PROCESS BIOCHEMISTRY
·
2023
·
IF 4.4
Xiaoxuan Cai,Miaoen Huang,Xixiang Huang,Huan Liu,Tianji Wang,Li Li,Weiguang Yang,Hui Luo,Yingnian Lu
The organism Sipunculus nudus L is a good resource for marine peptides because of its rich protein. However, the disadvantage of biological peptides is their short metabolic half-life, therefore, structural modification of peptides is necessary. In this research, the enzymatic hydrolysis of Sipunculus nudus L was performed using trypsin and then peptides were isolated and purified from the hydrolysates, four novel tripeptides were identified by LC-MS-MS as Ser-Arg-Pro (SRP, m/z = 358.39), Pro-Arg-Pro (PRP, m/z = 368.43), Arg-Pro-Ala (RPA, m/z = 342.39), and Lue-Pro-Lys (LPK, m/z = 356.46), their ACE inhibitory activity was investigated with IC 50 value as following 0.046, 0.42, 0.97, and 6.54 mM, respectively. Because of its highest ACE inhibitory activity, SRP was further selected to modify the chemical structure for a sustained release agent DSPE-PEG-SRP. Compared with 7 h of free SRP, the release of DSPE-PEG-SRP in vitro was approximately 120 h. The ACE inhibition rate of SRP released from the synthetic product and the raw material SRP was almost the same, molecular docking demonstrated that ACE inhibitory activity was attributed to the formation of hydrogen bonds between SRP and ACE active sites . In conclusion, DSPE-PEG-SRP, a novel sustained release agent, has great potential and is expected to be further applied to treat hypertension.
研究领域:
海洋生物学
·
药学
·
生物化学
✓ 已验证
产品关联
DOI
高影响因子
MOLECULAR PHARMACEUTICS
·
2015
·
IF 4.384
Yuan Sheng,Jinhui Xu,Yiwen You,Feifei Xu,Yun Chen
The extended use of doxorubicin (DOX) could be limited because of the emergence of drug resistance associated with its treatment. To reverse the drug resistance, two thiol-modified peptide sequences HAIYPRHGGC and THRPPMWSPVWPGGC were, respectively, conjugated to DOXO-EMCH, forming a maleimide bridge in this study (i.e., T10-DOX and T15-DOX). The structures and properties of peptide–DOX conjugates were characterized using 1H NMR, 13C NMR, mass spectrometry, and high-performance liquid chromatography. Their stability was also evaluated. By using MCF-7/ADR cells as an in vitro model system and nude mice bearing MCF-7/ADR xenografts as an in vivo model, the ability of these novel peptide–DOX conjugates to reverse drug resistance was accessed as compared with free DOX. As a result, the IC50 values for T10-DOX and T15-DOX significantly decreased (31.6 ± 1.6 μM and 27.2 ± 0.8 μM), whereas the percentage of apoptotic cell population increased (35.4% and 39.3%). The in vivo extent of inhibition was more evident in the mice groups treated with peptide–DOX conjugates (59.6 ± 8.99% and 46.4 ± 6.63%), which had DOX primarily accumulated in tumor. These conjugates also showed a longer half-life in plasma and cleared much more slowly from the body. Furthermore, T10-DOX may be more effective than T15-DOX with a higher efficacy and a lower side effect. Most importantly, evidence was provided to support the enhanced intracellular drug accumulation and the induction of lysosomal pathway of apoptosis underlying the drug resistance. As an endosomal/lysosomal marker, cathepsin D permealized the destabilized organelle membrane and was detected in the cytoplasm, leading to the activation of the effector caspase-3 in cell apoptosis. This report is among the first to demonstrate that peptide–DOX-like conjugates promote apoptosis through the initiation of the lysosomal pathway.
研究领域:
药理学
·
肿瘤学
·
化疗
·
药物递送系统
·
细胞生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
·
2013
·
IF 1.495
Weiqiang Lu,Junsheng Zhu,Shien Zou,Xi Li,Jin Huang
Human skin fibroblast collagenase also known as Matrix Metalloproteinase-1 (MMP-1) is a key enzyme in remodeling and degradation of extracellular matrix, and the inhibitors of human MMP-1 are effective drug candidates for the treatment of cancer. In this study, we report an improved method for high-level expression of soluble human MMP-1 catalytic domain (cd-MMP-1) in E.coli. The enzymatic activity is found maximum at pH 7.5 and temperature 40°C with a Km value of 13.02 µM. Effects of 17 structure-related flavonoids on MMP-1 activity are evaluated using a fluorescent assay, 6 inhibitors are identified with IC50 < 10 µM. Fisetin is the most active agent with an IC50 value of 1.35 µM and is identified as a mixed type inhibitor. Our improved soluble cd-MMP-1 expression method provides a basis for inhibitors identification and may be beneficial to discover novel anti-cancer agent targeting human MMP-1.
研究领域:
生物化学
·
分子生物学
·
药理学
·
癌症研究
✓ 已验证
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Jinpeng Liu,Mashooq Khan,Yong Wei,Anna Liu,Tao Wang,Qiongzheng Hu,Li Yu
High-throughput screening of lipase inhibitors is critical for developing anti-obesity drugs. Herein, a robust and versatile sensing platform based on the difference in the sliding time of droplets on liquid crystal-infused porous surfaces (LCIPS) is developed to detect lipase inhibitors. Lipase catalyzes glycerol trioleate (GT) to produce oleate that can adsorb at the solution/liquid crystal (LC) interface due to its high amphiphilicity, which impedes the sliding of droplets on LCIPS, resulting in an extended sliding time. However, in the presence of lipase inhibitors, the enzymatic hydrolysis of GT by lipase is inhibited, which shortens the sliding time. The screening of three lipase inhibitors, orlistat, cetilistat, and epigallocatechin gallate (EGCG), popular anti-obesity drugs, is demonstrated to have IC 50 values of 17.45 ± 3.15 nM, 72.15 ± 6.32 nM, and 2.19 ± 0.24 μM, respectively. This study provides a simple, inexpensive, portable, user-friendly, and instrument-free method for rapid, accurate, and high-throughput screening of lipase inhibitors. The LCIPS-based sensing platform is up-and-coming in developing advanced bioanalytical sensors.
研究领域:
药学
·
生物技术
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
TALANTA
·
2021
·
IF 6.057
Weiwei Tang,Andrew Gordon,Hui-ying Wang,Ping Li,Jun Chen,Bin Li
The development of in situ methods for the analysis and visualization of enzyme activity is of paramount importance in drug discovery, research, and development. In this work, the functionalized and array patterned indium tin oxide (ITO) glass slides were fabricated by non-covalent immobilization of amphipathic phospholipid-tagged peptides encompassing the thrombin cleavage site on steric acid-modified ITO slides. The fabricated peptide arrays provide 60 spots per slide, and are compatible with matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) measurement, free matrix peak interference, and tolerance to repeated aqueous washing. The peptide arrays were used for the investigation of thrombin activity and screening for its potential inhibitors. The thrombin activity and its Michaelis-Menten constant (K m ) for immobilized peptide substrate was determined using developed MALDI MS peptide array. To investigate the applicability and effectiveness of peptide arrays, the anti-thrombin activity of grape seed proanthocyanidins with different degrees of polymerization (DP) was monitored and visualized. MALDI MS imaging results showed that the fractions of proanthocyanidins with the mean DP of 4.61–6.82 had good thrombin inhibitory activity and their half-maximal inhibitory concentration (IC 50 ) were below 10 μg/mL. Therefore, the developed peptide array is a reliable platform for the discovery of natural thrombin inhibitors .
研究领域:
药物发现
·
分析化学
·
药学
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
·
2018
·
IF 0.824
Chan-Yuan YAO,Jin-Yi YANG,Zhen-Lin XU,Hong WANG,Hong-Tao LEI,Yuan-Ming SUN,Yuan-Xin TIAN,Yu-Dong SHEN
A specific polyclonal antibody was prepared based on a new hapten with stable structure. Based on this, an indirect competitive enzyme-linked immunosorbent assay (icELISA) was established for determination of tylosin residues in food and environmental samples. The experimental conditions were optimized as follows: the coating antigen and antibody were respectively diluted by 6000 times and 2000 times, and the competitive reaction time was 40 min. Under the optimized conditions, the method showed a detection limit of 0.07 ng mL −1 , an IC 50 of 1.39 ng mL −1 and a linear range of 0.17–11.0 ng mL −1 . The recoveries of spiked raw milk and water samples were ranged from 78.4% to 105.6%, with the RSDs of less than 15%. Good correlation between icELISA and HPLC method was obtained for spiked samples ( R 2 = 0.97). This method was suitable for the determination of tylosin residues in milk and water samples.
研究领域:
分析化学
·
食品科学
·
环境科学
✓ 已验证
产品关联
DOI
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2019
·
IF 2.931
Jianmin Chen,Qinglian Li,Yaling Ye,Ziyao Huang,Zhipeng Ruan,Nan Jin
Tyrosinase is the rate-limiting enzyme for controlling the production of melanin in the human body, and overproduction of melanin can lead to a variety of skin disorders. In this paper, the inhibitory kinetics of phloretin on tyrosinase and their binding mechanism were determined using spectroscopy, molecular docking, antioxidant assays and chromatography. The spectroscopic results indicate that phloretin reversibly inhibits tyrosinase in a mix-type manner through a multiphase kinetic process with the IC 50 of 169.36 μmol/L. It is shown that phloretin has a strong ability to quench the intrinsic fluorescence of tyrosinase mainly through a static quenching procedure, suggesting that a stable phloretin-tyrosinase complex is generated. Molecular docking results suggest that the dominant conformation of phloretin binds to the gate of the active site of tyrosinase. Moreover, the antioxidant assays demonstrate that phloretin has powerful antioxidant capacity and has the ability to reduce o -dopaquinone to l -dopa just like ascorbic acid . Interestingly, the results of spectroscopy and chromatography indicate that phloretin is a substrate of tyrosinase but also an inhibitor. The possible inhibitory mechanism is proposed, which will be helpful to design and search for tyrosinase inhibitors.
研究领域:
生物化学
·
药理学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
APPLIED ORGANOMETALLIC CHEMISTRY
·
2019
·Vol. 33
·期 10
·e5120
·
IF 3.259
Dongdong Li,Luyao Wang,Ernesto Yagüe,Linlin Dai,Xiumei Zhao,Zibo Yang,Shuang Zhi,Yunhui Hu
An innovative ternary copper(II) complex, [Cu(Cl-PIP)(Tyr)Cl] n , has been synthesized and characterized using infrared spectroscopy, elemental analysis and single-crystal X-ray diffraction analysis. X-ray crystallography indicates that the Cu atom is five-coordinated in a square-pyramidal configuration. The unit forms a one-dimensional chain along the crystallographic c -axis. The complex was screened for cytotoxicity against a panel of eight human cancer cell lines, namely MDA-MB-231, CAL-51, K562, HeLa, SGC-7901, A549, MCF-7 and SMMC-7721. The best anticancer activity was obtained with triple-negative breast cancer CAL-51 and MDA-MB-231 cell lines, with IC 50 values in the range 0.035–0.10 μM, and this was better than using carboplatin. The complex inhibits proteasomal chymotrypsin-like activity, and docking studies reveal its interaction with 20S proteasome. In addition, the complex causes accumulation of ubiquitinated proteins, induces apoptosis and inhibits cell proliferation, indicating its great potential as a novel therapy for triple-negative breast cancer.
研究领域:
癌症研究
·
化学
·
药理学
·
分子生物学
·
细胞生物学
✓ 已验证
产品关联
DOI
高影响因子
CHEMICAL RESEARCH IN CHINESE UNIVERSITIES
·
2017
·Vol. 33
·期 6
·876-881
·
IF 1.024
Su Ping,Chen Xiaonan,He Zhangjing,Yang Yi
Kanamycin is an aminoglycoside antibiotic used increasingly in human and veterinary medicine. However, kanamycin residues in food can cause serious side effects. Here we reported the preparation of polyclonal antibody and the development of an indirect competitive biotin-streptavidin-amplified-based enzyme-linked immunosorbent assay(BA-ELISA) that can sensitively and specifically detect kanamycin residues in milk and honey. The immunogen and coating antigen were synthesized by covalently linking kanamycin to carrier proteins using the carbodiimide method. The anti-kanamycin polyclonal antibodies were obtained from immunized rabbits. The key assay parameters were investigated and optimized. The results show that under optimum conditions, the limit of detection for kanamycin is 0.07 ng/mL and the IC 50 is 6.48 ng/mL. Cross-reactivity values of the antibody with four kanamycin analogues are all <1%. Trace amounts of kanamycin in milk and honey samples can be detected by this novel BA-ELISA method successfully with satisfactory recoveries of 91.0%—103.3%. The developed protocol was also validated against liquid chromatography-mass spectrometry, returning a significant correlation. These results indicate that BA-ELISA is a viable option for monitoring kanamycin residues in milk and honey.
研究领域:
免疫学
·
分析化学
·
食品科学
·
兽医学
✓ 已验证
产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2023
·
IF 8.2
Qiuyang Huang,Xiaoling Zang,Zhiwei Zhang,Hang Yu,Baoyan Ding,Zhuangzhuang Li,Simin Cheng,Xin Zhang,Mustafa R.K. Ali,Xue Qiu,Zhihua Lv
The discovery of new anti-cancer drugs targeting the PD-1/PD-L1 pathway has been a research hotspot in recent years. In this study, biological affinity ultrafiltration (BAU), UPLC-HRMS, molecular dynamic (MD) simulations and molecular docking methods were applied to search for endogenous active compounds that can inhibit the binding of PD-L1 to PD-1. We screened dozens of potential cancer related endogenous compounds. Surprisingly, cyclic adenosine monophosphate (cAMP) was found to have a direct inhibitory effect on the PD-1/PD-L1 binding with an in vitro IC 50 value of about 36.4 ± 9.3 μM determined by homogeneous time-resolved fluorescence (HTRF) assay. cAMP could recover the proliferation of Jurkat T cells co-cultured with DU-145 cells and may suppress PD-L1 expression of DU-145 cells. cAMP was demonstrated to bind and induce PD-L1 dimerization by FRET assay, and also predicted by MD simulations and molecular docking. The finding of cAMP as a potential inhibitor directly targeting the PD-1/PD-L1 interaction could advance our understanding of the activity of endogenous compounds regulating PD-L1.
研究领域:
肿瘤学
·
药理学
·
分子生物学
·
细胞生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY
·
2022
·
IF 7.088
Zhendong Song,Linlin Lou,Guangjin Fan,Lu Liu,Yang Ge,He Liu,Albert S.C. Chan,Xiaolei Zhang,Xiao-Feng Xiong
Oncogene KRAS plays predominant roles in human cancers by regulating cell proliferation , differentiation, and migration. Recent progress revealed that directly target KRAS G12C with allosteric inhibitors that covalently bind to the switch Ⅱ pocket is feasible. Herein, series of pyrrolo[2,3- d ]pyrimidine derivatives were designed and synthesized through systematic structural optimization, leading to the discovery of compound 2-(( S )-1-acryloyl-4-(2-((( 2R , 7aS )-2-fluorohexahydro-1 H -pyrrolizin-7a(5 H )-yl)methoxy)-7-methyl-6-(8-methylnaphthalen-1-yl)-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)piperazin-2-yl)acetonitrile ( 50 ) with high KRAS/SOS1 inhibitory potency (IC 50 = 0.21 μM) and strong anti-proliferation activities on cancer cells harboring KRAS p.G12C . Compound 50 also exhibited satisfactory selectivity , moderate pharmacokinetic characters, and good anticancer effects in vivo . Meaningfully, the identification of these compounds highlights the necessity of an appropriate conformational constraint for acquiring the applicable binding pose in the cryptic pocket of KRAS, and the results support efforts toward design of KRAS inhibitors with novel skeleton and binding mechanism could be beneficial for targeting the acquired drug resistance.
研究领域:
肿瘤学
·
药理学
·
药物化学
✓ 已验证
DOI
PubMed
高影响因子
BMC BIOLOGY
·
2025
·Vol. 23
·期 1
·1-17
·
IF 4.5
He Yi,Zhang Yan,Liu Minghao,Li Jiaying,Li Wannan,Han Weiwei
Background Dipeptidyl peptidase-4 (DPP4) is considered a crucial enzyme in type 2 diabetes (T2D) treatment, targeted by inhibitors due to its role in cleaving glucagon-like peptide-1 (GLP-1). In this study, a novel DPP4 inhibitor screening strategy was developed, which significantly improved screening accuracy. Results In this study, a DPP4 inhibitor screening method was developed, integrating receptor-based ConPLex, ligand-based KPGT, and molecular docking to enhance screening accuracy. Using this approach, four potential drugs were identified from the FDA database, achieving a 100% hit rate. Among these, Isavuconazonium demonstrated the highest inhibitory activity (IC 50 = 6.60 µM). Furthermore, a user-friendly server, DPP4META, was established to predict IC 50 values for DPP4 inhibitors. The binding and dissociation mechanisms of these drugs with DPP4 were further examined through Gaussian accelerated Molecular Dynamics (GaMD) and ligand Gaussian accelerated Molecular Dynamics (LiGaMD), revealing strong correlations with IC 50 values. Additionally, a Python-based toolkit, pymd, was developed to facilitate protein-compound binding analysis. Conclusions Our study offers a robust approach and valuable insights for the development of DPP4 inhibitors, providing an effective means to investigate the binding and dissociation mechanisms between proteins and compounds.
研究领域:
计算生物学
·
结构生物学
·
药学
·
生物信息学
·
糖尿病研究
·
人工智能辅助药物发现
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）