Design, in silico evaluation, and in vitro verification of new bivalent Smac mimetics with pro-apoptotic activity.

Bivalent Smac mimetics have been shown to possess binding affinity and pro-apoptotic activity similar to or more potent than that of native Smac, a protein dimer able to neutralize the anti-apoptotic activity of an inhibitor of caspase enzymes, XIAP, which endows cancer cells with resistance to anticancer drugs. We design five new bivalent Smac mimetics, which are formed by various linkers tethering two diazabicyclic cores being the IAP binding motifs. We built in silico models of the five mimetics by the TwistDock workflow and evaluated their conformational tendency, which suggests that compound 3, whose linker is n-hexylene, possess the highest binding potency among the five. After synthesis of these compounds, their ability in tumour cell growth inhibition and apoptosis induction displayed in experiments with SK-OV-3 and MDA-MB-231 cancer cell lines confirms our prediction. Among the five mimetics, compound 3 displays promising pro-apoptotic activity and deserves further optimization.

Aortic smooth muscle TRPV4 channels regulate vasoconstriction in high salt-induced hypertension.

Recent studies have focused on the contribution of vascular endothelial transient receptor potential vanilloid 4 (TRPV4) channels to hypertension. However, in hypertension, TRPV4 channels in vascular smooth muscle remain unexplored. In the present study, we performed wire myograph experiments in isolated aortas from endothelial cell specific TRPV4 channel knockout (TRPV4EC-/-) mice to demonstrate that GSK1016790A (a specific TRPV4 channel agonist) triggered aortic smooth muscle-dependent contractions from mice on a normal-salt diet, and the contractions were enhanced in high-salt diet (HSD) mice. Intracellular Ca2+ concentration ([Ca2+]i) and Ca2+ imaging assays showed that TRPV4-induced [Ca2+]i was significantly higher in aortic smooth muscle cells (ASMCs) from HSD-induced hypertensive mice, and application of an inositol trisphosphate receptor (IP3R) inhibitor markedly attenuated TRPV4-induced [Ca2+]i. IP3R2 expression was enhanced in ASMCs from HSD-induced hypertensive mice and the contractile response induced by TRPV4 was inhibited by the IP3R inhibitor. Whole-transcriptome analysis by RNA-seq and western blot assays revealed the involvement of interferon regulatory factor 7 (IRF7) in TRPV4-IRF7-IP3R2 signaling in HSD-induced hypertension. These results suggested that TRPV4 channels regulate smooth muscle-dependent contractions in high salt-induced hypertension, and this contraction involves increased [Ca2+]i, IP3R2, and IRF7 activity. Our study revealed a considerable effect of TRPV4 channels in smooth muscle-dependent contraction in mice during high-salt induced hypertension.

High power, twin-band mid-infrared PPMgLN optical parametric oscillator pumped at 1.679 µm.

We report on a high-power, twin-band mid-infrared (MIR) periodically poled MgO-doped ${{\rm LiNbO}_3}$LiNbO3 (PPMgLN) optical parametric oscillator (OPO) pumped by a near-infrared pulsed OPO with 20 kHz of repetition rate. The pump source has a central wavelength of 1.679 µm and a linewidth of 0.12 nm. The MIR OPO can be tuned from 2.74 to 2.80 µm for signal and from 4.34 to 4.19 µm for idler through raising the oven temperature from 30°C to 200°C, respectively. Under 100°C oven temperature, the OPO yielded maximum total output power of 15.4 W (2.76 µm signal plus 4.29 µm idler) with single-pass pumping configuration. The beam quality ${M^2}$M2 was measured to be $ \lt {2.5}$<2.5 for signal and $ \lt {4}$<4 for idler.

The TwistDock workflow for evaluation of bivalent Smac mimetics targeting XIAP.

Purpose: Mimetics based on Smac, the native inhibitor of XIAP, are promising drug-candidates for the treatment of cancer. Bivalent Smac mimetics inhibit XIAP with even higher potency than monovalent mimetics, but how to optimize the linker that tethers the two monovalent binding motifs remains controversial. Methods: To construct an ensemble of bivalent complex structures for evaluating various linkers, we propose herein a workflow, named TwistDock, consisting of steps of monovalent docking and linker twisting, in which the degrees of freedom are sampled focusing on the rotation of single bonds of the linker. Results: The obtained conformations of bivalent complex distribute randomly in the conformational space with respect to two reaction coordinates introduced by the linker, which are the distance of the two binding motifs and the dihedral angle of the two planes through the linker and each of the binding motifs. Molecular dynamics starting from 10 conformations with the lowest enthalpy of every complex shows that the conformational tendency of the complex participated by compound 9, one of the compounds with the largest binding affinity, is distinct from others. By umbrella sampling of the complex, we find its global minimum of the free energy landscape. The structure shows that the linker favors a compact conformation, and the two BIR domains of XIAP encompass the ligand on the opposite sides. Conclusion: TwistDock can be used in fine-tuning of bivalent ligands targeting XIAP and similar receptors dimerized or oligomerized.

110 W 1678 nm laser based on high-efficiency optical parametric interactions pumped by high-power slab laser.

This Letter presents a high-efficiency optical parametric amplifier pumped by a high-power slab laser with approximate uniform rectangular distribution. By optimizing the overlapping, spectrum matching, and pulse synchronization for the pump and signal lasers, output power of 110.8 W at 1678 nm with corresponding conversion efficiency of 32.3% was achieved in addition to sufficient usage of the effective area in MgO doped periodically poled lithium niobate crystal. It could also provide a designable and tunable wavelength of the amplified laser in a wide infrared region.

High-power and widely tunable mid-infrared optical parametric amplification based on PPMgLN.

We report on a high-power and widely tunable optical parametric amplifier (OPA) based on PPMgLN and pumped by a pulsed 1.064 µm MOPA laser. The operating wavelength of the OPA system is continuously tunable from 2.68 to 3.07 µm by adjusting the temperature of PPMgLN crystals, with average output power varying from 74.6 to 66.7 W for 310 W of pump power, corresponding to an optical-to-optical conversion efficiency of ∼22.8% at 2.68 µm and ∼20.5% at 3.07 µm, respectively. Output beam quality factor (M2) of the OPA was measured to be <4 over the whole tuning range.

High-power, narrow-bandwidth mid-infrared PPMgLN optical parametric oscillator with a volume Bragg grating.

We report on a high-power, narrow spectral bandwidth 2.907 µm PPMgLN optical parametric oscillator (OPO) pumped by a 1.064 µm pulsed Nd:YAG MOPA laser source. Free-running operation of the OPO exhibits maximum average output power of 71.6 W at 2.907 µm with a slope efficiency of 26.7%. Broad 2.907 µm spectral bandwidth of the free-running OPO was suppressed from ~9 nm to less than 0.7 nm by using a VBG as one cavity mirror. The maximum average power was 51.7 W at 2907.55 nm for the spectrum-narrowed OPO, corresponding to a slope efficiency of 22.5%. Continuously tunable ranges of ~8 nm around 2.907 µm had been achieved via adjusting the temperatures of the VBG and PPMgLN accordingly.

Crystal Structures of PI3Kα Complexed with PI103 and Its Derivatives: New Directions for Inhibitors Design.

The phosphatidylinositol 3-kinase (PI3K) signaling pathway plays important roles in cell proliferation, growth, and survival. Hyperactivated PI3K is frequently found in a wide variety of human cancers, validating it as a promising target for cancer therapy. We determined the crystal structure of the human PI3Kα-PI103 complex to unravel molecular interactions. Based on the structure, substitution at the R1 position of the phenol portion of PI103 was demonstrated to improve binding affinity via forming a new H-bond with Lys802 at the bottom of the ATP catalytic site. Interestingly, the crystal structure of the PI3Kα-9d complex revealed that the flexibility of Lys802 can also induce additional space at the catalytic site for further modification. Thus, these crystal structures provide a molecular basis for the strong and specific interactions and demonstrate the important role of Lys802 in the design of novel PI3Kα inhibitors.

Multi-armed poly(L-glutamic acid)-graft-polypropyleneinime as effective and serum resistant gene delivery vectors.

A new series of multi-armed MP-g-PPI dendrimers were synthesized by polymerization of BLG-NCA using G2.0PPI as macromolecular initiator and subsequent aminolysis with G1.0PPI or G2.0PPI. The chemical structure and composition of the MP-g-PPI dendrimers were characterized by Fourier transform infrared spectroscopy (FT-IR) and nuclear magnetic resonance spectroscopy ((1)H NMR). The MP-g-PPI showed a great ability to combine with pDNA to form complexes, which protect the pDNA from nuclease degradation. Dynamic light scattering (DLS) measurement illustrated that the sizes of complexes were in range of 111-219 nm. The transmission electron microscope (TEM) and atomic force microscope (AFM) observation showed that the morphology of these complexes was spherical. The MTT assay demonstrated that cytotoxicity of the MP-g-PPI was lower than that of PEI 25K. The in vitro transfection test indicated that MP-g-PPI gene vectors displayed relative high transfection efficiency than that of PEI 25K and Lipofectamine 2000 in serum-containing medium. Furthermore, MP-g-PPI at the weight ratio of 7.5 displayed better serum-resistant capability than that of PEI 25K and Lipofectamine 2000. The above facts revealed that multi-armed MP-g-PPI dendrimers may be promising gene vectors with low cytotoxicity, high transfection efficiency and serum-resistant ability.

2-(4-Chlorophenyl)-2-oxoethyl 4-benzamidobenzoate derivatives, a novel class of SENP1 inhibitors: Virtual screening, synthesis and biological evaluation.

Prostate cancer is one of the most prevalent types of malignant cancers in men and has a high mortality rate among all male cancers. Previous studies have demonstrated that Sentrin/SUMO-specific protease 1 (SENP1) plays an important role in the occurrence and development of prostate cancer, and has been identified as a novel drug target for development of small molecule drugs against prostate cancer. In this paper, we used virtual screening and docking to identify compound J5 as a novel lead compound inhibiting SENP1, from SPECS library. We further investigated the SAR (structure-activity relationship) of the benzoate substituent of compound J5, and discovered compounds 8d and 8e as better small molecule inhibitors of SENP1. Both compounds are the high potent SENP1 small molecule inhibitors discovered up to date, and further lead optimization may lead to a series of novel anti-SENP1 agents. Further SAR studies are in process and will be reported in due course.

Bivalent Smac mimetics with a diazabicyclic core as highly potent antagonists of XIAP and cIAP1/2 and novel anticancer agents.

Nonpeptidic, bivalent Smac mimetics designed based upon monovalent Smac mimetics with a diazabicyclic core structure bind to XIAP, cIAP1, and cIAP2 with low to subnanomolar affinities and are highly effective in antagonizing XIAP in cell-free functional assays. They efficiently induce the degradation of cIAP1 and cIAP2 in cancer cells at concentrations as low as 1 nM, activate caspase-3 and -8, and cleave PARP at 3-10 nM. The most potent compounds in the series have IC(50) of 3-5 nM in inhibition of cell growth in both MDA-MB-231 and SK-OV-3 cell lines and are promising lead compounds for the development of a new class of cancer therapy.

A potent and orally active antagonist (SM-406/AT-406) of multiple inhibitor of apoptosis proteins (IAPs) in clinical development for cancer treatment.

We report the discovery and characterization of SM-406 (compound 2), a potent and orally bioavailable Smac mimetic and an antagonist of the inhibitor of apoptosis proteins (IAPs). This compound binds to XIAP, cIAP1, and cIAP2 proteins with K(i) of 66.4, 1.9, and 5.1 nM, respectively. Compound 2 effectively antagonizes XIAP BIR3 protein in a cell-free functional assay, induces rapid degradation of cellular cIAP1 protein, and inhibits cancer cell growth in various human cancer cell lines. It has good oral bioavailability in mice, rats, non-human primates, and dogs, is highly effective in induction of apoptosis in xenograft tumors, and is capable of complete inhibition of tumor growth. Compound 2 is currently in phase I clinical trials for the treatment of human cancer.

Therapeutic potential and molecular mechanism of a novel, potent, nonpeptide, Smac mimetic SM-164 in combination with TRAIL for cancer treatment.

Smac mimetics are being developed as a new class of anticancer therapies. Because the single-agent activity of Smac mimetics is very limited, rational combinations represent a viable strategy for their clinical development. The combination of Smac mimetics with TNF-related apoptosis inducing ligand (TRAIL) may be particularly attractive because of the low toxicity of TRAIL to normal cells and the synergistic antitumor activity observed for the combination. In this study, we have investigated the combination synergy between TRAIL and a potent Smac mimetic, SM-164, in vitro and in vivo and the underlying molecular mechanism of action for the synergy. Our study shows that SM-164 is highly synergistic with TRAIL in vitro in both TRAIL-sensitive and TRAIL-resistant cancer cell lines of breast, prostate, and colon cancer. Furthermore, the combination of SM-164 with TRAIL induces rapid tumor regression in vivo in a breast cancer xenograft model in which either agent is ineffective. Our data show that X-linked IAP (XIAP) and cellular IAP 1 (cIAP1), but not cIAP2, work in concert to attenuate the activity of TRAIL; SM-164 strongly enhances TRAIL activity by concurrently targeting XIAP and cIAP1. Moreover, although RIP1 plays a minimal role in the activity of TRAIL as a single agent, it is required for the synergistic interaction between TRAIL and SM-164. This study provides a strong rationale to develop the combination of SM-164 and TRAIL as a new therapeutic strategy for the treatment of human cancer.

High-efficiency mid-infrared optical parametric oscillator based on PPMgO:CLN.

The experimental results of a high-efficiency mid-IR laser are presented on a quasi-phase-matched single-resonated optical parametric oscillator in PPMgO:CLN pumped by a 1064 nm laser of an elliptical beam. The pump source was an acousto-optical Q-switched cw-diode-side-pumped Nd:YAG laser. The beam polarization matched the e-ee interaction in PPMgO:CLN. When the crystal was operated at 110 degrees C and the pump power was 104 W with a repetition rate of 7 kHz, average output powers of 16.7 W at 3.84 microm and 46 W at 1.47 microm were obtained. The slope efficiency of the 3.84 microm laser with respect to the pump laser was 19.1%. The M(2) factors of the 3.84 mum laser were 2.03 and 5.89 in the parallel and perpendicular directions, respectively.

Potent, orally bioavailable diazabicyclic small-molecule mimetics of second mitochondria-derived activator of caspases.

A series of small-molecule Smac mimetics containing a diazabicyclic core structure have been designed, synthesized, and evaluated. The most potent compound (6) binds to XIAP, cIAP-1, and cIAP-2 with K(i) values of 8.4, 1.5, and 4.2 nM, respectively, directly antagonizes XIAP in a cell-free functional assay and induces cIAP-1 degradation in cancer cells. It inhibits cell growth with an IC(50) value of 31 nM, effectively induces apoptosis in the MDA-MB-231 cancer cell line, and has a good oral bioavailability.

Design of small-molecule peptidic and nonpeptidic Smac mimetics.

Smac/DIABLO is a protein released from mitochondria into the cytosol in response to apoptotic stimuli. Smac promotes apoptosis at least in part through antagonizing inhibitor of apoptosis proteins (IAPs), including XIAP, cIAP-1, and cIAP-2. Smac interacts with these IAPs via its N-terminal AVPI binding motif. There has been an enormous interest in academic laboratories and pharmaceutical companies in the design of small-molecule Smac mimetics as potential anticancer agents. This task is particularly challenging because it involves targeting protein-protein interactions. Nevertheless, intense research has now generated potent, specific, cell-permeable small-molecule peptidomimetics and nonpeptidic mimetics. To date, two types of Smac mimetics have been reported, namely, monovalent and bivalent Smac mimetics. The monovalent compounds are designed to mimic the binding of a single AVPI binding motif to IAP proteins, whereas the bivalent compounds contain two AVPI binding motif mimetics tethered together through a linker. Studies from several groups have clearly demonstrated that both monovalent and bivalent Smac mimetics not only enhance the antitumor activity of other anticancer agents but also can induce apoptosis as single agents in a subset of human cancer cell lines in vitro and are capable of achieving tumor regression in animal models of human cancer. In general, bivalent Smac mimetics are 100-1000 times more potent than their corresponding monovalent Smac mimetics in induction of apoptosis in tumor cells. However, properly designed monovalent Smac mimetics can achieve oral bioavailability and may have major advantages over bivalent Smac mimetics as potential drug candidates. In-depth insights on the molecular mechanism of action of Smac mimetics have been provided by several independent studies. It was shown that Smac mimetics induce apoptosis in tumor cells by targeting cIAP-1/-2 for the rapid degradation of these proteins, which leads to activation of nuclear factor kappaB (NF-kappaB) and production and secretion of tumor necrosis factor alpha (TNFalpha). TNFalpha promotes formation of a receptor-interacting serine-threonine kinase 1 (RIPK1)-dependent caspase-8-activating complex, leading to activation of caspase-8 and -3/-7 and ultimately to apoptosis. For the most efficient apoptosis induction, Smac mimetics also need to remove the inhibition of XIAP to caspase-3/-7. Hence, Smac mimetics induce apoptosis in tumor cells by targeting not only cIAP-1/-2 but also XIAP. The employment of potent, cell-permeable, small-molecule Smac mimetics has yielded important insights into the regulation of apoptosis by IAP proteins. To date, at least one Smac mimetic has been advanced into clinical development. Several other Smac mimetics are in an advanced preclinical development stage and are expected to enter human clinical testing for the treatment of cancer in the near future.

[Bioassay on oviposition repellency of non-preferable plant extracts against citrus red mite Panonychus citri].

The oviposition repellency of the alcohol extracts from 50 species of non-preferable plants and azadirachtin against citrus red mite (Panonychus citri) was determined using laboratory bioassays. In choice tests, the extracts from 42 of the 50 plant species and 1% azadirachtin (2000 x) significantly reduced oviposition 1d after treatment. The repellency effect of the extracts from the 42 plant species was better than that of azadirachtin. Mikania micrantha extract had the best result, with an Interference Index of Population Control (IIPC) of 0.087 1 d after treatment, and significantly reduced oviposition for 9 d, while the extracts from Sesbania cannabina, Allium tuberosum, Paederia scandens, Duranta erecta and Dicranopteris pedata also had good effects, with an oviposition repellency of over 70% 1 d after treatment, and significantly reduced oviposition for 4-6 d. The effect became weaker as time went on. None of the extracts showed significant oviposition attraction.

A concise method for the preparation of peptide and arginine-rich peptide-conjugated antisense oligonucleotide.

Peptide-oligonucleotide conjugates were synthesized using two strategies: a mimetic signal peptide-conjugated oligonucleotide was assembled stepwise on CPG by using 2,2-dimethyl-3-hydroxypropionic acid as a linker. To solve the precipitation problem in the coupling reaction caused by the electrostatic interaction of arginine-rich peptides and oligonucleotide, oligonucleotides were absorbed on an anion-exchange resin, and then the on-resin fragment was applied for the conjugation with arginine-rich peptide. The peptide-antisense oligonucleotides showed permeability to the cell membrane of HepG-2 cells.