Aberrant activity of the receptor tyrosine kinases MET, AXL, and FGFR1/2/3 has been associated with tumor progression in a wide variety of human malignancies, notably in instances of primary or acquired resistance to existing or emerging anticancer therapies. This study describes the preclinical characterization of S49076, a novel, potent inhibitor of MET, AXL/MER, and FGFR1/2/3. S49076 potently blocked cellular phosphorylation of MET, AXL, and FGFRs and inhibited downstream signaling in vitro and in vivo. In cell models, S49076 inhibited the proliferation of MET-and FGFR2-dependent gastric cancer cells, blocked METdriven migration of lung carcinoma cells, and inhibited colony formation of hepatocarcinoma cells expressing FGFR1/2 and AXL. In tumor xenograft models, a good pharmacokinetic/pharmacodynamic relationship for MET and FGFR2 inhibition following oral administration of S49076 was established and correlated well with impact on tumor growth. MET, AXL, and the FGFRs have all been implicated in resistance to VEGF/VEGFR inhibitors such as bevacizumab. Accordingly, combination of S49076 with bevacizumab in colon carcinoma xenograft models led to near total inhibition of tumor growth. Moreover, S49076 alone caused tumor growth arrest in bevacizumab-resistant tumors. On the basis of these preclinical studies showing a favorable and novel pharmacologic profile of S49076, a phase I study is currently underway in patients with advanced solid tumors.

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Receptor tyrosine kinases (RTKs) are key regulators of a multitude of cell processes, including survival, proliferation, migration, invasion and angiogenesis. Aberrant activity of certain of these receptor tyrosine kinases has been associated with tumor progression in a wide variety of human malignancies, making them promising drug targets for cancer therapy. Although, in some instances, specific inhibition of just one of these RTKs suffices for inhibition of tumor progression, in the majority of cases, targeting more than one RTK could be required for therapeutic efficacy. These RTKs include MET, the receptor for hepatocyte growth factor (HGF), the fibroblast growth factor receptor, FGFR, and AXL. Dysregulation of MET activity, due to its overexpression or mutation, or overexpression of its ligand, has been consistently associated with aggressive phenotype, resistance to certain anti-cancer therapies and poor outcome. The FGFRs have a well-documented role in tumor angiogenesis, and, more recently, have been implicated directly in tumor cell survival, proliferation and metastasis in a wide range of tumor types. AXL is one of a family of three tyrosine kinase receptors (TAM family) involved in the pathogenesis of several human cancers and has also recently incited interest as a cancer therapeutic target. S 49076 is a novel, potent, ATP-competitive tyrosine kinase inhibitor of MET, FGFR1/2/3 and AXL. S 49076 blocks autophosphorylation of these RTKs and their downstream signaling in cells with IC50 values of between 1 and 200 nM depending on the target and cell line. Furthermore, in kinase binding assays, S 49076 also binds to clinically-relevant MET mutated isoforms and a number of other kinases implicated in cancer pathology at concentrations of less than 100 nM. S 49076 is not, however, a potent inhibitor of VEGFR2. Although VEGFR2 is implicated in tumor angiogenesis, it also plays a major physiological role in the maintenance of vascular tone, and its inhibition by other RTK inhibitors has limited dosing of these molecules in the clinic. The unique inhibition profile of S 49076 may allow inhibition of oncogenic RTKs potentially without toxic effects encountered by VEGFR2 inhibitors. In vitro, S 49076 inhibits the proliferation of MET- and FGFR2-dependent gastric cancer cells, blocks MET-driven migration of lung carcinoma cells and inhibits colony formation of AXL-overexpressing hepatocarcinoma cells. In vivo, oral administration of S 49076 inhibits > 80% MET autophosphorylation in subcutaneous GTL-16 human gastric carcinoma tumors at 3 mg/kg. In both this GTL-16 model and in the MET-dependent U87-MG human glioblastoma model, S 49076 inhibits > 80% tumor growth at 6 mg/kg/day. S 49076 also inhibits FGFR2 autophosphorylation, downstream signaling and tumor growth in FGFR2-dependent SNU-16 gastric tumors, and tumor growth in LS-174T colon carcinoma. Based on these preclinical studies showing a favorable and novel pharmacological profile and the potential of S 49076 as innovative anticancer agent, a phase I study is planned soon to evaluate an oral formulation of S 49076 in patients with advanced solid tumors. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr A238.

Protein kinase inhibitors constitute a class of anticancer agents with demonstrated clinical efficacy. However, the clinical benefit of these agents is often limited to a subset of patients with specific genomic lesions in their tumor cells rendering them sensitive to inhibition of one or more target kinases. It is therefore becoming increasingly important to identify biomarkers of patient stratification to enable personalized care. For those inhibitors targeting just one kinase this stratification strategy is relatively straightforward, although the number of patients who will benefit from these highly selective inhibitors alone is in general severely limited. In the majority of cases, the simultaneous targeting of more than one kinase, possibly by the same drug, will be required for therapeutic efficacy. However, for multi-target kinase drugs, patient stratification is considerably more complex and is becoming a major challenge during clinical development. S 49076 is a novel, potent, ATP-competitive tyrosine kinase inhibitor with activity demonstrated in cell and animal assays against the receptor tyrosine kinases MET, FGFR1/2/3 and AXL, all of which are associated with progression of a wide range of human malignancies. In kinase binding assays, S 49076 also binds to clinically-relevant MET mutated isoforms and 32 other kinases, many of which have also been implicated in cancer pathology. In the present study, the antitumor activity of S 49076 was assessed in a panel of 53 patient-derived tumors of diverse origin growing in three-dimensional in vitro culture. Sixteen (30%) of these tumors were sensitive to S 49076 at 1 μM or less. With the aim to define molecular signatures of sensitivity or resistance, all 53 tumors were characterized for expression, activation and mutation of all of the known target kinases using protein, mRNA and DNA analysis techniques. On an individual basis, no clear relationship between expression level or activity of each target kinase and sensitivity or resistance to S 49076 could be established. However, analysis of microarray data using z-score transformation enabled a clear link to emerge between the sum of the z-scores of all potential target kinases (z-score) and tumor response to S 49076. Moreover, when the presence of RAS or PIK3CA activating mutations was taken into account, the predictive power of the analysis (in particular the positive predictive value for sensitivity) was even more striking. Overall, 91% (29/32) of tumors with a z-score less than 0 were resistant to S 49076, whilst 85% (11/13) of tumors with a z-score greater than 0 and with no known mutations in RAS or PIK3CA were sensitive. Six out of the 8 tumors with a z-score greater than 0 but with mutations in RAS or PIK3CA were resistant. In vivo evaluation of the response to S 49076 of several of the sensitive tumors is in progress in xenograft studies. Together, these data demonstrate the potential power of molecular profiling and, in particular, transcriptomic analyzes in the stratification of patient tumors for treatment with a multi-kinase inhibitor. It is hoped that similar analysis of patient tumors during clinical trials of S 49076 will ultimately contribute to the progress towards a more personalized approach to cancer patient care. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr A240.