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Preclinical Evaluation of AMG 900, a Novel Potent and Highly Selective Pan-Aurora Kinase Inhibitor with Activity in Taxane-Resistant Tumor Cell Lines Marc Payton1, 5, Tammy L. Bush1, Grace Chung1, Beth Ziegler1, Patrick Eden1, Patricia McElroy2,

Sandra Ross2, Victor J. Cee3, Holly L. Deak3, Brian L. Hodous3, Hanh Nho Nguyen3, Philip R.

Olivieri3, Karina Romero3, Laurie B. Schenkel3, Annette Bak4, Mary Stanton4, Isabelle Dussault1,

Vinod F. Patel3, Stephanie Geuns-Meyer3, Robert Radinsky1, and Richard L. Kendall1

Departments of 1Oncology Research, 2Hematology Research, 3Medicinal Chemistry, and

4Pharmaceutics, Amgen Inc., Thousand Oaks, California 91320 or Cambridge, Massachusetts 02142

5To whom correspondence should be addressed:

Amgen Inc., One Amgen Center Drive, Mailstop 14-1-B, Thousand Oaks, CA 91320-1789.

Phone: (805) 447-6697, Fax: (805) 375-8368

Email: [email protected]

Running title: AMG 900, a Novel Pan-Aurora Kinase Inhibitor

Keywords: AMG 900, aurora kinase, multidrug resistance, P-gp, antimitotic

Published OnlineFirst on October 8, 2010 as 10.1158/0008-5472.CAN-10-3001

Research. on February 3, 2019. © 2010 American Association for Cancercancerres.aacrjournals.org Downloaded from

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Abstract

In mammalian cells, the aurora kinases (aurora-A, -B, and -C) play essential roles in regulating cell

division. Aurora-A and -B expression is elevated in a variety of human cancers and is associated with

high proliferation rates and poor prognosis, making them attractive targets for anticancer therapy. AMG

900 is an orally bioavailable, potent and highly selective pan-aurora kinase inhibitor that is active in

taxane-resistant tumor cell lines. In tumor cells, AMG 900 inhibited autophosphorylation of aurora-A and -

B as well as phosphorylation of histone H3 on Ser10, a proximal substrate of aurora-B. The predominant

cellular response of tumor cells to AMG 900 treatment was aborted cell division without a prolonged

mitotic arrest, which ultimately resulted in cell death. AMG 900 inhibited the proliferation of 26 tumor cell

lines at low nanomolar concentrations, including cell lines resistant to the antimitotic drug paclitaxel and to

other aurora kinase inhibitors (AZD1152, MK-0457, and PHA-739358). Furthermore, AMG 900 was

active in an AZD1152-resistant HCT116 variant cell line that harbors an aurora-B mutation (W221L). Oral

administration of AMG 900 blocked the phosphorylation of histone H3 in a dose-dependent manner and

significantly inhibited the growth of HCT116 tumor xenografts. Importantly, AMG 900 was broadly active

in multiple xenograft models representing five tumor types, including three multidrug-resistant xenograft

models. AMG 900 has entered clinical evaluation in adult patients with advanced cancers and has the

potential to treat tumors refractory to anticancer drugs such as the taxanes.





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Introduction

Somatic cell division is a complex and highly coordinated process that ensures faithful

segregation of duplicated chromosomes into two daughter cells. Deregulation of the cell cycle is a

hallmark of cancer, characterized by uncontrolled proliferation and defects in chromosome segregation.

Antimitotic drugs that block tumor cell division are a proven intervention strategy in the treatment of

cancer. However, the clinical benefits of classical antimitotic drugs may be hampered by development of

multidrug resistance (MDR) and collateral damage to non-dividing cells including for example, peripheral

neuropathy (1). Aurora kinases are essential mitotic regulators and their potential role in tumorigenesis

makes them attractive targets for anticancer therapy (2-4).

In mammalian cells, the aurora family of serine/threonine protein kinases is comprised of three

paralogous genes (aurora-A, -B, and -C). Aurora-A and -B are essential regulators of mitotic entry and

progression, whereas aurora-C function is primarily restricted to male meiosis during spermatogenesis (5-

10). Aurora-A can function as an oncogene and is amplified in a subset of human tumors. Aurora-A and

-B expression is frequently elevated in human cancers and is associated with advanced clinical staging

(11).

The mitotic checkpoint, also referred to as the spindle assembly checkpoint (SAC), is a

surveillance mechanism responsible for controlling proper alignment, microtubule-kinetochore

attachments, and segregation of duplicated chromosomes (12). In tumor cells, genetic depletion or

pharmacological inhibition of aurora-A results in abnormal spindle formation and SAC activation. In

constrast, depletion or inhibition of aurora-B inactivates the SAC, resulting in aborted cell division without

a mitotic arrest. Importantly, dual suppression of aurora-A and -B phenocopies the effects of inhibiting

aurora-B alone (13, 14). The silencing of the SAC leads to an accumulation of tumor cells that contain 4N

DNA content in the G1-phase of the cell cycle. Continued suppression of aurora-B activity leads to further

rounds of genome replication without division, a process referred to as endoreduplication, which

ultimately results in tumor cell death (15, 16). This mechanism of action is distinct from that of

microtubule-binding antimitotic drugs (e.g. taxanes, vinca alkaloids, and epothilones) in that death of





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tumor cells is primarily driven by continued cell-cycle progression rather than by SAC activation and

prolonged cell arrest in mitosis (16, 17).

Drug resistance is a major problem limiting the efficacy of many current anticancer therapies.

The underlying mechanisms of clinical resistance to microtubule-binding agents are multifactorial and not

fully understood (18). In cultured tumor cells, two prominent mechanisms of resistance to the taxanes are

overexpression of drug efflux pumps and tubulin modifications (19-21). One strategy to overcome

susceptibility to the effects of MDR would be to design a novel antimitotic drug candidate whose activity is

not influenced by drug efflux, mediated by ATP-binding cassette (ABC) transporters such as P-

glycoprotein/P-gp (ABCB1) and BCRP (ABCG2) (22). Moreover, a small molecule inhibitor that is

equipotent against two essential mitotic kinases may reduce the potential for resistance driven by target-

modifying mutations (23, 24).

Currently, a variety of ATP-competitive inhibitors that target one or more of the aurora kinases

and that have varying degrees of kinase specificity are in early clinical development (25). This report

describes the preclinical activities of AMG 900, an orally bioavailable, potent and selective pan-aurora

kinase inhibitor with activity in MDR tumor cell lines. In contrast to paclitaxel and three well-characterized

aurora kinase inhibitors (AZD1152, MK-0457, and PHA-739358), AMG 900 showed uniform potency

across tumor cell lines, including P-gp and BCRP expressing cell lines. Furthermore, AMG 900 was

active in an HCT116 cell line adapted to grow in the presence of AZD1152. This HCT116 variant cell line

carries a missense mutation in one allele of the aurora-B gene resulting in an amino acid substitution

(W221L) in its activation loop. In vivo, AMG 900 blocks the phosphorylation of histone H3, a proximal

substrate of aurora-B (26) and inhibits the growth of multiple tumor xenografts, including three MDR

xenograft models resistant to paclitaxel or docetaxel. Our data provides compelling evidence that AMG

900 may be active in tumors resistant to taxanes and three other well-characterized inhibitors that target

aurora-B. AMG 900 is presently under clinical evaluation in adult patients with advanced cancers.

Materials and Methods





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Chemistry. AMG 900 N-(4-((3-(2-amino-4-pyrimidinyl)-2-pyridinyl)oxy)phenyl)-4-(4-methyl-2-thienyl)-1-phthalazinamine) was

synthesized at Amgen (WO 2007087276). The molecular structures have been disclosed for the following compounds: paclitaxel

and docetaxel (18), MLN8054 (25), MK-0457 (4), AZD1152 (25), and PHA-739358 (25).

Cell lines. Tumor cell lines were obtained from the American Type Culture Collection (ATCC) unless otherwise specified. Cell lines

were grown under recommended conditions. The HCT116 p21+ and p21- cell lines were obtained from Johns Hopkins University

under a licensing agreement. The CAL51 cell line was obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen

(DSMZ, Germany). The MCF-7 p53+ and p53- cell lines were generated as previously described (27). The MDA-MB-231-PTX and

NCI-H460-PTX cell lines were established by growing the cells in the presence of increasing concentrations of paclitaxel over a

period of 6 months. The HCT116 AZD1152-resistant cell line was established by growing the cells in the presence of AZD1152 at

80 nmol/L. The histone H2B-GFP (BD Biosciences) HeLa cell line was established at Amgen.

Animals. All experimental procedures were conducted in accordance with Institutional Animal Care and Use Committee and U.S.

Department of Agriculture regulations. Four to six-week-old female athymic nude mice (Harlan Sprague Dawley) were housed in

sterilized cages and maintained under aseptic conditions. Eight-week-old female BDF1 mice (Charles River Laboratories) were

housed five per filter-capped cage under pathogen-free conditions. The laboratory housing the animals provided alternating light

and dark cycles (12 hours each) and met the standards of the Association for Assessment and Accreditation of Laboratory Animal

Care specifications. Food, water, and nutritional supplements were offered ad libitum. All drugs were administered based on the

individual body weight of each mouse.

Western blot analysis. Cell lysates were processed for Western blot analysis as previously described (27). Immunoblots were

probed with the following antibodies: anti-aurora-A, anti-aurora-B (BD Biosciences), anti-p-aurora-A Thr288, anti-p-aurora-B Thr232,

anti-cleaved caspase-7 Asp198, and anti-total p53 (Cell Signaling), anti-β-actin (Sigma), and anti-p21Cip1 (Santa Cruz). Next, the

immunoblots were probed with either anti-rabbit or anti-mouse IgG (Vectastain kit, Vector Laboratories) and the protein bands were

detected using the Lightning-Enhanced Chemiluminescence Kit from PerkinElmer.

Details regarding enzyme assays, nocodazole block, cell imaging assays, colony formation assays, flow cytometry assays, aurora

kinase gene analysis, pharmacodynamic assays, tumor xenograft models, neutrophil assessment study, and statistical analyses are

described in the Supplementary Materials and Methods.

Results

AMG 900 is a potent and highly selective pan-aurora kinase inhibitor. The discovery of a

novel class of ATP-competitive phthalazinamine small molecule inhibitors of aurora kinases led to the

identification of AMG 900 (S. Geuns-Meyer, manuscript in preparation). AMG 900 inhibits the enzyme

activity of all three aurora kinase family members with IC50 values ≤ 5 nmol/L (Fig. 1A). To determine the





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specificity of AMG 900 across the kinome, a panel of 26 kinases was screened, and only p38α and TYK2

enzymes were inhibited (> 50%) at concentrations less than 500 nmol/L (Fig. 1A). Further screening was

performed against a panel of 353 distinct kinases using an ATP-site dependent competition binding assay

(28). AMG 900 exhibited low nanomolar binding affinity for aurora kinases as well as interactions (Kd < 50

nmol/L) with DDR1, DDR2, and LTK receptor tyrosine kinases (Fig. 1A).

To investigate the inhibition of aurora kinase activity in cells by AMG 900, the levels of

phosphorylated aurora-A Thr288, aurora-B Thr232, and histone H3 Ser10 (p-histone H3) were determined

using Western blotting and quantitative cell imaging techniques. In HeLa cells, AMG 900 inhibited

autophosphorylation of aurora-A and -B in a concentration-dependent manner (Fig. 1B). MLN8054

(aurora-A inhibitor) and AZD1152 (aurora-B inhibitor) selectively blocked the autophosphorylation of

either aurora-A Thr288 or aurora-B Thr232, respectively. In the cell imaging assay, HeLa cells treated with

AMG 900 for 6 hours showed a concentration-dependent decrease in p-aurora-A Thr288 and p-histone H3

immunofluorescence staining with IC50 values of 6.5 and 8.2 nmol/L, respectively (Fig. 1C). Next, the

effects of AMG 900 on chromosome dynamics and cell-cycle progression were examined by live-cell

imaging of HeLa cells that express a histone H2B-GFP fusion protein. The top image panel of Figure 1D

shows a representative DMSO-treated cell entering and exiting mitosis to yield two daughter cells. In

contrast, a representative AMG 900-treated cell entering mitosis failed to properly congress

chromosomes to the metaphase plate, which subsequently resulted in aborted cell division (bottom panel

of Fig. 1D). These data demonstrate that AMG 900 does not induce a prolonged cell arrest in mitosis,

consistent with inhibition of aurora-B and SAC silencing.

Effects of AMG 900 on proliferating cells are consistent with inhibition of aurora-B activity.

Suppression of aurora-B activity in mitosis is known to induce polyploidy in tumor cells as a consequence

of cytokinesis failure. To confirm this, we first determined the concentration of AMG 900 that completely

suppressed p-histone H3 in HCT116 cells (Fig. 2A). Treatment of HCT116 cells with 50 nmol/L of AMG

900 for 48 hours resulted in polyploidy and suppressed the formation of colonies after cell replating (Fig.

2B and 2C). Several reports have established that inhibition of aurora-B activates the post-mitotic G1-

checkpoint mediated by the p53 tumor suppressor protein (29, 30). Consistent with these findings, AMG





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900 induced a significant increase in p53 and p21Cip1 proteins that correlated with a corresponding

decrease in 5-bromo-2'-deoxyuridine (BrdUrd) incorporation, a direct measure of DNA synthesis

(Supplementary Fig. S1). To assess the kinetics of cell death induction by AMG 900, HCT116 cells were

treated with compound for 48 hours, washed twice, and then cultured in complete media lacking

compound for 0, 24, 48, and 72 hours. Treatment with AMG 900 induced a time-dependent increase in

apoptosis measured by the induction of cleaved caspase-7 (Fig. 2D). Other mechanisms of cell death

(e.g. mitotic catastrophe, multipolar cell division, and giant-cell necrosis) may also contribute to the loss of

tumor cell viability after treatment with AMG 900 (unpublished observations, 31). Consistent with the

highly selective profile of AMG 900, Jurkat cells treated with compound for 48 hours over a 5000-fold

concentration range (0.0012 to 5 μmol/L) maintained a stable polyploidy phenotype, indicating that the

dominant phenotype of AMG 900 inhibition is through inhibition of aurora-B (data not shown). AMG 900

had no obvious toxic effects on non-cycling human foreskin fibroblast cells up to a concentration of 25

μmol/L; however it did inhibit cell-cycle progression (without endoreduplication) and induced cell death in

proliferating human bone marrow mononuclear cells at low nanomolar concentrations as expected with

an on-mechanism effect of this agent (data not shown).

AMG 900 blocks the proliferation of multiple human tumor cell lines, including cell lines

resistant to paclitaxel, AZD1152, MK-0457, and PHA-739358. To investigate the antiproliferative

effects of AMG 900, a panel of 26 cell lines from diverse tumor types were treated with compound for 24

hours, washed twice in complete media lacking AMG 900, and then cultured for an additional 48 hours.

Concentration-response curves and corresponding cell count EC50 values were established for each cell

line (Fig. 3A and 3B). The results show that AMG 900 inhibited cell proliferation with EC50 values ranging

from 0.7 to 5.3 nmol/L. Importantly, four of these AMG 900-sensitive cell lines (HCT-15, MES-SA-Dx5,

769P, and SNU449) are resistant to paclitaxel and other anticancer agents (32-34). To further investigate

the activity of AMG 900 in MDR cells, four tumor cell lines expressing P-gp, were compared with three

non-P-gp expressing parental cell lines. As shown in Figure 3C, in a colony formation assay, there was a

profound decrease in the number of detectable colonies in all of the cell lines treated with AMG 900 at 5

or 50 nmol/L. By contrast, paclitaxel failed to inhibit colony formation in all four of the P-gp expressing





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tumor cell lines at concentrations effective in the parental cell lines. Intrigued by this potentially important

finding, we sought to understand if this was a common property of aurora kinase inhibitors. We therefore

evaluated three well-characterized aurora kinase inhibitors (AZD1152, MK-0457, and PHA-739358) in a

subset of MDR tumor cell lines expressing either P-gp or BCRP drug efflux transporters. Interestingly,

AMG 900 inhibited p-histone H3 or induced polyploidy across all the cell lines tested irrespective of P-gp

or BCRP status with uniform potency (IC50 or EC50 values ranging from 2 to 3 nmol/L). By contrast, the

other aurora kinase inhibitors were less potent in one or more of the MDR cell lines compared with the

matched sensitive tumor cell lines (Table 1). Inhibition of P-gp and BCRP drug pumps with GF120918

(21) sensitized HCT-15 and RPMI-8226 cells to AZD1152, suggesting that the loss of potency in the MDR

cell lines was due to drug efflux (data not shown).

To investigate alternative mechanisms of resistance to aurora kinase inhibitors (35), HCT116

cells were adapted to grow in the presence of AZD1152. The activity of AMG 900 was then evaluated in

the HCT116 parental and AZD1152-resistant cell lines. The cellular ≥ 4N DNA content EC50 values for

AMG 900 were 2 and 5 nmol/L compared to 34 and 672 nmol/L for AZD1152, respectively

(Supplementary Fig. S2A). AMG 900 inhibited the colony formation in both HCT116 cell lines at

concentrations ≥ 5 nmol/L, whereas the variant subline was insensitive to AZD1152 at 50 nmol/L

(Supplementary Fig. S2B). Both of the HCT116 cell lines were equally sensitive to paclitaxel and were

negative for P-gp and BCRP expression (Supplementary Fig. S2C). Interestingly, the HCT116 variant

subline harbors a missense mutation in one allele of the aurora-B gene (TGG TTG; W221L), whereas

no mutations were detected in the aurora-A and -C genes. These results suggest that AMG 900

maintains activity in tumor cells carrying a heterozygous mutation in aurora-B that may be responsible for

resistance to AZD1152. Somatic point mutations at the analogous residue adjacent to the invariant DFG

motif have been found in epidermal growth factor receptor and BRAF genes (L858R and L596R,

respectively) in primary human tumors (36, 37). Studies are underway to more fully characterize the

biological effects of this aurora-B mutation.

AMG 900 inhibits the phosphorylation of histone H3 and suppresses the growth of human

tumor xenografts in vivo. To confirm that AMG 900 inhibits aurora-B activity in vivo, mice bearing





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established HCT116 tumors were administered a single oral dose of vehicle alone or AMG 900 at 3.75,

7.5, or 15 mg/kg. As shown in Figure 4A and 4B, administration of AMG 900 resulted in significant dose-

dependent inhibition of p-histone H3 in tumors compared with the vehicle-treated control group (P ≤

0.008). Furthermore, the degree of p-histone H3 suppression in tumors correlated with plasma drug

concentrations. AMG 900 also inhibited p-histone H3 in mouse bone marrow cells at comparable doses

and plasma drug concentrations (data not shown). In a separate multiple dose study using the HCT116

tumor xenograft model, AMG 900 induced a marked increase in the percentage of tumor cells with ≥ 4N

DNA content (Supplementary Fig. S3).

Next, we compared the inhibition of aurora-B activity, as measured by the degree and duration of

p-histone H3 inhibition, with suppression of tumor growth in vivo. Mice bearing established HCT116

tumors were orally administered vehicle alone or AMG 900 at 3.75, 7.5, or 15 mg/kg twice daily (b.i.d.) for

2 consecutive days per week for 3 weeks. As shown in Figure 4C, intermittent administration of AMG 900

resulted in a dose-dependent inhibition of tumor growth compared with the vehicle-treated control group

(3.75 mg/kg (40%), 7.5 mg/kg (64%), 15 mg/kg (75%), P ≤ 0.0001). The main adverse effects observed

with AMG 900 at the highest dose were moderate body weight loss (average of 7%, Supplementary Fig.

S4A) with transient and reversible myelosuppression (data not shown). An alternative schedule based on

daily dosing was also tested. Mice with established HCT116 tumors were orally administered vehicle

alone or AMG 900 at 1.5, 2.25, or 3 mg/kg b.i.d. for 3 weeks (Fig. 4D). This continuous schedule with

lower doses of AMG 900 resulted in significant tumor growth inhibition when compared with the vehicle-

treated control group (2.25 mg/kg; 51% inhibition, P = 0.0019) and 3 mg/kg; 74% inhibition, P < 0.0001).

Daily dosing with AMG 900 was well-tolerated with no adverse effect on body weight (Supplementary Fig.

S4B), although a decrease in neutrophil counts was observed at the end of the study (data not shown).

The effect of AMG 900 on tumor growth was further evaluated in a panel of human xenografts

from five different tumor types (breast, colon, lung, pancreatic, and uterine), including three MDR

xenograft models. Mice bearing established tumors were orally administered AMG 900 at 15 mg/kg b.i.d.

for two consecutive days per week or at 3 mg/kg b.i.d. every day for the duration of the study. As shown

in Figure 5A, AMG 900 exhibited significant antitumor activity in all nine xenograft models tested (50 to





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97% tumor growth inhibition (TGI) compared with the vehicle-treated control group, *P < 0.005, **P <

0.0005). Importantly, AMG 900 was active in the MES-SA-Dx5 (84% TGI, P < 0.0001) and NCI-H460-

PTX (66% TGI, P < 0.0001) xenograft models that were resistant to either docetaxel or paclitaxel

administered at their respective maximum tolerated doses (Fig. 5B and 5C). Together, these in vivo data

provide evidence that AMG 900 inhibits the activity of aurora-B in HCT116 tumors and suppresses the

growth of multiple xenografts that represent diverse tumor types. Notably, our data indicate that

AMG 900 has the potential to treat tumors resistant to standard-of-care antimitotic drugs.

Since inhibition of aurora-B activity is known to cause myelosuppression (11, 25), studies were

performed to examine the effects of PEGylated-granulocyte colony-stimulating factor (SD/02)

administration on AMG 900-induced neutropenia. Mice were treated with AMG 900 at 15 mg/kg b.i.d for

two consecutive days alone or with SD/02 at 1 mg/kg one day subsequent to the first day of AMG 900

treatment. Administration of AMG 900 alone showed a marked reduction in neutrophils by day 4 after

treatment followed by a full recovery of neutrophils by day 8. Mice administered SD/02 one day after the

initiation of AMG 900 treatment showed a significant decrease in the duration of neutropenia compared to

AMG 900 treatment alone (days 6, 7, and 8 (P < 0.0001) and day 11 (P < 0.0018), Supplementary Fig.

S5).

Discussion

Targeting the structural components of the mitotic machinery is a proven intervention strategy in the

treatment of cancer. Microtubule-binding agents such as the taxanes and vinca alkaloids are highly

active against a variety of human cancer types, although multidrug resistance and peripheral neuropathy

remain persistent problems. Consequently, there is a need for novel antimitotic drugs that target

nonmicrotubule proteins such as the mitotic kinases (38, 39). In the past decade, aurora-A and -B have

gained prominence as essential regulators of somatic cell division and represent promising mitotic targets

for anticancer therapy.

In this report, we have described the preclinical activities of AMG 900, a novel potent and highly

selective pan-aurora kinase inhibitor. The predominant response of tumor cells to AMG 900 treatment





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was aborted cell division without a protracted mitotic arrest resulting in polyploidy-specific lethality. AMG

900 induces p53 and p21Cip1 proteins, which may act to regulate the rate of endoreduplication.

Remarkably, AMG 900 is active in all of the tested tumor cell lines at low nanomolar concentrations,

suggesting that it can inhibit the proliferation of tumor cells irrespective of genomic alterations or tumor

origin. It is possible that the underlying profile of genetic and epigenetic modifications in tumor cells may

play an important role in determining the fate of remnant cells that survive AMG 900 treatment. Because

cells must pass through G2-M phase of the cell cycle in order for AMG 900 to inhibit aurora kinase activity,

other factors such as mitotic and proliferation indices may impact tumor cell response. Further efforts will

be required to define potential susceptibility markers to help identify the most AMG 900-vulnerable tumors

as well as determining if combinatorial approaches lead to durable tumor regression.

Despite the ongoing debate regarding the relevance of drug efflux-mediated clinical resistance,

the consistent potency of AMG 900 against tumor cells irrespective of P-gp or BCRP status may

circumvent a mechanism by which tumor cells could otherwise escape the activity of this experimental

agent. Furthermore, we found that AMG 900 is active in an AZD1152-resistant cell line harboring a

missense mutation in aurora-B resulting in an amino acid substitution (W221L) in its kinase domain

activation loop. A recent report has shown that other aurora-B mutant alleles (Y156H, G160V) located in

the ATP-binding pocket confer resistance to multiple aurora inhibitors (35). It will be of interest to test

whether AMG 900 inhibits aurora-B activity in these catalytic domain mutants.

Consistent with our in vitro observations, AMG 900 inhibits the phosphorylation of histone H3 and

growth of multiple human tumor xenograft models using either intermittent or continuous dosing

schedules. Most relevant to our goal of developing an antimitotic agent that is effective against drug

resistant tumors, AMG 900 shows promising antitumor activity in tumor xenografts resistant to taxanes.

As anticipated, treatment of mice at efficacious doses of AMG 900 leads to a transient loss of body weight

and reversible myelosuppression.

In summary, AMG 900 is an orally bioavailable, potent and highly selective pan-aurora kinase

inhibitor with activity in taxane-resistant tumor cell lines. Together these features distinguish AMG 900

from other antimitotic drugs as well as three other aurora kinase inhibitors (AZD1152, MK-0457, and





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PHA-739358). These key attributes contribute to the profile of this attractive clinical candidate; AMG 900

has entered phase 1 evaluation in adult patients with advanced cancers.

Acknowledgements

The authors would like to thank our Aurora team colleagues in the Departments of Lead Discovery,

Oncology and Hematology Research, Pathology, Toxicology, Medicinal Chemistry, Pharmacokinetics and

Drug Metabolism, and Pharmaceutics for their contributions and to Julie Bailis, Greg Friberg, Robert

Loberg, and Glenn Begley for their critical reading of this manuscript. We also gratefully acknowledge the

excellent technical assistance by Brian Belmontes, Jacob Corcoran, Tibor Gyuris, Scott Heller,

Raheemuddin Khaja, and Ji-Rong Sun.

Figure 1. AMG 900 is a potent and selective inhibitor of aurora-A, -B and -C. A, chemical structure

and enzyme selectivity profile of AMG 900. B, Western blotting of cell lysates from HeLa cells

synchronized in mitosis and treated with DMSO, AMG 900 at the indicated concentrations, MLN8054

(aurora-A inhibitor) at 500 nmol/L, or AZD1152 (aurora-B inhibitor) at 1000 nmol/L for 3 hours. C,

immunofluorescence imaging assay (ArrayScan VTi, Cellomics) was used to determined the inhibition of

aurora kinase activity in HeLa cells. The concentration-response curves were calculated based on the

decrease in the fluorescence intensity of p-histone H3 Ser10 () and p-aurora-A Thr288 () as a

percentage of the DMSO control (POC). IC50 values represent two independent experiments run in

duplicate (error bars, ±SD). D, representative live-cell imaging analysis of histone H2B-GFP HeLa cells

treated with DMSO or AMG 900 at 50 nmol/L. Cells were imaged over a period of 75 minutes (lower-right

hand corner of each image denotes the time in minutes).

Figure 2. AMG 900 induces polyploidy and apoptosis in vitro in the human colon carcinoma

HCT116 cell line. A, representative images of cells treated with DMSO or AMG 900 at the indicated

concentrations for 6 hours. Cells were stained with p-histone H3 antibody (red) and DAPI (blue). B,





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DNA content profiles of cells treated with DMSO or AMG 900 at 50 nmol/L for 48 hours. Cells were

stained with propidium iodide and evaluated by flow cytometry. C, cells were treated with DMSO or AMG

900 at 50 nmol/L for 48 hours and replated in complete media lacking inhibitor. After 10 days of growth,

cells were stained with crystal violet and imaged (triplicate wells). D, cells were treated with DMSO or

AMG 900 at 50 nmol/L for 48 hours, washed in complete media, and then cultured in complete media

lacking inhibitor for the indicated time intervals. Western blotting documenting the protein levels of

cleaved caspase-7 and β-actin loading control.

Figure 3. AMG 900 inhibits cell proliferation and colony formation in human tumor cell lines at low

nanomolar concentrations, including MDR cell lines resistant to paclitaxel. A and B, fluorescence-

based cell count imaging assay (ArrayScan VTi) was performed on a panel of 26 tumor cell lines treated

with AMG 900 (concentration range 0.3 to 156 nmol/L). Cells were treated for 24 hours, washed twice,

and incubated in complete media lacking inhibitor for 48 hours. A, representative concentration-

response curve of PC3 cells treated with AMG 900. B, cell lines rank ordered by sensitivity to AMG 900

and color coded by tumor origin. C, colony formation assay was performed in three paclitaxel-sensitive

cell lines (parental) and in four MDR cell lines (P-gp expressing). Cells were treated with paclitaxel or

AMG 900 at the indicated concentrations for 48 hours and replated in complete media lacking inhibitor.

After the DMSO-treated cells reached confluence, the cells were stained with crystal violet and imaged

(duplicate wells).

Table 1. AMG 900 exhibits uniform potency in human tumor cell lines expressing P-gp and BCRP

resistant to one or more aurora kinase inhibitors

Figure 4. AMG 900 inhibits phosphorylation of histone H3 and the growth of HCT116 tumor

xenografts in vivo. A and B, mice bearing established tumors were orally administered a single dose of

vehicle alone or AMG 900 at 3.75, 7.5, or 15 mg/kg. Tumors were collected at 3, 6, or 9 hours after

treatment (n = 4 per time point per dose) and the level of p-histone H3 was determined by fluorescence-

based microscopy. Representative images of sectioned tumors stained with p-histone H3 antibody (red)





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and DAPI (blue). B, bar graph representing the pharmacodynamic-pharmacokinetic profile of AMG 900

in mice. The number of p-histone H3 positive cells per tumor image field (8 image fields per column,

mean +SD) and concentration of AMG 900 in mouse plasma (, mean ±SD). The control represents the

combined 3 and 6 hour vehicle-treated tumors (16 image fields per column, mean +SD). Statistically

significant inhibition of p-histone H3 compared with the vehicle-treated control is denoted by an asterisk

(*P ≤ 0.008). C and D, mice bearing established HCT116 tumors were orally administered vehicle alone

() and AMG 900 b.i.d. either intermittently at 3.75 mg/kg (), 7.5 mg/kg (), or 15 mg/kg () for 2

consecutive days per week for 3 weeks (C) or every day at 1.5 mg/kg (), 2.25 mg/kg (), or 3 mg/kg

() every day for 3 weeks (D). Tumor volumes (cubic mm) are represented as mean ±SE (n = 10).

Statistically significant tumor growth inhibition compared with the vehicle-treated control is denoted by an

asterisk (*P ≤ 0.0001).

Figure 5. AMG 900 inhibits the growth of multiple human tumor xenografts in vivo, including

multidrug resistant models. A, mice bearing established tumor xenografts were orally administered

AMG 900 at 15 mg/kg b.i.d. for 2 consecutive days per week for 3 weeks or at 3 mg/kg b.i.d. every day for

3 weeks. The maximum percentage of tumor growth inhibition (TGI) is reported for each xenograft model.

Statistically significant tumor growth inhibition compared with the vehicle-treated control is denoted by

asterisks (*P < 0.005, **P < 0.0005). B and C, mice bearing established multidrug resistant MES-SA-Dx5

or NCI-H460-PTX tumor xenografts were orally administered vehicle alone (), AMG 900 at 15 mg/kg

b.i.d. () for 2 consecutive days per week or at 3 mg/kg b.i.d. () every day. As a control, mice were

administered docetaxel intraperitoneal (i.p.) at 30 mg/kg () once per week or paclitaxel i.p. at 12.5

mg/kg () 5 days per week. Tumor volumes (cubic mm) are represented as mean ±SE (n = 10).

Statistically significant tumor growth inhibition compared with the vehicle-treated control is denoted by an

asterisk (*P ≤ 0.0019).

References

1. Quasthoff S, Hartung HP. Chemotherapy-induced peripheral neuropathy. J Neurol 2002;249:9-17.

2. Keen N, Taylor S. Aurora-kinase inhibitors as anticancer agents. Nat Rev Cancer 2004;4:927-36.





15

3. Keen N, Taylor S. Mitotic drivers - Inhibitors of the aurora B kinase. Cancer Met Rev 2009;28:185-95.

4. Carvajal RD, Tse A, Schwartz GK. Aurora kinases: new targets for cancer therapy. Clin Cancer Res 2006;12: 6869-75.

5. Barr AR, Gergeley, F. Aurora-A: The maker and breaker of spindle poles. J Cell Science 2007;120:2987-96.

6. Macurek L, Lindqvist A, Lim D, et al. Polo-like kinase-1 is activated by aurora A to promote checkpoint recovery. Nature

2008;455:119-23.

7. Ditchfield C, Johnson VL, Tighe A, et al. Aurora B couples chromosome alignment with anaphase by targeting BubR1,

Mad2, and Cenp-E to kinetochores. J Cell Biol 2003;161:267-80.

8. Steigemann P, Wurzenberger C, Schmitz MHA, et al. Aurora B-mediated abscission checkpoint protects against

tetraploidization. Cell 2009;136:473-84.

9. Liu D, Vader G, Vromans MJM, et al. Sensing chromosome bi-orientation by spatial separation of aurora B kinase from

kinetochore substrates. Science 2009;323:1350-53.

10. Kimmins S, Crosio C, Kotaja N, et al. Differential functions of the aurora-B and aurora-C kinases in mammalian

spermatogenesis. Mol Endocrinol 2007;21:726-39.

11. Gautschi O, Heighway J, Mack PC, Purnell PR, Lara PN, Gandara DR. Aurora kinases as anticancer drug targets. Clin

Cancer Res 2008;14:1639-48.

12. Musacchio A, Salmon ED. The spindle-assembly checkpoint in space in time. Nat Rev Mol Cell Biol 2007;8: 379-93.

13. Hauf S, Cole RW, LaTerra S, et al. The small molecule hesperadin reveals a role for aurora B in correcting kinetochore–

microtubule attachment and in maintaining the spindle assembly checkpoint. J Cell Biol 2003;161: 281-94.

14. Yang H, Burke T, Dempsey J, et al. Mitotic requirement for aurora A kinase is bypassed in the absence of aurora B

kinase. FEBS Letters 2005;579:3385-91.

15. Girdler F, Gascoigne KE, Eyers PA, et al. Validating aurora B as an anti-cancer drug target. J Cell Sci 2006; 119:3664-

75.

16. Weaver BA, Cleveland DW. Decoding the links between mitosis, cancer, and chemotherapy: The mitotic checkpoint,

adaptation, and cell death. Cancer Cell 2005;8:7-12.

17. Gascoigne KE, Taylor SS. Cancer cells display profound intra- and interline variation following prolonged exposure to

antimitotic drugs. Cancer Cell 2008;14:111-22.

18. Perez EA. Microtubule inhibitors: Differentiating tubulin-inhibiting agents based on mechanism of action, clinical activity,

and resistance. Mol Cancer Ther 2009;8:2086-95.

19. Yin S, Bhattacharya R, Cabral F. Human mutations that confer paclitaxel resistance. Mol Cancer Ther 2010; 9:327-35.

20. McGrogan BT, Gilmartin B, Carney DN, McCann A. Taxanes, microtubules, and chemoresistant breast cancer. Biochim et

Biophys Acta 2008;1785:96-132.

21. Szakács G, Paterson JK, Ludwig JA, Booth-Genthe C, Gottesman MM. Targeting multidrug resistance in cancer. Nat Rev

Drug Disc 2006 5:219-34.





16

22. Guo J, Anderson MG, Tapang P, et al. Identification of genes that confer tumor cell resistance to the aurora B kinase

inhibitor, AZD1152. Pharmacogenomics J 2009;9:90-102.

23. Carter TA, Wodicka LM, Shah NP, et al. Inhibition of drug-resistant mutants of ABL, KIT, and EGF receptor kinases.

PNAS 2005;102:11011-16.

24. Yun C-H, Boggon TJ, Li Y, et al. Structures of lung cancer-derived EGFR mutants and inhibitor complexes: mechanism of

activation and insights into differential inhibitor sensitivity. Cancer Cell 2007;11:217–27.

25. Lapenna S, Giordano A. Cell cycle kinases as therapeutic targets for cancer. Nat Rev Drug Disc 2009;8:547-66.

26. Crosio C, Fimia GM, Loury R, et al. Mitotic phosphorylation of histone H3: spatio-temporal regulation by mammalian

aurora kinases. Mol Cell Bio 2002;22:874-85.

27. Payton M, Chung G, Yakowec P, et al. Discovery and evaluation of dual CDK1 and CDK2 inhibitors. Cancer Res

2006;66:4299-4308.

28. Karaman MW, Herrgard S, Treiber DK, et al. A quantitative analysis of kinase inhibitor selectivity. Nature Biotech

2008;26:127-32.

29. Gizatullin F, Yao Y, Kung V, Harding MW, Loda M, Shapiro GI. The Aurora kinase inhibitor VX-680 induces

endoreduplication and apoptosis preferentially in cells with compromised p53-dependent post-mitotic checkpoint function.

Cancer Res 2006;66:7668-77.

30. Nair JS, Ho AL, Tse AN, et al. Aurora B kinase regulates the postmitotic endoreduplication checkpoint via

phosphorylation of the retinoblastoma protein at serine 780. Mol Biol Cell 2009;20:2218-28.

31. Vakifahmetoglu H, Olsson M, Zhivotovsky B. Death through a tragedy: mitotic catastrophe. Cell Death and Diff

2008;15:1153-62.

32. Harker WG, Sikic BI. Multidrug (pleiotropic) resistance in doxorubicin-selected variants of the human sarcoma cell line

MES-SA. Cancer Res 1985;45:4091-96.

33. Szakacs G, Annereau J-P, Lababidi S, et al. Drug sensitivity and resistance: Profiling ABC transporter genes in cancer

cells. Cancer Cell 2004;6:129-37.

34. Gyorffy B, Surowiak P, Kiesslich O, et al. Gene expression profiling of 30 cancer cell lines predicts resistance towards 11

anticancer drugs at clinically achieved concentrations. Int J Cancer 2006;118:1699-1712.

35. Girdler F, Sessa F, Patercoli S, et al. Molecular basis of drug resistance in aurora kinases. Chemistry & Biology

2008;15:552-62.

36. Davies H, Bignell GR, Cox C, et al. Mutations of the BRAF gene in human cancer. Nature 2002;417:949-54.

37. Paez JG, Janne PA, Lee JC, et al. EGRF mutations in lung cancer: Correlation with clinical response to Gefitinib therapy.

Science 2004;304:1497-1500.

38. Jackson JR, Patrick DR, Dar MM, Huang PS. Targeted anti-mitotic therapies: Can we improve on tubulin agents? Nat Rev

Cancer 2007;7:107-17.





17

39. Luo J, Solimini NL, Elledge SJ. Principles of cancer therapy: oncogene and non-oncogene addication. Cell 2009;136:823-

37.

40. Freeman D, Bush T, Ogbagabriel S, et al. Activity of panitumumab alone or with chemotherapy in non-small cell lung

cancinoma cell lines expressing mutant epidermal growth factor receptor. Mol Cancer Ther 2009;9:1536-46.




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Published OnlineFirst October 8, 2010.Cancer Res Marc N. Payton, Tammy L. Bush, Grace Chung, et al. Taxane-Resistant Tumor Cell LinesSelective Pan-Aurora Kinase Inhibitor with Activity in Preclinical Evaluation of AMG 900, a Novel Potent and Highly

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