Lysosome-targeted Aza-BODIPY photosensitizers for anti-cancer photodynamic therapy.

Developing effective near-infrared (NIR) photosensitizers (PSs) has been an attractive goal of photodynamic therapy (PDT) for cancer treatment. In this study, we synthesized N, N-diethylaminomethylphenyl-containing Aza-BODIPY photosensitizers and comprehensively investigated their photophysical/photochemical properties, as well as cell-based and animal-based anti-tumor studies. Among them, BDP 1 has strong NIR absorption at 680 nm and higher singlet oxygen yield in PBS which showed favorable pH-activatable and lysosome-targeting ability. BDP 1 could be easily taken up by tumor cells and showed negligible dark activity (IC50 > 50 µM), however strong phototoxicity upon exposure to light irradiation. The acceptable fluorescence emission from BDP 1 allowed convenient in vivo fluorescence imaging for organ distribution studies in mice. After PDT treatment with upon single time PDT treatment at the beginning using relatively low light dose (54 J/ cm2), BDP 1 (2 mg/kg, 0.1 mL) was found to have strong efficacy to inhibit tumor growth and even to ablate off tumor without causing body weight loss. Therefore, pH-activatable and lysosome-targeted PS may become an effective way to develop potent PDT agent.

Access to Pyridinyl or Pyridinium Aza-BODIPYs with Tunable Near-Infrared Fluorescence through ICT from 4-Pyridinyl Pyrroles.

Aza-dipyrromethene boron difluoride (Aza-BODIPY) dyes have attracted much interest in recent decades. In this manuscript, we provide a facile way to access pyridine-substituted pyrroles. A series of 1/7-pyridinyl and pyridinium near-infrared (NIR) Aza-BODIPYs with/without rigidity were constructed and their spectroscopic properties were extensively investigated. These pyridinyl Aza-BOIDIPYs displayed slight bathochromic shifts and maintained bright NIR emission. Pyridinium Aza-BODIPYs showed increased bathochromic shift, however, the molar extinction coefficients and fluorescence quantum yields were decreased owing to ICT effect. The impacts followed the order of pyridin-4-ium>pyridin-2-ium>pyridin-3-ium, which was in consistence with their ICT strength. MO calculation was performed to provide possible explanation to the red-shifted absorption/emission, and apparent charge separation in pyridin-4-ium Aza-BODIPYs. The pyridinium Aza-BODIPY localized in lysosome and potentially avoided harmful ''alkalinizing effects'' of traditional lysosome-targeting fluorescent dyes containing amine moiety. We are working on construction of pyridinyl and pyridinium Aza-BODIPY photosensitizers against microbials or tumors.

Water-soluble near-infrared fluorescent heptamethine dye for lymphatic mapping applications.

The identification of sentinel lymph node (SLN) is an important method for prognostic evaluation and minimally invasive staging of metastatic tumors. Here, we report a series of near-infrared fluorescent heptamethylamine dyes (series A, B and C) with central cycloalkene ring modified by tyrosine or N-Boc tyrosine via ether linkage. N-Boc tyrosine/tyrosine modification provided enhanced absorption coefficient and fluorescence quantum yield in DMSO, however with slight hypsochromic shift compared to the mother dyes in DMSO. In PBS, series A and B were found to be more fluorescent than ICG and showed brighter images. Compound A1 was found to exhibit the most favorable imaging performance among all the dyes investigated and was selected for in vivo sentinel lymph node mapping experiments in mice. A1 showed faster response and stronger fluorescence emission than FDA-approved ICG. The lymph node tracing with A1 could be assisted by MB staining. Ex vivo imaging of harvested organs indicated that similar metabolic characteristics of A1 and ICG. Overall, A1 is advantageous over ICG and is very promising for non-invasive lymph node imaging.

Goal-driven method for decoding the configuration of coherent wave groups required for the generation of arbitrary-order vortex lattices.

Together, the number of waves, wave vectors, amplitudes, and additional phases constitute the coherent wave group configuration and determine the pattern of the interference field. Identifying an appropriate wave group configuration is key to generating vortex lattices via interferometry. Previous studies have approached this task by first assigning the four elements, then calibrating the vortex state of the interference field. However, this method has failed to progress beyond generating third-order vortex lattices, which are insufficient for some practical applications. Therefore, this study proposes a method for determining the proper wave group configurations corresponding to arbitrary-order vortex lattices. We adopt a goal-driven approach: First, we set a vortex lattice as the target field and model it, before decomposing the target field into a sum of multiple harmonics using Fourier transforms. These harmonics constitute the wave group required to generate the target vortex lattice. As vortex lattices of any order can be set as the target field, the proposed method is compatible with any mode order. Simulations and experiments were conducted for fourth- and fifth-order vortex lattices, thus demonstrating the effectiveness of the proposed method.

Circ_GRN Promotes the Proliferation, Migration, and Inflammation of Vascular Smooth Muscle Cells in Atherosclerosis Through miR-214-3p/FOXO1 Axis.

ABSTRACT: Dysfunction of vascular smooth muscle cells (VSMCs) assumes a fundamental part in the pathogenesis of atherosclerosis (AS). Circular RNA granulin precursor (circ_GRN) was identified to promote the proliferation and invasion of human VSMCs (HVSMCs) in an in vitro AS model. However, the underlying mechanisms remain unclear. Levels of circ_GRN, microRNA (miR)-214-3p, and forkhead box protein O1 (FOXO1) were detected using quantitative real-time polymerase chain reaction and Western blot assays. The proliferation, migration, and inflammatory response of HVSMCs were evaluated by using flow cytometry, colony formation, Cell Counting Kit-8, Western blot, transwell assays, and enzyme-linked immunosorbent assay, respectively. The binding interaction between miR-214-3p and circ_GRN or FOXO1 was detected by dual-luciferase reporter assay. In this study, we found that circ_GRN was elevated in the serum of AS and oxidized low-density lipoprotein (ox-LDL)-induced HVSMCs. The in vitro AS model was established by exposing HVSMCs to ox-LDL, and we found circ_GRN knockdown reversed ox-LDL-evoked cell proliferation, migration, and inflammation. In a mechanical study, miR-214-3p directly bound to circ_GRN or FOXO1, and circ_GRN could regulate FOXO1 expression by competitively binding to miR-214-3p. Importantly, we demonstrated that miR-214-3p inhibition attenuated the protective effects of circ_GRN knockdown on ox-LDL-induced HVSMCs; besides that, miR-214-3p overexpression abolished ox-LDL-triggered HVSMC proliferation, migration, and inflammation, which were counteracted by FOXO1 upregulation. In conclusion, circ_GRN promoted the proliferation, migration, and inflammation of HVSMCs through miR-214-3p/FOXO1 axis in ox-LDL-induced AS model in vitro, suggesting the potential involvement in an AS process, which provided a potential candidate for future clinic intervention in AS.

Discovery of novel hydroxyamidine derivatives as indoleamine 2,3-dioxygenase 1 inhibitors with in vivo anti-tumor efficacy.

Indoleamine 2,3-dioxygenase 1 (IDO1) is closely associated with immune escape in many tumor tissues, and is considered to be a valuable therapeutic target in cancer immunotherapy. In this study, the modification of amino sidechain was performed with the hydroxyamidine core kept intact to optimize lead compound Epacadostat. 19 new compounds with hydrazide, thietane or sulfonamide moiety as polar capping group in sidechain were prepared and their IDO1 inhibitory activities were evaluated. Sulfonamide 3a showed potent IDO1 inhibition in both enzymatic and cellular assays with the IC50 value of 71 nM and EC50 value of 11 nM, respectively. Furthermore, in vivo Lewis lung cancer (LLC) allograft studies of 3a indicated that it handicapped the tumor growth with similar efficacy to Epacadostat. Molecular docking demonstrated that the change of polar capping group affords influence on the orientation of amino ethylene side chain and forms new hydrogen bonding.

Simplified unobscured optics design for a diffractive telescope.

An unobscured optical system provides a good solution for a high-performance diffractive telescope. However, the unobscured diffractive telescope, such as an off-axis three-mirror diffractive telescope (OTDT), suffers from the complex calculation of initial parameters, strict tolerance, and fabrication of aspheric mirror diffractive optical elements (DOE). In this paper, a simplified unobscured design method, which is effective in achieving compact and loose tolerance, is proposed. Combining the deflection of an optical path and Schupmann achromatic theory, a novel unobscured diffractive telescope is rapidly converted from a coaxial transmissive system. Not only is the loose tolerance obtained, but the fabrication of an aspheric mirror DOE is avoided. The designed system, with a focal length of 400 mm, $f$f-number of 5, and working waveband of 612.8-652.8 nm, is analyzed. The results show that the imaging quality approached the diffraction limit within a field of view of ${0.04}^\circ \times {0.02}^\circ $0.04∘×0.02∘. Compared with the OTDT, the designed system has great advantages in design step, DOE fabricating, and system alignment. It also provides a reference for unobscured diffractive telescope development.

Multi-beams engineered to increase patterns of vortex lattices by employing zero lines of the coherent non-diffracting field.

The number of zero lines of the real and imaginary parts of the optical vortex (OV), both are the same as the topological charge (TC), and all of these lines intersect at one point where the phase singularity is. Furthermore, zero crossings distribute regularly on the transverse plane of the OV lattice. Zero lines of the real and imaginary parts of the non-diffracting fields without OV that generated by multi-waves interference are periodic but coincident. We stack two groups of these kind of zero lines which can be regarded as a set of zero lines of the real part and a set of zero lines of the imaginary part respectively, to satisfy the cross state of zero lines of an OV lattice. Then two groups of multi-waves corresponding to the two fields can be obtained. The expected OV lattice that generated by the two groups of engineered waves interference together is validated through both numerical simulations and experiments.

Analysis and optimization of the Glass pattern realized by a microlens array.

It is well known that the superposition of two identical random dot patterns may give rise to a particular form of moiré effect known as a Glass pattern. By integrating a microlens array and a micropattern array and applying a small geometrical transformation, it is possible to show the parallactic or the orthoparallactic motion effect when varying the viewing angle. In this paper, we study the effect of the structural parameters of a Glass pattern optical system on its image quality. The position of each elemental pattern in the micropattern array is derived based on a revised model that considers the finite observation distance. The image quality is evaluated by the ray analysis on the observation plane and the reconstructed image plane. Rays from different pixels of the micropattern array are evaluated by the spot size. The depth of the Glass pattern in image space is also analyzed. In addition, a Glass pattern optical system is simulated and optimized by ZEMAX software. An optimal result is obtained that validates the theoretical analysis. Our study can find potential application for designing Glass pattern optical systems or optimization of those imaging systems based on microlens arrays.

Synthesis and evaluation of novel dimethylpyridazine derivatives as hedgehog signaling pathway inhibitors.

We report herein the design and synthesis of a series of structural modified dimethylpyridazine compounds as novel hedgehog signaling pathway inhibitors. The bicyclic phthalazine core and 4-methylamino-piperidine moiety of Taladegib were replaced with dimethylpyridazine and different azacycle building blocks, respectively. The in vitro Gli-luciferase assay results demonstrate that the new scaffold still retained potent inhibitory potency. Piperidin-4-amine moiety was found to be the best linker between pharmacophores dimethylpyridazine and fluorine substituted benzoyl group. Furthermore, the optimization of 1-methyl-1H-pyrazol and 4-fluoro-2-(trifluoromethyl)benzamide by different aliphatic or aromatic rings were also investigated and the SAR were described. Several new derivatives were found to show potent Hh signaling inhibitory activity with nanomolar IC50 values. Among these compounds, compound 11c showed the highest inhibitory potency with an IC50 value of 2.33 nM, which was comparable to the lead compound Taladegib. In vivo efficacy of 11c in a ptch+/-p53-/- mouse medulloblastoma allograft model also indicated encouraging results.

Properties of dynamic image displacements based on microstructure.

The superposition structure of dynamic moiré images contains a layer of micropattern array and a corresponding layer of microlens array. Dynamic moiré images can be observed without special eyeglasses and will shift with variation of the viewing angle. In this work, it is described as a presentation of a variety of the moiré. We chiefly investigate the motion effects of dynamic moiré images. Analytical models to describe the structures and methods to evaluate the displacement of dynamic moiré images are proposed. Experiments are conducted to validate the formula of the displacement. The results of this study are helpful to design dynamic moiré images, which can be applied for potential applications in anti-counterfeiting films, packaging, and authentication.

Bioimaging of Intravenous Polymeric Micelles Based on Discrimination of Integral Particles Using an Environment-Responsive Probe.

One of the biggest challenges in bioimaging of nanoparticles is how to identify integral particles from bulk signals of probes. Signals of free probes are always mistakenly counted into total signals of particles. In this study, in vivo fate of intravenous polymeric micelles (PMs, mPEG2.5k-PDLLA2.5k) was explored using a highly sensitive near-infrared environment-responsive fluorescent probe. This probe is able to emit fluorescence when embedded in nanocarriers but quench spontaneously and absolutely upon release into water, based on the aggregation-caused quenching effect, which means that the interference generated by free probes can be completely diminished. Analysis of blood-borne fluorescence reveals rapid clearance of PMs from blood following a tricompartmental pharmacokinetic model. Live imaging shows pervasive distribution of PMs throughout the body, and a tendency of accumulation to extremities with fluorescence density 3-5 times higher than the trunk. Ex vivo examination reveals that most PMs are found in vital organs following an order of lung > liver > spleen > heart > kidney in concentration, but an order of liver > lung > spleen > heart ≈ kidney in total amount. The distribution to other organs and tissues is even lower, and to brain, negligible. It is concluded that the biodistribution of PMs to vital organs and extremities warns of potential toxicity and can be translated to explain the toxicity of its commercial counterpart with similar chain lengths.

Controlling Release of Integral Lipid Nanoparticles Based on Osmotic Pump Technology.

PURPOSE: To achieve controlled release of integral nanoparticles by the osmotic pump strategy using nanostructured lipid carriers (NLCs) as model nanoparticles. METHODS: NLCs was prepared by a hot-homogenization method, transformed into powder by lyophilization, and formulated into osmotic pump tablets (OPTs). Release of integral NLCs was visualized by live imaging after labeling with a water-quenching fluorescent probe. Effects of formulation variables on in vitro release characteristics were evaluated by measuring the model drug fenofibrate. Pharmacokinetics were studied in beagle dogs using the core tablet and a micronized fenofibrate formulation as references. RESULTS: NLCs are released through the release orifices of the OPTs as integral nanoparticles. Near zero-order kinetics can be achieved by optimizing the influencing variables. After oral administration, decreased C max and steady drug levels for as long as over 24 h are observed. NLC-OPTs show an oral bioavailability of the model drug fenofibrate similar to that of the core tablets, which is about 1.75 folds that of a fast-release formulation. CONCLUSION: Controlled release of integral NLCs is achieved by the osmotic pump strategy.

Synthesis and evaluation of novel benzylphthalazine derivatives as hedgehog signaling pathway inhibitors.

We report herein the design and synthesis of a series of novel benzylphthalazine derivatives as hedgehog signaling pathway inhibitors. Gli-luciferase assay demonstrated that changing piperazine ring of Anta XV to different four, five or six-membered heterocyclic building blocks afforded significant influences on Hh pathway inhibition. In particular, compound 10e with piperidin-4-amine moiety was found to possess 12-fold higher Hh inhibitory activities comparing to the lead compound in vitro. In vivo efficacy of 10e in a ptch(+/-)p53(-/-) mouse medulloblastoma allograft model also indicated encouraging results.

Environment-responsive aza-BODIPY dyes quenching in water as potential probes to visualize the in vivo fate of lipid-based nanocarriers.

Environment-responsive near-infrared (NIR) aza-BODIPY dyes capable of fluorescence quenching in water were explored to visualize the in vivo fate of model lipid-based nanocarriers, solid lipid nanoparticles (SLNs). The water-quenching effect of the dyes was confirmed to be sensitive and remained stable for at least 24h. In vitro lipolysis measured by fluorescence quenching completed within 20min, which was in correlation with alkaline compensation results. In vivo live imaging indicated predominant digestion of SLNs within 2h and complete digestion within 4h, which correlated well to in vitro data. Rekindling of quenched dyes by mixed micelles was observed in vitro, but not in vivo. In sharp contrast, SLNs encapsulating another NIR dye DiR showed persistent fluorescence both in vitro and in vivo despite significant lipolysis. It was envisaged that water-quenching fluorescence dyes can be used as probes to monitor the in vivo fate of lipid-based nanocarriers. FROM THE CLINICAL EDITOR: Lipid-based drug delivery systems can provide an excellent nanocarrier platform for the delivery of poorly water-soluble drugs. Nonetheless, the mechanism of oral absorption and subsequent kinetics is poorly understood. In this article, the authors studied the novel use of near-infrared (NIR) aza-BODIPY dyes to visualize the fate of these lipid-based nanocarriers. The positive finding means that this approach may be useful for in-vivo monitoring of lipid-based nanocarriers.

Tyrosine kinase inhibitor Thiotanib targets Bcr-Abl and induces apoptosis and autophagy in human chronic myeloid leukemia cells.

Chronic myeloid leukemia (CML) is characterized by abnormal Bcr and Abl genes and enhanced tyrosine kinase activity. Anti-CML therapy has been much improved along with the applications of tyrosine kinase inhibitors (TKIs) which selectively target Bcr-Abl and have a cytotoxic effect on CML. Recently, four-membered heterocycles as "compact modules" have attracted much interest in drug discovery. Grafting these small four-membered heterocycles onto a molecular scaffold could probably provide compounds that retain notable activity and populate chemical space otherwise not previously accessed. Accordingly, a novel TKI, Thiotanib, has been designed and synthesized. It selectively targets Bcr-Abl, inducing growth inhibition, cell cycle arrest, and apoptosis of CML cells. Meanwhile, the compound Thiotanib could also induce autophagy in CML cells. Interestingly, inhibition of autophagy promotes Thiotanib-induced apoptosis with no further activation of caspase 3, while inhibition of caspases did not affect the cell survival of CML cells. Moreover, the compound Thiotanib could inhibit phosphorylation of Akt and mTOR, increase beclin-1 and Vps34, and block the formation of the Bcl-2 and Beclin-1 complex. This indicates the probable pathway of autophagy initiation. Our results highlight a new approach for TKI reforming and further provide an indication of the efficacy enhancement of TKIs in combination with autophagy inhibitors.

Compound Z526 alleviates chemotherapy-induced cachectic muscle loss by ameliorating oxidative stress-driven protein metabolic imbalance and apoptosis.

Chemotherapy is one of the primary and indispensable intervention against cancers though it is always accompanied by severe side effects especially cachexia. Cachexia is a fatal metabolic disorder syndrome, mainly characterized by muscle loss. Oxidative stress is the key factor that trigger cachectic muscle loss by inducing imbalance in protein metabolism and apoptosis. Here, we showed an oral compound (Z526) exhibited potent alleviating effects on C2C12 myotube atrophy induced by various chemotherapeutic agents in vitro as well as mice muscle loss and impaired grip force induced by oxaliplatin in vivo. Furthermore, Z526 also could ameliorate C2C12 myotube atrophy induced by the combination of chemotherapeutic agents with conditioned medium of various tumor cells in vitro as well as mice muscle atrophy of C26 tumor-bearing mice treated with oxaliplatin. The pharmacological effects of Z526 were based on its potency in reducing oxidative stress in cachectic myocytes and muscle tissues, which inhibited the activation of NF-κB and STAT3 to decrease Atrogin-1-mediated protein degradation, activated the AKT/mTOR signaling pathway to promote protein synthesis, regulated Bcl-2/BAX ratio to reduce Caspase-3-triggered apoptosis. Our work suggested Z526 to be an optional strategy for ameliorating cachexia muscle atrophy in the multimodality treatment of cancers.

ADAM17 is overexpressed in non-small cell lung cancer and its expression correlates with poor patient survival.

The purpose of this study was to assess ADAM17 expression and to explore its contribution to the non-small cell lung cancer (NSCLC). Real-time quantitative reverse transcriptase-polymerase chain reaction was conducted to detect ADAM17 mRNA expression. In addition, ADAM17 expression was analyzed by immunohistochemistry in 124 clinicopathologically characterized NSCLC cases. The correlation of ADAM17 expression with patients' survival rate was assessed by Kaplan-Meier and Cox regression. The expression levels of ADAM17 mRNA and protein in NSCLC tissues were both significantly higher than those in non-cancerous tissues. In addition, high expression of ADAM17 was significantly correlated with tumor grade (P=0.026), tumor size (P=0.001), clinical stage (P=0.016), and lymph node metastases (P<0.001). Furthermore, multivariate analysis suggested that tumor grade, tumor size, clinical stage, lymph node metastases, and ADAM17 expression were independent prognostic indicators for NSCLC. Our data suggest for the first time that the increased expression of ADAM17 in NSCLC is associated significantly with aggressive progression and poor prognosis. ADAM17 may be an important molecular marker for predicting the carcinogenesis, progression, and prognosis of NSCLC.

High expression of JMJD6 predicts unfavorable survival in lung adenocarcinoma.

The upregulated expression of JMJD6 was observed in various human cancers. However, little was known about JMJD6 expression and its clinicopathological significance in lung adenocarcinoma. The aim of this study was to investigate the expression and significance of JMJD6 in lung adenocarcinoma progression and prognosis. The levels of JMJD6 mRNA and protein in lung adenocarcinoma specimens and corresponding non-tumorous lung tissues were evaluated by quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) and Western blot. In order to investigate the correlations between JMJD6 and the clinicopathological features of lung adenocarcinoma, the expression of JMJD6 in 154 patients with lung adenocarcinoma was detected by immunohistochemistry. By qRT-PCR and Western blot, the relative expression levels of JMJD6 mRNA and protein were significantly higher in lung adenocarcinoma tissues than in corresponding non-tumorous lung tissues (P < 0.001). Immunohistochemical staining revealed that high JMJD6 expression was closely correlated with tumor size (P = 0.005), pathological grade (P = 0.003), pT status (P = 0.012), pN status (P = 0.003), and pleural invasion (P < 0.001). Moreover, the results of Kaplan-Meier analysis indicated that a high expression level of JMJD6 resulted in a significantly poor prognosis of lung adenocarcinoma patients. Multivariate analysis showed that the status of JMJD6 expression was an independent prognostic factor for lung adenocarcinoma patients. Our results showed that JMJD6 plays a key role in lung adenocarcinoma and therefore may provide an opportunity for developing a novel therapeutic target as well as a prognostic marker in lung adenocarcinoma.

Four-membered heterocycles-containing 4-anilino-quinazoline derivatives as epidermal growth factor receptor (EGFR) kinase inhibitors.

We report herein the design and synthesis of novel azaspirocycle or azetidine substituted 4-anilinoquinazoline derivatives. The EGFR inhibitory activities and in vitro antitumor potency of these newly synthesized compounds against two lung cancer cell lines HCC827 and A549 were evaluated. Most of the target compounds possess good inhibitory potency. In particular, compounds 21g with 2-oxa-6-azaspiro[3.4]octane substituent was found to possess higher EGFR inhibitory activities and similar antitumor potency comparing to the lead compound gefitinib with improved water solubility.