Entrectinib [N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide]
is an oral small molecule inhibitor of TrkA (IC50 = 1 nM), TrkB
(IC50 = 2, 3 nM) and TrkC (IC50 = 5 nM), as well
as ROS1 (IC50 = 7 nM) and ALK (IC50 = 12 nM),
with high potency and selectivity. Entrectinib has demonstrated potent
pharmacological activity in preclinical studies and has the potential to be
first-in-class against the Trk family of kinases [1, 5].
 |
| Entrectinib: 2D and 3D Structure |
Entrectinib was being developed as Anaplastic Lymphoma Kinase (ALK)
inhibitor where studies showed that it induces tumor regression in mouse models
of NPM-ALK–driven lymphoma and EML4-ALK-driven NSCLC; it also has activity
against the Crizotinib-resistant ALK mutants L1196M and
C1156Y. Entrectinib also crosses the blood-brain barrier, inhibits
tumor growth, and prolongs survival in mice with intracranially injected
NCI-H2228 EML4-ALK cells [2].
Due to the structural similarity between ALK and ROS1 kinases, the activity of Entrectinib on ROS1 was evaluated in biochemical and cellular assays. The compound was found to be a potent, ATP competitive inhibitor of ROS1 (IC50 = 7 nM). This biochemical potency translates into inhibition of the proliferation of HCC-78, a NSCLC cell line characterized by ROS1-driven activated signaling, due to the presence of the SCL34A2-ROS1 fusion gene. To further characterize the compound in cells, an activated form of ROS1 was expressed in Ba/F3 cells generating an IL3-independent, ROS1-driven tumor model. NMS-E628 strongly inhibits the proliferation of Ba/F3-ROS1 cells in vitro with confirmed dose-dependent decrease of ROS1 phosphorylation. When tested in vivo, the compound was found to induce complete tumor regression after 10-day continuous treatment in nude mice bearing established Ba/F3-ROS1 tumors [3].
Important timelines in the history of Entrectinib:
a: In Nov 2009, Nerviano Medical Sciences revealed
NMS-E628, a small molecule inhibitor of anaplastic lymphoma kinase with
antitumor efficacy in ALK-dependent lymphoma and non-small cell lung cancer
models at AACR-NCI-EORTC International Conference.
b: In Nov 2011, Nerviano Medical Sciences revealed
NMS-E628 also potently inhibits ROS1 and induces tumor regression in ROS-driven
models at American Association Cancer Society Annual Meeting.
c: In Nov 2013 Nerviano Medical Sciences signed a license
agreement with Ignyta Inc granting the American biotech company exclusive
global development and marketing rights to RXDX-101 and RXDX-102.
d: In 2014 at ASCO Annual Meeting Nerviano
Medical Sciences/Ignyta Inc added pan-TRK activity to the pre-existing ALK
and ROS1 activities.
Dosages and Approvals:
On December 29, 2014, Ignyta, Incorporated announced that the FDA had granted Orphan Drug status and a Rare Pediatric Disease designation for Entrectinib for the treatment of neuroblastoma.
On December 29, 2014, Ignyta, Incorporated announced that the FDA had granted Orphan Drug status and a Rare Pediatric Disease designation for Entrectinib for the treatment of neuroblastoma.
On Feburary 05, 2015 Ignyta announced that the Food and Drug Administation
(FDA) has granted Orphan Drug designation to Entrectinib for
the treatment of TrkA-positive, TrkB-positive, TrkC-positive, ROS1-positive and
ALK-positive non-small cell lung cancer (NSCLC).
Entrectinib is currently in Phase I/II clinical trials with activity
against TrkA as well as TrkB, TrkC, ALK and ROS1 proteins (encoded by NTRK1,
NTRK2, NTRK3, ALK and ROS1 genes, respectively). Entrectinib is
indicated for patients with advanced, refractory, relapsed or metastatic
solid tumors who have exhausted standard treatment options and who exhibit
molecular alterations in any of the target genes.
The activity of Entrectinib is as follows:
IC50 (TRKA enzyme assay) = 1 nM
IC50 (TRKB enzyme assay) = 2 nM; 3 nM [5]
IC50 (TRKC enzyme assay) = 5 nM
IC50 (ROS1 enzyme assay) = 7 nM
IC50 (ALK enzyme assay) = 12 nM; 55 nM [4]
IC50 (IGF1R enzyme assay) = 263 nM
IC50 (AURKA enzyme assay) = 338 nM
IC50 (TRKB enzyme assay) = 2 nM; 3 nM [5]
IC50 (TRKC enzyme assay) = 5 nM
IC50 (ROS1 enzyme assay) = 7 nM
IC50 (ALK enzyme assay) = 12 nM; 55 nM [4]
IC50 (IGF1R enzyme assay) = 263 nM
IC50 (AURKA enzyme assay) = 338 nM
Summary
Common Name: Entrectinib
Synonyms: RXDX-101;
RXDX 101; RXDX101; NMS-E628; NMSE628; NMS E628
IUPAC Name: N-(5-(3,5-difluorobenzyl)-1H-indazol-3-yl)-4-(4-methylpiperazin-1-yl)-2-((tetrahydro-2H-pyran-4-yl)amino)benzamide
CAS Number: 1108743-60-7
SMILES:O=C(NC1=NNC2=C1C=C(CC3=CC(F)=CC(F)=C3)C=C2)C4=CC=C(N5CCN(C)CC5)C=C4NC6CCOCC6
Mechanism of Action: Kinase Inhibitor; Multi-Kinase Inhibitor; pan-TRK Kinase
Inhibitor; ALK Kinase Inhibitor; ROS1 Kinase Inhibitor
Indication: Various
Cancers; Anti-tumor Agents
Development Stage: Phase II
Company: Nerviano
Medical Sciences/Ignyta Inc
The Trk family of kinases, which include TrkA, TrkB and TrkC, are high affinity receptors for the neurotrophin family of nerve growth factors. Deregulated kinase activities of Trk family members due to chromosome rearrangements, gene mutations, splicing variants and overexpression have been shown to be associated with tumorigenesis and poor prognosis in a number of cancer types. Particularly, several chromosomal rearrangements involving TrkA have been reported in lung, colorectal, papillary thyroid, glioblastoma, melanoma and other cancers, and are believed to be the key oncogenic driver in these tumors. Therefore oncogenic Trk may represent a promising therapeutic target in Trk-driven tumors.
In KM-12, a human colorectal cancer cell line driven by constitutively active
TrkA fusion TPM3-NTRK1, Entrectinib exhibited in vitro anti-proliferative
activity with an IC50 of 17 nM, accompanied by
inhibition of TrkA phosphorylation and concomitant inactivation of downstream
effectors, PLCgamma1, AKT and ERK, as well as cell cycle arrest and apoptosis.
In mice bearing KM- 12 xenografts, treatment
with Entrectinib resulted in tumor regression and durable stasis
under either intermittent or continuous dosing regimens, accompanied by
sustained intratumoral inhibition of phospho-TrkA and PLCgamma1. In these
studies, Entrectinib was well tolerated during the course of
treatment [1].
J Med Chem 2016, 59(7), 3392-3408: The article reports synthesis of Entrectinib and its analogues. Pre-clinical data is also reported.
Intermediate:
Final Synthesis:
Identifications:
Entrectinib Synthesis
J Med Chem 2016, 59(7), 3392-3408: The article reports synthesis of Entrectinib and its analogues. Pre-clinical data is also reported.
Intermediate:
Final Synthesis:
Identifications:
 |
| 1H NMR (Estimated) for Entrectinib |
Experimental: 1H NMR (400.5 MHz, DMSO-d6) δ ppm 1.29-1.41 (m, 2H) 1.89-1.97 (m, 2H) 2.24 (s,
3 H) 2.41-2.48 (m, 4H) 3.23-3.29 (m, 4 H) 3.49 (ddd, J = 11.7, 10.2, 2.3 Hz, 2
H) 3.62-3.72 (m, 1 H) 3.80(ddd, J = 11.7, 3.8, 3.8 Hz, 2 H) 4.04 (s, 2 H) 6.13
(d, J = 2.1 Hz, 1 H) 6.23 (dd, J = 9.0, 2.2 Hz, 1 H) 6.93-7.04 (m, 3 H) 7.25
(dd, J = 8.7, 1.5 Hz, 1 H) 7.40 (d, J = 8.7 Hz, 1 H) 7.48 (s, 1 H) 7.80 (d, J =
9.0 Hz, 1 H) 8.29 (d, J = 7.6 Hz, 1 H) 10.07 (s, 1 H) 12.62 (s, 1 H).
References:
1. Anderson, D.; et. al. Inhibition of
Trk-driven tumors by the pan-Trk inhibitor RXDX-101. Euro J Cancer 2014, 50(supp
6),101. (FMO only)
2. Ardini, E.; et. al. Abstract
A243: Characterization of NMS-E628, a small molecule inhibitor of
anaplastic lymphoma kinase with antitumor efficacy in ALK-dependent lymphoma
and non-small cell lung cancer models Mol Cancer Ther 2009, 8(12
suppl), A244. (FMO only)
3. Ardini, E.; et. al. Abstract 2092: The
ALK inhibitor NMS-E628 also potently inhibits ROS1 and induces tumor regression
in ROS-driven models . Cancer Res 2013, 73(8 suppl),
2092. (free copy)
4. Lombardi, B. A.; et. al. Substituted indazole
derivatives active as kinase inhibitors. WO2009013126A1
5. Menichincheri, M.; et. al. 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. J Med Chem 2016, 59(7), 3392-3408. (FMO only)
