In vitro cytotoxicity, pharmacokinetics, tissue distribution, and metabolism of small-molecule protein kinase D inhibitors, kb-NB142-70 and kb-NB165-09, in mice bearing human cancer xenografts

Actin turnover in dendritic spines influences spine development, morphology, and plasticity, with functional consequences on learning and memory formation. In nonneuronal cells, protein kinase D (PKD) has an important role in stabilizing F-actin via multiple molecular pathways. Using in vitro models of neuronal plasticity, such as glycine-induced chemical long-term potentiation (LTP), known to evoke synaptic plasticity, or long-term depolarization block by KCl, leading to homeostatic morphological changes, we show that actin stabilization needed for the enlargement of dendritic spines is dependent on PKD activity. Consequently, impaired PKD functions attenuate activity-dependent changes in hippocampal dendritic spines, including LTP formation, cause morphological alterations in vivo, and have deleterious consequences on spatial memory formation. We thus provide compelling evidence that PKD controls synaptic plasticity and learning by regulating actin stability in dendritic spines.

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In-vitro Cytotoxicity of Wrightia pubescens (Blanco) Merr., Aphanamixis polystachya (Wall.) Parker, and Platymitra arborea (Blanco) against selected human cancer cell lines

International Journal of Biosciences (IJB) ◽

10.12692/ijb/11.5.204-213 ◽

2017 ◽

Vol 11 (5) ◽

pp. 204-213

Keyword(s):

Cell Lines ◽

Cancer Cell ◽

Human Cancer ◽

Cancer Cell Lines ◽

In Vitro Cytotoxicity ◽

Human Cancer Cell ◽

Human Cancer Cell Lines

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Six New neo-Clerodane Diterpenoids from Aerial Parts of Scutellaria barbata and Their Cytotoxic Activities

Planta Medica ◽

10.1055/a-0638-8255 ◽

2018 ◽

Vol 84 (17) ◽

pp. 1292-1299 ◽

Cited By ~ 9

Author(s):

Guo-Chun Yang ◽

Jia-Hui Hu ◽

Bing-Long Li ◽

Huan Liu ◽

Jia-Yue Wang ◽

...

Keyword(s):

Cell Lines ◽

Human Cancer ◽

In Vitro Cytotoxicity ◽

Cytotoxic Activities ◽

Scutellaria Barbata ◽

Human Cancer Cell Lines ◽

Aerial Parts ◽

Ic50 Values ◽

Clerodane Diterpenoids

AbstractSix new neo-clerodane diterpenoids (1–6), scutebatas X – Z, A1-C1, along with twelve known ones (7–18) were obtained via the phytochemical investigation of the aerial parts of Scutellaria barbata. Their structures were established by detailed spectroscopic analysis. The absolute configurations of 1 and 2, as the representative members of this type, were identified based on a circular dichroic exciton chirality method. Moreover, in vitro cytotoxicity of compounds 1–6 were evaluated against three human cancer cell lines (SGC-7901, MCF-7, and A-549) using the MTT method. Compound 6 showed cytotoxic activities against all the three cell lines with IC50 values of 17.9, 29.9, and 35.7 µM, respectively.

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New tools for carbohydrate sulfation analysis: heparan sulfate 2-O-sulfotransferase (HS2ST) is a target for small-molecule protein kinase inhibitors

Biochemical Journal ◽

10.1042/bcj20180265 ◽

2018 ◽

Vol 475 (15) ◽

pp. 2417-2433 ◽

Cited By ~ 8

Author(s):

Dominic P. Byrne ◽

Yong Li ◽

Krithika Ramakrishnan ◽

Igor L. Barsukov ◽

Edwin A. Yates ◽

...

Keyword(s):

Protein Kinase ◽

Heparan Sulfate ◽

Small Molecule ◽

Kinase Inhibitors ◽

Kinase Inhibitor ◽

Protein Kinase Inhibitors ◽

Fluorescent Substrate ◽

Raf Kinase ◽

Shift Analysis

Sulfation of carbohydrate residues occurs on a variety of glycans destined for secretion, and this modification is essential for efficient matrix-based signal transduction. Heparan sulfate (HS) glycosaminoglycans control physiological functions ranging from blood coagulation to cell proliferation. HS biosynthesis involves membrane-bound Golgi sulfotransferases, including HS 2-O-sulfotransferase (HS2ST), which transfers sulfate from the cofactor PAPS (3′-phosphoadenosine 5′-phosphosulfate) to the 2-O position of α-l-iduronate in the maturing polysaccharide chain. The current lack of simple non-radioactive enzyme assays that can be used to quantify the levels of carbohydrate sulfation hampers kinetic analysis of this process and the discovery of HS2ST inhibitors. In the present paper, we describe a new procedure for thermal shift analysis of purified HS2ST. Using this approach, we quantify HS2ST-catalysed oligosaccharide sulfation using a novel synthetic fluorescent substrate and screen the Published Kinase Inhibitor Set, to evaluate compounds that inhibit catalysis. We report the susceptibility of HS2ST to a variety of cell-permeable compounds in vitro, including polyanionic polar molecules, the protein kinase inhibitor rottlerin and oxindole-based RAF kinase inhibitors. In a related study, published back-to-back with the present study, we demonstrated that tyrosyl protein sulfotranferases are also inhibited by a variety of protein kinase inhibitors. We propose that appropriately validated small-molecule compounds could become new tools for rapid inhibition of glycan (and protein) sulfation in cells, and that protein kinase inhibitors might be repurposed or redesigned for the specific inhibition of HS2ST.

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