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Zoledronate-loaded aluminum salt nanovaccines amplify cellular immune response by enhancing cross-presentation

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Nano Research 2025年第1期18卷 559-571页

作者： Chunting He Penghui He Xue Tang Shuting Bai Ming Qin Yunting Zhang Zhaofei Guo Guangsheng Du Xun Sun Key Laboratory of drug-Targeting and drug Delivery System of the Education Ministry West China School of PharmacySichuan UniversityChengdu 610041China

Being a Th2 stimulator,classic aluminum salt-based adjuvants only stimulate weak cellular immune responses that are required for vaccination against intracellular viruses or cancerous *** a thirdgeneration bisphosphonate,zoledronate(ZOL)can enhance antigen crosspresentation by inhibiting key enzymes of the mevalonate ***,we developed the subunit antigen ovalbumin(OVA)and ZOL co-loaded aluminum hydroxide nanoparticles(APN-OVA-ZOL)and investigated their capacity for inducing cellular immune responses against the *** results showed that the developed nanovaccines could successfully encapsulate OVA and ZOL,and enabled efficient lymph node *** by the mevalonate pathway inhibition effect of ZOL,APN-OVA-ZOL significantly promoted *** a result,APN-OVA-ZOL induced robust cellular immunity,including the activation of T and B *** a EG7-OVA tumor-bearing murine model,APN-OVA-ZOL significantly inhibited the tumor growth and prolonged mice *** work provided a strong empirical foundation indicating that zoledronate-loaded aluminum salt nanovaccines had a strong potency for cancer immunotherapy.

关键词： zoledronate(ZOL) mevalonate pathway antigen cross-presentation cellular immune responses

Research advances in the detection of miRNA

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Journal of Pharmaceutical Analysis 2019年第4期9卷 217-226页

作者： Jiawei Ye Mingcheng Xu Xueke Tian Sheng Cai Su Zeng Institute of drug metabolism and Pharmaceutical Analysis Zhejiang Province Key Laboratory of Anti-Cancer Drug ResearchZhejiang University

MicroRNAs (miRNAs) are a family of endogenous, small (approximately 22 nucleotides in length), noncoding, functional RNAs. With the development of molecular biology, the research of miRNA biological function has attracted significant interest, as abnormal miRNA expression is identified to contribute to serious human diseases such as cancers. Traditional methods for miRNA detection do not meet current demands. In particular, nanomaterial-based methods, nucleic acid amplification-based methods such as rolling circle amplification (RCA), loop-mediated isothermal amplification (LAMP), strand-displacement amplification (SDA) and some enzyme-free amplifications have been employed widely for the highly sensitive detection of miRNA. MiRNA functional research and clinical diagnostics have been accelerated by these new techniques. Herein, we summarize and discuss the recent progress in the development of miRNA detection methods and new applications. This review will provide guidelines for the development of follow-up miRNA detection methods with high sensitivity and specificity, and applicability to disease diagnosis and therapy.

关键词： MicroRNAs Detection method Amplification strategies Nanomaterial-based methods Nucleic acid amplification-based methods

Screening and verifying potential NTCP inhibitors from herbal medicinal ingredients using the LLC-PK1 cell model stably expressing human NTCP

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Chinese Journal of Natural Medicines 2016年第7期14卷 549-560页

作者： SHEN Zhuo-Wei LUO Meng-Yue HU Hai-Hong ZHOU Hui JIANG Hui-Di YU Lu-Shan ZENG Su Institute of drug metabolism and Pharmaceutical Analysis Zhejiang Province Key Laboratory of Anti-Cancer Drug ResearchCollege of Pharmaceutical Sciences Zhejiang University

NTCP is specifically expressed on the basolateral membrane of hepatocytes, participating in the enterohepatic circulation of bile salts, especially conjugated bile salts, to maintain bile salts homeostasis. In addition, recent studies have found that NTCP is a functional receptor of HBV and HDV. Therefore, it is important to study the interaction between drugs and NTCP and identify the inhibitors/substrates of NTCP. In the present study, a LLC-PK1 cell model stably expressing human NTCP was established, which was simple and suitable for high throughput screening, and utilized to screen and verify the potential inhibitors of NTCP from 102 herbal medicinal ingredients. The results showed that ginkgolic acid(GA)(13 : 0), GA(15 : 1), GA(17 : 1), erythrosine B, silibinin, and emodin have inhibitory effects on NTCP uptake of TCNa in a concentration-dependent manner. Among them, GA(13 : 0) and GA(15 : 1) exhibited the stronger inhibitory effects, with IC_(50) values being less than 8.3 and 13.5 mmol·L^(-1), respectively, than the classical inhibitor, cyclosporin A(CsA)(IC_(50) = 20.33 mmol·L^(-1)). Further research demonstrated that GA(13 : 0), GA(15 : 1), GA(17 : 1), silibinin, and emodin were not substrates of NTCP. These findings might contribute to a better understanding of the disposition of the herbal ingredients in vivo, especially in biliary excretion.

关键词： Herbal medicinal ingredients Human NTCP Inhibitor Transport Biliary excretion

pH-triggered cancer-targeting polymers:From extracellular accumulation to intracellular release

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Nano Research 2023年第4期16卷 5155-5168页

作者： Rizwan Ullah Khan Jinning Shao Jia-Yu Liao Linghui Qian Institute of drug metabolism and Pharmaceutical Analysis Zhejiang Province Key Laboratory of Anti-Cancer Drug ResearchCollege of Pharmaceutical SciencesCancer Center&Hangzhou Institute of Innovative MedicineZhejiang UniversityHangzhou 310058China

Stimuli-responsive polymers are promising to achieve targeted delivery,improved stability during circulation,and controlled release of therapeutic and diagnostic *** them,pH-responsive polymeric nanocarriers have attracted significant attention as pH varies in different body fluids(e.g.,stomach,intestine,and colon)and intracellular organelles(e.g.,endosome,lysosome,and mitochondria)to maintain homeostasis,while distinctive pH changes are also found in certain pathological *** example,the extracellular environment of the tumor is acidic,which can be employed to drive selective *** the internalization process,since most nanocarriers enter cells upon endocytosis where a drop of pH from 6.5 to 5.0 can occur from endosome to lysosome,pH-sensitive groups have been developed for enhanced cargo *** this review,both non-covalent and covalent interactions responsive to pH changes are introduced,with a focus on the structure-property relationship and their applications in cancer targeting and endosomal escape.

关键词： cancer targeting pH-responsive structure-property relationship charge shifting acid-labile linkage

Mechanism for ginkgolic acid(15：1)-induced MDCK cell necrosis： Mitochondria and lysosomes damages and cell cycle arrest

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Chinese Journal of Natural Medicines 2017年第5期15卷 375-383页

作者： YAO Qing-Qing LIU Zhen-Hua XU Ming-Cheng HU Hai-Hong ZHOU Hui JIANG Hui-Di YU Lu-Shan ZENG Su Institute of drug metabolism and Pharmaceutical Analysis Zhejiang Province Key Laboratory of Anti-Cancer Drug Research College of Pharmaceutical Sciences Zhejiang University Hangzhou 310058 China

Ginkgolic acids(GAs), primarily found in the leaves, nuts, and testa of ginkgo biloba, have been identified with suspected allergenic, genotoxic and cytotoxic properties. However, little information is available about GAs toxicity in kidneys and the underlying mechanism has not been thoroughly elucidated so far. Instead of GAs extract, the renal cytotoxicity of GA(15 : 1), which was isolated from the testa of Ginkgo biloba, was assessed in vitro by using MDCK cells. The action of GA(15 : 1) on cell viability was evaluated by the MTT and neutral red uptake assays. Compared with the control, the cytotoxicity of GA(15 : 1) on MDCK cells displayed a time-and dose-dependent manner, suggesting the cells mitochondria and lysosomes were damaged. It was confirmed that GA(15 : 1) resulted in the loss of cells mitochondrial trans-membrane potential(ΔΨm). In propidium iodide(PI) staining analysis, GA(15 : 1) induced cell cycle arrest at the G0/G1 and G2/M phases, influencing on the DNA synthesis and cell mitosis. Characteristics of necrotic cell death were observed in MDCK cells at the experimental conditions, as a result of DNA agarose gel electrophoresis and morphological observation of MDCK cells. In conclusion, these findings might provide useful information for a better understanding of the GA(15 : 1) induced renal toxicity.

关键词： Ginkgolic acids(15:1) Cytotoxicity Mechanism Necrosis

Recent progress in the molecular imaging of therapeutic monoclonal antibodies

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Journal of Pharmaceutical Analysis 2020年第5期10卷 397-413页

作者： Kaifeng He Su Zeng Linghui Qian Institute of drug metabolism and Pharmaceutical Analysis Zhejiang Province Key Laboratory of Anti-Cancer Drug ResearchCollege of Pharmaceutical SciencesZhejiang UniversityHangzhou310058China

Therapeutic monoclonal antibodies have become one of the central components of the healthcare system and continuous efforts are made to bring innovative antibody therapeutics to patients in *** is equally critical to acquire sufficient knowledge of their molecular structure and biological functions to ensure the efficacy and safety by incorporating new detection approaches since new challenges like individual differences and resistance are *** techniques for determining antibody disposition including plasma drug concentration measurements using LC-MS or ELISA,and tissue distribution using immunohistochemistry and immunofluorescence are now complemented with molecular imaging modalities like positron emission tomography and near-infrared fluorescence imaging to obtain more dynamic information,while methods for characterization of antibody’s interaction with the target antigen as well as visualization of its cellular and intercellular behavior are still under *** progress in detecting therapeutic antibodies,in particular,the development of methods suitable for illustrating the molecular dynamics,is described here.

关键词： Therapeutic monoclonal antibodies Molecular structure Biological function Molecular imaging

Recent advances and perspectives of nucleic acid detection for coronavirus

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Journal of Pharmaceutical Analysis 2020年第2期10卷 97-101页

作者： Minzhe Shen Ying Zhou Jiawei Ye Abdu Ahmed Abdullah AL-maskri Yu Kang Su Zeng Sheng Cai Institute of drug metabolism and Pharmaceutical Analysis Zhejiang Province Key Laboratory of Anti-Cancer Drug ResearchCollege of Pharmaceutical SciencesZhejiang UniversityHangzhouZhejiang310058China

The recent pneumonia outbreak caused by a novel coronavirus(SARS-CoV-2)is posing a great threat to global public ***,rapid and accurate identification of pathogenic viruses plays a vital role in selecting appropriate treatments,saving people's lives and preventing *** is important to establish a quick standard diagnostic test for the detection of the infectious disease(COVID-19)to prevent subsequent secondary *** chain reaction(PCR)is regarded as a gold standard test for the molecular diagnosis of viral and bacterial infections with high sensitivity and *** nucleic acid amplification is considered to be a highly promising candidate method due to its fundamental advantage in quick procedure time at constant temperature without thermocycler opera-tion.A variety of improved or new approaches also have been *** review summarizes the currently available detection methods for coronavirus nucleic *** is anticipated that this will assist researchers and clinicians in developing better techniques for timely and effective detection of coro-navirus infection.

关键词： Coronavirus Nucleic acid detection PCR-Based methods Isothermal nucleic acid amplification-based methods Microarray-based methods

The metabolism and hepatotoxicity of ginkgolic acid(17:1) in vitro

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Chinese Journal of Natural Medicines 2018年第11期16卷 829-837页

作者： YAO Qing-Qing LI Li XU Ming-Cheng HU Hai-Hong ZHOU Hui YU Lu-Shan ZENG Su Institute of drug metabolism and Pharmaceutical Analysis Zhejiang Province Key Laboratory of Anti-Cancer Drug Research College of Pharmaceutical Sciences Department of Pharmacy & Geriatrics Institute of Zhejiang Province Zhejiang Hospital

Pharmacological activities and adverse side effects of ginkgolic acids(GAs), major components in extracts from the leaves and seed coats of Ginkgo biloba L, have been intensively studied. However, there are few reports on their hepatotoxicity. In the present study, the metabolism and hepatotoxicity of GA(17:1), one of the most abundant components of GAs, were investigated. Kinetic analysis indicated that human and rat liver microsomes shared similar metabolic characteristics of GA(17:1) in phase I and II metabolisms. The drug-metabolizing enzymes involved in GA(17:1) metabolism were human CYP1 A2, CYP3 A4, UGT1 A6, UGT1 A9, and UGT2 B15, which were confirmed with an inhibition study of human liver microsomes and recombinant enzymes. The MTT assays indicated that the cytotoxicity of GA(17:1) in HepG2 cells occurred in a time-and dose-dependent manner. Further investigation showed that GA(17:1) had less cytotoxicity in primary rat hepatocytes than in HepG2 cells and that the toxicity was enhanced through CYP1 A-and CYP3 A-mediated metabolism.

关键词： Ginkgolic acid (17 : 1) Cytotoxicity Liver microsomes Recombinant enzyme Hepatotoxicity

Significance and challenges of stereoselectivity assessing methods in drug metabolism

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Journal of Pharmaceutical Analysis 2016年第1期6卷 1-10页

作者： Zhuowei Shen Chuang Lv Su Zeng Institute of drug metabolism and Pharmaceutical Analysis Zhejiang Province Key Laboratory of Anti-Cancer Drug Research College of Pharmaceutical Sciences Zhejiang University Biogen Idec Cambridge

Stereoselectivity in drug metabolism can not only influence the pharmacological activities, tolerability, safety, and bioavailability of drugs directly, but also cause different kinds of drug-drug interactions. Thus, assessing stereoselectivity in drug metabolism is of great significance for pharmaceutical research and development （R＆D） and rational use in clinic. Although there are various methods available for assessing stereoselectivity in drug metabolism, many of them have shortcomings. The indirect method of chro- matographic methods can only be applicable to specific samples with functional groups to be derivatized or form complex with a chiral selector, while the direct method achieved by chiral stationary phases （CSPs） is expensive. As a detector of chromatographic methods, mass spectrometry （MS） is highly sen- sitive and specific, whereas the matrix interference is still a challenge to overcome. In addition, the use of nuclear magnetic resonance （NMR） and immunoassay in chiral analysis are worth noting. This review presents several typical examples of drug stereoselective metabolism and provides a literature-based evaluation on current chiral analytical techniques to show the significance and challenges of stereo- selectivity assessing methods in drug metabolism.

关键词： Enantiomer Chiral chromatography Capillary electrophoresis Mass spectrometry NMR Immunoassay