The past decade has witnessed significant success with personalized. Targeting ROS1 with Anaplastic Lymphoma Kinase Inhibitors

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1 PATHWAY OF THE MONTH Targeting ROS1 with Anaplastic Lymphoma Kinase Inhibitors A Promising Therapeutic Strategy for a Newly Defined Molecular Subset of Non Small-Cell Lung Cancer Leow Pay Chin, PhD,* Ross A. Soo, MD,* Richie Soong, PhD,* Sai-Hong I. Ou, MD, PhD Abstract: Chromosomal rearrangements involving the ROS1 receptor tyrosine kinase have been described in a variety of human malignancies including non small-cell lung cancer (NSCLC), cholangiocarcinoma and glioblastoma multiforme. Recently, clinicopathologic characteristics of c-ros oncogene 1, receptor tyrosine kinase (ROS1)-rearranged NSCLC patients have been described. Furthermore, anaplastic lymphoma kinase inhibitor, novel class of drugs targeting this tyrosine kinase receptor is currently under clinical trial in this molecular subset of NSCLC patients. This review will focus on the current knowledge of ROS1 rearrangements in NSCLC, methods to detect ROS1 rearrangement, and targeting ROS1-rearranged NSCLC patients with anaplastic lymphoma kinase inhibitors. Key Words: Non -small-cell lung cancer, Targeted therapy, ROS1, Anaplastic lymphoma kinase inhibitors, Crizotinib. (J Thorac Oncol. 2012;7: ) The past decade has witnessed significant success with personalized cancer therapy, and this paradigm shift has led to tremendous improvements in clinical outcomes in patients with molecularly defined subsets of tumors. Notably, the discovery of a variety of molecular and genetic alternations in non small-cell lung cancer (NSCLC) has provided the opportunity for targeted therapy, such as epidermal growth factor receptortyrosine kinase inhibitors (gefitinib and erlotinib) for activating EGFR mutations in NSCLC, and anaplastic *Cancer Science Institute of Singapore, National University of Singapore, Singapore; Department of Internal Medicine, Division of Hematology/ Oncology, Chao Family, Comprehensive Cancer Center, California; and Department of Haematology-Oncology, National University Cancer Institute, National University Health System. Disclosure: Sai-Hong I. Ou is on the speaker bureau, acts a consultant for Pfizer, and has stock ownership in ARIAD pharmaceuticals. The other authors declare no conflicts of interest. Address for correspondence: Ross A. Soo, Department of Haematology- Oncology, National University Cancer Institute, National University Health System, 1E Kent Ridge Road, NUHS Tower Block Level 7, Singapore ross_soo@nuhs.edu.sg Copyright 2012 by the International Association for the Study of Lung Cancer ISSN: /12/ lymphoma kinase (ALK) inhibitor (crizotinib) for NSCLC harboring ALK gene fusion. The c-ros oncogene 1 (ROS1) receptor tyrosine kinase (RTK) has recently emerged as a potentially relevant therapeutic target in NSCLC. Here, our review focuses on the oncogenic role of aberrant ROS1 in NSCLC and targeting ROS1-rearranged NSCLC patients with ALK inhibitors. ROS1 RECEPTOR STRUCTURE AND FUNCTION v-ros1 is an oncogene product of the avian sarcoma RNA tumor virus UR2. 1 Wild-type ROS1 is located on chromosome 6 and encodes full-length ROS1, a 2,347 amino acid transmembrane tyrosine kinase (TK) receptor consisting of an extracellular ligand-binding domain composed of nine repeated fibronectin-like motifs, a short transmembrane domain, and an intracellular TK domain. 2 Although very little is known of the extracellular domain function of this orphan receptor, the structural combination of ROS1 suggests its ability to directly couple extracellular adhesion-mediated events to tyrosine phosphorylation-based intracellular signaling. Using the Multiple Sequence Comparison by Log-Expression software, it has been found that there is a 49% amino acid homology between human ROS and ALK within the kinase domain 3 and 77% identity at the adenosine triphosphate (ATP)-binding site. 4 Dysregulated ROS1 may occur as a result of ROS1 gene fusion, overexpression, or mutations. Aberrant ROS1 kinase activity leads to activated downstream signaling of several oncogenic pathways including the phosphoinositide-3 kinase/v-akt murine thymoma viral oncogene homolog 1 (AKT)/mechanistic target of rapamycin (serine/ threonine kinase) (mtor), signal transducers and activators of transcription-3, RAS-MAPK/ERK, vav 3 guanine nucleotide exchange factor 1 (VAV3) and Src-homology 2 domain-containing phosphatase (SHP)-1 and -2 pathways (Fig. 1). 5,6 With the activation of ALK fusion proteins, the downstream signaling pathways are similar to ROS1 RTK activation with the involvement of the RAS-MAPK/ERK, Janus kinase 3-signal transducers and activators of transcription-3, and phosphoinositide-3 kinase /Akt pathways, which control cell proliferation, survival, and cell cycling. 7,8 However, unlike the echinoderm microtubule-associated Journal of Thoracic Oncology Volume 7, Number 11, November

2 Chin et al. Journal of Thoracic Oncology Volume 7, Number 11, November 2012 FIGURE 1. ROS1 signaling pathways. ROS1 engages multiple signalling pathways to exert its transformation activity. PLCγ, MAP kinase, IRS1 (signalling molecule to PI3K), VAV3, STATS, cytoskeleton and cell-to-cell interaction (β-integrin, tensin, α-, β-, δ-catenin, N-catenin) proteins can interact with ROS1, and phospohorylation of these proteins by ROS1 can lead to activation of the respective oncogenic pathways. ROS autophosphorylation sites are docking sites for the tyrosine phosphatases SHP-1 and SHP, Src-homology 2 domain-containing phosphatase; ROS1, c-ros oncogene 1, receptor tyrosine kinase c-ros oncogene 1, receptor tyrosine kinase; PLCγ, phospholipase C, gamma; MAP, mitogen-activated protein kinase; IRS1, insulin receptor substrate 1; VAV3, vav 3 guanine nucleotide exchange factor 1; STATS, signal transducers and activators of transcription-3; PDK1, pyruvate dehydrogenase kinase, isozyme 1; AKT (v-akt murine thymoma viral oncogene homolog 1); mtor, mechanistic target of rapamycin (serine/threonine kinase); P13K, phosphoinositide-3 kinase; and RHO, rhodopsin. protein-like 4 (EML4)-ALK gene fusion in which the coiled-coil domain within echinoderm microtubule-associated protein-like 4 mediates the constitutive dimerization resulting in aberrant constitutive activity, 9 protein dimerization domains have not been described in ROS1 fusion receptor tyrosine kinases. 5 TABLE 1. ROS1 Fusion Variants Described in Non Small-Cell Lung Cancer Fusion Variant Chromosomal Translocation Lung Tumor Type ROS1 Exon Breakpoint Partner Exon Breakpoint Reference CD74-ROS1 t(5;6) (q32;q22) Non small-cell lung cancer EZR-ROS1 Inv(6) (q22;q25.3) Adenocarcinoma LRIG3-ROS1 t(6;12) (q22;q14.1) Adenocarcinoma SDC4-ROS1 t(6;20) (q22;q12) Adenocarcinoma SLC34A2-ROS1 t(4;6) (p15.2;q22) HCC78 cell line TPM3-ROS1 t(1;6) (q21.2;q22) Adenocarcinoma , Copyright 2012 by the International Association for the Study of Lung Cancer

3 Journal of Thoracic Oncology Volume 7, Number 11, November 2012 Targeting ROS1 with ALK Inhibitors ROS1 Fusion Genes To date, seven distinct ROS1 gene fusions have been identified in solid tumors including fused in glioblastoma (FIG)-ROS1, CD74-ROS1, solute carrier family 34 (sodium phosphate), member 2 (SLC34A2)-ROS1, tropomyosin 3 (TPM3)-ROS1, syndecan 4 (SDC4)-ROS1, ezrin (EZR)-ROS1, leucine-rich repeats, and immunoglobulin-like domains 3 (LRIG3-ROS1) all of which encoded the same cytoplasmic portion of ROS1 TK domain. The break point of ROS1 with EZR is exon 34, TPM3, FIG, and LRIG3 is exon 35 whereas for CD74, SDC4, and SLC34A2 are exons 32 and 34. (Table1). All of the breakpoints in ROS1 allow the resulting fusion to retain the ROS1 kinase domain whereas with SDC4-ROS1, the ROS1 transmembrane domain is also retained. 3 More recently, Govindan et al. 9a have identified a novel ROS1 fusion, KDELR2 (endoplasmic reticulum protein retention receptor 2) in a never smoker patient with NSCLC. FIG-ROS1 fusion was first described in glioblastoma multiforme 6,10 and subsequently in 8.7% of cholangiocarcinoma tumors. 11 The clinicopathologic characteristics of patients with glioblastoma multiforme and cholangiocarcinoma harboring FIG-ROS1 is unknown. In NSCLC, SLC34A2-ROS1 and CD74-ROS1 were initially discovered in the HCC78 cell line and in a patient tumor, respectively. 12 Down-regulation of SLC34A2-ROS1 inhibited cellular proliferation in HCC78 cells 12 whereas more recently, the transforming ability of FIG-ROS1 and TPM3-ROS1, SDC4-ROS1, SLC34A2-ROS1, CD74-ROS1, LRIG-ROS1, and EZR-ROS1 was demonstrated in vivo by Gu et al. 11 and Takeuchi et al. 13 In a screen using fluorescence in situ hybridization (FISH) for ROS1 rearrangements, 1.7% of NSCLC patients (18 of 1073) harbored CD74-ROS1 or SLC34A2-ROS1 fusions, and ROS1 fusions were more likely in younger patients, never smokers with adenocarcinoma subtype. 14 Of the 18 cases reported in this study, no fusion partner was identified in eight samples. Takeuchi et al. 13 have reported ROS1 fusion genes in 0.9% NSCLC (13 of 1476) and 1.2% of patients (13 of 1116) with adenocarcinomas. Four novel ROS1 gene fusions were reported: TPM3-ROS1, SDC4-ROS1, EZR-ROS1, and LRIG3-ROS1. 13 In one case, no fusion partner was identified. In a study of East Asian never smokers with lung adenocarcinoma, 1% (2 of 202) harbored ROS1 fusion. 15 Using immunohistochemistry, and confirming the findings by FISH and reverse transcriptase polymerase chain reaction (), Rimkunas et al. 16 reported ROS1 was overexpressed in 1.6% of all lung NSCLC (9 of 556) and 3.3% of lung adenocarcinoma (8 of 246) from Chinese patients. The pattern of cellular localization of ROS1 protein was variable with diffuse cytoplasm, perinuclear aggregates, and membranous and vesicular localization reported. It is interesting that, this is the first study to report a case of FIG-ROS1 in NSCLC. On the basis of the studies described above, it can be stated that ROS1 fusions represent a unique molecular subset of NSCLC with no overlap with other described oncogene drivers, and are rare with a frequency of 0.7% to 1.7% (Table 2) as compared with EGFR mutations (6% 33%) 17 and ALK fusion (4%). 18 There is overlap in the clinicopathologic characteristics between ROS1 and ALK rearrangement in TABLE 2. Frequency of ROS1 Fusions in Non Small-Cell Lung Cancer Study Rikova et al. 12 Bergethon et al. 14 Li et al. 15 Takeuchi et al. 13 Rimkunas et al. 16 Method for Screening And Confirmation Phosphoproteomics FISH (break-apart) Direct sequencing FISH (break-apart) FISH (fusion-specific) IHC screen Study Population (Sample Size) Number of ROS1 Fusions (%) Chinese (150) 1 (0.7%) Total (1073) Asian (45) Non-Asian (942) Not available (86) 18 (1.7%) 5 (11.1%) 13 (1.4%) 0 (0%) Chinese (202) a 2 (1. 0%) Japanese (1476) 13 (0.9%) Chinese (556) 9 (1.6%) a All were adenocarcinoma, never smokers FISH, fluorescence in situ hybridization; IHC, immunohistochemistry; : reverse transcriptase polymerase chain reaction. NSCLC patients, namely in younger patients with a history of never smoking and adenocarcinoma subtype. ROS1 Overexpressions and Mutations Microarray analysis of NSCLC demonstrated significantly elevated ROS1 expressions in 20% to 30% of cases, and elevated ROS1 expression was found to be part of a molecular signature for lung adenocarcinoma subtype ROS1 mutations have been reported in NSCLC 22,23 and interestingly, they were in nonadenocarcinoma histologic subtypes. ROS1 somatic mutations were also reported in very low frequencies in cancers of the colon, ovary, breast, upper aerodigestive tract, and skin. 22,24 26 The significance of these mutations is unknown. Overexpression of ROS1 has been reported in 33% to 56 % of glioblastoma and meningeal tumors METHODS OF DETECTION FOR ALTERED ROS1 The first ROS1 translocations in human NSCLC (SLC34A2-ROS1 and CD74-ROS1) were discovered through a global survey of phosphotyrosine kinase signaling in 41 NSCLC cell lines and more than 150 NSCLC tumor samples using an immunoaffinity phosphoproteomic profiling strategy. 12 More recently, a ROS1 break-apart FISH was used to screen for ROS1 rearrangements. 14 As FISH provides no information about the translocation partner, rapid amplification of complementary DNA assays and fusion-specific combined with Sanger sequencing of the polymerase chain reaction products have been applied to identify ROS1 fusion variants in ROS1-positive screens. Validation of ROS1 fusions after their initial discovery was mostly performed using Immunohistochemistry with the D4D6 rabbit monoclonal antibody (Cell Signaling, Danvers, MA) that recognizes amino acids at the carboxyl terminus of human ROS1 has recently been reported to detect ROS1 fusions identified by ROS1 FISH analysis with high sensitivity and no false-positive identification. 16 ROS1 was not detectable using D4D6 in normal lung Copyright 2012 by the International Association for the Study of Lung Cancer 1627

4 Chin et al. Journal of Thoracic Oncology Volume 7, Number 11, November 2012 tissues, although in rare cases, non-neoplastic cells, such as macrophages and bronchial epithelial cells, displayed expression. This is in comparison to a previous report according to which high levels of ROS1 are expressed in lung normal tissue. 5 As only 25% of cases (138 of 556) underwent ROS1 FISH analysis in this study, the false-negative rate of this assay remains to be clarified. Results from this study by Rimkunas et al. 16 suggest ROS1 immunohistochemistry may be a useful method for screening for the low frequency of patients with NSCLC harboring ROS1 fusions. Advantages of an immunohistochemistry assay include a more rapid turnaround time and analysis can be performed on a small amount of tissue. In contrast, FISH requires more specialized equipment, is more costly, and signal fadeout occurs over time. In addition, nextgeneration sequencing 31,32 and chromogenic in situ hybridization 33,34 approaches may have further advantages in detecting ROS1 rearrangement, as shown in ALK-rearranged NSCLC. Targeting ROS1-Rearranged NSCLC Patients with ALK Inhibitors Given the high homology in the kinase domains of ROS1 and ALK, ALK inhibitors were tested efficacious TABLE 3. IC 50 of Selected ROS1 Tyrosine Kinase Inhibitors Compound IC 50 Reference Crizotinib 1.7 nm 36 AP nm 37 WZ nm 35 TAE nm 35 KIST nm 38 KIST nm 39 against ROS1-positive cell lines and tumors (Table 3). In preclinical studies, TAE684, an ALK inhibitor, demonstrated in vitro activity against HCC78 cell lines, 35 attenuated phosphorylation of downstream ROS1 signaling, and induced apoptosis in FIG-ROS positive BaF cells. 11 In silico modeling suggests the drug amino acid interactions between TAE684 and ROS1 kinase domain is as strong as with the ALK kinase domain. 3 A phase I trial (NCT ) of crizotinib was amended to include ROS1-positive advanced-stage NSCLC patients. Preliminary results in this subset of 14 evaluable patients reported a response rate of 57% with a disease control rate FIGURE 2. ROS1 fusion variants described in non small-cell lung cancer. The breakpoints of ROS1 are located at exons 32 (e32), 34 (e34), and 35 (e35). All the ROS1 variants harbor the kinase domain whereas the exon 32 breakpoint allows the resulting fusion variants to harbor the TM. In the fusion partners, orange and dark green represent coiled-coil and transmembrane domains, respectively. TM, transmembrane domain; and PD2, Copyright 2012 by the International Association for the Study of Lung Cancer

5 Journal of Thoracic Oncology Volume 7, Number 11, November 2012 Targeting ROS1 with ALK Inhibitors TABLE 4. Clinical Trials Targeting ROS1 Fusions Trial Compound Sponsor Phase Primary Endpoint NCT Crizotinib Pfizer I Safety, dose, pharmacokinetics NCT AP26113 Ariad Pharmaceuticals I/II Recommended phase II dose, overall response rate NCT ASP3026 Astellas Pharma Inc I Safety and tolerability NCT NCT AZD1480 AstraZeneca I Safety and tolerability accessed July 9, of 79% at 8 weeks. 4 Other early-phase studies that include patients with tumors harboring ROS1 fusions of second-generation ALK inhibitors such as AP26113 (NCT ) and ASP3026 (NCT ) are ongoing (Table 4). Perspective, Future Direction and Conclusion Although rare, the evidence supports ROS1 fusions as a valid therapeutic target in a molecularly defined subset of NSCLC patients. Results from NCT are eagerly awaited. Future work will include testing the sensitivity of the ROS1 fusion variants to crizotinib. Further issues to be addressed include the optimal method to detect clinically relevant ROS1, exact definition of ROS1-positive tumors for treatment stratification and diagnosis, characterization of downstream signaling pathways in ROS1 fusion tumors, and understanding the resistance to ROS1 RTK inhibitors. REFERENCES 1. Shibuya M, Hanafusa H, Balduzzi PC. Cellular sequences related to three new onc genes of avian sarcoma virus (fps, yes, and ros) and their expression in normal and transformed cells. J Virol 1982;42: Nagarajan L, Louie E, Tsujimoto Y, Balduzzi PC, Huebner K, Croce CM. The human c-ros gene (ROS) is located at chromosome region 6q16 6q22. Proc Natl Acad Sci USA 1986;83: Ou SH, Tan J, Yen Y, Soo RA. ROS1 as a druggable receptor tyrosine kinase: lessons learned from inhibiting the ALK pathway. Expert Rev Anticancer Ther 2012;12: Shaw AT, Camidge DR, Engelman JA, et al. Clinical activity of crizotinib in advanced non-small cell lung cancer (NSCLC) harboring ROS1 gene rearrangement. J Clin Oncol 2012;30:(suppl; abstr 7508). 5. Acquaviva J, Wong R, Charest A. The multifaceted roles of the receptor tyrosine kinase ROS in development and cancer. Biochim Biophys Acta 2009;1795: Charest A, Wilker EW, McLaughlin ME, et al. ROS fusion tyrosine kinase activates a SH2 domain-containing phosphatase-2/phosphatidylinositol 3-kinase/mammalian target of rapamycin signaling axis to form glioblastoma in mice. Cancer Res 2006;66: Chiarle R, Simmons WJ, Cai H, et al. Stat3 is required for ALK-mediated lymphomagenesis and provides a possible therapeutic target. Nat Med 2005;11: Chiarle R, Voena C, Ambrogio C, Piva R, Inghirami G. The anaplastic lymphoma kinase in the pathogenesis of cancer. Nat Rev Cancer 2008;8: Soda M, Takada S, Takeuchi K, et al. A mouse model for EML4-ALKpositive lung cancer. Proc Natl Acad Sci USA 2008;105: a. Govindan R, Ding L, Griffith M, et al. Genomic landscape of non-small cell lung cancer in smokers and never-smokers. Cell 2012;150: Charest A, Lane K, McMahon K, et al. Fusion of FIG to the receptor tyrosine kinase ROS in a glioblastoma with an interstitial del(6)(q21q21). Genes Chromosomes Cancer 2003;37: Gu TL, Deng X, Huang F, et al. Survey of tyrosine kinase signaling reveals ROS kinase fusions in human cholangiocarcinoma. PLoS ONE 2011;6:e Rikova K, Guo A, Zeng Q, et al. Global survey of phosphotyrosine signaling identifies oncogenic kinases in lung cancer. Cell 2007;131: Takeuchi K, Soda M, Togashi Y, et al. RET, ROS1 and ALK fusions in lung cancer. Nat Med 2012;18: Bergethon K, Shaw AT, Ou SH, et al. ROS1 rearrangements define a unique molecular class of lung cancers. J Clin Oncol 2012;30: Li C, Fang R, Sun Y, et al. Spectrum of oncogenic driver mutations in lung adenocarcinomas from East Asian never smokers. PLoS ONE 2011;6:e Rimkunas V, Crosby K, Kelly M, et al. Analysis of receptor tyrosine kinase ROS1 positive tumors in non-small cell lung cancer: identification of a FIG-ROS1 fusion. Clin Cancer Res 2012;18: Shigematsu H, Gazdar AF. Somatic mutations of epidermal growth factor receptor signaling pathway in lung cancers. Int J Cancer 2006;118: Solomon B, Varella-Garcia M, Camidge DR. ALK gene rearrangements: a new therapeutic target in a molecularly defined subset of non-small cell lung cancer. J Thorac Oncol 2009;4: Bhattacharjee A, Richards WG, Staunton J, et al. Classification of human lung carcinomas by mrna expression profiling reveals distinct adenocarcinoma subclasses. Proc Natl Acad Sci USA 2001;98: Garber ME, Troyanskaya OG, Schluens K, et al. Diversity of gene expression in adenocarcinoma of the lung. Proc Natl Acad Sci USA 2001;98: Bild AH, Yao G, Chang JT, et al. Oncogenic pathway signatures in human cancers as a guide to targeted therapies. Nature 2006;439: Kan Z, Jaiswal BS, Stinson J, et al. Diverse somatic mutation patterns and pathway alterations in human cancers. Nature 2010;466: Davies H, Hunter C, Smith R, et al. Somatic mutations of the protein kinase gene family in human lung cancer. Cancer Res 2005;65: Ruhe JE, Streit S, Hart S, et al. Genetic alterations in the tyrosine kinase transcriptome of human cancer cell lines. Cancer Res 2007; 67: Cancer Genome Atlas Research Network. Integrated genomic analyses of ovarian cancer. Nature 2011;474: Durinck S, Ho C, Wang NJ, et al. Temporal dissection of tumorigenesis in primary cancers. Cancer Discov 2011;1: Mapstone T, McMichael M, Goldthwait D. Expression of platelet-derived growth factors, transforming growth factors, and the ros gene in a variety of primary human brain tumors. Neurosurgery 1991;28: Watkins D, Dion F, Poisson M, Delattre JY, Rouleau GA. Analysis of oncogene expression in primary human gliomas: evidence for increased expression of the ros oncogene. Cancer Genet Cytogenet 1994; 72: Zhao JF, Sharma S. Expression of the ROS1 oncogene for tyrosine receptor kinase in adult human meningiomas. Cancer Genet Cytogenet 1995;83: Girish V, Sachdeva N, Minz RW, Radotra B, Mathuria SN, Arora SK. Bcl2 and ROS1 expression in human meningiomas: an analysis with respect to histological subtype. Indian J Pathol Microbiol 2005;48: Maher CA, Kumar-Sinha C, Cao X, et al. Transcriptome sequencing to detect gene fusions in cancer. Nature 2009;458: Levin JZ, Berger MF, Adiconis X, et al. Targeted next-generation sequencing of a cancer transcriptome enhances detection of sequence variants and novel fusion transcripts. Genome Biol 2009;10:R115. Copyright 2012 by the International Association for the Study of Lung Cancer 1629

6 Chin et al. Journal of Thoracic Oncology Volume 7, Number 11, November Kim H, Yoo SB, Choe JY, et al. Detection of ALK gene rearrangement in non-small cell lung cancer: a comparison of fluorescence in situ hybridization and chromogenic in situ hybridization with correlation of ALK protein expression. J Thorac Oncol 2011;6: Yoshida A, Tsuta K, Nitta H, et al. Bright-field dual-color chromogenic in situ hybridization for diagnosing echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase-positive lung adenocarcinomas. J Thorac Oncol 2011;6: McDermott U, Iafrate AJ, Gray NS, et al. Genomic alterations of anaplastic lymphoma kinase may sensitize tumors to anaplastic lymphoma kinase inhibitors. Cancer Res 2008;68: Lovly CM, Heuckmann JM, de Stanchina E, et al. Insights into ALKdriven cancers revealed through development of novel ALK tyrosine kinase inhibitors. Cancer Res 2011;71: Katayama R, Khan TM, Benes C, et al. Therapeutic strategies to overcome crizotinib resistance in non-small cell lung cancers harboring the fusion oncogene EML4-ALK. Proc Natl Acad Sci USA 2011;108: Park BS, El-Deeb IM, Yoo KH, et al. Design, synthesis and biological evaluation of new potent and highly selective ROS1-tyrosine kinase inhibitor. Bioorg Med Chem Lett 2009;19: El-Deeb IM, Park BS, Jung SJ, et al. Design, synthesis, screening, and molecular modeling study of a new series of ROS1 receptor tyrosine kinase inhibitors. Bioorg Med Chem Lett 2009;19: Copyright 2012 by the International Association for the Study of Lung Cancer