereas its isomeric metabolite (scutellarein-6-O-glucuronide,
S-6-G) dominates in plasma. A preliminary study in rats also revealed a low
bioavailability of S-7-G, as well as a high plasma concentration of S-6-G. Therefore,
the present study tried to explore the possible causes of the unusual pharmacokinetics
of scutellarin in humans through investigating the absorption and disposition of S-7-G
in rats. After oral administration to rats, S-7-G was largely hydrolyzed in the intestinal
tract and was absorbed as aglycone. While passing through the intestinal wall,
aglycone was extensively glucuronidated into S-7-G and S-6-G (about 20:1), which
subsequently entered the mesenteric blood (about 15:1). However, because S-7-G
exhibited more rapid uptake in hepatocytes, was glucuronidated at a 2.7-fold higher
rate in the liver and was excreted in greater amounts through bile and urine than S-6-G,
the S-7-G/S-6-G ratio eventually declined to around 1.5:1 in the systemic circulation.
Findings revealed that S-7-G cannot be absorbed directly; S-7-G and S-6-G in the
body were mostly generated from aglycone in the intestinal wall; a larger amount of
S-7-G than S-6-G entered the mesenteric blood at the absorption stage, but the gap
between them shrank quickly mainly due to the higher hepatic first-pass elimination
of S-7-G. These findings in rats are of great value as reference for the further study to
accurately interpret the pharmacokinetics of S-7-G in humans.
OPB-06
水溶性青蒿素衍生物 SM934 在 MRL/lpr 小鼠狼疮模型中疗效及作用机
制的研究
何世君，侯立飞，李鑫，杨扬，何佩岚，朱峰华，周宇，唐炜，左建平1,2
中国科学院上海药物研究所，免疫药理实验室，
摘要
目的：青蒿素类衍生物 SM934 具有显著的免疫抑制及免疫调节活性。本研究
观察了 SM934 在自身免疫性疾病狼疮小鼠模型（MRL/lpr，雌性）中的疗效作用
及机理。方法：运用 12 周龄和 16 周龄的雌性 MRL/lpr，分别口服灌胃给药 4 周
和 8 周的不同剂量的 SM934，检测药物对 MRL/lpr 的体重、蛋白尿、血清抗体和
肾脏病理变化等指标的影响作用；同时运用免疫学研究方法，探讨了 SM934 治疗
干预狼疮小鼠病程进展的疗效作用机制。结果：（1） SM934 能抑制 TCR 信号介
导的 CD4+ T 细胞活化，IFN-γ 和 IL-17 的分泌，同时抑制初始 CD4+ T 细胞向 Th1
及 Th17 型辅助 T 细胞的分化，而不影响向调节性辅助 T 细胞（Treg）的分化。
（2）SM934 能缓解 MRL/lpr 小鼠的疾病进程，改善蛋白尿及肾脏损伤，降低血
清尿素氮、IFN-γ 及抗双链 DNA 水平，减小异常增大的脾脏体积及致病性双阴
性 T 细胞（CD3+B220+CD4-CD8- T 细胞）比例。
（3）SM934 给药后能提高 MRL/lpr 小
鼠体内的调节性 T 细胞比例，抑制辅助性致病性 Th1 及 Th17 细胞比例，并对调
控淋巴细胞存活、增殖、分化以及细胞因子分泌的关键信号蛋白 STAT1、STAT3、
STAT5 的磷酸化具有显著抑制作用。结论：证实了 SM934 对 MRL/lpr 狼疮小鼠的
疾病进程具有显著的干预和治疗作用，阐明了 SM934 能通过抑制 Th1 及 Th17 型
免疫反应缓解自身免疫疾病进程的疗效机制。
OPB-07
Novel Mucus-penetrating Liposomes with Enhanced Cellular Uptake
for Oral Drug Delivery
Xiuying Li, Yong Gan*
Abstracts
The aim of the study was to investigate intestinal mucus penetrating property and
intestinal cellular uptake of two types of Pluronic F127 modified liposomes. The two
types of Pluronic F127 (F127) modified liposomes, i.e. F127-inlaid liposomes and
F127-adsorbed liposomes, were prepared by a thin-film hydration method followed
by extrusion, in which courmarin-6 was loaded as fluorescence labels. A modified
Franz-diffusion cell loaded with intestinal mucus of rats was used to study the
diffusion efficiency of the two types of F127 liposomes through the rat intestinal
mucus. Cell uptake studies were conducted in Caco-2 cells and analyzed using
confocal laser scanning microcopy (CLSM) and also flow cytometry (FACS). The
results showed that the diffusion efficiency of the two types of F127 modified
liposomes through intestinal mucus were 5-7 folds higher than that of unmodified
liposomes. As compared to unmodified liposomes Pluronic F127-inlaid liposomes
exhibited significantly higher cellular uptake of courmarin-6, while Pluronic
F127-adsorbed liposomes exhibited decreased cellular uptake. Moreover the two
types of F127 modified liposomes revealed different cellular uptake mechanisms in
Caco-2 cell lines. These results suggested that Pluronic F127-inlaid liposomes with
improved intestinal mucus penetrating ability and enhanced cellular uptake efficiency
could be a potential carrier candidate for oral drug delivery.
Keywords: Pluronic F127; Mucus penetrating particles; Liposome; Oral drug
delivery
OPB-08
杏仁核突触骨架重塑调节吗啡
戒断负性情绪记忆的分子机制研究
刘瑶，刘景根*
摘要
药物成瘾表现为对毒品的强烈渴求和强迫性用药，以及撤药后难以忍受的戒
断症状。成瘾性药物戒断所产生的负性情绪记忆是导致强迫性用药的主要原因，
会引起相关脑区发生诸如突触传递和神经结构可塑性等适应性变化。
我们实验室采用巴普洛夫条件位置厌恶模型（Conditioned Place Aversion,
CPA），主要研究吗啡戒断负性情绪记忆中突触可塑性的分子机制。前期研究结果
表明，杏仁核和海马脑区的突触骨架 actin 的结构可塑性变化参与了吗啡戒断负
性记忆的形成。然而，突触骨架 actin 重塑调节这一负性记忆形成的机制还不清
楚。
本文对 CPA 模型中大鼠杏仁核脑区突触结构可塑性进行了进一步研究，通过
蛋白免疫印记杂交、免疫组织荧光、核团定位注射、慢病毒 RNA 干扰、BS3 交联
等技术手段，主要探讨了吗啡戒断引起的负性情绪记忆形成中，突触骨架 actin
重塑对 Arc 转运到突触后膜的作用，以及 Arc 蛋白通过调控 AMPA 受体的内吞，
促进吗啡戒断后负性情绪记忆的形成与巩固的分子机制。
具体的研究结果如下：
1. Actin 重排的发生依赖于 NMDA 受体及 RhoA-ROCK 信号通路的激活。
CPA 模型的形成过程涉及到细胞骨架 actin 重排事件的发生，即 F-actin 含
量增加。杏仁核核团注射 NMDA 受体拮抗剂 D-AP5 或 RhoA-ROCK 抑制剂 Y27632
均能抑制 F-actin 的增加，行为学结果也显示模型配对前核团给予 Y27632 能干
扰 CPA 模型的形成。
2. 细胞骨架 actin 重排是即刻早期基因 Arc 转运到突触活性部位所必须的。
条件化和非条件化 CPA 模型中均能引起 Arc 总蛋白水平的增加，而仅在条件
化 CPA 模型中发生了 Arc 蛋白的突触转运表达和 actin 重排事件。使用 NMDA 受
体阻断剂 D-AP5 能同时干扰 Arc 在总蛋白和突触水平的表达；使用 Y27632、
latrunculin A 等干扰 actin 骨架的重排能阻止 Arc 向突触活性部位的转运表达，
而不会影响总蛋白水平 Arc 蛋白的合成。
3. Arc 蛋白通过调控 AMPA 受体的内吞来影响吗啡戒断负性记忆的形成。
CPA 模型形成过程中存在 AMPA 受体的内吞现象。运用慢病毒 RNA 干扰技术
抑制 Arc 蛋白表达能阻断 AMPA 受体这一内吞过程的发生，并能干扰 CPA 模型的
形成。
4. GluR2 亚基依赖的 AMPA 受体内吞参与了吗啡戒断负性记忆的形成。
杏仁核核团注射 AMPA 受体阻断剂 CNQX 能干扰 CPA 模型的形成，作用于 AMPA
受体 GluR2 亚型 C 末端的特异性多肽 Tat-GluR23Y 能同时阻断杏仁核 GluR1 和
GluR2 亚型 AMPA 受体的内吞，并能干扰 CPA 模型的形成。
综上所述，我们主要探讨了杏仁核脑区突触 actin 的重排调节吗啡戒断负性
情绪记忆的分子机制。本研究首次揭示了急性 CPA 模型形成过程中，突触骨架
actin 重塑的机制及其对即刻早期基因 Arc 向突触转运的重要作用；并研究了 Arc
蛋白对谷氨酸 AMPA 受体内吞的调控，以及 AMPA 受体的内吞对吗啡戒断负性情绪
记忆形成的重要作用。
OPB-09
LGH00168 inhibits tumor growth via mitoROS-NF-
-CHOP
pathway
Yan-min Peng1, Yi-ming Ma1, Qiong-hua Zhu, An-hui Gao, Yu-bo ZHOU, Jia LI*
The National Center for Drug Screening, Shanghai Institute of Materia Medica, Chinese Academy
of Sciences,Shanghai 201203, China
Abstracts
Aim：To identify novel CHOP activator and to further evaluate its anti-cancer effect in
vitro and in vivo. Methods：High-throughput screening (HTS) based on the CHOPpromoter luciferase assay was used to identify novel small-molecule CHOP activator.
A panel of tumor cell lines, such as Hela, HepG2, and so on, was used to evaluate the
cell survival and growth effects. A549 was used in the mechanism study of
LGH00168. The in vivo efficacy of LGH00168 was evaluated in a BALB/c athymic
mouse model. Results：LGH00168 activated CHOP in a time and dose dependent
manner, and it showed beneficial inhibitory effects on tumor survival and growth in
vivo and in vitro. The mechanism of LGH00168 was proved to be through
mitoROS-NF-
-CHOP
pathway.
Conclusion:
Taken
together,
using
high-throughput screening, a novel compound LHG00168 was identified to be CHOP
activator, which showed inhibitory effects on tumor growth in vivo and in intro. These
results demonstrate the potential of small-molecule CHOP activators as a promising
therapeutic approach for the treatment of cancer.
1
Yan-min Peng and Yi-ming Ma contributed equally to this work.
OPB-10
Paeoniflorin Ameliorate Ischemia by Transactivation of Epidermal
Growth Factor Receptor Mediated by Adenosine A1 Receptor
Min Zhong and Linyin Feng*
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road,
Shanghai 201203, China
Abstracts
Aim: Paeoniflorin (PF) could ameliorate brain ischemic/reperfusion injury in rats.
The neuroprotective effects of PF might be mediated by adenosine A1 receptor (A1R).
It is necessary to investigate its mechanism and important for us to develop
neuroprotecive drug against stroke. Methods: In HEK293 cells stably transfected
with A1R (HEK293/A1R) and primary cultured rat cortical neurons, we treated
serum-starved cells with different concentration of PF and detected downstream
signaling pathways by Western Blot. Receptor interactions were detected by
co-immunoprcipitation
and
immunofluorescence
assays.
Oxygen
glucose
deprivation/reperfusion (OGD/R) model was applied to investigate survivability of
neurons. Results: In HEK293/A1R cells, PF activated PI3K/Akt and MAPK/ERK
signaling pathways beginning at 5min while EGFR (Tyr 1068) was also
phosphorylated. A1R antagonist DPCPX and selective Epidermal Growth Factor
Receptor (EGFR) kinase inhibitor AG1478 could abolish ERK1/2 and Akt
phosphorylation which inferred that A1R and EGFR would be involved in signaling
cascade. Furthermore,
A1R and EGFR co-localized on the surface of HEK293/A1R
cells. In HEK293/A1R cells, inhibitor of Matrix Metalloprotease (MMP) BiPs but not
inhibitor of Src kinase PP2 could inhibit ERK1/2 phosphorylation induced by PF. In
cultured cortical neurons, PF protected neurons from OGD/R injury while it
phosphorylated both Akt and ERK1/2.Conclusion: Taken together, PF could promote
cultured neurons survival in OGD/R injury. The mechanism of its neuroprotective
effect was that PF transactivated EGFR signaling cascade which activated both
PI3K/Akt and ERK1/2 signaling pathways mediated by A1R.
Keywords:
Paeoniflorin,
Adenosine
transactivation, ischemic/reperfusion injury
A1
receptor,
EGFR,
neuroprotecion,
OPB-11
基于 LDLR/PCSK9 新靶点的新型化合物降脂作用研究
马艳玲，王逸平*
摘要
背景：低密度脂蛋白受体 LDLR 是肝脏回收摄取低密度脂蛋白 LDL 的主要受体，
其表达水平和血浆胆固醇水平密切相关。PCSK9 蛋白是最近几年才发现的与高
脂血症相关的一个蛋白，分泌性的 PCSK9 蛋白与 LDL 共同被 LDLR 受体摄取入
肝，促使后者在溶酶体内降解，减少其再循环利用率，从而降低细胞表面的
LDLR。实验方法： 我们利用体外的 HepG2 细胞培养，研究化合物在 LDL uptake
中的作用, 用 Western Blot, Real-time PCR 等实验手段研究蛋白和基因的表达变
化。体内采用高脂饮食诱导脂质代谢紊乱的金黄地鼠模型腹腔或口服给药方式评
估化合物的体内降脂效果。结果： ZB 系列化合物通过体外 HepG2 细胞 LDL 吞
噬实验证实时间依赖性和剂量依赖性地增加 LDL uptake。细胞表面的 LDLR 水
平显著上调，这种作用经证实是通过增加 LDLR 基因稳定性和蛋白稳定性共同
实现的。ZB 系列化合物通过降低 PCSK9 基因转录进而影响 PCSK9 蛋白的细胞
内和细胞外的蛋白水平，最后作用于 LDLR，导致后者蛋白稳定性显著上调。上
述作用依赖或部分依赖于于 ERK 通路激活。体内药学学评价证实 ZB 系列中的
ZBH2，ZBM30 两个化合物腹腔注射或口服给药可以降低高脂模型金黄地鼠的血
脂水平，降低其肝脏脂质，同时改善其肝功能指标。结论： 实验证实 ZB 系列
化合物是机制新颖的潜在降脂化合物。
OPB-12
小分子化合物提高诱导性多能干细胞诱导效率
陈国芳，谢欣*
摘要
诱导性的多能干细胞（Induced Pluripotent Stem Cells，iPSCs）即在小
鼠的成纤维细胞上过表达 Oct4, Sox2, Klf4 和 c-Myc 四种转录因子而得到具有
大部分干细胞特性的细胞。近年来，随着基因转染诱导体细胞重编程为干细胞的
成功，干细胞研究进入了一个新的阶段。iPS 细胞研究中存在一些问题，诱导效
率低和逆转录病毒的安全性问题。于是采用化学生物学的策略，将小分子化合物
用于干细胞的自我更新、定向分化以及体细胞重编程等方面的研究，可以避免采
用四种逆转录病毒诱导产生多能干细胞过程中出现的癌变问题。小分子化合物通
过调节干细胞自我更新和分化相关的信号通路能够显著提高重编程效率。
我们通过建立稳定的 iPS 药物筛选体系，在大批的化合物库中筛选能够提
高 iPS 效率、减少外源转录因子，加快 iPS 进程的小分子化合物，并通过进一步
的深入实验研究小分子化合物对于 iPS 的作用机制和 iPS 的重编程机制。通过筛
选，得到了效果较好的小分子化合物 CGF-1，不但能极大提高 ips 效率，同时能
减少外源转录因子的插入，仅在一因子 OCT4 与该化合物存在下即能诱导 ips 细
胞。目前正在进一步研究其促进 ips 效率、加快重编程的分子机制。
OPB-13
Arctigenin efficiently enhanced sedentary mice treadmill endurance
Xuan Tang1,#, Jingjing Zhuang2,#, Jing Chen1,*, Liang Yu1, Lihong Hu1,*, Hualiang
Jiang1, Xu Shen1,2,*
1
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy
of Sciences, Shanghai 201203, China. 2School of Pharmacy, East China University of Science and
Technology, Shanghai 200237, China.
Abstracts
Aim: To investigate the effect of the natural product arctigenin on sedentary mice
endurance and explore its potential underlying mechanisms. Methods: Mice
endurance was evaluated by fatigue test. The effects of arctigenin on AMPK
phosphorylation and downstream genes expression in vitro and in vivo were studied
using Western blot and real-time PCR. Results: The natural product arctigenin from
the traditional herb Arctium lappa L. (Compositae) strongly increased AMPK
phosphorylation and subsequently up-regulated its downstream pathway in both
H9C2 and C2C12 cells. It was discovered that arctigenin phosphorylated AMPK via
calmodulin-dependent protein kinase kinase (CaMKK) and serine/threonine kinase
11(LKB1)-dependent pathways. Mice treadmill based in vivo assay further indicated
that administration of arctigenin improved efficiently mice endurance as reflected by
the increased fatigue time and distance, and potently enhanced mitochondrial
biogenesis and fatty acid oxidation (FAO) related genes expression in muscle tissues.
Conclusion: Our results thus suggested that arctigenin might be used as a potential
lead compound for the discovery of the agents with mimic exercise training effects to
treat metabolic diseases.
OPB-14
新型小分子抗 HBV 候选新药的作用机理研究
杨莉，童贤崑，石丽萍，王桂凤，张仰明，冯春兰，何佩岚，
许怡玢，唐炜，南发俊，左建平
中国科学院上海药物研究所，免疫药理实验室，新药筛选中心
上海浦东张江祖冲之路 555 号 201203
摘要
慢性乙肝病毒（HBV）感染是严重危害人类健康的全球性感染疾病。在中
国，乙肝病毒引起的肝脏疾病位列全国十大死因之一，严重影响了人们正常的工
作和生活。目前，上市的抗乙肝病毒药物主要分为免疫调节剂（如干扰素 ）
和核苷类似物两大类，但是干扰素的低应答率和毒副作用以及核苷类似物的高耐
药突变率等问题仍需要治疗应对。因此，靶向 HBV 聚合酶以外其他作用分子靶
点的新型抗乙肝病毒药物亟待开发。我们在抗 HBV 药物研究中发现和确立了一
个新型小分子抗 HBV 候选新药（NZ-4，异噻氟定）已完成新药临床前研究，正
在申报新药临床研究。NZ-4 抗 HBV 作用和机理的研究结果显示，NZ-4 对 HBV
的复制具有强烈的抑制活性，同时对核苷类药物耐药 HBV 病毒株具有同等抑制
效应，与核苷类抗 HBV 药物的联合用药具有协同增效作用。NZ-4 具有新颖的抗
HBV 作用机制，NZ-4 对 HBV 聚合酶无抑制活性，不抑制 HBV-RNA 的转录，
不影响病毒蛋白的表达，分布和分泌，而是通过干扰病毒核壳体的正常组装来发
挥抗病毒作用。NZ-4 能阻断 pgRNA 与病毒核心蛋白结合，从而抑制 pgRNA 包
装进入核壳体，产生不含病毒核酸的空载病毒颗粒，而失去了 pgRNA 模板的核
壳体，无法完成逆转录过程。
OPB-15
Connexin 43 as Stabilizer of Astrocytes in Stroke-like Milieu
Facilitates Neuronal Recovery Mediated by Inhibiting Upregulation
of Ephrin-A4
Leyu Wu, Linying Feng*
Abstracts
Aim：Astrocytes, taken as the dominant type of glia in central nervous system, would
take place raft of pathological changes during stroke. Amongst these deleterious
alterations, the formation of glia scar and other factors prohibit neurites growth could
disturb neuronal recovery. Despite there are myriad of researches have been
accomplished to attenuate the pathological alteration of astrocytes, aiming to promote
neurites protrusion and neuronal recovery, the explicit molecular stabilizers in
astrocytes were still unknown. Methods：Oxygen Glucose Deprivation /Reperfusion
(OGD/RP) model was utilized to simulate pathological process of stroke. The whole
project was composed of in vitro (primary culture of astrocytes &neuron) and ex vivo
(brain slice culture) experiments. Results: After the treatment of OGD/RP, connexin
43 in ASR was internalized and degraded, leading to the calcium overloading and
upregulation of Ephrin-A4 in protein level. Moreover, the increased intracellular
concentration of calcium might activate CaMKII/CREB and Calcineurin /NFAT,
inducing the expression of Ephrin-A4.This postulation could be testified by
administration of KN-93 (inhibitor of CaMKII) and cyclosporine A (inhibitor of
Calcineurin). Overexpression of connexin 43 and administration of ZBB-1 (analogue
of triptolide, which could inhibit the downregulation of connexin 43) abolished the
above phenomena. Immature neurons stretched shorter neurites and revealed retard
maturation when plated with OGD/RP model astrocytes treated by OGD/RP model.
Conclusion: OGD/RP incurs internalization and degradation of connexin 43, bringing
on calcium overloading. Therein, CaMKII/CREB and Calcineurin/NFAT pathway
were activated, leading to upregulated expression of Ephrin-A4, which would hamper
neuronal recovery.
OPB-16
An Examination of the Potential Effect of Lipids on the First-Pass
Metabolism of the Lipophilic Drug Anethol Trithione
Hongzhen Yu and Yong Gan*
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road,
Zhangjiang Hi-Tech Park, Shanghai, 201203, P. R. China
Abstracts
In this study, an examination of the potential effect of lipids on the first-pass
metabolism of anethol trithione (ATT) was investigated. ATT is metabolized rapidly
and extensively in liver into 4-hydroxy-anethole trithione (ATX), which was
confirmed using the rat intestinal perfusion with the mesenteric cannulation model.
Male Sprague–Dawley rats were orally administered of the lipid-based formulations
(prepared by medium chain triglycerides (MCT)), the cyclodextrin formulation and
the suspension formulation, respectively. For 6.75 mg/kg groups, ATX/ATT area
under the plasma concentration-time curve (AUC) ratio decreased by 87% and 76%
after administration of the MCT-based formulations and the cyclodextrin formulation,
when compared with the suspension formulation (p < 0.05), respectively; for 2.25
mg/kg groups, it decreased by 53% in the MCT group when compared with the
cyclodextrin group (p < 0.05). The saturation of pre-system metabolism of ATT was
observed after administration of the MCT-based formulations and the cyclodextrin
formulation, likely as a result of enhanced absorption and therefore presentation of
higher drug concentrations to liver, when compared with the suspension formulation.
A trend toward lower systemic metabolite to parent ratios was evident after
administration of the lipid formulations, when compared with the cyclodextrin
formulation; however, this was not statistically significant. Further studies on the
potential for lipids to inhibit hepatic metabolism are therefore warranted.
OPB-17
肝脏细胞色素 P450 降低马兜铃酸引起的肾脏 DNA 加合物形成和基因
突变频率
邢国振 戚新明 姚军 武元峰 陈敏 宫丽崑 顾军 能美健彦 栾洋 任进*
中国科学院上海药物研究所
摘要
马兜铃酸（aristolochic acid, AA）是马兜铃酸肾病和巴尔干地方性肾病的主
要致病因素。一般认为 AA 除可引起肾损伤外，还可与 DNA 形成加合物，诱发
突变和致癌。体外研究表明细胞色素 P450（CYP）可以代谢活化 AA 与 DNA
形成加合物，而 CYP 在体内是否参与 AA-DNA 加合物形成和致突变过程迄今未
见报道。首先本研究应用肝脏细胞色素 P450 特异敲除小鼠（Hepatic CYP
Reductase Null mouse, HRN）在肝脏 CYP 诱导、正常和缺失三种情况下比较
了 AAI 的血浆药动学、组织分布以及肾脏 AAI-DNA 加合物形成。结果显示，与
肝脏 CYP 功能正常的野生型小鼠（WT）比较，经肝脏 CYP1A 诱导剂 β-萘黄酮
（β-naphthoflavone，BNF）预处理后的 WT 小鼠 AAI 消除加快，组织内 AAI
原形和还原产物浓度减少，肾脏 AAI-DNA 加合物含量明显减少（p=0.0002）；
HRN 小鼠 AAI 消除减慢，组织内 AAI 原形和还原产物浓度增加，肾脏 AAI-DNA
加合物含量显著增加（p=0.0004）。研究中只在 HRN 小鼠肝脏内检测到 AAI-DNA
加合物。为进一步探索研究肝脏 CYP 在体内对 AAI 肾脏致突变中的作用，本研
究将 HRN 小鼠与 gpt delta 转基因小鼠杂交，建成肝脏细胞色素 P450 缺陷型
gpt delta 转基因小鼠（HRN/gpt）。利用新建小鼠模型研究发现灌胃给予 AAI 15
mg/kg，每 w 1 次，连续 4 w 后，与野生型小鼠（WT/gpt）相比 HRN/gpt 小鼠
肾脏 gpt 基因点突变频率明显增加（p=0.003）。基因序列比对发现 WT/gpt 和
HRN/gpt 小鼠的点突变类型主要是 AT→TA。综上，本研究应用小鼠模型首次证
明体内肝脏 CYP 降低 AAI 引起的肾脏 DNA 加合物形成和基因突变频率。本研
究结果为深入开展马兜铃酸毒性机制研究和中药安全性评价提供新思路。
OPB-18
(+)-Rutamarin as a dual activator of both GLUT4 translocation and
expression efficiently ameliorates glucose homeostasis in mice
Xin-gang Yao1,3Yu Zhang1,3, Haitao Zhang1,3, , Hong Shen1, Jing Chen1, Chenjing Li1,
Lili Chen1, Mingyue Zheng1, Jiming Ye2, Lihong Hu1*, Xu Shen1* and Hualiang
Jiang1*
1
Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute
of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Zhangjiang Hi-Tech
Park, Shanghai, 201203, P. R. China 2School of Medical Sciences, University of New South Wales,
Sydney, NSW, Australia. 3These authors contributed equally to this work.
Abstracts
Aim: To discovery activators on GLUT4 translocation or expression for anti-T2DM.
Methods: For discovery of the potent dual activators on both GLUT4 translocation
and expression, we constructed an In Cell Analyzer 1000 based platform, and the lab
in-house natural product library (~5,000) was screened. The determined activator was
subsequently applied for relevant pathways and targets exploration. Finally, we
evaluated the effects of the compound on glucose homeostasis and insulin sensitivity
in diet-induced obese (DIO) mice. Results: The natural product (+)-rutamarin (Rut)
was discovered as an efficient dual activator on both GLUT4 translocation and
expression. Rut induced GLUT4 translocation through PI3 kinase-AKT/PKB pathway
by functioning as a protein tyrosine phosphatase 1B (PTP1B) inhibitor, and increased
GLUT4 expression as a selective agonist of retinoid X receptor 
). By using
molecular modeling and crystallographic approaches, we studied the binding modes
of Rut to these two targets at atomic levels. Rut-treated 3T3-L1 adipocytes exhibited
much enhanced insulin-induced glucose uptake, while DIO mice based assay further
confirmed the significant Rut-induced improvement of glucose homeostasis and
insulin sensitivity in vivo. Conclusion: Our results thereby indicated that Rut could
serve as an attractive lead compound for anti-T2DM drug discovery, and also a
potential chemical probe for addressing GLUT4 associated pathway.
OPB-19
Compound yhhu981, derivated from resveratrol, regulates fatty acid
metabolism homeostasis by AMPK activation
Hongliang Zeng, Ying Leng*
Abstracts
Aim: To investigate effects of the newly identified resveratrol analogue, compound
yhhu981, on fatty acid metabolism and to find out the possible underlying mechanism.
Methods: Differentiated C2C12 cells and HepG2 cells were pretreated with or
without various inhibitors, followed by compound yhhu981 treatment, fatty acid
oxidation or fatty acid synthesis was measured. Ob/ob mice fed with high-fat diet
were administrated with compound yhhu981 once daily, plasma non-esterified free
fatty acid (NEFA), triglyceride (TG), total cholesterol (TC), gastrocnemius TG and
liver TG content were assessed at the end of treatment. AMP-activated protein kinase
(AMPK) and acetyl coenzyme A carboxylase (ACC) phosphorylation levels were
determined. Results: In vitro studies showed that compound yhhu981 could promote
fatty acid oxidation in differentiated C2C12 cells and prevent fatty acid synthesis in
HepG2 cells. It increased AMPK and ACC phosphorylation level, and upregulated
fatty acid oxidation related genes expression in skeletal muscle cells. The yhhu981
enhancement of fatty acid oxidation and suppression of hepatic fatty acid synthesis
were prevented by inhibitors of AMPK, while inhibitors for SIRT1 partially inhibited
this effect. In vivo studies showed that ob/ob mice plasma NEFA and gastrocnemius
TG accumulation were markedly decreased by 3 weeks of yhhu981 administration,
while plasma TG, TC and liver TG content were decreased but without significantly
difference. Conclusion: These results indicated that compound yhhu981 could
regulate fatty acid metabolism homeostasis by increasing fatty acid oxidation and
reducing lipids accumulation possibly by activation of AMPK.
OPB-20
CCR6 小分子拮抗剂对自身免疫性疾病 EAE 的治疗性作用
张菲菲，谢欣*
中国科学院上海药物研究所
摘要
人类多发性硬化症(Multiple selerosis , MS)是一种神经退行性自身免疫
性疾病，临床表现为视力下降或失明及四肢无力甚至瘫痪的症状。该疾病的产生
主要是由于炎症细胞 Th1 及 Th17 进入中枢神经系统引起神经细胞轴突脱髓鞘病
变。目前针对该疾病的治疗药物主要有抑制 T 细胞增殖类药物如干扰素，米托蒽
醌等，抑制 T 细胞迁移类药物如那伐他珠单抗及抑制炎症因子产生的药物如他汀
类等。但这些药物存在着许多缺陷，如多为免疫抑制剂或者需要口服等，因此，
研发出一批针对特异性靶点的小分子化合物将使 MS 患者的用药选择翻番，且口
服药也是人们最为期盼的治疗手段。
Th17 被认为在 MS 中发挥非常重要的作用，而 Th17 特异性高表达趋化因子
受体 CCR6，预示着 CCR6 对 Th17 细胞向炎症部位的迁移有重大贡献。所以，我
们以 CCR6 为靶点，筛选其小分子拮抗剂，以期抑制 Th17 细胞向中枢神经系统的
迁移，进而缓解疾病的发展。
目前，我们已经筛选到一个小分子化合物能够抑制 CCR6 的功能，并且在模
拟 MS 疾病的小鼠模型实验性自身免疫性脑脊髓炎(Experimental Autoimmune
Encephalomyelitis,EAE)中也可以非常有效的缓解 EAE 的病情。
OPB-21
A novel compound Activates AMP-activated Protein Kinase with
beneficial effects for type 2 diabetes
Li-Na Zhang, Lie Xu, Jing-ya Li*, Jia Li*
The National Center for Drug Screening, Shanghai Institute of Materia Medica, Chinese Academy
of Sciences,Shanghai 201203, China
Abstracts
Aim：To discover novel AMPK activator and to further evaluate the compound in
vitro and in vivo. Methods：Using a homogeneous scintillation proximity assay to
identify new small-molecule AMPK activator. L6 myotubes，Hela and rat primary
hepatocytes were used in cell level evaluation. The in vivo efficacy of ZLN024 was
evaluated in a BKS db/db diabetic mouse model with a dose of 15 mg·kg-1·d-1 for 6
weeks. Results：ZLN024 allosterically stimulated active AMPK heterotrimers and
inactive AMPK α1 subunit truncations. It has no effect on 2 or S. pombe 
truncations. It activated AMPK in L6 myotubes without increasing AMP:ATP ratio. It
also activated AMPK in primary hepatocytes and Hela cells. ZLN024 stimulated
glucose uptake and fatty acids oxidation in L6 myotubes. It decreased glucose output
in primary hepatocytes and mRNA level of glucose production genes. ZLN024 also
inhibited FAS, mtGPAT and HMGR genes expression. Treatment of db/db mice with
ZLN024 reduced blood glucose and improved glucose tolerance; reduced liver weight,
triglycerides and total cholesterol content. It also mildly decreased anabolic genes
expression in liver, meanwhile increased catabolic genes expression. Conclusion: A
novel compound was discovered to be AMPK activator and show beniffical effect for
type 2 diabetes. These results demonstrate small-molecule activators of AMPK
provide a novel approach to treat metabolic syndrome.
OPB-22
MiR-10b and miR-149 mediate tumor angiogenesis and growth
Xiaokun Shen, Jie Li, Kan Ding*
Abstracts
Aim: MicroRNAs (miRNAs) are short noncoding RNAs involved in gene expression
regulation under physiological and pathological situations. More and more evidence
showed that miRNAs were implicated in tumor angiogenesis and tumor growth.
Elucidation the specific roles of miRNAs in the regulation of genes involved in tumor
angiogenesis and tumor growth is proving to be extremely significant in
understanding tumor development. The aim of this study is to investigate the function
of miR-10b and miR-149 in tumor angiogenesis and growth. Methods: Microarray
and in situ hybridization were employed to analyze abnormal expression of miRNAs,
and miRNA qRT-PCR was used to verify these changes. Dual luciferase report assay
and western blotting were employed to confirm the miRNAs’ target. Scratch wound
migration assay, transwell assay, tube formation on matrigel assay and angiogenesis
in vivo model were used to assess miRNAs’ effect on angiogenesis. MTT, FACS and
PI/Annex-V assays were used to determine effect of miRNAs on tumor growth and
apoptosis. Results: Firstly, we show that miR-10b is down-regulated by
antiangiogenetic heparin’s stimulation in human microvascular endothelial cells
(HMEC-1). Further, overexpression of miR-10b induces HMEC-1 cell migration, tube
formation and angiogenesis, and downregulates homeobox D10 (HoxD10) expression
via direct binding of miR-10b to the putative 3’UTR of HoxD10. Using quartz crystal
microbalance (QCM) analysis, we also show that heparin binds to thrombin, thereby
inhibiting thrombin induced expression of Twist and miR-10b. Interestingly, we find
that heparin attenuates miR-10b expression and induces HoxD10 expression in vivo to
inhibit angiogenesis and impairs the growth of MDA-MB-231 tumor xenografts.
These results provide insight into the molecular mechanism by which heparin and
thrombin
regulate
angiogenesis
(Shen
X,
et
al.
J
Biol
Chem.
2011,
286:26616-27).Secondly, by analyzing abnormal expression of miRNAs gliomas
tissure, we find that miR-149 was lowly expressed in human brain but up-regulated in
both low-grade and high-grade gliomas. We also showed that overexpression of
miR-149 by lentivirus vector promoted glioma cell growth and resisted to apoptosis
induced by cisplatin. Further study indicated miR-149 facilitated G1/S-phase
transition. This effect could be reversed by anti-miR-149 in glioma cells. We
documented interactions between miR-149 and the 3'UTR of Caspase-2. These
cellular responses are mediated by reduced expression of direct target Caspase-2,
since siRNA for Caspase-2 also caused these alterations. Furthermore, specific
inhibition of miR-149 with antisense oligonucleotides led to elevated levels of
Caspase-2 and therefore reduced glioma cell growth by arresting cell cycle or/and
inducing cell apoptosis. Finally, miR-149 overexpression in a mouse model of human
glioma resulted in significant tumor growth. In vivo study also indicated that
Caspase-2 expression was downregulated by miR-149 in tumor tissue. Altogether, our
experiments validated an important role of miR-149 in gliomagenesis, revealed a
novel mechanism of miR-149-mediated regulation, and suggested the possibility of its
future use as a therapeutic target in gliomas. Conclusion: MiR-10b induces tumor
angiogenesis by targeting HoxD10. MiR-149 targets caspase-2 to promotes glioma
cell growth and resists to apoptosis.
OPB-23
Suppressing the mitochondrial permeability of oligomeric A1-42 by
huperzine A contributes to the ameliorating effect on mitochondrial
abnormality
Ling Yang, Haiyan Zhang*
Abstract
Interestingly, recent studies indicate that interfering mitochondrial function
might be involved in the beneficial profiles of huperzine A, an active herbal medicine
for the treatment of Alzheimer's disease (AD) in China. As an extension, the current
study for the first time demonstrates the protective effect of huperzine A on
oligomeric -amyloid1-42 (A1-42) induced reduction in cell survival, mitochondrial
membrane potential, ATP level, and overproduction in reactive oxygen species in
cultured primary cortical neurons (cellular level). Moreover, the ameliorating effect of
huperzine A is further validated on oligomeric A1-42 induced adenosine
5’-triphosphate (ATP) reduction on percoll-purified brain cortical mitochondria
(subcellular level). Our in-depth study indicates that the above-mentioned protective
effects by huperzine A are associated with the improvement of the activities of
respiratory chain complexes, especially complex II-III and complex IV, which may
attribute to the blockage of oligomeric A1-42 from penetrating into mitochondria.
These results shed more light on a potential direct target of huperzine A on isolated
mitochondria, which may be largely different from its well-recognized specific
inhibition on acetylcholinesterase (AChE). Above information would provide crucial
information for better understanding the beneficial profiles of this drug and
discovering novel potential pharmacological target for AD therapy.
OPB-24
Extinction of morphine withdrawal-induced conditioned place
aversion requires CREB-dependent chromatin regulation of BDNF
transcription in the rat ventromedial prefrontal cortex via activation
of ERK signaling pathway
Wei-Sheng WANG1, Shuo KANG1,, Wen-Tao LIU2, Mu LI1, Yao LIU1, Chuan
YU1,Jing-Gen LIU1 *
1
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese
Academy of Sciences, Shanghai 201203, China
2
School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210046, China
Abstracts
The drug-associated conditioned stimulus(CSs) come to influence drug-seeking and
relapse behaviour through the memories they evoke. The animals undergo extinction
training (no drug ) will decrease response rates and extinction entails new learning
rather than forgetting. In the model of conditioned place aversion (CPA) of acute
morphine-dependent rats, we found that lesions of ventral medial prefrontal cortex
(vmPFC) impaired the consolidation of extinction of CPA. In addition, we found that
both extracellular signal-regulated kinases (ERK) pathway and histone acetylation in
vmPFC are involved in consolidation of extinction and also found that Trichostatin A,
a histone deacetylase (HDAC) inhibitor,can facilitate the extinction of CPA. We
further found that manipulation of vmPFC ERK activity by its antagonist U0126 and
agonist D-cycloserine not only differentially influenced the behavior of CPA rats but
also altered histone acetylation. Furthermore, we found that extinction of CPA
increased BDNF exon I but not exon IV in the vmPFC, We further found that the
increasement is accompanied by a significant increase in histone H3 acetylation and
pCREB around the BDNF exon I gene promoter。We also found that inhibition of trk
B by intra-vmPFC injections of K252a significantly disrupted the consolidation of
extinction. Therefore, our findings suggest that histone acetylation is mediated by
ERK pathway and the increasement of BDNF protein expression in vmPFC are
required for the consolidation of CPA extinction and the tyrosine kinase inhibitor
K252a prevented the consolidation of CPA.
Keywords: extinction; vmPFC; ERK; histone acetylation; BDNF
OPB-25
Altered cytochrome P450 expression in hepatocyte-specific Dicer1
knockout mouse livers
Lingling Miao (Dr. Jin Ren’s lab)
Abstracts
Background:Cytochrome P450 (CYP) is
a
superfamily of hemethioloate
monooxygenase enzymes that are involved in the oxidative metabolism of a number
of endogenous and exogenous compounds such as steroids, drugs, carcinogens and
mutagens. CYP enzymes exhibit high interindividual variability that is not completely
explained by known environmental and genetic factors. To further understand this
variability, we hypothesized that microRNAs may regulate CYP expression.
Methods:Dicer1, an enzyme essential for the processing of microRNAs, was
disrupted in hepatocytes using a conditional knockout mouse model to elucidate the
consequences of loss of microRNAs. Albumin-Cre and Dicer1loxP/loxP mice were
crossed to obtain hepatocyte-specific Dicer1 knockout mice (Albumin-Cre;
Dicer1loxP/loxP mice).
Human hepatoma PLC/PRF/5 cells which endogenously
expressed CYP2E1 were used for the in vitro experiments. CYP mRNA and protein
expression were detected by real-time quantitative reverse transcription-PCR and
western blot. Results:We analyzed expression of four Cyps (Cyp2e1, Cyp1a2,
Cyp3a11, Cyp2b10) in Dicer1-deficient and control mouse livers at four time points
(3, 4, 5, 6 weeks after birth). Cyp2e1 mRNA and protein levels were reduced in
3-week-old Dicer1-deficient liver, a time point at which Dicer1 is efficiently
eliminated. The reduction of Cyp2e1 expression was progressive with age. Decrease
of Cyp1a2 and Cyp3a11 expression was only seen at the 5 and 6 weeks time points
(mRNA and protein level). Cyp2b10 expression was also not changed at the first two
time points, while increased dramatically at 5 and 6 weeks after birth. Knockdown of
DICER1 in human hepatoma PLC/PRF/5 cells lead to decrease of CYP2E1 mRNA
and protein expression. Conclusions:As expression of Cyp2e1 changed in the early
stage after Dicer1 was knocked out in mouse livers, consistent with the result that
CYP2E1 expression was changed in DICER1 knockdown human cells, we concluded
that CYP2E1 could be regulated by DICER1, highly possible through microRNAs.
More interestingly, DICER1 deficiency lead to down-regulation of CYP2E1,
indicating that CYP2E1 might be positively regulated by a microRNA that could be
processed by DICER1, or negatively regulated by a microRNA which was
up-regulated by DICER1 deficiency.
OPB-26
Bioreducible poly (β-amino esters)/shRNA complex nanoparticles for
efficient RNA delivery
Yin Qi, Li Yaping*
Abstracts
Aim: To design new non-viral vectors with high transfection efficiency and low
toxicity for RNA interference (RNAi) mediating gene silencing.
Methods: Three novel bioreducible poly (β-amine esters) (PAEs) with different
amino monomers in the main chain were designed and synthesized by Michael
addition polymerization. The ability of the PAEs to condense shRNA and the
physical-chemical characteristics of the PAEs/shRNA complex nanoparticles (PAENs)
were investigated. In vitro cellular uptake, RNAi and cytotoxicity experiments were
carried out in HEK-293 cells and U-87 MG cells. The biodistribution and in vivo
RNAi efficiency of PAENs in BALB/c mice bearing U-87 MG-GFP tumor were
studied.
Results: All PAEs could condense shRNA into complex nanoparticles with particle
size (60–200 nm) and positive surface charges (N+10 mV). The PAENs were stable
under the extracellular physiological condition, while it would degrade in the
reductive environment due to the cleavage of the disulfide bonds in the PAEs main
chain. PAENs could achieve efficient cellular uptake and EGFP silencing in HEK-293
cells and U-87 MG cells with low cytotoxicity. The high accumulation of PAENs in
tumor and high silencing efficiency of intratumor EGFP expression occurred when
PAENs were intravenously injected into BALB/c mice bearing U-87 MG-GFP tumor.
Conclusion: The PAENs could be a promising non-viral vector for efficient RNA
delivery.
Keywords: RNA interference; non-viral; poly (β-amine esters); bioreducible
OPC-01
Knowledge-Based Scoring Functions in Drug Design: 2. Can the
Knowledge Base Be Enriched?
Qiancheng Shen, Bing Xiong, Mingyue Zheng, Xiaomin Luo, Cheng Luo, Xian
Liu,Yun Du, Jing Li, Weiliang Zhu, Jingkang Shen, and Hualiang Jiang
Abstracts
Aim: Fast and accurate predicting of the binding affinities of large sets of diverse
protein-ligand complexes is an important, yet extremely challenging, task in drug
discovery. The development of knowledge-based scoring functions exploiting
structural information of known protein-ligand complexes represents a valuable
contribution to such a computational prediction. Methods: In our approach, the
functions and atom types of PMF04 were inherited to implicitly capture binding
effects that are hard to model explicitly, and a novel iteration device was designed to
gradually tailor the initial potentials. Results: We evaluated the performance of the
resultant IPMF with a diverse set of 219 protein-ligand complexes and compared it
with seven scoring functions commonly used in computer-aided drug design,
including GLIDE, AutoDock4, VINA, PLP, LUDI, PMF, and PMF04. While the
IPMF is only moderately successful in ranking native or near native conformations, it
yields the lowest mean error of 1.41 log Ki/Kd units from measured inhibition
affinities and the highest Pearson's correlation coefficient of Rp2 0.40 for the test set.
Conclusion: These results corroborate our initial supposition about the role of
“enriched” knowledge base. With the rapid growing volume of high-quality structural
and interaction data in the public domain, this work marks a positive step toward
improving the accuracy of knowledge-based scoring functions in binding affinity
prediction.
Keywords: Protein-ligand Interaction; Scoring Function; PMF
OPC-02
鉴定吡咯里西啶生物碱新型谷胱甘肽结合型代谢物
陈美霞，李亮，钟大放，陈笑艳*
摘要
吡咯里西啶生物碱是一类具有四氢吡咯环结构的生物碱，广泛分布于 6000
多种植物中。这类化合物在体内经肝脏代谢生成具有亲电活性的吡咯代谢中间
体，与蛋白质和 DNA 发生共价结合导致毒性，活性中间体也可与谷胱甘肽形成结
合物。本实验采用 UPLC/Q-TOF MS 系统研究了大鼠体内吡咯里西啶生物碱代谢生
成的谷胱甘肽结合型代谢物。大鼠腹腔注射大黄橐吾碱后，收集 0-24 h 胆汁和
尿。在大鼠肝微粒体中检测到三个单谷胱甘肽结合物和两个双谷胱甘肽结合物。
合成代谢物对照品并鉴定结构。在大鼠胆汁中，谷胱甘肽结合物生成情况与微粒
体中一致，其中主要的单谷胱甘肽结合物其色谱行为与已鉴定的 7 位谷胱甘肽结
合物（7-G-D）不符，而对应于 9 位谷胱甘肽结合物（9-G-D），这是实验中新发
现的一个代谢物。另外两个次要的单谷胱甘肽结合物则对应于 7(R)-G-D 和
7(S)-G-D。该结果与文献中大鼠肝微粒体和胆汁中主要单谷胱甘肽结合物为
7-G-D 的报道不一致。在大鼠尿中仅检测到 7(R)-G-D 和 7(S)-G-D 两个结合物，
而未检测到相应的 9-G-D。采用倒千里光碱和野百合碱进行大鼠体内实验，谷胱
甘肽结合物生成情况与大黄橐吾碱一致。本实验研究了大鼠胆汁和尿中吡咯里西
啶生物碱谷胱甘肽结合型代谢物，并发现了一种新型的谷胱甘肽结合型代谢物，
为吡咯里西啶生物碱致毒机制研究提供了更丰富的信息。
*
Corresponding author: Tel./Fax: + 86 21 50800738; Email: xychen@mail.shcnc.ac.cn
OPC-03
海洋天然产物 Bengamides 类似物的设计与合成
张润涛 南发俊*
中国科学院上海药物研究所
摘要
基于海洋天然产物 Bengamides 广泛而优异的抗肿瘤活性，以 Novatis 开
发的活性小分子化合物 LAF389 为研究起点，通过合理药物设计，合成了一系列
结 构 新 颖 活 性 优 异 的 Bengamides 类 化 合 物 ， 并 在 此 基 础 上 总 结 得 到 了
Bengamides 类化合物的构效关系。
OPC-04
Synthesis and c-Met Kinase Inhibition of 3,5-Di- and 3,5,7-TriSubstituted Quinolines: Identification of 3-(4-Acetylpiperazin-1-yl)5-(3-nitrobenzylamino)- 7- (trifluoromethyl)quinoline as a Novel
Anticancer Agent2
Yuanxiang Wang,†,§ Jing Ai,‡,§ Yi Chen‡, Lu Wang,‡ Gang Liu,† Meiyu Geng,‡,Ao
Zhang†*
Abstracts
By using an improved synthetic strategy, a series of 3,5-di- and 3,5,7-tri- substituted
quinolines were readily prepared. Among the 3,5-di-substituted quinoline subseries,
some compounds showed pronounced c-Met activity with IC50 values of 140, 54 and
110 nM respectively. Remarkable enhancement was achieved by introducing a
C7-trifluoromethyl group into these quinolines and seven 3,5,7-tri-substituted
quinolines were identified as the most potent c-Met inhibitors in current study
showing IC50 values less than 1.0 nM. Compound 21B showed the most promising
overall PK profile, and therefore was selected for further characterization both in vitro
and in vivo. Compound 21B effectively inhibited the proliferation of multiple human
cancer cell lines. At doses of 100 mg/kg, compound 21B showed statistically
significant tumor growth inhibition (63-65%) in both NIH-3T3-TPR-Met and U-87
2
Yuanxiang Wang, Jing Ai, Yi Chen, Lu Wang, Gang Liu, Meiyu Geng, Ao Zhang. J. Med. Chem. 2011, 54, 2127–2142.
MG human gliobastoma xenograft models, and no significant body weight loss was
observed. These results clearly indicated that compound 21B is a novel potent and
selective c-Met inhibitor, and its favorable in vitro and in vivo profiles warrant for
further investigation.
OPC-05
Design of Novel Sulfur-Based Olefin Ligands for Asymmetric
Catalysis
Wei-Yi Qi, Ting-Shun Zhu, Ming-Hua Xu*
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road,
Shanghai 201203, China
Email: xumh@mail.shcnc.ac.cn
Abstracts
Chiral olefins have recently attracted considerable attention as new steering ligands
and evoked great interest from the chemical society1. Nevertheless, despite the fact
that heteroatom-olefin hybrid ligands generally exhibit increased coordination ability
to transition-metals, their development and use in asymmetric catalysis is far less
extensive than that of diene ligands, only rare examples of pnictogen-based olefin2
analogues have been disclosed in the literature. On the other hand, catalytic
asymmetric transformations applying sulphur-based ligands have been more
intensively investigated in recent years. Compared to the phosphorus- or
nitrogen-containing ligands, chiral sulphur ligands present advantages of easy
synthesis, high stability, good coordination ability and special S-stereogenic control.
Therefore, it is reasoned that a structurally proper olefin containing heteroatomic
sulphur could possibly act as a previously unexplored sulphur-olefin hybrid class
ligand, which should offer new opportunities for asymmetric applications.
Fig. 1- Chiral Sulfoxide-Olefin Ligand Design
Herein, we disclose a new class of sulfoxide-olefin hybrid ligands featured with a
chiral t-butyl sulfinyl group attached to a trans-stilbene structure in 1,2-fasion3. These
ligands can be easily prepared in short steps and are air stable. Preliminary
investigation of the activity and selectivity of these ligands in Rh-catalyzed
1,4-addition of arylboronic acids to α,β- unsaturated ketones exhibit promosing results.
By changing different substituted groups in ligands and screening the reaction
conditions, the product can be obtained with up to quantitative yield and 98% ee.
References:
1. For leading reviews on chiral diene ligands in asymmetric catalysis, see: (a) Glorius, F.
Angew. Chem., Int. Ed. 2004, 43, 3364. (b) Defieber, C.; Grützmacher, H.; Carreira, E. M.
Angew. Chem., Int. Ed. 2008, 47, 4482. (c) Shintani, R.; Hayashi, T. Aldrichim. Acta. 2009,
42, 31.
2. For leading references on pnictogen-based olefin ligands, see: (a) Maire, P.; Deblon, S.;
Breher, F.; Geier, J.; Böhler, C.; Rüegger, H.; Schönberg, H.; Grützmacher, H. Chem.- Eur.
J. 2004, 10, 4198. (b) Duan, W.-L.; Iwamura, H.; Shintani, R.; Hayashi, T. J. Am. Chem.
Soc. 2007, 129, 2130. (c) Liu, Z.; Du, H. Org. Lett. 2010, 12, 3054. (d) Hahn, B. T.; Tewes,
F.; Fröhlich, R.; Glorius, F. Angew. Chem., Int. Ed. 2010, 49, 1143.
3. Qi, W.-Y.; Zhu, T.-S.; Xu, M.-H. Org. Lett. 2011, 13, 3410.
OPC-06
苦楝子（Melia azedarach）化学成分及活性的研究
刘洪兵，岳建民
摘要
Three limonoids (1–3), and two triterpenes (4) and (5), along with twelve known
compounds, were isolated from the seeds of Melia azedarach. Their structures were
established on the basis of extensive spectroscopic analysis. Compound 3 showed
moderate antimicrobial activity.
OPC-07
基于配体结构的 Grb2-SH2 抑制剂的结构优化
智英, 彭电, 龙亚秋*
（中国科学院上海药物研究所新药研究国家重点实验室，上海，201203）
摘要
Grb2－SH2 是一个 25 KDa 的接头蛋白，由于在活化致癌的 Ras 的信号传导
途径中起关键作用，因此已成为抗肿瘤药物设计的重要靶标。我们从噬菌体库展
示得到的非磷酸环肽出发，基于配体与蛋白相互作用的结构特征，开展了结构优
化研究，前期的构效关系研究显示氮端以大的疏水性基团保护较氮端裸露的化合
物活性高。我们以此为基础，同时基于 Grb2-SH2 与配体相互作用的结构特征，
设计合成了一系列的非磷酸 Grb2-SH2 拟肽抑制剂，以考察氮端保护基对活性的
影响，以及 X+1 以不同的氨基酸诱导形成的 β-折叠对 X0 位置氨基酸的结构要求。
从中发现化合物 LSG-105 对人乳腺癌细胞株 MDA-MB-453 表现出非常好的抑制活
性(IC50=9.22 μM)，我们推测可能是因为 Ac6c 诱导的 β-转角使得 α,α-双取
代氨基酸的酸性边链能够与 Grb-SH2 蛋白的结合口袋相结合，这就弥补了关键氨
基酸 Tyr 的缺失。这一猜想从计算机模拟的 LSG-105 与 Grb-SH2 蛋白的结合模式
中得到支持。
我们基于配体结构的构效关系研究产生了低电荷的拟肽 Grb2-SH2 抑制剂，
在磷酸酪氨酸甚至酪氨酸缺失的前提下依然具有优秀的 Grb2-SH2 结合活性以及
肿瘤细胞增殖抑制活性，为发展具有生物相容性和临床应用前景的 Grb2-SH2 抑
制剂提供了新颖的先导化合物和结合模式。
OPC-08
思茅红椿中的化学成分研究
刘佳，岳建民*
摘要
Ten limonoids, toonacilianins AJ, and two norlimonoids, toonacilianins K and L,
together with seven known compounds were isolated from the stems of Toona ciliata
var. henryi (Meliaceae). Their structures were elucidated by spectroscopic analysis.
Two compounds showed strong cytotoxic activities.
OPC-09
Catalytic Mechanism Investigation of Lysine-Specific Demethylase 1
(LSD1): A computational study
Xiangqian Kong, Shisheng Ouyang, Zhongjie Liang, Junyan Lu, Fei Ye, Bairong
Shen, Donghai Li, Mingyue Zheng, Keqin Kathy Li, Cheng Luo and Hualiang Jiang
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy
of Sciences, 555 Zu Chongzhi Road, Shanghai 201203, China
Abstracts
Lysine-specific demethylase 1 (LSD1), the first identified histone demethylase, can
steadily regulate the transcription level of related genes. It participates in a broad
spectrum of biological processes and is of high importance in cell reprogramming,
embryonic development and cell proliferation. Till now, the catalytic mechanism for
the rate-limiting reductive half-reaction in demethylation remains controversial. By
employing a combined computational approach included molecular modeling,
molecular dynamics (MD) simulations and quantum mechanics/molecular mechanics
(QM/MM) calculations, the catalytic mechanism and key residues involved in
demethylation were characterized in details. Our data indicated that the reductive
half-reaction of LSD1 employed the direct hydride transfer mechanism. Our findings
from this study shed light on the discovery of novel mechanism-based modulator and
are beneficial to unveil the crucial role of LSD1 in cell programming and
reprogramming.
Keywords LSD1; Catalytic mechanism; Hydride transfer; Molecular Dynamics
simulations; QM/MM
Figure. The potential energy surface of the hydride transfer reaction in LSD1
demethylation and the corresponding minimum reaction path.
Reference
1． Shi Y, Lan F, Matson C, Mulligan P, Whetstine JR, et al. Histone demethylation mediated
by the nuclear amine oxidase homolog LSD1. Cell, 2004, 119: 941-953.
2． Stavropoulos P, Blobel G, Hoelz A. Crystal structure and mechanism of human Lysine
specific demethylase-1. Nature Structural & Molecular Biology, 2006, 13: 626-632
3． Forneris F, Battaglioli E, Mattevi A, Binda C. New roles of flavoproteins in molecular cell
biology: histone demethylase LSD1 and chromatin. FEBS Journal , 2009, 276: 4304-4312.
4． Foster CT, Dovey OM, Lezina L, Luo JL, Gant TW, et al. Lysine-specific demethylase 1
regulates the embryonic t ranscriptome and CoREST stability. Molecular and Cellular
Biology, 2009, 30: 4851-4863.
5． Kong XQ, Ouyang SS, Liang ZJ, Lu JY, Ye F, Shen BR, Li DH, Zheng MY, Li KQ, Luo C
and Jiang HL,,XXXXXXXXX, PLoS ONE, revised.
__________________________
*This work was supported by grants from the State Key Program of Basic Research of China grant
(2009CB918502), the National Natural Science Foundation of China grants (20972174, 21021063 and
91029704), Shanghai Committee of Science and Technology grants (10410703900), and the Chinese
Academy of Sciences (XDA01040305).
Phone: 86-21-50806600; Fax: 86-21-50807088; E-mail: cluo@mail.shcnc.ac.cn (Cheng Luo).
OPC-10
Design and Synthesis of First-in-Class Small Molecule RhoA
Inhibitors: A New Promising Therapy for Cardiovascular Diseases?
Enguang Feng, Jing Deng, Sheng Ma, Yan Zhang, Xiaofeng Liu, Honglin Li, Huang
Huang, Jin Zhu, Weiliang Zhu, Xu Shena, Liyan Miao, Hong Liu, Hualiang Jiang, Jian
Li
Abstract
RhoA is a member of Rho GTPases, a sub-group of the Ras superfamily of small
GTP-binding proteins. The RhoA/ROCK inhibitors have emerged as a new promising
treatment for cardiovascular diseases. RhoA, as an important regulator of diverse
cellular signaling pathways, play significant roles in cytoskeletal organisation,
transcription and cell-cycle progression. However, to date, RhoA inhibitors are
macromolecules, and to our knowledge, small molecular-based inhibitors have not
been reported. In this study, a series of first-in-class small molecular RhoA inhibitors
have been discovered by using structure-based virtual screening in conjunction with
chemical synthesis and bioassay. Virtual screening of ~200,000 compounds, followed
by SPR-based binding affinity assays resulted in three compounds with binding
affinities to RhoA at micro-molar level (compounds 1-3). Compound 1 was selected
for the further structure modifications in considering binding activity and synthesis
ease. Fourty one new compounds (1, 12a-v, 13a–h and 14a–j) were designed and
synthesized accordingly. It was found that eight (12a, 12j, 14a, 14b, 14d, 14e, 14g
and 14h) showed high RhoA inhibition activities with IC50s of 1.24 to 3.00 µM.
Pharmacological assay indicated that two compounds (14g and 14h) demonstrated
noticeable vasorelaxation effects against PE-induced contraction in thoracic aorta
artery rings and served as good leads for developing more potent cardiovascular
agents.
O2N
N
H
N
N
NH
O
O
S
OH
O
OH
O
OH
OH
NH
NH
N
N
O
N
H
O
1 ( KD = 5.18 µM)
O
2 (KD = 13.1 µM)
N
N
H
3 (KD = 4.47 µM)
Structures of 3 binders (or hits) of RhoA selected from the candidates by virtual
screening and SPR-based binding affinity assay.
Keywords: RhoA inhibitors, cardiovascular, quinoxalines, Glide;
Reference
1. Pérez, L.; Danishefsky, S. J. Chemistry and biology in search of antimetastatic agents. ACS
Chem. Biol. 2009, 3, 159–162.
2. Etienne-Manneville, S.; Hall, A. Rho GTPases in cell biology. Nature 2002, 420, 629.
3. Chang, H. R.; Wang, C. J. The suppressive effect of Rho kinase inhibitor, Y-27632, on
oncogenic Ras/RhoA induced invasion/migration of human bladder cancer TSGH cells.
Chem.-Biol Interact. 2010, 183, 172–180.
OPC-11
Structural revision of periplocosides and periperoxides, natural
immunosuppressive agents from the genus Periploca
Luoyi Wang, Weimin Zhao
Abstracts
The structures of a series of peroxy function containing pregnane glycosides isolated
from Periploca sepium and P. forrestii were revised to be orthoester group bearing
ones using 2D NMR techniques as well as chemical transformations and X-ray
crystallographic diffraction analysis. The orthoester function appears to be essential
structural feature for the immunosuppressive activity.
Keywords: Periploca sepium; Periploca forrestii; Immunosuppressive; Pregnane
glycoside; Structural revision
OPC-12
Solvent-free synthesis of δ-carbolines/carbazoles from
3-nitro-2-phenylpyridines/2-nitrobiphenyl derivatives using DPPE as
a reducing agent
Haixia Peng, Xuxing Chen, Yanhong Chen, Qian He, Yuyuan Xie, Chunhao Yang *
Abstracts
A green and efficient preparation of functionalized δ-carbolines/carbazoles via
reductive ring closure by 1,2-bis(dipenylphosphino)ethane under solvent-free
conditions is described. The starting materials
3-nitro-2-phenylpyridines/2-nitrobiphenyl derivatives are readily prepared through
Suzuki-Miyaura cross-coupling reaction from commercially available compounds.
And the polar by-product ethane-1,2-diylbis(diphenylphosphine oxide) is easily
removed from the relatively polar reaction mixture. Various substituted
δ-carbolines/carbazoles are obtained in acceptable yields. It is particularly worth
mentioning that substrates with electron-withdrawing groups (EWG) also give the
desired products in good yield.
Keyword: Solvent-free δ-Carbolines Carbazoles Cadogen cyclization DPPE
OPC-13
霉酚酸衍生物的设计合成与免疫抑制活性研究
陈炜，段文虎*
摘要
霉酚酸(MPA)是由短密青霉菌发酵代谢产生的具有高度取代的邻羟基苯甲酸
内酯天然产物。霉酚酸具有抑制淋巴细胞的能力，但其生物利用度较低，可能是
在胃肠腔中的络合，狭窄的吸收窗，吸收前的代谢等引起的。霉酚酸酯和霉酚酸
钠是霉酚酸的两个重要衍生物，临床上主要用于治疗器官移植的排异反应。我们
通过前药的设计思路，对可能引起副作用的酚羟基进行保护，制备了酚羟基碳酸
酯类衍生物，大部分化合物的免疫抑制活性与霉酚酸相当甚至增强，细胞毒性降
低，其中 MPA-48 的部分体外实验较 MMF 表现更优。
OPC-14
基于螯合作用机理的新型 HIV-1 整合酶抑制剂
的发现和构效关系研究
张凤华，黄少胥，樊兴，龙亚秋*
（中国科学院上海药物研究所新药研究国家重点实验室，上海，201203）
摘要
HIV-1 整合酶是临床有效的抗艾药物新靶标。我们基于整合酶催化作用需要
金属离子作为辅因子的原理，设计以儿茶酚--水杨酸结构母核通过螯合作用抑制
酶的链转移和 3’-切断反应，获得了同时对 3’-P 和 ST 有抑制活性的新结构整合酶
抑制剂 LNZ-90。对这个化合物进行结构优化，合成了一系列活性良好的化合物，
其中 LNZ-91 ST, IC50: 5μM, 但这类化合物对 3’-P 的抑制活性没有很大的突破。
为进一步提高化合物 3’-P 抑制活性，我们从 Merck 公司的 L-870,810 出发，
将赖氨酸、苯丙氨酸和色氨酸这几个热点氨基酸引入 1,6-二氮杂萘环。成功得到
了多个对 3’-P 抑制活性良好的化合物，其中 NLH-29 ST, IC50 : 0.4μM，3’-P, IC50:
2μM。由于与现有抑制剂机理不同，这为开发新型的 HIV-1 IN 抑制剂提供了很好
的结构基础。
OPC-15
Metabolism and Excretion of Morinidazole in Humans:
Identification of Diastereoisomeric Morpholine N+-Glucuronides
Catalyzed by UDP-Glucuronosyltransferase 1A9
Ruina Gao, Dafang Zhong, Liang Li, Cen Xie, Xingxing Diao and Xiaoyan Chen
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China (R. G., D.
Z., L. L., C. X., X. D., X. C.)
Abstracts
Morinidazole (1-(2-methyl-5-nitro-1H-imidazol-1-yl)-3-morpholinopropan-2-ol),
is a chiral 5-nitroimidazole class antimicrobial agent. The objectives of the present
study were to determine the metabolism and excretion of morinidazole in humans and
to identify enzyme responsible for the major clearance pathway. Plasma and urine
samples were collected for metabolite profiling before and after an intravenous drip
infusion of 500 mg morinidazole. A total of 10 metabolites were detected by
UPLC/Q-TOF mass spectrometry. Major metabolites were isolated and purified from
human urine and the structures were confirmed by MS and 2D-NMR.
Glucuronidation of the parent drug followed by renal excretion was the major
elimination pathway, accounting for 35% of the dose. The metabolic pathway
displayed regio- and stereoselectivity. Unexpectedly, the nitrogen of morphline ring
was glucuronidated, rather than the hydroxyl group of the aliphatic side chain to form
S-morinidazole glucuronide (M8-1) and S-entiomer glucuronide (M8-2). The system
exposure of M8-2 was 6 times higher than that of M8-1. The mechanisms of
N+-glucuronidations were investigated using human liver microsomes (HLMs) and
recombinant UDP glucuronosyltransferases in the presence of chemical inhibitor.
Results indicated that N+-glucuronidation of morinidazole was catalyzed mainly by
UGT1A9. A kinetic study showed that N+-glucuronidation of racemic morinidazole in
HLMs and in UGT1A9 both obeyed the typical Michaelis-Menten plot. HLMs
catalyzed M8-1 and M8-2 formation at a ratio of 1:11.3 with similar
affinity. In
conclusion, morinidazole and its metabolites were eliminated primarily by renal
excretion in humans. The major metabolites were two diastereoisomeric
N+-glucuronides, and UGT1A9 played an important role for N+-glucuronidation.
OPC-16
Fragment-based prediction of skin sensitization using recursive
partitioning
Jing Lua, Mingyue Zhenga, Yong Wanga, Qiancheng Shena, Xiaomin Luoa,
Hualiang Jianga,b, Kaixian Chena
Abstracts
Skin sensitization is an important toxic endpoint in the risk assessment of
chemicals. In this paper, structure-activity relationships (SAR) analysis was
performed on the skin sensitization potential of 357 compounds with local lymph
node assay (LLNA) data. Structural fragments were extracted by GASTON
(GrAph/Sequence/Tree extractiON) from the training set. Eight fragments with
accuracy significantly higher than 0.73 (p<0.1) were retained to make up an indicator
descriptor fragment. The fragment descriptor and eight other physicochemical
descriptors closely related to the endpoint were calculated to construct the recursive
partitioning tree (RP tree) for classification. The balanced accuracy of the training set,
test set I, and test set II in the leave-one-out model were 0.846, 0.800, and 0.809,
respectively. The results highlight that fragment-based RP tree is a preferable method
for identifying skin sensitizers. Moreover, the selected fragments provide useful
structural information for exploring sensitization mechanisms, and RP tree creates a
graphic tree to identify the most important properties of skin sensitization. They can
provide some instructions for designing of drugs with lower sensitizability.
Keywords Skin sensitization. LLNA. SAR. Fragment. Recursive partitioning tree.
Substructure mining algorithm
OPC-17
基于海洋天然产物 Scleritodermin A 中的 ACT 结构单元的新型 PTP1B
抑制剂的设计合成及构效关系研究
陈悦婷，南发俊*
摘要
以天然产物 Scleritodermin A 中的 ACT 结构单元为起点，运用多样性导向合
成（DOS: diversity-oriented synthesis）策略进行“类天然产物”化合物库的设计
与合成。发现了一类具有良好活性的 PTP1B 抑制剂，部分化合物能提高 CHO/hIR
细胞的 IR 磷酸化水平，并促进 L6 细胞的葡萄糖摄取。
OPC-18
Discovery and Preclinical Profile Research of TPN729
Guang-hui TIAN1, Zhen WANG1, Zheng LIU2, Xiao-jun YANG2, Jian-feng LI1,
Jin SUO1, Rong-xia ZHANG1, Xiang-rui JIANG1 and Jing-shan SHEN1*
1
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, No. 555, Zu chong zhi
Road, Shanghai 201203, P.R.China
2Topharman
Shanghai Co., Ltd. 1088 Chuansha Road, Pudong, Shanghai 201209, P.R.China
*Corresponding
E-mail: jsshen@mail.shcnc.ac.cn
Abstracts
Aim: To seach drugable PDE5 inhibitors with higher selectivities. Methods:
Hundreds
of
compounds
with
flavonoid,
phenylquinazoline
and
pyrazolopyrimidinone scaffolds were designed and synthesized. The in vitro
inhibitory activities against PDE5 and PDE6 were done. In vivo pharmacologic
profile was also assessed in electrostimulation ED and PAH rat. Preliminary
pharmacokinetic studies on 8 selected compounds including absorption, distribution,
metabolism, and excretion were performed in rats and dogs, respectively.
Results: TPN729 was finally set as a qualified drug candidate for ED and PAH
treatment by complete evaluation of drug-like properties from 8 selected
pyrazolopyrimidinone derivatives. The most favorable salt and polymorph form was
obtained by salts selection and polymorphs screening studies. Process research and
optimization was done and batches of TPN729 were got for the purpose of full
preclinical research. Single dose toxicity studies in mice, three-month repeat-dose
toxicity studies in rats and dogs, in vitro and in vivo genotoxicity, and reproductive
and development toxicity studies in rats were conducted.The results from PK/PD and
safety evaluation proved that TPN729 is worthy to go into next stage of clinical
research for the indications of ED and PAH. Conclusion: TPN729 is a novel, orally
available PDE5 inhibitor with high selectivity, low toxicity and superior
pharmacokinetic profiles. These properties of TPN729 indicate that it is a potential
drug candidate in the treatment of ED and PAH.
Keywords: PDE5 inhibitors, TPN729, drug candidate, preclinical research, ED, PAH
OPC-19
Highly Diastereoselective Friedel-Crafts reaction of Arenes with
N-tert-Butanesulfinylimino Ester: Efficient Synthesis of Optically
Active α-Arylglycines
Yi Li, Du-Ming Ji and Ming-Hua Xu*
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road,
Shanghai 201203, People’s Republic of China.
Abstracts
As an important class of non-proteinogenic amino acids, α-arylglycine and its
derivatives that exist in various biologically natural and unnatural products have
attracted much attention1. Therefore, lots of attempts to develop efficient
methodologies for the asymmetric synthesis of these valuable optically active
compounds have been performed2. Despite the fact that the strategies involving
asymmetric catalysis2b-e, h, enzymatic resolution3and chiral auxiliary-mediated
induction2f, g, i have been employed, far fewer methods capable of the efficient
synthesis of enantiomerically enriched α-arylglycines were reported.
Recently, we have successfully developed a highly diastereoselective
Friedel-Crafts alkylation of arenes with chiral N-tert-butanesulfinylimino ester in the
presence of catalytic amount of Lewis acid at room temperature. It not only allows
exceptionally mild and efficient access to various enantiomerically enriched
α-arylglycines in good yields and up to > 99% de but also enables the further
synthesis of more challenging Friedel-Crafts dialkylation product in a moderate yield
with an extremely high diastereo- and enantioselectivity (anti : syn > 99 : 1, >99%
ee).
O
1
R
S
LA
N
H
COOR2
O
Friedel-Crafts
S
NH
2
R OOC
alkylation
R1
up to 91% yield, >99% de
O
δ-
O
N LA
O
H
δ-
δMeO
S
Re
OMe
TS-1 (favored)
O
S
LA
N
δMeO
O
δδ-
H
Si
OMe
O
TS-2 (disfavored)
Keywords: α-arylglycine, Friedel-Crafts reaction
References
1. Hodgson DRW, Sanderson JM. Chem Soc Rev 2004; 33: 422.
2. (a) Williams RM, Hendrix JA. Chem Rev 1992; 92: 889. (b) Krueger CA, Kuntz KW,
Dzierba CD, Wirschun WG, Gleason JD, Snapper LM, Hoveyda AH. J Am Chem Soc 1999;
121: 4284. (c) Sigman MS, Vachal P, Jacobsen EN. Angew Chem Int Ed 2000; 39: 1279. (d)
Saaby S, Fang X, Gathergood N, Jørgensen KA. Angew Chem Int Ed 2000; 39: 4114. (e)
Shirakawa S, Berger R, Leighton JL. J Am Chem Soc 2005; 127: 2858. (f) Beenen MA,
Weix DJ, Ellman JA. J Am Chem Soc 2006; 128: 6304. (g) Dai H, Lu X. Org Let 2007; 9:
3077. (h) Enders D, Seppelt M, Beck T. Adv Synth Catal 2010; 352: 1413. (i) Reddy LR,
Gupta AP, Liu Y. J Org Chem 2011; 76: 3409.
3. Scott JW. Topics in Stereochemistry John Wiley & Sons Inc New York 2007; Vol. 19: pp
209.
OPC-20
Molecular Basis of NDM-1, a new Antibiotic Resistance Determinant1
Zhongjie Lianga, Lianchun Lia, Yuanyuan Wanga, Limin Chen, Xiangqian Kong, Lefu
Lan, Mingyue Zheng, Cai-Guang Yang, Hong Liu, Xu Shen, Cheng Luo*, Keqin
Kathy Li*, Kaixian Chen and Hualiang Jiang*
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy
of Sciences, 555 Zu Chongzhi Road, Shanghai 201203, China
State Key Laboratory of Medical Genomics, Shanghai Institute of Hematology, Rui Jin Hospital
Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China
Abstracts
The New Delhi metallo-β-lactamase (NDM-1) was first reported in 2009 in a
Swedish patient. A recent study reported that Klebsiella pneumonia NDM-1 positive
strain or Escherichia coli NDM-1 positive strain was highly resistant to all antibiotics
tested except tigecycline and colistin. These can no longer be relied on to treat
infections and therefore, NDM-1 now becomes potentially a major global health
threat.
In this study, we performed modeling studies to obtain its 3D structure and
NDM-1/antibiotics complex. It revealed that the hydrolytic mechanisms are highly
conserved. In addition, the detailed analysis indicates that the more flexible and
hydrophobic loop1, together with the evolution of more positive-charged loop2 leads
to NDM-1 positive strain more potent and extensive in antibiotics resistance
compared with other MBLs. Furthermore, through biological experiments, we firstly
revealed the molecular basis for antibiotics catalysis of NDM-1 on the enzymatic
level. We found that NDM-1 enzyme was highly potent to degrade carbapenem
antibiotics, while mostly susceptible to tigecycline, which was later proved to be a
competitive inhibitor for NDM-1. Meanwhile, the mutagenesis experiments, including
D124A, C208A, K211A and K211E, which displayed down-regulation on meropenem
catalysis, proved the accuracy of our model.
At present, there are no effective antibiotics against NDM-1 positive pathogen.
Our study will provide clues to investigate the molecular basis of extended antibiotics
resistance of NDM-1 and then accelerate the search for new antibiotics against
NDM-1 positive strain in clinical studies.
Keywords NDM-1; Molecular model; antibiotics resistance
(A)
(B)
Fig .1 (A) Cartoon representation of the overall structure of NDM-1. (B) Detailed
binding mode of antibiotics meropenem with NDM-1.
(A)
(B)
Fig .2 (A) Seven antibiotics were hydrolyzed by NDM-1 protein. The enzymatic
reactions were dynamically monitored by the degradation of the antibiotics. (B)
Compare with the wild type enzyme, the point mutants C208A, D124A, K211A and
K211E totally disrupt the hydrolytic activity to meropenem; while mutations of loop1
hardly affect the NDM-1 hydrolytic activity.
Reference
6． Liang Z, Lianchun Li, Yuanyuan Wang, Limin Chen, Xiangqian Kong, Yao Hong, Lefu Lan,
Mingyue Zheng, Cai-Guang Yang, Hong Liu, Xu Shen, Cheng Luo, Keqin Kathy Li,
Kaixian Chen and Hualiang Jiang, PLOS ONE, in press.
__________________________
*This work was supported by grants from National Natural Science Foundation of China (20972173,
20972174, 90913010 and 91029704), the 973 project (2009CB918502), and the Council of Shanghai
Municipal Government of S&T (10410703900),
Phone: 86-21-50806600; Fax: 86-21-50807088; E-mail: hljiang@mail.shcnc.ac.cn (Hualiang Jiang).
OPC-21
Combinatorial Approach to Diversity-oriented Fluorescence Library
Based on the Novel Fluorescent Core Skeletons
Diliang Guo, Hong Liu*
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy
of Sciences, 555 Zu Chong Zhi Road, Shanghai, P. R. China
Department of Medicinal Chemistry, China Pharmaceutical University, 24 Tong jia Xiang,
Nanjing, P. R. China
Abstract
Fluorescent probes with high sensitivity and ease of handling have been extensively
employed in biological science, clinical diagnosis, and drug discovery. The
development of fluorescent probes, particularly synthetic small molecular fluorescent
probes with various core skeletons, such as fluorescein, BODIPY, and rhodamine, has
received special attention.1 Although significant progress has been made in this field,
there is still a great demand for novel fluorescent core skeletons with flexible
synthetic strategies and good photophysical properties for bioimaging. By making use
of combinatorial chemistry, we can derivatize fluorogenic scaffolds, tune their
photophysical spectra and adjust their properties (from cell permeability to quantum
yields) to generate diversity-oriented fluorescence libraries of potential probes.2 In our
ongoing efforts to develop convenient and efficient approaches to the synthesis of
biologically active heterocyclic compounds,3 we have designed and synthesized an
18-member fluorescent library based on the 2-iminocoumarin-3-carboxamide core
skeleton, which can undergo a dramatic change in fluorescence properties simply by
changing the substituents at the two variation points of the fluorophore. Furthermore,
the dyes can permeate the living cell membranes with low cytotoxicity and
accumulate preferentially in the mitochondria, thus possessing potential utility in
intracellular imaging.4
References
1. Goncalves, M. S. T. Chem. Rev. 2009, 109, 190-212.
2. Vendrell M., Lee J-. S., Chang Y-. T. Curr. Opin. Chem. Biol. 2010, 14, 383-389.
3.
(a) Guo D. L., Huang H., Zhou Y. et al. Green Chem. 2010, 12, 276-281. (b) Zhang X., Ye
D. J., Sun H. F. et al. Green Chem. 2009, 11, 1881-1888.
4.
Guo D. L., Chen T., Ye D. J. et al. Org. Lett. 2011, 13, 2884-2887.
OPC-22
Structural AND Theoretical STUDIES INDICATE DYNAMIC
switchings of Staphylococcus aureus CLPP1
Fei Ye, Jie Zhang, Lefu Lan, Caiguang Yang, Cheng Luo* and Hualiang Jiang*
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy
of Sciences, 555 Zu Chong Zhi Road, Zhangjiang Hi-Tech Park, Shanghai, 201203, P. R. China
Abstracts
The highly conserved ClpP protease is a potential biological target against bacteria.
It consists of two heptameric rings that enclose a large chamber. ClpP might undergo
switching between an active extended state required for substrate degradation and an
inactive compact state allowing product release. Here, we present the wild type ClpP
structures in two distinct states from Staphylococcus aureus. One strucutre is closely
similar to those solved ClpP structures in extended states; the other is novel, in which
the handle domain kinks to take a compressed conformation. Based on structural
analysis and Molecular Dynamics simulations, we propose a mechanism that the
dynamic conformational change of SaClpP is mediated by electrostatic interaction and
hydrogen bonding network between Arg171 and Asp170 from apposing rings. The
handle domain controls the degradative products exit the chamber by dynamic
conformational switching from extended state to compressed state. Our work provides
critical information for understanding the dynamics of enzymatic cycle of the ClpP
family proteins in general and light up the road for the future anti-bacteria drug
discovery studies.
Keywords: ClpP; crystal structure; structural switchings; Molecular Dynamics simulations;
Arg171_Asp170 network
Fig. 1. R171_D170 network keeps ring-ring
Fig. 2. Rearrangement of catalytic triad after degradation
interlocking and handle helix extended
causes destruction of R171_D170 network
Reference
1． Zhang J.1, Ye F. 1, Lan L., Jiang H., Luo C., Yang C. J. Biol. Chem.,revised.
2． Gribun A., Kimber M. S., Ching R., Sprangers R., Fiebig K. M., Houry, W. A. J. Biol. Chem. 2005, 280,
16185.
3． Kimber M. S., Yu A. Y., Borg M., Leung E., Chan H. S., and Houry W. A. Structure. 2010, 18, 798
__________________________
*This work was supported by grants from National Natural Science Foundation of China (20972173,
20972174, 90913010 and 91029704), and the Council of Shanghai Municipal Government of S&T
(10410703900), the 973 project (2009CB918502),
OPC-23
川楝的化学成分研究
张益，叶阳
中国科学院上海药物研究所天然药物化学研究室（50806600-3322）
摘要
通过对川楝树皮和果实系统的化学成分研究，共分离鉴定了 65 个化合物，
主要包括柠檬苦素和三萜类，其中 35 个为新化合物。我们发现从中得到的新柠
檬苦素化合物结构类型多样，包括 prolimonoids， meliacin，C-19/C-29 bridged
acetals，trichinlin，以及 ring C-seco limonoids 等。三萜类化合物中 24 位羟基位
于柔性边连，ROESY 谱等波谱方法在此并不可靠，我们通过测定四氯化碳溶液
中邻羟基化合物与 Pr(FOD)3 形成络合物的 CD 谱的方法和 Mosher 法确定了 24
位的绝对构型。我们还发现，川楝树皮和果实中化学成分相同的主要为杀虫活性
较强的 C-19/C-29 bridged acetals 柠檬苦素，而且含量较高，这与它们均作为传统
中药中驱虫药使用的记载是一致的，但二者化学成分也有差异，树皮中三萜类化
合物较多，且变化较小，而果实中柠檬苦素类化合物较多，而且类型多样，结构
复杂。我们对分离得到的一些柠檬苦素类和三萜类化合物进行了细胞毒活性测
试，从中未发现活性较好的化合物。
关键词：川楝、三萜、柠檬苦素、CD 谱、Mosher 法、细胞毒活性
OPC-24
Velocity-scaling optimized replica exchange molecular dynamics of
proteins in a hybrid explicit/implicit solvent
Jinan Wang，Weiliang Zhu，Guohui Li，Ulrich H. E. Hansmann
Abstracts
Aim: To develop a new scheme for replica exchange molecular dynamics of proteins
in explicit solvent that minimizes the number of required replicas using velocity
rescaling. Methods: Our approach relies on a hybrid method where the protein
evolves at each temperature in an explicit solvent, but replica exchange moves utilize
an implicit solvent term. The two terms are coupled through the velocity rescaling.
Results: We test the efficiency of this approach for a common test case, the trp-cage
protein and found that our new method leads to a significant increase of the replica
exchange rates. It has been demonstrated that the number of replicas could be reduced
four times (in respect to the classical exchange scheme within canonical ensemble)
without any worsening of the simulation results. Conclusion: Our scheme can reduce
the number of replicas in the replica exchange molecular dynamics of proteins in
explicit solvent and could be useful in the protein folding.
Keywords: replica exchange molecular dynamics; protein folding;
OPC-25
维生素 B1 在绿色有机合成中的应用
雷敏，胡立宏*
摘要
绿色化学是近十几年才产生和发展起来的一个新兴的化学分支，它以“原
子经济性”为原则，研究如何在生成目的产物的过程中充分利用原料，并减少有
害物质的释放。我们课题组研究了以廉价、易得、无毒的维生素 B1 为催化剂的
有机反应，成功合成了酰胺甲基萘酚，1,2-二氢萘并[1,2-e][1,3]噁嗪-3-酮和
14-氢二苯并[a,j]氧杂蒽衍生物。我们还发现维生素 B1 对 Mannich 反应，氨基
甲酰化反应，Biginelli 型反应，Hantzsch 反应均有良好的催化效果，而且催
化剂容易回收，可重复使用不失活。
OPC-26
One-pot Tandem Reactions of 3-(1-Alkynyl)chromones to Generate
Diversified Scaffolds
Yang Liu, Youhong Hu*
Abstracts
Tandem reactions provide an efficient way to generate molecular complexity from
readily
accessible
intermediates.
We
focused
on
functionalized
3-(1-alkynyl)chromones to generate natural product-like scaffolds by cascade
reactions.
And
a
series
of
One-pot
tandem
reactions
of
functionalized
3-(1-alkynyl)chromones promoted by base without a transition metal catalyst and
inert atmosphere have been developed: Functionalized xanthones were obtained via a
novel
base-promoted
1,3-dicarbonyl
tandem
compounds
reaction
under
from
mild
3-(1-alkynyl)chromones
reaction
conditions;
By
with
replacing
1,3-dicarbonyl compounds with acetonitriles, functionalized amino-substituted
xanthones
could
also
be
obtained
under
microwave
irradiation;
Using
2-(2-bromophenyl) acetonitrile and its various similar substrates, a novel functional
polycyclic chromones library could be build up rapidly; 3C-xanthone-linked
3C-chromone scaffold, which is a nature-product-like scaffold has been obtained via
the dimerization of 3-(1-alkynyl)chromones and 2-methyl-3-(1-alkynyl)chromones;
By replacing 3-(1-alkynyl)chromones with electron-deficient chromone-fused dienes,
a novel benzo[α]xanthone scaffold could be obtained with the formation of three new
C-C bonds and one C-O bond.
We have generated these different nature-product-like molecules libraries from
different tandem processes which involves multiple reactions, such as Michael
addition, cyclization, 1, 5-H shift, 1, 2-addition, 8pi-cyclization and elimination. The
biological evaluations of these compounds are under investigation.
OPC-27
N-Propylnoraporphin-11-yl 5-(1,2-Dithiolan-3-yl) pentanoate: A
Dopamine D2 and Serotonin 5-HT1A Dual-Agonist with
Anti-Parkinsonian Effect
Na Ye, Hai Zhang, Xuechu Zhen, and Ao Zhang*
Synthetic Organic and Medicinal Chemistry Laboratory (SOMCL), State Key Laboratory of Drug
Research, Shanghai Institute of Materia Medica (SIMM), Chinese Academy of Sciences, Shanghai,
China 201203
Abstracts
Parkinson’s disease (PD) is an age-related progressive neurodegenerative
disorder and is the second most common debilitating neurodegenerative disease. To
find safe anti-PD therapies, the D2 and 5-HT1A receptor dual agonist profile have
attracted more attention, with several drugs recently progressing to the clinic.
By analyzing the tetracyclic skeleton of aporphine analogues, we found there is a
5-HT1A binding site existing in this long-standing scaffold for the D2 receptor agonists.
Therefore, we synthesized a series of new aporphine analogues bearing a C-, N-, or
O-linkage at the C11 position. Lipoic ester (-)-15 was identified as a full agonist at the
dopamine D2 and serotonin 5-HT1A receptors with Ki values of 174 and 66 nM,
respectively. It elicited antiparkinsonian action on Parkinsin’s disease (PD) rats with
minor dyskinesia. Chronic use of (-)-15 reduced L-DOPA-induced dyskinesia (LID)
without attenuating the antiparkinsonian effect. These results suggest that 5-HT1A and
D2 dual-receptor agonist (-)-15 may present a novel candidate drug in the treatment of
PD and LID.
Keywords: D2 and 5-HT1A receptor dual-agonists, aporphine
References:
1. Liu, Z.; Zhang, H.; Ye, N.; Zhang, J.; Zhen, X.; Zhang, A. J. Med. Chem. 2010, 53, 1319-28.
2. Ye, N.; Wu, Q.; Zhu, L.; Zhen, X.; Zhang, A. Bioorg. Med. Chem. 2011, 19, 1999-2008.
3. Zhang, H.; Ye, N.; Liu, Z.; Gao, B.; Zhen, X.; Zhang, A. J. Med. Chem. 2011, 54, 4324-38.
OPC-28
三唑并三嗪类 c-Met 酪氨酸激酶抑制剂的研究
陈方，段文虎*
摘要
受体型蛋白酪氨酸激酶 c-Met 通过与其配体 HGF 相互作用，在肿瘤的发生
发展过程中发挥重要的作用，阻断 HGF/c-Met 的信号转导是治疗肿瘤的重要策
略。我们设计合成了七十余个三唑并三嗪类小分子 c-Met 酪氨酸激酶抑制剂，该
类化合物在分子水平、细胞水平对 c-Met 均有很好的抑制活性，其中三十余个化
合物在分子水平对 c-Met 抑制活性 IC50 小于 1.0 nM。特异性 c-Met 抑制剂 I-19
(Simm530) 能显著抑制 c-Met 活化介导的肿瘤细胞增殖和由 HGF 诱导的细胞
（NCI-H441/MDCK）运动能力，并且能显著抑制 U-87MG 裸小鼠移植瘤的生长
（图一），具有较好的成药前景，其药代动力学性质和毒性在进一步的研究中。
**
***
**
**
**
图一、化合物 I-19 (Simm530) 对人神经胶质瘤 U-87MG 裸小鼠移植瘤的生长抑制
*
图二、化合物 I-19 (Simm530) 对人神经胶质瘤 U-87MG 荷瘤裸小鼠体重影响
关键词： 三唑并三嗪 c-Met 抑制剂 抗肿瘤