Discovery of Entrectinib: A New 3-Aminoindazole As a Potent Anaplastic Lymphoma Kinase (ALK), c-ros Oncogene 1 Kinase (ROS1), and Pan-Tropomyosin Receptor Kinases (Pan-TRKs) inhibitor.

Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase responsible for the development of different tumor types. Despite the remarkable clinical activity of crizotinib (Xalkori), the first ALK inhibitor approved in 2011, the emergence of resistance mutations and of brain metastases frequently causes relapse in patients. Within our ALK drug discovery program, we identified compound 1, a novel 3-aminoindazole active on ALK in biochemical and in cellular assays. Its optimization led to compound 2 (entrectinib), a potent orally available ALK inhibitor active on ALK-dependent cell lines, efficiently penetrant the blood-brain barrier (BBB) in different animal species and highly efficacious in in vivo xenograft models. Moreover, entrectinib resulted to be strictly potent on the closely related tyrosine kinases ROS1 and TRKs recently found constitutively activated in several tumor types. Entrectinib is currently undergoing phase I/II clinical trial for the treatment of patients affected by ALK-, ROS1-, and TRK-positive tumors.

The TPM3-NTRK1 rearrangement is a recurring event in colorectal carcinoma and is associated with tumor sensitivity to TRKA kinase inhibition.

The NTRK1 gene encodes Tropomyosin-related kinase A (TRKA), the high-affinity Nerve Growth Factor Receptor. NTRK1 was originally isolated from a colorectal carcinoma (CRC) sample as component of a somatic rearrangement (TPM3-NTRK1) resulting in expression of the oncogenic chimeric protein TPM3-TRKA, but there has been no subsequent report regarding the relevance of this oncogene in CRC. The KM12 human CRC cell line expresses the chimeric TPM3-TRKA protein and is hypersensitive to TRKA kinase inhibition. We report the detailed characterization of the TPM3-NTRK1 genomic rearrangement in KM12 cells and through a cellular screening approach, the identification of NMS-P626, a novel highly potent and selective TRKA inhibitor. NMS-P626 suppressed TPM3-TRKA phosphorylation and downstream signaling in KM12 cells and showed remarkable antitumor activity in mice bearing KM12 tumors. Finally, using quantitative reverse transcriptase PCR and immunohistochemistry (IHC) we identified the TPM3-NTRK1 rearrangement in a CRC clinical sample, therefore suggesting that this chromosomal translocation is indeed a low frequency recurring event in CRC and that such patients might benefit from therapy with TRKA kinase inhibitors.

Crystal structures of anaplastic lymphoma kinase in complex with ATP competitive inhibitors.

Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase involved in the development of several human cancers and, as a result, is a recognized target for the development of small-molecule inhibitors for the treatment of ALK-positive malignancies. Here, we present the crystal structures of the unphosphorylated human ALK kinase domain in complex with the ATP competitive ligands PHA-E429 and NVP-TAE684. Analysis of these structures provides valuable information concerning the specific characteristics of the ALK active site as well as giving indications about how to obtain selective ALK inhibitors. In addition, the ALK-KD-PHA-E429 structure led to the identification of a potential regulatory mechanism involving a link made between a short helical segment immediately following the DFG motif and an N-terminal two-stranded beta-sheet. Finally, mapping of the activating mutations associated with neuroblastoma onto our structures may explain the roles these residues have in the activation process.

Geochemistry and magmatc evolution of explosive tephras et3 and et4 from Arenal Volcano, Costa Rica

An outcrop at El Cruce, 5.7 km from the vent of Arenal volcano, exposes thick sequences of two recent tephras, ET4 (silicic) and, immediately above, ET3 (mafic). The ET4 to ET3 transition is abrupt, with no soil between the layers. Therefore, they appear to be part of one eruptive phase. A petrological model is inferred from the detailed stratigraphic record of these eruptions. The ET4 sequence can be modeled by crystal fractionation via gravity settling. ET4 is most mafic at the top of the section. The silicic tephra at the bottom of this unit is aphyric. Compatible elements increase and incompatible elements decrease moving down through the magma chamber. The percentage of phenocrysts increases through the top. Models of crystal fractionation, using the most mafic and phenocryst enriched tephra as the parent magma, produce the most silicic tephra by removal of 37

phenocrysts. The ET3 sequence shows the opposite trend of that seen in the ET4 sequence, becoming less enriched in incompatible elements down through the section. Some crystal sorting, followed by the removal of a silicic melt from the top of the magma chamber, could generate the reversed trend for the ET3 sequence.(AU)

Fluid - volcano interaction in an active stratovolcano : The crater lake system of Poás volcano, Costa Rica

Seismic and geochemical data collected at Poás volcano, Costa Rica, since 1978 suggest that temperature and chemical variations recorded in subaerial fumaroles and the crater lake are related to episodic release of heat and volatiles associated with hydrofracturing of the upper margin of the shallow magma body. Power outputs associated with these events approach 600 MW and are superimposed on a baseline energy flux of approximately 200 MW. Finally, analysis of heat and water budgets for the crater lake over the period 1978-1989 indicates that the recent decline and disappearance of the lake was caused by variations in summit rainfall combined with an increase in subsurface heat flow to the lake in 1987 - 1988. The increase in heat flow is related to continued degassing of high - temperature volatiles following a hydrofracturing / magma ascent episode that ocurred in 1986 (AU)

Síntesis geovulcanológica del Arenal (Costa Rica) : 20 años de contínua actividad eruptiva (1968-1988)

El Volcán Arenal es un pequeño y joven estrato volcán (1.633 m.s.n.m; 12.5 km3) andesítico-basáltico (muy pobre en potasio y rico en alúmina) ubicado en medio de las dos principales cordilleras volcánicas de Costa Rica y de un quimismo diferente al de ellos. Desde 1968 hasta el presente (20 años), se ha producido su reciente período eruptivo (después de carecer de registros históricos) con la efusión de lavas piroclastos. Resaltan la ciclicidad petrológica de sus productos, la constante y variada actividad sísmica del actual período, así como la dinámica de las coladas de lava. Esto le ha dado el carácter de "laboratorio vulcanológico". El presente trabajo es una síntesis de las principales características de este interesante volcán y de su evolución en el tiempo. Se evidencia cómo la dinámica eruptiva de un volcán puede variar bastante en un corto intervalo (AU)