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Fibroblast growth factor 2 (FGF2) is closely involved in a variety of tumors, including gastric cancer (GC). FGF2 inhibitors exert good antitumor activity, but no FGF2 inhibitor has been employed for clinical use. To obtain a low-toxicity, stable peptidomimetic (called P29) target to FGF2, the affinity between P29 and FGF2 was detected by surface plasmon resonance. The stability of P29 was measured by high performance liquid chromatography. MTT assay and transwell assay were used to access the proliferative and invasive ability of GC cells, respectively. Western blot assay and flow cytometric analysis were applied to study the mechanism of P29. P29 possessed high affinity with FGF2 and a longer half-life in vitro. P29 suppressed the FGF2-induced proliferation of GC cells. It also inhibited the phosphorylation of FRS2, ERK1/2, and AKT triggered by FGF2 in GC. In addition, P29 blocked GC cell transformation from the G1/G0 phase to the S phase and weakened the invasive capability of GC cells. In this paper, we present a novel FGF2 inhibitor that could exert improved anticancer effect in GC in vitro.

Background: Telaprevir is a protease inhibitor used in the treatment of hepatitis C virus infection. Analytical methods for telaprevir should separate the compound from its R-diastereomer VRT-127394, which is 30-fold less active. The objective of this work was to develop liquid chromatography-tandem mass spectrometer (LC-MS/MS) assays for telaprevir both in plasma and in dried blood spot (DBS), capable of stabilizing the equilibrium and chromatographically separating the 2 epimers., Methods: Human plasma was acidified with formic acid and frozen within 1 hour after collection to stabilize the equilibrium between the 2 telaprevir diastereomers ex vivo in plasma. After protein precipitation, the sample was analyzed with LC-MS/MS. For the DBS assay, sampling paper was impregnated with citric acid solution to achieve stabilization of the epimers on the sampling paper. DBS samples were extracted before LC-MS/MS analysis. LC-MS/MS analysis comprised online solid-phase extraction and separation on a C18 column, with the mass spectrometer operating in TurboIonSpray-negative ionization mode and performing multiple reaction monitoring., Results: The assays were linear over the concentration range of 0.1-10 mg/L in plasma and DBS. Accuracies ranged from 97% to 106% in plasma and from 93% to 99% in DBS. Within- and between-day coefficients of variation were <7.9% in plasma and <9.3% in DBS. Human whole blood samples with hematocrit values of 27%-47% gave reproducible quantitation results in the DBS assay, and spot volume did not affect results of the DBS assay either. Acidified plasma with telaprevir was stable for 5 hours at 20°C, and telaprevir on impregnated DBS paper was stable for at least 3 months at 4°C or at 20°C., Conclusions: An assay was developed and validated for the determination of telaprevir in human plasma, separating telaprevir from its R-diastereomer VRT-127394. In addition, a DBS assay was developed, which avoids immediate centrifuging, acidification, and freezing of patient samples to stabilize the equilibrium between the 2 telaprevir diastereomers.

RGD (Arg-Gly-Asp) tripeptide was identified as the minimal active core peptide sequence common to adhesive proteins. In this paper, the authors report preparation of RGD containing peptide albumin conjugate (RGD ALB), and its effects on cellular adhesive function in vitro. RGD ALB was prepared via a coupling reaction of albumin with pentapeptide (GRGDS; Gly-RGD-Ser) by water soluble carbodiimide. Bovine endothelial cells (ECs) adhered to and spread well on a surface coated with RGD ALB, whereas few ECs adhered on surfaces coated with GRGESP-albumin conjugate (GRGESP peptide with little cell attachment activity; false control) and albumin. Cellular behavior, such as adhesion, spreading, growth, and migration, on surfaces coated with GRD-ALB, fibronectin (FN), and vitronectin (VN) were quantitatively examined. Adherent cell number on RGD ALB coated surfaces was larger than on those coated with FN and VN. Cell morphology on RGD ALB coated surfaces was similar to that on FN coated surfaces. The cell growth and migration activities on RGD ALB coated surfaces were almost equal to those coated with FN and VN. Thus, RGD ALB was found to promote cell adhesion, migration, and growth as effectively as fibronectin. This indicates that an artificial adhesive protein, simply derived with bioactive peptidyl ligand, can find versatile applications in fields in which cellular events play a critical role--for example, extracellular matrices and wound healing promotion aids.