A phase Ia dose-escalation trial of Ametumumab (a fully human monoclonal antibody against epidermal growth factor receptor) in patients with advanced solid malignancies.

Background: Epidermal growth factor receptor (EGFR) is a well-known target for cancer treatment. However, the authorized anti-EGFR monoclonal antibodies generally cause several toxic effects, especially severe cutaneous toxicities as well as infusion reactions, and the clinical indications are limited. Here we developed Ametumumab, a fully human recombinant anti-EGFR monoclonal antibody. Objectives: To assess the safety, tolerability, pharmacokinetics (PK), and immunogenicity of Ametumumab. Design: A first-in-human phase Ia dose escalation study of Ametumumab in patients with advanced solid malignancies. Methods: An open-label, first-in-human dose escalation study was done in 22 patients with advanced malignancies who received six ascending dosages ranging from 75 to 750 mg/m2. Following a single dosage and a 28-day dose-limiting toxicity (DLT) monitoring period, patients were given repeated doses weekly. Blood samples were taken to determine the PK parameters of Ametumumab and anti-drug antibody concentrations. Every 8 weeks, radiographic tumor evaluations were conducted. Results: In this trial, no DLT was observed, and the maximum tolerated dose was not reached at doses up to 750 mg/m2. There were no severe adverse events but mild and moderate adverse effects, such as headache, proteinuria, and rash. Single-dose PK results demonstrated a straightforward linear relationship with dosage escalation. The medication concentrations accumulated and attained steady-state after four rounds of injections. It was calculated that 10 patients with disease control would be observed in the 22 evaluable patients. The disease control rate was 45.5%. Conclusion: The Ametumumab was well tolerated and safe in patients with advanced solid malignancies, exhibiting minimal immunogenicity, a long half-life, high levels of drug exposure in the blood, and preliminary effectiveness. Registration: The trial was registered with CTR20170343 on 10 April 2017, The China Center for Drug Evaluation.

Hybridization chain reaction-based aptameric system for the highly selective and sensitive detection of protein.

We introduce here a novel assay for the detection of platelet-derived growth factor BB (PDGF-BB) via hybridization chain reaction (HCR) based on an aptameric system, where stable DNA monomers assemble only upon exposure to a target PDGF-BB aptamer. In this process, two complementary stable species of biotinylated DNA hairpins coexist in solution until the introduction of initiator aptamer strands triggers a cascade of hybridization events that yields nicked double helices analogous to alternating copolymers. In detail, the aptamer firstly opens the hairpins in the solution, creating long concatemers, and then reacts with the antibody captured PDGF-BB on the well surface. Moreover, several experimental conditions including different PDGF-BB aptamers, the spacer length of the selected aptamer and hairpin, etc. are investigated and optimized. Our results show that the coupling of HCR to aptamer triggers for the amplification detection of PDGF-BB achieves a better performance in the fluorescence detection of PDGF-BB as compared to the traditional antibody-antigen-aptamer assays. Upon modification, the approach presented herein could be extended to detect other types of targets. We believe such advancements will represent a significant step towards improved diagnostics and more personalized medical treatment and environmental monitoring.

Instantaneous derivatization technology for the simultaneous and homogeneous detection of multiple double-stranded PCR amplicons.

Herein we report on the development of instantaneous derivatization technology for the homogeneous and simultaneous detection of multiple PCR amplicons specific to the Hepatitis B Virus (HBV) by using three carriers: magnetic beads, polystyrene beads, and thermo-sensitive poly-N-isopropylacrylamide (PNIP). Briefly, PCR amplicons are labeled with digoxin, biotin or FITC via the modified up-stream primers respectively. After PCR amplification, the immunoreactions occur between a mixture of three target PCR amplicons and three modified carriers with anti-digoxin antibody, streptavidin or anti-FITC antibody in a single vessel, and then each carrier is separated from the others under different conditions based on their physio-chemical attributes. And then direct CL detection proceeds via the instantaneous derivatization reaction between intrinsic guanine nucleobases and 3,4,5-trimethoxylphenylglyoxal (TMPG). This new protocol directly measures the double-stranded DNA and therefore does not require a denaturing step, thus offering an enhanced sensitivity due to the absence of competitive hybridization, i.e., the detection limit had a 20-fold improvement on the conventional PCR measurement. Additionally, by comparison of previous guanine based detection formats, this protocol is easy to be used for the detection of any guanine containing targets without the use of guanine-free or inosine-substituted capture probes. Overall, the proposed technique takes the advantages of sensitivity, high-speed and cost-effectivity, which provides a promising alternative for the analysis of multiple PCR targets in a variety of clinical, environmental, and biodefense fields.

Mechanism of three inhibitors of TACE in blocking the converting of pro-TNF alpha into sTNF alpha.

The effects of inhibitors of TNF alpha converting enzyme (TACE) on TNF alpha secretion were studied to develop an approach to interfere inflammation processes. The HL-60 cell lines were stimulated in vitro with LPS intravenously for different time to establish the cellular model of inflammation and simultaneously induce in vivo inflammation animal model by LPS The cytotoxic effects of soluble TNF alpha were checked using MTT colorimetric method to determine the rate of cell proliferation. The level of expression of TACE was detected by using RT-PCR, FCM and immuno-histochemical technique respectively. It was found Chinese medicine Reduqing (RDQ) could inhibit the transcription of TNF alpha mRNA induced by LPS stimulation (P < 0.01, compared with the control). The antioligodeoxyribonucleotide (anti-ODN) of TNF alpha mRNA could inhibit 78.9% of TNF alpha secretion. The mimic peptides of TACE substrates with hydroxamine group showed potency in vivo and in vitro against converting of pro-TNF alpha. It was concluded that all the three types of TACE inhibitors can regulate the expression of TACE at different levels and inhibit sTNF alpha secretion, indicating TACE is a novel target for inflammation therapy.