Bioanalysis of Targeted Nanoparticles in Monkey Plasma via LC-MS/MS.

This work represents the first reporting of a comprehensive bioanalytical GLP methodology detailing the mass spectrometric quantitation of PF-05212384 dosed as a targeted polymeric encapsulated nanoparticle (PF-07034663) to monkeys. Polymeric nanoparticles are a type of drug formulation that enables the sustained release of an active therapeutic agent (payload) for targeted delivery to specific sites of action such as cancer cells. Through the careful design and engineering of the nanoparticle formulation, it is possible to improve the biodistribution and safety of a given therapeutic payload in circulation. However, the bioanalysis of nanoparticles is challenging due to the complexity of the nanoparticle drug formulation itself and the number of pharmacokinetic end points needed to characterize the in vivo exposure of the nanoparticles. Gedatolisib, also known as PF-05212384, was reformulated as an encapsulated targeted polymeric nanoparticle. The bioanalytical assays were validated to quantitate both total and released PF-05212384 derived from the encapsulated nanoparticle (PF-07034663). Assay performance calculated from quality control samples in three batch runs demonstrated intraday precision and accuracy within 10.3 and 12.2%, respectively, and interday precision and accuracy within 9.1 and 8.5%, respectively. This method leveraged automation to ease the burden of a laborious and complicated sample pretreatment and extraction procedure. The automated method was used to support a preclinical safety study in monkeys in which both released and total PF-05212384 concentrations were determined in over 1600 monkey plasma study samples via LC-MS/MS.

Mailuoning prevents high-glucose-mediated human umbilical vein endothelial cells apoptosis.

OBJECTIVE: To investigated the role of Mailuoning in the prevention of high-glucose-mediated cell apoptosis in human umbilical vein endothelial cells (HUVEC) and the mechanisms involved. METHODS: MTT assay was used to investigate cell viability, western blot was used to investigate protein expression, and flow cytometric detection technology was used to detect cell apoptosis. RESULTS: Exposure of HUVEC to high glucose (50 mM) significantly suppressed cell viability and increased cell apoptosis compared with normal glucose (11 mM) (all P < 0.05). However, Mailuoning prevented high-glucose-induced HUVEC apoptosis in dose-dependent manner. Further studies indicated that Mailuoning suppressed high-glucose-induced p38 mitogen-activated protein kinase phosphorylation, but had no effect on extracellular signal-regulated kinase 1/2 and Akt phosphorylation. CONCLUSION: Mailuoning can prevent high-glucose-induced HUVEC apoptosis by suppressing p38 activation.

Escin sodium induces apoptosis of human acute leukemia Jurkat T cells.

Escin sodium has been used in the clinic as an antioedematous, antiexudative and vasoprotective agent for many years and has shown excellent tolerability. However, little is known about its anticancer activity. This is a report for the first time that escin sodium exerts a cytotoxic effect on human acute leukemia Jurkat T cells via the induction of apoptosis rather than cell cycle arrest. Escin sodium activated the initiator caspase-8, -9, and the effector caspase-3, degraded poly (ADP-ribose) polymerase (PARP) and attenuated the expression of Bcl-2. In addition, escin sodium inhibited the growth of cancer cells in a selective manner with Jurkat cells most sensitive to it. Taken together, the data show that escin sodium possesses potent apoptogenic activity toward human acute leukemia Jurkat T cells.

Immunological regulation of Chinese herb Guizhi Fuling Capsule on rat endometriosis model.

AIM OF THE STUDY: To investigate the immunological regulation of Guizhi Fuling Capsule (GZFLC) on rat endometriosis. MATERIALS AND METHODS: Twenty-seven rats, in which endometriotic implants were induced by transplanting autologous uterine tissue to the peritoneum, were randomly divided into three groups equally: (1) the GZFLC group of low dose (480 mg/kg/day); (2) the GZFLC group of high dose (1,920 mg/kg/day); and (3) the model group(saline solution). Another 10 rats were treated as sham operation group. After rats were treated for four weeks, we examined the alterations of implants volume, the percentage of CD4(+) T lympholeukocyte, the activity of NK cell and the expression of cytokines (MCP-1 and ICAM-1) on each group. RESULTS: Statistical analysis showed that posttreatment volumes were significantly reduced compared with pretreatment in GZFLC groups, whereas there was no significant change in the model group. The percentage of CD4(+) T lympholeukocyte and the activity of NK cell in GZFLC groups significantly increased to the level of the sham group compared with the model. RT-PCR and immunohistochemistry showed that the endometria of the sham operation and treatment groups were similar on expression level of MCP-1 and ICAM-1. CONCLUSIONS: GZFLC plays an important role in the regression of endometriotic implants by immunological regulation in the rat model.