Cenisertib [(1S,2S,3R,4R)-3-((5-fluoro-2-((3-methyl-4-(4-methylpiperazin-1-yl)phenyl) amino)pyrimidin-4-yl)amino)bicyclo[2.2.1]hept-5-ene-2-carboxamide]
is an orally available, small molecule pan-Aurora inhibitor currently in clinical
phase studies. Cenisertib has been reported to inhibit Aurora A, B, and C with
IC50 values of 4, 4.8, and 6.8 nM, respectively and to inhibit Abl,
FLT1 (fms-related tyrosine kinase), and FLT3 (FMS-related tyrosine kinase 3)
oncogenic kinases. Cenisertib has potent anti-proliferative activity against
many cell types accompanying unique phenotypic changes such as enlarged cell
size, endoreduplication and apoptosis [1,2].
An excess amount of ATP reduced the level of inhibition of Aurora kinase A by Cenisertib in vitro, indicating that Cenisertib is an ATP competitive inhibitor. The compound was tested in a panel of in vitro kinase assays using diVerent recombinant kinases. Aurora kinase A (%Inhibition = 100%), Abelson murine leukemia viral oncogene homolog 1 (ABL1, %Inhibition = 92%), FLT1 (VEGFR1, %Inhibition = 96%), and FLT3 (%Inhibition = 99%) were more than 90% inhibited by 100 nM of Cenisertib. The cyclin dependent kinases, CDK1, CDK2 and CDK6 were not inhibited at this concentration by Cenisertib [2]. Moreover, the compound inhibited VEGFR2 (KDR) with %inhibition value of 75% at 100 nM concentration.
Inhibition of Aurora kinase B was confirmed by an image-based Aurora kinase-dependent assay detecting intracellular phosphorylation of histone H3 serine 10. Cenisertib reduced the phosphorylation during mitosis with an average EC50 of 14 nM. The selectivity of Cenisertib was assessed using a series of cell-based kinase assays, which measured the activation of kinases using antibodies against phosphorylated sites of specific substrates. Cenisertib exhibited potent inhibition of FLT3 activation mediated by the ITD mutation and VEGF165-induced VEGFR2 activation. Inhibition of Aurora kinase activity in tumor cells had at least 10-fold selectivity over inhibition of AMPK, AXL, AKT, and SAPK activity.
An excess amount of ATP reduced the level of inhibition of Aurora kinase A by Cenisertib in vitro, indicating that Cenisertib is an ATP competitive inhibitor. The compound was tested in a panel of in vitro kinase assays using diVerent recombinant kinases. Aurora kinase A (%Inhibition = 100%), Abelson murine leukemia viral oncogene homolog 1 (ABL1, %Inhibition = 92%), FLT1 (VEGFR1, %Inhibition = 96%), and FLT3 (%Inhibition = 99%) were more than 90% inhibited by 100 nM of Cenisertib. The cyclin dependent kinases, CDK1, CDK2 and CDK6 were not inhibited at this concentration by Cenisertib [2]. Moreover, the compound inhibited VEGFR2 (KDR) with %inhibition value of 75% at 100 nM concentration.
Inhibition of Aurora kinase B was confirmed by an image-based Aurora kinase-dependent assay detecting intracellular phosphorylation of histone H3 serine 10. Cenisertib reduced the phosphorylation during mitosis with an average EC50 of 14 nM. The selectivity of Cenisertib was assessed using a series of cell-based kinase assays, which measured the activation of kinases using antibodies against phosphorylated sites of specific substrates. Cenisertib exhibited potent inhibition of FLT3 activation mediated by the ITD mutation and VEGF165-induced VEGFR2 activation. Inhibition of Aurora kinase activity in tumor cells had at least 10-fold selectivity over inhibition of AMPK, AXL, AKT, and SAPK activity.
Cenisertib was designed and developed based on an
image-based phenotypic screen by Rigel.
The activity of Cenisertib is
as follows:
IC50 (AURKA enzyme assay) = 4.0 nM
IC50 (AURKA enzyme assay) = 4.8 nM
IC50 (AURKC enzyme assay) = 6.8 nM
Common Name: Cenisertib
Synonyms: R763;
R 763; R-763; AS703569; AS 703569; AS-703569; MSC1992371A
IUPAC Name: (1S,2S,3R,4R)-3-((5-fluoro-2-((3-methyl-4-(4-methylpiperazin-1-yl)phenyl) amino)pyrimidin-4-yl)amino)bicyclo[2.2.1]hept-5-ene-2-carboxamide
CAS Number: 871357-89-0; 1145859-64-8 (benzoate)
SMILES: O=C([C@H]1[C@](C2)([H])C=C[C@]2([H])[C@H]1NC3=NC(NC4=CC=C(N5CCN(C)CC5)C(C)=C4)=NC=C3F)N
Mechanism of Action: Kinase Inhibitor; pan-Aurora Inhibitor
Indication: Various Cancers; Anti-Tumor Therapy
Development Stage: Phase I
Company: Rigel
In
a preclinical phase, the in vivo efficacy of Cenisertib was tested in MiaPaCa-2,
adriamycin-resistant tumor, MOLT-4, and MV4-11 xenograft models. Significant
reduction in tumor volumes did not occur in the MiaPaCa-2 xenograft model, but
histological regression and reduction in histone H3 phosphorylation (Ser10) was
observed. In contrast, tumor volumes were significantly reduced in
adriamycin-resistant tumors. Treatment of the MOLT-4 xenograft model resulted
in a 5-10% reduction in the total number of bone marrow cells. The percentages
of leukemia cells were significantly reduced, whereas control groups were not
affected. In the MV4-11 xenograft, Cenisertib induced pronounced anticancer activity
in a dose-dependent manner. For a dose of 20 mg/kg/day, undetectable levels of
tumors were seen in 17% of animals. Increased life span was observed in all
treated groups, whereas all control mice died early [2,3].
The effect of Cenisertib on intracellular phosphorylation of specific substrates was measured in ELISA format assays using anti-phosphoprotein antibodies. The effect of Cenisertib on a variety of cellular activities was analyzed in order to check the selectivity of the compound. The compound selectively inhibited AURKB and FLT3 with nearly the same potency, EC50 = 0.014, 0.011 uM, respectively. It showed nearly 50 times preference against AKT (EC50 = 0.713 uM), 100 times against EGFR (EC50 = 1.491 uM) and more than 500 times against JNKs (EC50 = 7.429 uM). Only VEGFR was inhibited with 2 fold selectivity, EC50 = 0.027 uM [2].
R763/ AS703569 did not show potent inhibition of immune cell activation mediated by many stimuli tested, while it potently inhibited tumor growth. The ability of R763/AS703569 to inhibit tumor growth was examined in colony forming assays using 64 different human tumor xenografts in vitro. R763/AS703569 was active against various tumor types. The mean EC50 value was 0.09 u M. A majority of lung, breast, and renal tumors had EC50 values below the mean value.
A phase I dose-escalation study of Cenisertib was carried out in patients with hematologic malignancies. Patients received escalating doses either on days 1-3 and 8-10 (n = 36) or on days 1-6 (n = 39) of a 21-day cycle. The maximum tolerated doses were 37 and 28 mg/m2/day, respectively. Dose-limiting toxicities included severe neutropenia with infection and sepsis, mucositis/stomatitis, and diarrhea. Complete responses occurred in 3 patients. Four disease-specific expansion cohorts then received the dose and schedule dictated by the escalation phase but the study was prematurely discontinued due to hematologic and gastrointestinal toxicity at clinically effective doses [4].
References:
1. Romanelli, A.; et. al. Inhibiting aurora kinases reduces tumor growth and suppresses tumor recurrence after chemotherapy in patient-derived triple-negative breast cancer xenografts. Mol Cancer Ther 2012, 11(12), 2693-2703.
2. Goulaouic McLaughlin, J.; et. al. Preclinical characterization of Aurora kinase inhibitor R763/AS703569 identified through an image-based phenotypic screen. J Cancer Res Clin Oncol 2010, 136(1), 99-113.
3. Kollareddy, M.; et. al. Aurora kinase inhibitors: Progress towards the clinic. Invest New Drugs 2012, 30(6), 2411-2432.
4. Graux, C.; et. al. A phase I dose-escalation study of MSC1992371A, an oral inhibitor of aurora and other kinases, in advanced hematologic malignancies. Leuk Res 2013, 37(9), 1100-1106.
The
endoreduplication cycle induced by Cenisertib was irreversible even after
the compound was withdrawn from the culture. Oral administration of Cenisertib demonstrated marked inhibition of tumor growth in xenograft models of
pancreatic, breast, colon, ovarian, and lung tumors and leukemia. An acute
myeloid leukemia cell line MV4-11, which carries a FLT3 internal tandem
duplication mutation, is particularly sensitive to Cenisertib in vivo [2]. Cenisertib appeared to induce
endoreduplication within 48 h as evidenced by the accumulation of 4n and 8n
cells. Colo205, HeLa, and MiaPaCa-2 underwent apoptosis after 48 h. In this
study, Colo205, MiaPaCa-2, HeLa, and MV4-11 cells were observed to be most
sensitive to Cenisertib (IC50 = 2 to 8 nM), but primary proliferating
cells were also sensitive despite having higher IC50 values (IC50
= 31 to 160 nM) [2,3]. Moreover, Cenisertib was also tested in a slow growing and adriamycin-resistant tumor (NCI-ADR) xenograft model in nude mice. Tumor growth was signiWcantly suppressed by the treatment with Cenisertib (7 and 10 mg/kg day) in comparison with the control group.
The effect of Cenisertib on intracellular phosphorylation of specific substrates was measured in ELISA format assays using anti-phosphoprotein antibodies. The effect of Cenisertib on a variety of cellular activities was analyzed in order to check the selectivity of the compound. The compound selectively inhibited AURKB and FLT3 with nearly the same potency, EC50 = 0.014, 0.011 uM, respectively. It showed nearly 50 times preference against AKT (EC50 = 0.713 uM), 100 times against EGFR (EC50 = 1.491 uM) and more than 500 times against JNKs (EC50 = 7.429 uM). Only VEGFR was inhibited with 2 fold selectivity, EC50 = 0.027 uM [2].
R763/ AS703569 did not show potent inhibition of immune cell activation mediated by many stimuli tested, while it potently inhibited tumor growth. The ability of R763/AS703569 to inhibit tumor growth was examined in colony forming assays using 64 different human tumor xenografts in vitro. R763/AS703569 was active against various tumor types. The mean EC50 value was 0.09 u M. A majority of lung, breast, and renal tumors had EC50 values below the mean value.
A phase I dose-escalation study of Cenisertib was carried out in patients with hematologic malignancies. Patients received escalating doses either on days 1-3 and 8-10 (n = 36) or on days 1-6 (n = 39) of a 21-day cycle. The maximum tolerated doses were 37 and 28 mg/m2/day, respectively. Dose-limiting toxicities included severe neutropenia with infection and sepsis, mucositis/stomatitis, and diarrhea. Complete responses occurred in 3 patients. Four disease-specific expansion cohorts then received the dose and schedule dictated by the escalation phase but the study was prematurely discontinued due to hematologic and gastrointestinal toxicity at clinically effective doses [4].
References:
1. Romanelli, A.; et. al. Inhibiting aurora kinases reduces tumor growth and suppresses tumor recurrence after chemotherapy in patient-derived triple-negative breast cancer xenografts. Mol Cancer Ther 2012, 11(12), 2693-2703.
2. Goulaouic McLaughlin, J.; et. al. Preclinical characterization of Aurora kinase inhibitor R763/AS703569 identified through an image-based phenotypic screen. J Cancer Res Clin Oncol 2010, 136(1), 99-113.
3. Kollareddy, M.; et. al. Aurora kinase inhibitors: Progress towards the clinic. Invest New Drugs 2012, 30(6), 2411-2432.
4. Graux, C.; et. al. A phase I dose-escalation study of MSC1992371A, an oral inhibitor of aurora and other kinases, in advanced hematologic malignancies. Leuk Res 2013, 37(9), 1100-1106.
