Crizotinib

A simple, précised, accurate method was developed for the estimation of Crizotinib by RP-HPLC technique. Chromatographic conditions used are stationary phase BDS 250 × 4.6 mm, 5 µ. Mobile phase buffer: Acetonitrile in the ratio of 60:40 and flow rate was maintained at 1 ml/min, detection wave length was 267 nm, column temperature was set to 30°C and diluents was methanol: water. System suitability parameters were studied by injecting the standard five times and results were well (50:50), conditions were finalized as optimized method under acceptance criteria. Linearity study was carried out between 25% to 150% levels, r2 value was found to be as 0.999. Precision was found to be 1.26 for repeatability and 0.93 for intermediate precision. LOD and LOQ are 0.080 µg/ml. By using above method assay of marketed formulation was carried out 100.24% was present.

Research targeting gene fusion events with drugs, such as Crizotinib, has provided exciting promise for personalized treatment of cancer patients. Until recently, identifying these fusions has been a challenge due to the computational resources required. With the advent of cloud computing, the ability to create compute clusters with nearly unlimited compute and storage resources has become feasible, and has enabled us to identify fusions in a more rapid and cost-effective fashion. Gene fusions are the result of chromosomal rearrangements that ultimately lead to expression of a fusion protein with oncogenic properties. There are several well-described examples where targeted inhibition of the resulting fusion protein can produce dramatic clinical responses. The EML4-ALK fusion is estimated to occur in 3-5% of non-small cell lung carcinoma (NSCLC) and the ALK inhibitor Crizotinib has been approved for treatment of fusion positive patients. Crizotinib has inhibitory activity not only against ALK, but also inhibits ROS1, RON and MET and LTK tyrosine kinases.
Here we present the cloud computing framework used to process more than 7000 RNA-Seq samples from the TCGA project. This framework includes systematic methods developed to identify and characterize both known and novel oncogenic gene fusion events. These methods also include extensive filters to remove false positive fusions and help identify driver fusion events. An exon expression imbalance algorithm is also used to leverage RNA-Seq exon-level expression data for each patient to provide secondary evidence of true positive fusion events.
These approaches have enabled us to confirm the presence of EML4-ALK fusions in lung and colorectal cancer. We have also identified novel ALK and ROS1 fusions in several other cancer types, thus potentially broadening the scope of therapeutic opportunity for inhibitors like Crizotinib. Using cell line RNA-Seq data from the CCLE project, we were also able to identify cell lines harboring these fusions. With the growing availability of next-generation sequencing, such analyses can support hypothesis- driven development of targeted therapies.

Recent findings from studies [KEYNOTE 10, 24, 189, 407 (pembrolizumab); Check Mate-17,57,227 (nivolumab); IM power 131,150,OAK (atezolizumab)] using checkpoint inhibitors as a monotherapy as well as in combination of chemotherapy has demonstrated improved outcome in patients with advanced NSCLC without actionable mutation driver and also showed a tolerable toxicity profile and durable response. Based on analysis of studies performed in the first line management of advanced NSCLC, pembrolizumab is preferred for patients without actionable driver mutation. Pembrolizumab should be used as a monotherapy in patients with PD-L1 expression ≥ 50%. In others, it should be added to chemotherapy. For patients with actionable driver mutation, osimertinib for sensitizing EGFR mutation is preferred over afatinib, gefitinib, erlotinib as a first line therapy. For patients with ALK rearrangement alectinib is preferred over crizotinib restricting use of crizotinib as first line therapy to patients with ROS1 rearrangement. Dabrafenib + trametinib have been found effective in patients with BRAFV600E mutations.