Yuangang Zu

Review camptothecin : Current perspectives

The review provides a detailed discussion of recent advances in the medicinal chemistry of camptothecin, a potent antitumor agent that targets topoisomerase I. Thousands of CPT derivatives have been synthesized. Two of them, Topotecan and Irinotecan, are commercially approved for use in clinic as antitumor agents while more are still in clinic trials. This review summarizes the current status of the modern synthetic approaches to CPT, the mechanism of action of CPT, the structure-activity relationship(SAR), a number of novel CPT analogs and their biologic activity. There is a systematic evaluation of A-, B- and E-ring- modified camptothecins reported recently.

Medicinal Chemistry of Paclitaxel and its Analogues

Paclitaxel belongs to the most successful anticancer drugs developed and utilised during the past two decades. Nevertheless, the development of resistance of tumor cells and severe side effects in the patients require further improvement of the drug. In this review, we provide a detailed overview of the state-of-the-art in the medicinal chemistry of paclitaxel and its analogues. A number of strategies have been explored to obtain sufficient amounts of paclitaxel for clinical use from natural resources. Semi-synthesis from its precursor, 10-deacetylbaccatin III, which can be extracted from Taxus leavesturned out as the most appropriate method for commercial production. So far, many paclitaxel derivatives have been synthesized, and their effect on microtubules stabilization and cytotoxicity were investigated in terms of structure-activity relationships (SAR). One of them, docetaxel, was approved as a more potent anticancer agent than paclitaxel towards a variety of tumor types. This review summarizes current possibilities to harvest sufficient amount of drugs from natural sources, including the production of taxanes in bioreactors and synthetic approaches for paclitaxel and its analogues, their mechanism of action and structure-activity relationships. In addition, future developments and perspectives for this class of compounds are outlined.

Chemical composition of the SFE-CO2 extracts from Cajanus cajan (L.) Huth and their antimicrobial activity in vitro and in vivo

The in vitro and in vivo antimicrobial activities of SFE-CO₂(supercritical fluid extraction) extracts and ethanol extracts from Cajanus cajan (L.) Huth were investigated. The flavonoid compounds orientin, vitexin, isovitexin, pinostrobin and the stilbene cajaninstilbene acid were detected in SFE-CO₂ extracts by HPLC-DAD. In vitro antimicrobial activities of the extracts were evaluated against eight microbial strains (the bacteria Staphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Proteus vulgaris, Pseudomonas aeruginosa, Escherichia coli; and the fungi Aspergillus niger and Candida albicans). A marked inhibitory effect of the SFE extracts was observed against Staphylococcus epidermidis, Staphylococcus aureus and Bacillus subtilis. The IC(50) of SFE-CO₂ extracts ranged from 0.0557 mg/ml to 0.0689 mg/ml consisting of cancer (MCF-7 (0.0557 mg/ml)) as well as non-cancer (BHK-21 (0.0641 mg/ml), RAW264.7 (0.0689 mg/ml) and Vero (0.0625 mg/ml)) cells. Flow cytometry (FCM) was used to analyze death rate of the most sensitive strain (Staphylococcus aureus) caused by the SFE extracts. Additionally, the whole cell proteins of Staphylococcus aureus were analyzed by SDS-PAGE to detect if there were changes in protein patterns. In vivo antimicrobial activity was studies in mice that had been inoculated with Staphylococcus aureus. The potential mechanism of antimicrobial activity in vivo was studied by histopathology.

The conformational analysis and proton transfer of neuraminidase inhibitors: a theoretical study

With the aid of density functional calculations, it was revealed that neuraminidase (NA) inhibitors (Scheme 1) exist exclusively in the neutral form. However, the docking and molecular dynamics simulations indicated that the zwitterionic, rather than neutral, isomers are the active form in NA-receptors. The neutral and zwitterionic isomers of BA (Compound 7 in Scheme 1) are quite different in structure and is therefore expected to show distinct bioactivities. With the addition of four water molecules, the geometry of the neutral isomer (nBA4) is induced to resemble the zwitterion and remains rather stable throughout the proton transfer process (nBA4 --> zBA4); in addition, the energy difference between the zwitterionic vs. the neutral isomer is lowered from 24.76 to 2.54 kcal mol(-1). The zwitterion is the predominant isomer in aqueous solution, consistent with the conformational preference in NA-receptors. The proton transfer process of nBA4 --> zBA4 is divided into two elementary steps. Step 1, rather than step 2, plays a decisive role, owing to the larger energy barrier; however, step 1 is not assisted by solvent water, even if it is not water-suppressed. Accordingly, the proton transfer process that leads to the formation of zwitterions is not facilitated in aqueous solution, albeit they may be more stable than the neutral isomers. It is thus suggested that the designed antiviral inhibitors should be facile to transform into the zwitterionic form in aqueous solution. In this way, the oral bioavailability of the antiviral drugs can be improved.

Binding of Phytoestrogens to Rat Uterine Estrogen Receptors and Human Sex Hormone-binding Globulins

The interaction of phytoestrogens with the most important binding sites of steroid hormones, i.e. sex hormone-binding globulin and estrogen receptors, was investigated. Relative binding affinities and association constants for 21 compounds among them isoflavones, flavones, flavonols, flavanones, chalcones and lignans were determined. The lignan nordihydroguaiaretic acid weakly displaced 17β-[3H]-estradiol from estrogen receptor and Scatchard analysis suggests non-conformational changes. Compounds from Glycyrrhiza glabra, liquiritigenin and isoliquiritigenin, showed estrogenic affinities to both receptors. 18β-Glycyrrhetinic acid displaced 17β-[3H]-estradiol from sex hormone-binding globulin but not from the estrogen receptor. Phytoestrogens compete with 17β-estradiol much stronger than with 5α-dihydrotestosterone for binding to sex hormone-binding globulin.

Preparative enrichment and separation of astragalosides from Radix Astragali extracts using macroporous resins

The enrichment and separation of astragalosides I-IV (AGs I-IV) were studied on eight macroporous resins in the present study. SA-3 resin offered the best adsorption and desorption capacities for AGs I-IV than other resins. The models of adsorption kinetics were investigated in order to elucidate the mechanism of adsorption. The pseudo-second-order model was the better choice than the pseudo-first-order model to describe the adsorption behavior of AGs I-IV onto SA-3 resin. The equilibrium experimental data were well fitted to Langmuir and Freundlich isotherms. SA-3 resin adsorption chromatography tests were carried out to optimize the separation process of AGs I-IV from Radix Astragali extracts. With the optimum parameters for adsorption and desorption, the contents of AGs I-IV were 8.78-, 11.60-, 10.52- and 11.28-fold increased with the recovery yields being 65.88, 90.92, 84.25 and 94.17%, respectively. The preparative enrichment and separation of AGs I-IV from Radix Astragali extracts can be easily and effectively achieved by SA-3 resin adsorption chromatography. The developed methodology can also be referenced for the separation of other active constituents from herbal materials and manufacture of Radix Astragali products.

Sensitivity and resistance towards isoliquiritigenin, doxorubicin and methotrexate in T cell acute lymphoblastic leukaemia cell lines by pharmacogenomics

The development of drug resistance in cancer cells necessitates the identification of novel agents with improved activity towards cancer cells. In the present investigation, we compared the cytotoxicity of the chalcone flavonoide, isoliquiritigenin (ISL), with that of doxorubicin (DOX) and methotrexate (MTX) in five T cell acute lymphoblastic leukaemia (T-ALL) cell lines (Jurkat, J-Jhan, J16, HUT78 and Karpas 45). To gain insight into the molecular mechanisms which determine the response of T-ALL cells towards ISL, DOX and MTX, we applied array-based matrix comparative genomic hybridisation and microarray-based mRNA expression profiling and compared the genomic and transcriptomic profiles of the cell lines with their 50% inhibition (IC50) values for these three drugs. The IC50 values for ISL did not correlate with those for DOX or MTX, indicating that ISL was still active in DOX- or MTX-unresponsive cell lines. Likewise, the genomic imbalances of chromosomal clones and mRNA expression profile significantly correlating with IC50 values for ISL were different from thoses correlating with IC50 values for DOX and MTX. In conclusion, ISL represents a cytotoxic natural product with activity towards T-ALL cell lines. There was no cross-resistance between ISL and DOX or MTX, and the genomic and transcriptomic profiles pointed to different molecular modes of action of ISL as compared to DOX and MTX, indicating that ISL may be a valuable adjunct for cancer therapy to treat otherwise drug-resistant tumours.

Synthetic cajanin stilbene acid derivatives inhibit c-MYC in breast cancer cells

Abstract In the present study, we investigated the activity and modes of action of cajanin stilbene acid (CSA) and its derivatives in terms of cytotoxicity, gene expression profile, and transcription factor activity. XTT assays on MCF7 cells were performed upon treatment with CSA or derivatives. After the determination of IC50 values, gene expression profiling was performed with Agilent microarray experiments. Deregulated genes were determined with Chipster software, pathway and functional analyses were performed with Ingenuity pathway software. In order to identify the potential upstream regulators, MatInspector software was used to perform transcription factor binding motif search in the promoter regions of the deregulated genes. Molecular docking on MYC/MAX complex and reporter cell line experiments were performed to validate the MYC inhibitory activity of CSA and its derivatives. Two known MYC inhibitors: 10058-F4 and 10074-G5 were used as positive control. All compounds showed cytotoxicities in the micromolar range. Microarray analyses pointed to cell cycle, DNA damage, and DNA repair as mainly affected cellular functions. Promoter motif analysis of the deregulated genes further supported the microarray gene expression analysis results emphasizing the relevance of transcription factors regulating cell cycle and proliferation, with MYC as being the most pronounced one. Luciferase-based reporter cell line experiments and molecular docking studies yielded supportive results emphasizing the inhibitory activity of CSA and its derivatives on MYC. CSA and its derivatives are shown to be promising anticancer compounds with low toxicity. They inhibit MYC activity comparable to 10058-F4 and 10074-G5. Further studies are warranted to analyze the therapeutic applicability of these compounds in more detail.

Enhanced extraction efficiency of bioactive compounds and antioxidant activity from Hippophae rhamnoides L. by-products using a fast and efficient extraction method

ABSTRACT In this study, a novel method named high-speed homogenization coupled with ultrasound-assisted extraction (HSH-UAE) followed by high-performance liquid chromatography (HPLC) was employed and investigated for the extraction and determination of main flavonoids rutin, quercetin-3-O-glucoside, quercetin, kaempferol and isorhamnetin from Hippophae rhamnoides L. by-products. Optimal conditions for HSH-UAE, proposed by the single factor and the Box–Behnken design (BBD) tests coupled with response surface methodology (RSM), were 68% ethanol concentration, ultrasonic power 250 W, homogenate speed 9000 rpm, liquid/solid ratio 22 mL/g and extraction time 12 min. The extraction yields of five main flavonoids were significantly improved by HSH-UAE than UAE and high-speed homogenization-assisted extraction (HSHAE). Furthermore, the antioxidant activity of HSH-UAE extracts was assessed by 1,1-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing/antioxidant power (FRAP) assays with IC50 value of 0.091 mg/mL and 2.837 mmol FeSO4/g DW. Good linear ranges were obtained for five main flavonoids in the range of 2.5–500 μg/mL with the coefficient higher than 0.99. The recoveries of the five flavonoids were in the range of 97.87–99.26%. The limit of detection (LOD) and the limit of quantification (LOQ) for the main flavonoids were within the ranges of 0.28–0.46 and 0.89–1.74 μg/mL, respectively. The present results showed that the proposed method HSH-UAE could be developed as a fast and efficient method for extracting the active ingredients from medical edible plant materials for potential application.

First-principle conformational analysis of glycine residues in the αβ-tubulin dimer

Amino acids, especially glycine, have been extensively studied whereas their conformational behaviors in proteins are rather limited. In this work, all the polypeptides containing glycine residues are truncated from the αβ-tubulin dimer and refined with the partial optimization technique, where the backbone atoms of the previous and posterior residues to the glycine residues are fixed at the experimental Cartesian positions whereas the others fully relaxed. The combination of the polypeptide models and partial optimization technique is validated by twolayer ONIOM calculations, being effective to retain the local structures of proteins and meanwhile optimizing the concerned glycine residues towards energy minima. Owing to the lack of side chains, various types of hydrogen bonding interactions are detected in the glycine residues. The conformational analyses of proteins are mainly based on the dihedrals. Θ1(∠H5C2C3O4), Θ2(∠H6C2C3O4) and Θ3(∠N1C2C3O4) are the three key dihedrals to determine the glycine conformations, which were found to change synchronously and correlate with each other by four linear equations. It thus provided the first evidence of the correlation of intra-dihedrals for the amino acid residues in proteins. In addition, the three dihedrals of the gas-phase glycine conformations were also found to suit well with these linear equations, elucidating the rationality of using amino acids as the computational models of proteins.