In vivoand in vitrostructure and relationship analysis of andrographolide derivatives:Identifying AGL-2 as a novel anti-angiogenesis agent

OBJECTIVE To analyse structure and relationship of several andrographolide derivatives in multiple in vivo and in vitro angiogenesis assays,and to demonstrate a novel compound named AGL-2as a potential anti-angiogenesis agent.METHODS Human umbilical vein endothelial cells(HUVECs)in vitro and zebrafish(Danio rerio)in vivo models were used to screen and identify the anti-angiogenesis activities of six andrographolide derivatives;namely,AGL-1,AGL-2,AGS-72,AGS-72 a,AGS-79 and AGP-151.RESULTS AGL-2exhibited the strongest anti-angiogenic activity among all the derivatives in zebrafish model.Interestingly,another compound named AGS-72 showed stronger anti-angiogenic activity than AGL-2 in VEGF-induced HUVECs proliferation,migration,invasion and tube formation assays.In addition,AGL-2 was found to suppress the VEGF-induced VEGFR-2auto-phosphorylation and inhibit the activity of VEGFR-2 mediated signaling cascades in a dose-dependent manner.CONCLUSION AGL-2was demonstrated to be a promising anti-angiogenic agent among all the tested derivatives.The mechanism underlying the anti-angiogenic activity of AGL-2 probably involve VEGFR-2 signaling pathway.Even though,how some of chemical structure alterations result in discrepancy between in vivo and in vitro activities still remains to be resolved,this study shall provide new insight into how modification of the chemical structure of andrographolide affects this newly identified anti-angiogenesis activity.Meanwhile,AGL-2 can be exploited as a potential therapeutic agent for the treatment of angiogenesis-related diseases.

A Sensitive HPLC-MS/MS Analysis of Dencichine in Rat Plasma and Its Application to Pharmacokinetics

In order to quantitate dencichine in biological samples, a selective and sensitive method for the determination of dencichine in rat plasma based on high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed and validated. (l)-2-amino-3-(carboxymethylthio)propionic acid was used as the internal standard (I.S.). After a protein precipitation extraction with acetonitrile, dencichine and the I.S. were chromatographed on an Xterra MS-C18 column. The mobile phase was consisted of 20mmol/L ammonium acetate solution-acetonitrile (35:65, V/V) at a flow rate of 0.2 mL/min. The detection was performed on a triple quadrupole mass via electrospray ionization (ESI) source in the positive mode. The intra- and inter-day precision (relative standard deviation, R.S.D.) values of dencichine were below 6.7%. The extraction recoveries were up 85%. The lower limit of quantification was 20 ng/ml, which was sensitive enough to detect the analyte. The HPLC-MS/MS method was successfully applied to the pharmacokinetic study after an intravenous administration of dencichine in Sprague-Dawley (SD) rat.

Synthesis and Crystal Structure of(S)-N-(hexahydro-2-oxo-1H-azepin-3-yl)-3-phenyl-2-propenamide

The title compound（S）-N-（hexahydro-2-oxo-1H-azepin-3-yl）-3-phenyl-2-propenamide（C15H18N2O2, Mr = 258.31） was synthesized by the reaction of（S）-α-amino-ε-caprolactam with 3-phenyl-2- propenoyl chloride. Its chemical structure was determined by 1H NMR, 13C NMR, H RMS and X-ray single-crystal diffraction. The crystal belongs to the monoclinic system, space group P21/n with a = 14.957（3）, b = 4.884（1）, c = 18.630（4）, β = 95.91（3）o, V = 1353.7（5）3, Z = 4, Dc = 1.267 g/cm^3, μ = 0.085 mm-1, F（000） = 552, R = 0.0671 and wR = 0.1547 for 2473 observed reflections with I 〉 2σ（I）.

Synthesis and biological evaluation of 5'-phenyl-3'H-spiro-[indoline-3,2'-[1,3,4]oxadiazol]-2-one analogs

A series of 5＇-phenyl-3＇H-spiro[indoline-3,2＇-[l,3,4]thiadiazol]-2-one analogs were synthesized and their Bcl-2 protein inhibitory activities were studied. The lead compound was originally identified using a fluorescence polarization-based competitive binding assay. Among the 10 compounds investigated, I k showed good binding affinities to Bcl-xL and Mcl-l, with inhibition constants of 8.9 μmol/L and 3.4 μmol/L, respectively. While compound lc achieved tight binding affinities to Bcl-xL （Ki= 0.16 μmol/L）, has the potential to be a new lead compound.