Discovery of MDV6058 (PF-06952229), a selective and potent TGFßR1 inhibitor: Design, synthesis and optimization.

Compound 1 is a potent TGF-ß receptor type-1 (TGFßR1 or ALK5) inhibitor but is metabolically unstable. A solvent-exposed part of this molecule was used to analogue and modulate cell activity, liver microsome stability and mouse pharmacokinetics. The evolution of SAR that led to the selection of 2 (MDV6058 / PF-06952229) as a preclinical lead compound is described.

Ultra-Sensitive NT-proBNP Quantification for Early Detection of Risk Factors Leading to Heart Failure.

Cardiovascular diseases such as acute myocardial infarction and heart failure accounted for the death of 17.5 million people (31% of all global deaths) in 2015. Monitoring the level of circulating N-terminal proBNP (NT-proBNP) is crucial for the detection of people at risk of heart failure. In this article, we describe a novel ultra-sensitive NT-proBNP test (us-NT-proBNP) that allows the quantification of circulating NT-proBNP in 30 min at 25 °C in the linear detection range of 7.0-600 pg/mL. It is a first report on the application of a fluorescence bead labeled detection antibody, DNA-guided detection method, and glass fiber membrane platform for the quantification of NT-proBNP in clinical samples. Limit of blank, limit of detection, and limit of quantification were 2.0 pg/mL, 3.7 pg/mL, and 7 pg/mL, respectively. The coefficient of variation was found to be less than 10% in the entire detection range of 7-600 pg/mL. The test demonstrated specificity for NT-proBNP without interferences from bilirubin, intra-lipid, biotin, and hemoglobin. The serial dilution test for plasma samples containing various NT-proBNP levels showed the linear decrement in concentration with the regression coefficient of 0.980-0.998. These results indicate that us-NT-proBNP test does not suffer from the interference of the plasma components for the measurement of NT-proBNP in clinical samples.

A glass fibre membrane platform for ultra-sensitive detection of cardiac troponin T.

A glass fibre membrane platform that allows quantification of circulating cTnT with a LoD of 0.87 pg mL-1 is described. The proposed platform uses a glass fibre membrane, DNA-guided detection method, and antibody-conjugated fluorescent beads for the quantification of cTnT in the analytical detection range of 1-120 pg mL-1 at room temperature in 30 min. Glass fibre membranes were chemically modified to immobilize the oligonucleotide probes that catch a biomolecular complex (FB-dAB-cTnT-cAB-DNA) containing complementary oligonucleotides. There were no interferences from human cTnI, cTnC, skTnT, biotin, and hemoglobin (each 1 µg mL-1). The linearity in the serial dilution test of plasma samples indicates that this platform is highly applicable for regular health check-up to assess the risk of AMI and HF.

6 HCV Genotyping 9G test for HCV 1a, 1b, 2, 3, 4 and 6 (6a, 6f, 6i and 6n) with high accuracy.

According to EASL guidelines and WHO recommendations, the accurate detection of HCV genotypes such as HCV 1a, HCV1b, HCV 2, HCV 3, HCV 4, and HCV 6 (6a, 6f, 6i, 6n) is crucial for the efficient treatment of hepatitis C. HCV Genotyping 9G test allows simultaneous genotyping of HCV 1a, 1b, 2, 3, 4, and 6 (6a, 6f, 6i, and 6n) in clinical samples in 30min. The performance of the test was evaluated by comparison with sequence analysis. Serum samples (n=152) from HCV-infected patients (n=110) and healthy individuals (n=42) were processed under blinded codes. The k coefficient (kappa) values indicated high agreement between the HCV Genotyping 9G test and sequencing. The sensitivity and specificity of the test were 99.1% and 99.7%, respectively. The results indicate that HCV Genotyping 9G test is rapid, reliable, sensitive, and accurate for screening and genotyping of HCV in the clinical specimens.

6 HCV genotyping 9G test and its comparison with VERSANT HCV genotype 2.0 assay (LiPA) for the hepatitis C virus genotyping.

In this article, we describe the 6 HCV Genotyping 9G test and its evaluation by using clinical samples and plasmid DNA standards. In tests with 981 plasmid DNA standards, the 6 HCV Genotyping 9G test showed higher than 92.5% sensitivity and 99.4% specificity. The 6 HCV Genotyping 9G test was compared with the VERSANT HCV Genotype 2.0 assay (LiPA 2.0) for detection and discrimination of HCV genotypes in clinical samples. The results of both tests were verified by genomic sequencing. The 6 HCV Genotyping 9G test demonstrated a 100% agreement with the sequencing results, which was higher than LiPA 2.0. These results indicate that the 6 HCV Genotyping 9G test can be a reliable, sensitive, and accurate diagnostic tool for the correct identification of HCV genotypes in clinical specimens. 6 HCV Genotyping 9G test can genotype six HCV types in 1 PCR in 30min after PCR amplification. The 6 HCV Genotyping 9G test, thus provide critical information to physicians and assist them to apply accurate drug regimen for the effective hepatitis C treatment.

HBV/4DR 9G test and its comparison with INNO-LiPA HBV multi-DR test for the detection of drug-resistant Hepatitis B virus.

A significant proportion of patients with chronic Hepatitis B infection require antiviral therapy during their life time. The Antiviral therapy with lamivudine or adefovir or telbivudine has shown to be a major risk factor for selection of resistance. Eighty percent of patients showed a development of lamivudine-resistant strains after five years of treatment with lamivudine alone. Adefovir and telbivudine inhibit HBV with very high efficacy and have moderate incidences of drug resistance. Entecavir and tenofovir have been shown to have a higher barrier to resistance with rates of less than 1.5% after five years of treatment. The rtA181V, rtM204V/I, rtN236T and, rtM250V are high prevalent mutations found in the drug-resistant HBV strains. Therefore, for accurate treatment of HBV-infected patients, it is important to discriminate the drug-resistant HBV strains by using simple and accurate detection method. In this study, we describe the HBV/4DR 9G test and its evaluation by using clinical samples and plasmid DNA standards with a range of HBV mutation sites. In tests with 384 plasmid DNA standards, the HBV/4DR 9G test showed higher than 95% sensitivity and 98% specificity. The HBV/4DR 9G test was compared with the INNO-LiPA HBV Multi DR test for detection of drug-resistant HBV strains only in clinical samples. The plasma samples were collected from patients suspected with HBV drug-resistant strain infection. The results of both tests were cross-checked with the HBV DNA sequence analysis. The HBV/4DR 9G test demonstrated a good agreement with the sequencing results as compared to the INNO-LiPA HBV Multi-DR test. These results indicate that the HBV/4DR 9G test can be a reliable, sensitive, and accurate diagnostic tool for the detection of drug-resistant genotypes of HBV in clinical specimens. HBV/4DR 9G test can genotype 4 drug resistant HBV strains in 1 PCR. The HBV/4DR 9G test will help to minimize the risk of HBV patients from liver cancer.

Biomarker detection technologies and future directions.

Biomarkers play a vital role in disease detection and treatment follow-up. It is important to note that diseases in the early stage are typically treated with the greatest probability of success. However, due to various technical difficulties in current technologies for the detection of biomarkers, the potential of biomarkers is not explored completely. Therefore, the developments of technologies, which can enable the accurate detection of prostate cancer at an early stage with simple, experimental protocols are highly inevitable. This critical review evaluates the current methods and technologies used in the detection of biomarkers. The aim of this article is to provide a comprehensive review covering the advantages and disadvantages of the biomarker detection methods. Future directions for the development of technologies to achieve highly selective and sensitive detection of biomarkers for point-of-care applications are also commented on.

Immobilization techniques for microarray: challenges and applications.

The highly programmable positioning of molecules (biomolecules, nanoparticles, nanobeads, nanocomposites materials) on surfaces has potential applications in the fields of biosensors, biomolecular electronics, and nanodevices. However, the conventional techniques including self-assembled monolayers fail to position the molecules on the nanometer scale to produce highly organized monolayers on the surface. The present article elaborates different techniques for the immobilization of the biomolecules on the surface to produce microarrays and their diagnostic applications. The advantages and the drawbacks of various methods are compared. This article also sheds light on the applications of the different technologies for the detection and discrimination of viral/bacterial genotypes and the detection of the biomarkers. A brief survey with 115 references covering the last 10 years on the biological applications of microarrays in various fields is also provided.