Palbocicb Phase 2 Clinical Trial

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Articles The cyclin-dependent kinase 4/6 inhibitor palbociclib in combination with letrozole versus letrozole alone as first-line treatment of oestrogen receptor-positive, HER2-negative, advanced breast cancer (PALOMA-1/TRIO-18): a randomised phase 2 study Richard S Finn, John P Crown, Istvan Lang, Katalin Boer, Igor M Bondarenko, Sergey O Kulyk, Johannes Ettl, Ravindranath Patel, Tamas Pinter, Marcus Schmidt, Yaroslav Shparyk, Anu R Thummala, Nataliya L Voytko, Camilla Fowst, Xin Huang, Sindy T Kim, Sophia Randolph, Dennis J Slamon Summary Background Palbociclib (PD-0332991) is an oral, small-molecule inhibitor of cyclin-dependent kinases (CDKs) 4 and 6 with preclinical evidence of growth-inhibitory activity in oestrogen receptor-positive breast cancer cells and synergy with anti-oestrogens. We aimed to assess the safety and efficacy of palbociclib in combination with letrozole as firstline treatment of patients with advanced, oestrogen receptor-positive, HER2-negative breast cancer. Lancet Oncol 2015; 16: 25–35 Methods In this open-label, randomised phase 2 study, postmenopausal women with advanced oestrogen receptorpositive and HER2-negative breast cancer who had not received any systemic treatment for their advanced disease were eligible to participate. Patients were enrolled in two separate cohorts that accrued sequentially: in cohort 1, patients were enrolled on the basis of their oestrogen receptor-positive and HER2-negative biomarker status alone, whereas in cohort 2 they were also required to have cancers with amplification of cyclin D1 (CCND1), loss of p16 (INK4A or CDKN2A), or both. In both cohorts, patients were randomly assigned 1:1 via an interactive web-based randomisation system, stratified by disease site and disease-free interval, to receive continuous oral letrozole 2·5 mg daily or continuous oral letrozole 2·5 mg daily plus oral palbociclib 125 mg, given once daily for 3 weeks followed by 1 week off over 28-day cycles. The primary endpoint was investigator-assessed progressionfree survival in the intention-to-treat population. Accrual to cohort 2 was stopped after an unplanned interim analysis of cohort 1 and the statistical analysis plan for the primary endpoint was amended to a combined analysis of cohorts 1 and 2 (instead of cohort 2 alone). The study is ongoing but closed to accrual; these are the results of the final analysis of progression-free survival. The study is registered with the ClinicalTrials.gov, number NCT00721409. See Comment page 2 Findings Between Dec 22, 2009, and May 12, 2012, we randomly assigned 165 patients, 84 to palbociclib plus letrozole and 81 to letrozole alone. At the time of the final analysis for progression-free survival (median follow-up 29·6 months [95% CI 27·9–36·0] for the palbociclib plus letrozole group and 27·9 months [25·5–31·1] for the letrozole group), 41 progression-free survival events had occurred in the palbociclib plus letrozole group and 59 in the letrozole group. Median progression-free survival was 10·2 months (95% CI 5·7–12·6) for the letrozole group and 20·2 months (13·8–27·5) for the palbociclib plus letrozole group (HR 0·488, 95% CI 0·319–0·748; one-sided p=0·0004). In cohort 1 (n=66), median progression-free survival was 5·7 months (2·6–10·5) for the letrozole group and 26·1 months (11·2–not estimable) for the palbociclib plus letrozole group (HR 0·299, 0·156–0·572; one-sided p<0·0001); in cohort 2 (n=99), median progression-free survival was 11·1 months (7·1–16·4) for the letrozole group and 18·1 months (13·1–27·5) for the palbociclib plus letrozole group (HR 0·508, 0·303–0·853; one-sided p=0·0046). Grade 3–4 neutropenia was reported in 45 (54%) of 83 patients in the palbociclib plus letrozole group versus one (1%) of 77 patients in the letrozole group, leucopenia in 16 (19%) versus none, and fatigue in four (4%) versus one (1%). Serious adverse events that occurred in more than one patient in the palbociclib plus letrozole group were pulmonary embolism (three [4%] patients), back pain (two [2%]), and diarrhoea (two [2%]). No cases of febrile neutropenia or neutropenia-related infections were reported during the study. 11 (13%) patients in the palbociclib plus letrozole group and two (2%) in the letrozole group discontinued the study because of adverse events. Interpretation The addition of palbociclib to letrozole in this phase 2 study significantly improved progression-free survival in women with advanced oestrogen receptor-positive and HER2-negative breast cancer. A phase 3 trial is currently underway. Funding Pfizer. www.thelancet.com/oncology Vol 16 January 2015 Published Online December 16, 2014 http://dx.doi.org/10.1016/ S1470-2045(14)71159-3 See Online for podcast interview with Dennis Slamon University of California Los Angeles, Los Angeles, CA, USA (R S Finn MD, Prof D J Slamon MD); Irish Cooperative Oncology Research Group, Dublin, Ireland (Prof J P Crown MD); Orszagos Onkologiai Intezet, Budapest, Hungary (I Lang MD); Szent Margit Korhaz, Onkologia, Budapest, Hungary (K Boer MD); Dnipropetrovsk Medical Academy, City Multiple-Discipline Clinical Hospital 4, Dnipropetrovsk, Ukraine (Prof I M Bondarenko MD); Municipal Treatment and Prophylactic Institution, Donetsk, Ukraine (S O Kulyk MD); Technical University of Munich, Munich, Germany (J Ettl MD); Comprehensive Blood and Cancer Center, Bakersfield, CA, USA (R Patel MD); Petz Aladar Megyei Oktato Korhaz, Gyor, Hungary (T Pinter MD); University Hospital Mainz, Mainz, Germany (M Schmidt MD); Lviv State Oncologic Regional Treatment and Diagnostic Centre, Lviv, Ukraine (Y Shparyk MD); Comprehensive Cancer Centers of Nevada, Las Vegas, NV, USA (A R Thummala MD); Kyiv City Clinical Oncology Centre, Kyiv, Ukraine (N L Voytko MD); Pfizer Oncology, Milan, Italy (C Fowst MD); and Pfizer Oncology, San Diego, CA, USA (X Huang PhD, S T Kim BS, S Randolph MD) 25 Articles Correspondence to: Prof Dennis J Slamon, UCLA Oncology, Santa Monica, CA 90404, USA [email protected] See Online for appendix 26 Introduction Breast cancer is a molecularly diverse disease with several defined molecular subgroups. Clinically, however, three therapeutic groups are used: those classified as hormone receptor-positive (ie, oestrogen receptorpositive, progesterone receptor-positive, or both, with normal HER2 expression), those classified as HER2positive, as defined by HER2 gene amplification or overexpression (about 45% of these cancers can also have variable expression of oestrogen receptors, progesterone receptors, or both), and those classified as triple-negative by virtue of low or absent hormone receptors and the absence of the HER2 alteration.1 More than 1·5 million new breast cancers are reported worldwide each year, with roughly 60–65% of cases hormone receptor-positive, 20–25% HER2-positive, and 15–18% triple-negative.2 Hormonally directed drugs including anti-oestrogens have been the mainstay of treatment for women with oestrogen receptor-positive breast cancers. However, some of these cancers have de-novo resistance to this approach and even more will eventually develop acquired resistance to these treatments and recur. At that point, patients often receive chemotherapy that has little activity in this setting and that is often associated with clinically significant toxic effects. New classes of molecularly targeted drugs can affect the natural history of some subgroups of breast cancer such as HER2-positive disease.3 However, until recently the addition of such drugs to anti-oestrogens has not resulted in similar improvements in hormone receptorpositive disease. This situation changed with the approval of everolimus for aromatase inhibitor-resistant disease, which, when added to oestrogen blockade with exemestane, resulted in an improvement in investigatorassessed progression-free survival (hazard ratio [HR] 0·43, 95% CI 0·35–0·54; p<0·001) in oestrogen receptorpositive advanced disease.4 Dysregulation of the cell cycle is one of the defined hallmarks of cancer5 and several genetic alterations in key cell cycle regulatory proteins have been described in various cancers, including breast cancer.5,6 The cyclindependent kinases (CDKs) are a large family of serine threonine kinases that together with their regulatory protein partners, the cyclins, have a crucial role in the orderly and controlled progression through the cell cycle. Molecular changes in several of the genes controlling the cell cycle have been reported in various cancers, making them an attractive potential target for new treatments.7 So far, several CDK-targeted drugs have entered clinical development, but none have shown significant activity in solid tumours and several are associated with safety concerns.8 Palbociclib (PD-0332991) is a reversible, oral, smallmolecule inhibitor of cyclin dependent kinases 4 and 6 (CDK4/6).9 CDK4/6 and cyclin D have a crucial role in the regulation of the G1/S transition through regulation of the phosphorylation state of pRb. When hyperphosphorylation of pRb occurs, it causes release of transcription factors that then allow the transition from G1 to S phase and progression of the cell cycle.10 To investigate the therapeutic potential for palbociclib in breast cancer, we tested its growth inhibitory effects preclinically in a large panel of human breast cancer cell lines and identified potent activity in two therapeutic groups, those that were oestrogen receptor-positive and those that were HER2amplified.11 This activity was associated with a major blockade of pRb hyperphosphorylation, resulting in a G1 arrest in sensitive cells. We also noted that in combination with the anti-oestrogen drug tamoxifen, palbociclib had synergistic growth inhibitory activity as well as efficacy in a model of acquired tamoxifen resistance.11 We noted similar findings in HER2-amplified breast cancer cell lines with trastuzumab used in combination with palbociclib.11 Based on these data, we designed a clinical study to test the safety and efficacy of CDK4/6 inhibition by palbociclib in combination with anti-oestrogen drugs in oestrogen receptor-positive breast cancer. Initially, a single-arm, phase 1b study was done to assess the safety of palbociclib given with letrozole in patients with oestrogen receptorpositive, HER2-negative, advanced breast cancer and to determine a recommended phase 2 dose of the combination.12 The results suggested a dose and schedule consisting of oral palbociclib 125 mg once daily for 3 weeks followed by 1 week off treatment in a 28-day cycle, combined with the standard dose of oral letrozole 2·5 mg once daily. No drug–drug interactions were identified and the most common treatment-related adverse events were neutropenia, leucopenia, and fatigue. Based on these clinical data, we planned a randomised, open-label, phase 2 study to assess the safety and efficacy of the palbociclib and letrozole combination compared with letrozole alone in the firstline treatment of women with advanced oestrogen receptor-positive, HER2-negative breast cancer. Methods Study design and patients In this international, phase 2, multicentre, open-label randomised study (PALOMA-1/TRIO-18), postmenopausal women (aged 18 years or older) with oestrogen receptor-positive, HER2-negative, advanced breast cancer were recruited from 50 sites in 12 countries (Canada, France, Germany, Hungary, Ireland, Italy Russia, South Africa, South Korea, Spain, Ukraine, USA; appendix). Patients were enrolled in two separate cohorts that accrued sequentially: in cohort 1, patients were enrolled on the basis of their oestrogen receptor-positive and HER2-negative biomarker status alone, whereas in cohort 2 they were also required to have cancers with amplification of cyclin D1 (CCND1), loss of p16 (also known as INK4A or CDKN2A), or both. All patients were required to have either locally recurrent disease not amenable to surgery or evidence of metastatic disease. www.thelancet.com/oncology Vol 16 January 2015 Articles 400 patients assessed for eligibility (cohorts 1 and 2) Cohort 1 81 screened 15 excluded 11 did not meet inclusion criteria 4 met exclusion criteria Cohort 2 319 screened 220 excluded 203 did not meet inclusion criteria 13 met exclusion criteria 4 did not meet inclusion criteria and met exclusion criteria 66 randomly assigned 99 randomly assigned 34 allocated to palbociclib plus letrozole 33 received allocated intervention 1 did not receive allocated intervention 1 withdrew consent 32 allocated to letrozole 29 received allocated intervention 3 did not receive allocated intervention 3 withdrew consent 50 allocated to palbociclib plus letrozole 50 received allocated intervention 49 allocated to letrozole 48 received allocated intervention 1 did not receive allocated intervention 1 withdrew consent 26 discontinued intervention 8 because of adverse events 16 had objective progression or relapse 2 withdrew consent 28 discontinued intervention 1 because of adverse events 1 because of global deterioration of health status 22 had objective progression or relapse 2 withdrew consent 2 other reasons 38 discontinued intervention 3 because of adverse events 5 because of global deterioration of health status 26 had objective progression or relapse 1 died 3 withdrew consent 41 discontinued intervention 1 because of adverse events 2 because of global deterioration of health status 35 had objective progression or relapse 3 withdrew consent 34 analysed by intention to treat 32 analysed by intention to treat 50 analysed by intention to treat 49 analysed by intention to treat 165 in the total intention-to-treat population 84 in the palbociclib plus letrozole group 81 in the letrozole group Figure 1: Trial profile Oestrogen receptor status was determined by routine immunohistochemistry and HER2 status was assessed by either fluorescent in-situ hybridisation (FISH) or immunohistochemistry, with both eligibility markers reported at enrolling sites. Oestrogen receptor status was determined on the basis of either the original tissue staining or, if available, a biopsy from the recurrence. For enrolment into cohort 2 of the study, central laboratory determination of CCND1 amplification or p16 loss was required. CCND1 amplification was defined as a CCND1-to-chromosome enumeration probe (CEP) 11 ratio greater than 1·5 and p16 loss as a p16-to-CEP9 ratio less than 0·8. These cutoffs came from an internal analysis of British Columbia Cancer Foundation data from 778 breast cancer cases that showed the frequency of CCND1 amplification (more than three copies) and p16 loss (loss of heterozygosity) to be roughly 36% in patients with luminal B breast cancer. To establish definitions for the genomic changes used for cohort 2, we used a four-colour FISH assay (CCND1-to-CEP11 and p16-to-CEP9) to analyse 113 breast cancer samples from a tumour bank and, using a cutoff of CCND1-towww.thelancet.com/oncology Vol 16 January 2015 CEP11 greater than 1·5 or p16-to-CEP9 less than 0·8, we determined that 42 (37%) of 113 patients in this cohort had CCND1 amplification, CDKN2A loss, or both. No previous treatment for advanced disease was permitted and all patients were required to have measurable disease by Response Evaluation Criteria in Solid Tumors (RECIST, version 1.0) or bone-only disease with an Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1 and adequate organ function (as assessed by haematological and blood chemistry analyses). Patients were excluded from study if they had received letrozole as either neoadjuvant or adjuvant treatment within the 12 months before study entry, had received any previous treatment for advanced breast cancer, had brain metastasis, or had previously been treated with a CDK inhibitor. The study was done in accordance with the International Conference on Harmonization and Good Clinical Practice standards. Institutional review board approval was obtained from all participating institutions and patients provided written informed consent before the start of any study-specific screening procedures. 27 Articles Median age (years) Both cohorts Cohort 1 Palbociclib plus Letrozole letrozole (n=84) (n=81) Palbociclib plus letrozole (n=34) Letrozole (n=32) Cohort 2 Palbociclib plus letrozole (n=50) Letrozole (n=49) 63 (54–71) 64 (56–70) 66 (56–72) 64 (57–70) 62 (54–70) 63 (56–71) ECOG performance status 0 46 (55%) 45 (56%) 23 (68%) 20 (63%) 23 (46%) 25 (51%) 1 38 (45%) 36 (44%) 11 (32%) 12 (38%) 27 (54%) 24 (49%) Disease stage III 2 (2%) 1 (1%) 2 (6%) IV 82 (98%) 80 (99%) 32 (94%) 32 (100%) 0 Visceral 37 (44%) 43 (53%) 10 (29%) Bone only 17 (20%) 12 (15%) 7 (21%) Other (non-visceral) 30 (36%) 26 (32%) >12 months from adjuvant treatment to recurrence 25 (30%) ≤12 months from adjuvant treatment to recurrence or de-novo advanced disease 59 (70%) 0 1 (2%) 50 (100%) 48 (98%) 11 (34%) 27 (54%) 32 (65%) 6 (19%) 10 (20%) 6 (12%) 17 (50%) 15 (47%) 13 (26%) 11 (23%) 30 (37%) 10 (29%) 10 (31%) 15 (30%) 20 (41%) 51 (63%) 24 (71%) 22 (69%) 35 (70%) 29 (59%) 44 (52%) 37 (46%) 19 (56%) 17 (53%) 25 (50%) 20 (41%) None 44 (52%) 37 (46%) 19 (56%) 17 (53%) 25 (50%) 20 (41%) Chemotherapy 34 (40%) 37 (46%) 11 (32%) 14 (44%) 23 (46%) 23 (47%) Hormonal 27 (32%) 28 (35%) 11 (32%) 11 (34%) 16 (32%) 17 (35%) Tamoxifen 24 (29%) 24 (30%) 8 (24%) 8 (25%) 16 (32%) 16 (33%) Anastrozole 8 (10%) 11 (14%) 4 (12%) 5 (16%) 4 (8%) 6 (12%) Letrozole 2 (2%) 1 (1%) 0 0 2 (4%) 1 (2%) Exemestane 4 (5%) 2 (2%) 3 (9%) 1 (3%) 1 (2%) 1 (2%) Disease site* Disease-free interval* De-novo advanced disease only Previous systemic treatment Data are n (%) or median (IQR). ECOG=Eastern Cooperative Oncology Group. *Based on case report form data. Table 1: Baseline characteristics (intention-to-treat population) Randomisation and masking Patients were randomly allocated 1:1 to receive either palbociclib plus letrozole or letrozole alone. The investigator or other member of the research staff used an interactive web-based randomisation system to register and randomly assign patients with two stratifications factors: disease site (visceral, bone only, or other) and disease-free interval (greater than 12 months from the end of adjuvant treatment to recurrence vs 12 months or less from the end of adjuvant treatment to recurrence or de-novo metastatic disease). The randomisation system generated the random assignment of the two treatments in a block size of six for each of the stratification levels. Although this was an open-label study, the randomisation codes were only released at the time of interim and final analyses and crossover was not allowed at any time. Procedures Patients randomly allocated to letrozole received oral letrozole 2·5 mg once daily. Those allocated to palbociclib plus letrozole received the same dose of letrozole plus oral palbociclib 125 mg, given once daily for 3 weeks followed by 1 week off in 28-day cycles. Study treatment continued until disease progression, unacceptable toxic 28 effects, study withdrawal, or death. Dose interruptions and reductions were allowed for management of toxic effects (appendix). Tumour assessments were done locally at screening and every 8 weeks and consisted of CT or MRI scan of the chest, abdomen, and pelvis; radiography for bone lesions (when applicable); and clinical assessment of cutaneous disease. Bone scans were done at baseline and every 12 weeks. All patients with tumour responses were required to have response confirmation no less than 4 weeks after documentation of the initial response report. Assessment of adverse events included incidence, severity (graded by the National Cancer Institute Common Terminology Criteria for Adverse Events version 3.0), timing, seriousness, and relatedness to study drug. Haematological and blood chemistry analyses were done every 2 weeks for the first two treatment cycles and at the beginning of each cycle thereafter. Outcomes The primary endpoint was investigator-assessed progression-free survival, defined as the time from randomisation to radiological disease progression or death on study. Secondary efficacy endpoints were www.thelancet.com/oncology Vol 16 January 2015 Articles Both cohorts 90 80 Statistical analysis Figure 2: Progression-free survival (intention-to-treat population) HR=hazard ratio. www.thelancet.com/oncology Vol 16 January 2015 70 60 50 40 30 20 10 HR 0·488 (95% CI 0·319–0·748; one-sided p=0·0004) 0 0 Number at risk Palbociclib plus letrozole 84 Letrozole 81 4 8 12 16 20 24 28 32 36 40 67 48 60 36 47 28 36 19 28 14 21 6 13 3 8 3 5 1 1 Cohort 1 100 90 Progression-free survival (%) 80 70 60 50 40 30 20 10 HR 0·299 (95% CI 0·156–0·572; one-sided p<0·0001) 0 0 Number at risk Palbociclib plus letrozole 34 Letrozole 32 4 8 12 16 20 24 28 32 36 40 26 15 23 10 18 8 15 5 13 4 11 4 8 3 8 3 5 1 1 28 32 36 40 Cohort 2 100 90 80 Progression-free survival (%) We used a two-part study design (ie, two sequential cohorts) to allow us to assess both the activity of the palbociclib plus letrozole combination and to determine whether further patient selection on the basis of additional biomarkers (CCND1 or p16) was warranted. We planned to enrol 60 patients (30 per treatment group) into cohort 1 to provide initial safety and efficacy (progression-free survival) data in patients with oestrogen receptor-positive, HER2-negative, advanced breast cancer. In cohort 2, we planned to include 150 patients (75 per treatment group) who also had CCND1 gene amplification or loss of p16. Cohort 1 was intended to be an exploratory analysis, and the analysis of the primary endpoint was initially intended to be based on cohort 2 only. Assuming 114 progressionfree survival events in cohort 2 and using a one-sided α of 0·10, a sample size of 150 would have 80% power to detect an HR of 0·67 (palbociclib plus letrozole vs letrozole alone), including one futility interim analysis. This HR would represent a median progression-free survival of 9 months in the control group and 13·5 months in the experimental group.13 However, an unplanned interim analysis of cohort 1 based on 31 progression-free survival events was done when we noted that almost twice as many patients in the control group were coming off the study because of disease progression. This interim analysis showed clinically meaningful activity of the palbociclib plus letrozole combination compared with letrozole alone (HR 0·35, 95% CI 0·170·72, p=0·006). These preliminary results from cohort 1 also suggested that further patient selection based on CCND1 amplification or p16 loss was unlikely to further improve patient outcome over the use of oestrogen receptor and HER2 status alone (HR with CCND1 or p16 copy changes 0·37 [95% CI 0·10–1·40; p=0·13] vs HR with no CCND1 or p16 copy changes 0·19 [0·05–0·67; p=0·0045]). As a result, we stopped further enrolment into cohort 2 and amended the statistical analysis plan such that the primary endpoint would be analysed in cohorts 1 and 2 combined instead of cohort 2 alone. These study changes were made prospectively without any efficacy results from cohort 2 and were Palbociclib plus letrozole Letrozole 100 Progression-free survival (%) objective response (by RECIST version 1.0), clinical benefit (as defined by the sum of complete plus partial responses and stable disease for 24 weeks or more), duration of response, and overall survival. Additional secondary endpoints were safety and tissue and serum biomarker analyses. Finally, we also assessed patientreported outcomes using the modified Brief Pain Inventory (Short Form; mBPI-sf) done on day 1 of each treatment cycle; the mBPI-sf was used to capture whether palbociclib adds to the commonly reported adverse event seen with aromatase inhibitors (myalgias and joint pain). 70 60 50 40 30 20 10 HR 0·508 (95% CI 0·303–0·853; one-sided p=0·0046) 0 0 Number at risk Palbociclib plus letrozole 50 Letrozole 49 4 8 12 16 20 24 Time (months) 41 33 37 26 29 20 21 14 15 10 10 2 5 29 Articles Hazard ratio (95% CI) Palbociclib plus letrozole Letrozole Patients Events Patients Events All patients (intention-to-treat population) 84 41 81 59 0·488 (0·319–0·748) Interaction p value* Cohort 1 34 15 32 25 0·299 (0·156–0·572) 2 50 26 49 34 0·508 (0·303–0·853) <65 years 47 24 42 35 0·315 (0·184–0·539) ≥65 years 37 17 39 24 0·505 (0·269–0·948) 0·14 Age group (years) 0·34 Baseline ECOG performance status 0 46 21 45 31 0·434 (0·246–0·766) 1 38 20 36 28 0·398 (0·220–0·721) Visceral 37 21 43 34 0·547 (0·317–0·944) Bone Only 17 5 12 7 0·294 (0·092–0·945) Other 30 15 26 18 0·402 (0·200–0·808) 0·78 Disease site 0·44 Previous chemotherapy Yes 34 17 37 24 0·479 (0·255–0·898) No 50 24 44 35 0·397 (0·234–0·671) 0·75 Previous antihormonal therapy Yes 27 12 28 19 0·460 (0·222–0·956) No 57 29 53 40 0·397 (0·244–0·646) Yes 40 20 44 28 0·539 (0·302–0·962) No 44 21 37 31 0·341 (0·194–0·599) 0·88 Previous systemic therapy 0·36 Time from end of adjuvant treatment to disease recurrence ≤12 months (including de-novo presentation) 59 31 51 39 0·418 (0·259–0·674) >12 months 25 10 30 20 0·399 (0·185–0·858) ≤12 months (excluding de-novo presentation) 15 7 14 5 0·765 (0·232–2·523) 0·062 0·125 0·250 0·500 Favours palociclib plus letrozole 1·000 2·000 0·95 0·34 4·000 Favours letrozole Figure 3: Subgroup analysis for progression-free survival ECOG=Eastern Cooperative Oncology Group. *Two-sided p value. overseen and approved by the study steering committee. At the time enrolment was stopped, 165 patients had been randomly assigned (66 patients in cohort 1 and 99 patients in cohort 2). Based on the same original assumption that palbociclib plus letrozole would increase progression-free survival from 9 months to 13·5 months compared with letrozole alone, this sample size would have 80% power to detect an HR of 0·67 based on 114 progression-free survival events in the final analysis. A second interim analysis was added with these protocol amendments, which was to be done when about half of the expected number of progression-free survival events across both cohorts (ie, about 57 of 114 total events) had occurred. At the time of the second interim analysis, 61 events had occurred and the HR for progression-free survival for the entire intention-to-treat population was 0·37 (95% CI 0·21–0·63; one-sided p<0·0001). After these analyses were done, we noted a substantial fall in the event rate over time and therefore made another adjustment to the final analysis plan such that the final analysis of progression-free survival would 30 be done when 95 progression-free survival events had accumulated, giving greater than 98% power to detect an HR of 0·50 at a one-sided α of 0·10, or 75% power to detect an HR of 0·67. We adjusted the significance level for the final analysis for the interim analyses using the Lan-DeMets procedure with an O’Brien-Fleming stopping boundary. At the final analysis of progression-free survival, we used a gatekeeping procedure for hypotheses testing in a hierarchical approach to further control for family-wise errors. This process began with assessment of all randomly assigned patients (in cohorts 1 and 2 combined). If the null hypothesis was rejected, then the Holm procedure would be used to test the same hypotheses for cohorts 1 and 2 as two separate studies. Using this approach, we compared progression-free survival data between the treatment groups using a stratified log-rank test with stratification for site of disease, disease-free interval, and study cohort. We estimated the HR using the Cox proportional hazards regression model; the proportionality of hazards assumption was verified14 and the results were satisfactory. www.thelancet.com/oncology Vol 16 January 2015 Articles To explore the effect of prespecified baseline prognostic factors on progression-free survival, we did a multivariate analysis using the Cox regression model. The primary and secondary efficacy analyses were done in the intention-to-treat population. The safety analyses were done for all randomly assigned patients who received at least one dose of the study treatment. We only controlled the type I error for the analysis of primary endpoint, not for any of the secondary endpoints. We did seven prespecified sensitivity analyses for progression-free survival (unstratified analysis; analysis stratified per case report form data; including symptomatic deterioration as disease progression; including disease progression or death after 28 days of treatment discontinuation as disease progression; forced progression-free survival times to the planned assessment times; as-treated population analysis; and multivariate analysis). All statistical analyses were done with the SAS version 9.2 or later. The study is registered with the ClinicalTrials.gov, number NCT00721409. Role of the funding source The funder provided funding to the investigators for study design, conduct, treatment administration, and data collection. The study database was held by the funder. The steering committee that oversaw the conduct of the study consisted of the principal investigator (RSF), senior author (DJS), an independent statistician, two additional investigators, and three representatives from the funder (one clinician, one operations representative, and a statistician). The study steering committee was involved and in all discussions about study conduct. All authors had unrestricted access to the raw and final study data, and Palbociclib plus letrozole were responsible for data interpretation, preparation of the report, and the decision to submit for publication. The authors attest to study completeness and the accuracy of the data and data analysis. Results Between Dec 22, 2009, and May 12, 2012, 165 women were randomly assigned, 84 to receive letrozole plus palbociclib and 81 to receive letrozole alone (figure 1). Baseline demographic characteristics and established prognostic factors of the intention-to-treat population were generally well balanced, although there were slight imbalances in disease site, disease-free interval, and previous treatment (table 1). About half of the patients in each group had never received either adjuvant or neoadjuvant systemic treatment. Conversely, a third of patients in each group had received previous antihormonal treatment, with half of these individuals having previously received aromatase inhibitors. As of the data cutoff for the final analysis (Nov 29, 2013), median follow-up was 29·6 months (95% CI 27·9–36·0) for the palbociclib plus letrozole group and 27·9 months (25·5–31·1) for the letrozole group, with 19 (23%) of 84 patients in the palbociclib plus letrozole group and eight (10%) of 81 in the letrozole group remaining on treatment. At the time of the final analysis for progression-free survival, 41 progression-free survival events had occurred in the palbociclib plus letrozole group and 59 in the letrozole group. Median progressionfree survival was 20·2 months (95% CI 13·8–27·5) for the palbociclib plus letrozole group and 10·2 months (5·7–12·6) for the letrozole alone group (HR 0·488, 95% CI 0·319–0·748; one-sided p=0·0004; figure 2). For patients in cohort 1, median progression-free survival was 26·1 months (95% CI 11·2–not estimable [NE]) for the combination and 5·7 months (95% CI 2·6–10·5) for Letrozole Palbociclib plus letrozole Letrozole 100 Intention-to-treat population* 1 (1%) 1 (1%) Partial response 35 (42%) 26 (32%) Stable disease 37 (44%) 30 (37%) Stable disease ≥24 weeks 32 (38%) 20 (25%) Stable disease <24 weeks 5 (6%) 10 (12%) 3 (4%) 18 (22%) Progressive disease Indeterminate 8 (10%) 6 (7%) 1 (2%) 0 Patients with measurable disease† Complete response 80 70 Overall survival (%) Complete response 90 60 50 40 30 20 Partial response 35 (54%) 26 (39%) Stable disease 20 (31%) 22 (33%) 10 Progressive disease 2 (3%) 15 (23%) 0 Indeterminate 7 (11%) 3 (5%) Data are n (%). *n=84 in the palbociclib plus letrozole group, n=81 in the letrozole alone group. †n=65 in the palbociclib plus letrozole group, n=66 in the letrozole alone group. Table 2: Best overall response www.thelancet.com/oncology Vol 16 January 2015 HR 0·813 (95% CI 0·492–1·345; two-sided p=0·42) 0 Number at risk Palbociclib plus letrozole 84 Letrozole 81 4 8 12 16 80 76 78 74 73 67 68 64 20 24 Time (months) 65 59 47 37 28 32 36 40 44 35 23 22 14 17 12 7 5 2 1 Figure 4: Overall survival (intention-to-treat population) HR=hazard ratio. 31 Articles letrozole alone (HR 0·299, 95% CI 0·156–0·572; onesided p<0·0001; figure 2). For patients in cohort 2, median progression-free survival was 18·1 months (95% CI 13·1–27·5) for the combination and 11·1 months Palbociclib plus letrozole (n=83) Letrozole (n=77) Grade 1–2 Grade 3 Grade 4 Grade 1–2 Grade 3 19 (23%) 49 (59%) 14 (17%) 49 (64%) 16 (21%) 0 Neutropenia 17 (20%) 40 (48%) 5 (6%) 3 (4%) 1 (1%) Leucopenia 20 (24%) 16 (19%) 0 2 (3%) 0 0 Fatigue 30 (36%) 2 (2%) 1 (1%) 0 Anaemia 24 (29%) 4 (5%) 1 (1%) 4 (5%) 1 (1%) 0 Nausea 19 (23%) 2 (2%) 0 9 (12%) 1 (1%) 0 Arthralgia 18 (22%) 2 (3%) 0 Alopecia 18 (22%) Diarrhoea 14 (17%) 3 (4%) Hot flush 17 (21%) 0 Thrombocytopenia 12 (14%) 2 (2%) Decreased appetite 12 (14%) Dyspnoea 11 (13%) Nasopharyngitis 13 (16%) 0 0 Back pain 11 (13%) 0 1 (1%) Headache 12 (14%) 0 0 8 (10%) 0 0 Vomiting 12 (14%) 0 0 2 (3%) 1 (1%) 0 Any adverse event 1 (1%) 2 (2%) 17 (22%) Grade 4 0 0 10 (13%) NA 2 (3%) NA 0 8 (10%) 0 NA 9 (12%) 0 NA 0 1 (1%) 0 0 1 (1%) 0 5 (6%) 0 0 2 (2%) 0 5 (6%) 1 (1%) 0 8 (10%) 0 0 1 (1%) 0 NA 11 (14%) NA 0 Asthenia 9 (11%) 2 (2%) 0 3 (4%) 0 0 Bone pain 8 (10%) 1 (1%) 1 (1%) 3 (4%) 0 0 Constipation 10 (12%) 0 0 7 (9%) 0 0 Cough 10 (12%) 0 0 8 (10%) 0 0 Stomatitis 10 (12%) 0 0 2 (3%) 0 0 Epistaxis 9 (11%) 0 0 1 (1%) 0 0 Influenza 8 (10%) 1 (1%) 0 1 (1%) 0 0 Musculoskeletal pain 8 (10%) 1 (1%) 0 5 (6%) 0 0 Upper respiratory tract infection 8 (10%) 1 (1%) 0 2 (3%) 0 0 Dizziness 8 (10%) 0 0 3 (4%) 0 0 Peripheral neuropathy 8 (10%) 0 0 4 (5%) 0 0 Oropharyngeal pain 8 (10%) 0 0 1 (1%) 0 0 Pain in extremity 8 (10%) 0 0 6 (8%) 0 0 Data are n (%) unless otherwise specified. One (1%) grade 5 adverse event occurred in the palbociclib plus letrozole group (from disease progression) and none in the letrozole group. Additional grade 4 adverse events reported in the palbociclib plus letrozole group that are not shown in the table are neutropenia (five [6%]), pulmonary embolism (four [5%]), bone pain (one [1%]), chest pain (one [1%]), increased blood uric acid increased (1 [1%]), gangrene (one [1%]), and humerus fracture (one [1%]). No additional grade 4 adverse events were reported in the letrozole group. Additional grade 3 adverse events reported in the palbociclib plus letrozole group that are not shown in the table are abdominal pain (two [2%]), increased γ-glutamyl transferase (two [2%]), decreased white blood cell count (two [2%]), increased aspartate aminotransferase (one [1%]), pain (one [1%]), spinal pain (one [1%]), increase blood lactate dehydrogenase (one [1%]), joint stiffness (one [1%]), pneumonia (one [1%], wound (one [1%]), decreased blood potassium (one [1%]), cancer pain (one [1%]), ischaemic colitis (one [1%]), coronary artery disease (one [1%]), exertional dyspnoea (one [1%]), fallopian tube cancer (one [1%]), fractured sacrum (one [1%]), gastrointestinal disorder (one [1%]), granulocytopenia (one [1%]), hypermagnesaemia (one [1%]), intervertebral disc protrusion (one [1%]), nephrolithiasis (one [1%]), decreased neutrophil count (one [1%]), pathological fracture (one [1%]), periostitis (one [1%]), petechiae (one [1%]), renal disorder (one [1%]), staphylococcal bacteraemia (one [1%]), and urethral obstruction (one [1%]). Additional grade 3 adverse events reported in the letrozole group that are not shown in the table are pleural effusion (two [3%]), hypertension (one [1%]), chest pain (one [1%]), decreased haemoglobin (one [1%]), pain (one [1%]), spinal pain (one [1%]), dysphagia (one [1%]), hypocalcaemia (one [1%]), mucosal inflammation (one [1%]), neck pain (one [1%]), pelvic pain (one [1%]), cardiac failure (one [1%]), hip fracture (one [1%]), ileus (one [1%]), laceration (one [1%]), oesophageal achalasia (one [1%]), pneumothorax (one [1%]), and subcutaneous emphysema (one [1%]). Table 3: Most common all-cause adverse events that occurred in at least 10% of patients (safety population) 32 (7·1–16·4) for letrozole alone (HR 0·508, 95% CI 0·303–0·853; one-sided p=0·0046; figure 2). The effect of the combination treatment relative to letrozole alone was consistent across all demographic subgroups and patient baseline prognostic factors, apart from patients with disease recurrence 12 months or less from the end of adjuvant treatment, although this subgroup is limited by small numbers in both groups (figure 3). The results from prespecified sensitivity analyses were consistent with those of the main analysis (data not shown). Table 2 shows best responses to treatment. A greater proportion of patients in the palbociclib plus letrozole group than in the letrozole group had an objective response to treatment, both in the intention-to-treat population (36 [43%, 95% CI 32–54] vs 27 [33%, 23–45]; p=0·13) and in the population with measurable disease (36 [55%, 43–68] vs 26 [39%, 28–52]; one-sided p=0·047). Similarly, a greater proportion of patients in the intention-to-treat population achieved clinical benefit (68 [81%, 95% CI 71–89] vs 47 [58%, 47–69]; one-sided p=0·0009). The median duration of response for patients who had a complete or partial response was 20·3 months (95% CI 13·4–25·8) for the palbociclib plus letrozole group and 11·1 months (9·3–31·6) for the letrozole group. At the same time as the final progression-free survival analysis, we also assessed overall survival. Median overall survival was 37·5 months (95% CI 28·4–NE; 30 events) in the palbociclib plus letrozole group and 33·3 months (26·4–NE; 31 events) in the letrozole alone group (HR 0·813, 95% CI 0·492–1·345; two-sided p=0·42; figure 4). The most common adverse events reported for the palbociclib plus letrozole group were neutropenia, leucopenia, and fatigue (table 3). All 83 patients who received palbociclib plus letrozole had at least one adverse event, compared with 65 (84%) of 77 who received letrozole alone. Despite the increase in all grades of neutropenia and leucopenia with palbociclib plus letrozole, no cases of neutropenic fever were reported. Other adverse events (of any cause) that were increased in the palbociclib plus letrozole group included anaemia, nausea, arthralgia, and alopecia, but most of these were low grade (table 3). Of these adverse events, the difference between treatment groups was significant only for anaemia (two-sided p<0·0001) and alopecia (two-sided p=0·0002). Serious adverse events that occurred in more than one patient in the palbociclib plus letrozole group were pulmonary embolism (three [4%] patients), back pain (two [2%]), and diarrhoea (two [2%]). No serious adverse events occurred in more than one patient in the letrozole group. 27 (33%) patients in the palbociclib plus letrozole had dose interruptions because of adverse events, compared with only three (4%) patients in the letrozole group. In the combination group, 37 (45%) patients required a delay in the start of a subsequent treatment cycle because of an adverse event and 33 (40%) patients had a dose www.thelancet.com/oncology Vol 16 January 2015 Articles reduction. However, the mean relative dose intensity for palbociclib in the combination group was 94% (SD 26). Cycle delays and dose reductions are not applicable to the letrozole group. The main reason for study discontinuation in both treatment groups was disease progression (42 [50%] patients in the palbociclib plus letrozole group vs 57 [70%] patients in the letrozole group). 11 (13%) patients in the palbociclib plus letrozole group and two (2%) patients in the letrozole group discontinued the study because of an adverse event. Of these discontinuations, six (7%) patients in palbociclib plus letrozole group and two (2%) patients in the letrozole group discontinued because of treatment-related adverse events. One death occurred during the study in the palbociclib plus letrozole group because of disease progression; no treatment-related deaths occurred. We noted no significant differences in pain severity or the effect of pain on daily activities between the two treatment groups. Discussion The results of this open-label, phase 2 study show that patients with oestrogen receptor-positive, HER2-negative advanced breast cancer had significantly longer progression-free survival when treated with palbociclib and letrozole than when treated with letrozole alone. Additionally, the proportions of patients with an objective response and clinical benefit were greater in the combination group than in the letrozole alone group. The study was not powered to detect an overall survival advantage and few overall survival events had occurred at the time of this analysis; however, the initial data suggest no detrimental effect on overall survival with the addition of palbociclib in the first-line setting (panel). Hormone directed drugs have been the mainstay of treatment for advanced oestrogen receptor-positive breast cancer for more than four decades. Improvements in clinical outcomes have occurred with several drugs that target either specific hormone production (ie, ligands or the hormone receptor pathway), including tamoxifen, steroidal and non-steroidal aromatase inhibitors, and fulvestrant.15 Despite efforts to further improve clinical outcomes for patients with oestrogen receptor-positive breast cancer with drugs that target other pathways thought to have a role in the development of resistance to hormone drugs, most results have been largely disappointing, including efforts to target the HER1 and HER2 pathways, angiogenesis, and IGFR.16–18 Recently, however, targeting of mTOR, a crucial component of the PI3K pathway, with everolimus, used in combination with a steroidal aromatase inhibitor, resulted in improved progression-free survival, although not overall survival, in patients with oestrogen receptorpositive, advanced disease who had progressed on antihormonal treatment.4,19 Our findings need to be interpreted in the context of the limitations of the study design. Specifically, the study www.thelancet.com/oncology Vol 16 January 2015 is open-label and did not use central radiology review to prospectively assess the primary endpoint, but rather a retrospective, masked, independent review after accrual was completed. This analysis was limited by the fact that scans were obtained retrospectively and were not used to make on-treatment decisions. On-treatment decisions were made on the basis of scan reviews at the individual study sites. We noted some baseline imbalances based on the case report form data; however, sensitivity analyses including multivariate analysis to control for baseline factors consistently showed treatment benefit in the combination group across all demographic and clinical subgroups. These clinical results are supported by preclinical data11 that provide a clear biological rationale for the development of palbociclib in this patient population. As with most other molecularly targeted drugs in oncology, the greatest gains are often seen when rational and appropriate patient selection can be used prospectively. Preclinical data11 with palbociclib identified oestrogen receptor-positive breast cancer cells as one of the subtypes most sensitive to CDK4/6 inhibition, the other being HER2 amplification. Our results provide clinical validation of these preclinical data and support the further development of palbociclib for the oestrogen receptorpositive, HER2-negative subgroup of breast cancers. Genetic changes in cyclin D1 and p16 are known to occur in breast cancer and might have a role in the further selection of patients for treatment with a CDK4/6 inhibitor. In cohort 2, we investigated the potential for these genetic changes to be used to improve patient selection beyond use of oestrogen receptor-positive status alone. However, our results did not substantiate this hypothesis. This analysis confirmed that oestrogen receptor positivity is currently the best and most effective predictive marker for the identification of patients likely to respond to CDK4/6 inhibition. Further biomarker Panel: Research in context Systematic review We searched PubMed for reports of clinical trials published in English between Nov 1, 2004, and Nov 1, 2014, using the terms “breast cancer” and “cyclin-dependent kinase inhibitors”. We did not identify any studies that investigated cyclin-dependent kinase (CDK) inhibitors specifically in women with breast cancer, despite laboratory data suggesting a role for CDK inhibition in breast cancer.7,8 Interpretation Our results show that palbociclib, a novel, oral, inhibitor of CDK4/6, used in combination with letrozole resulted in longer progression-free survival than letrozole alone in postmenopausal women with advanced oestrogen receptor-positive, HER2-negative breast cancer. These results provide the first clinical data to show a role for CDK4/6 inhibition in breast cancer. The safety profile of the combination was predictable and manageable. Overall, our results suggest that palbociclib in combination with letrozole could become an important treatment option, substantially improving upon letrozole alone for the treatment of postmenopausal women with oestrogen receptor-positive, HER2-negative, advanced breast cancer. 33 Articles research designed to improve on oestrogen receptorpositive status as the selection biomarker is ongoing; however, in view of the large proportion of patients in our study who achieved a clinical benefit response (more than 80%), the benefit of additional biomarkers could be difficult to ascertain. Negative-selection biomarkers of resistance might be more easily identified and will also be assessed in ongoing and future molecular studies. One of the most important markers of sensitivity to palbociclib is the presence of an intact Rb pathway; however, since pRb loss is uncommon in oestrogen receptor-positive, HER2-negative breast cancers, it was not used as a prospective independent biomarker for patient selection in the present study. Potentially, relative amounts of pRb (rather than its presence or absence) in the various breast cancer subtypes might be of predictive importance and early preclinical data11 suggest that this possibility should be investigated. When comparing median progression-free survival in the letrozole alone groups, we noted a difference between cohorts 1 and 2, suggesting a potential predictive value for cyclin D1 gains, p16 loss, or both in determining response to letrozole alone. However, this finding could simply be an artifact of the sample size in cohort 1, so further study is necessary. Since the initiation of this study, additional laboratory findings have linked CDK 4/6 inhibition to endocrine sensitivity in oestrogen receptor-positive breast cancer.20,21 The Cancer Genome Atlas has been used to identify common changes in the Rb pathway in all breast cancer subtypes, including the luminal oestrogen receptorpositive, HER2-negative subgroup. However, our findings suggest that limiting patient selection to those with defined genetic changes in the Rb pathway might exclude a much larger group of patients that could benefit from CDK4/6 inhibition. Additionally, the results of a recent phase 2, single-arm study22 of palbociclib in patients with heavily pretreated advanced breast cancer showed singleagent activity in some patients with oestrogen receptorpositive, HER2-negative breast cancers. Our results also provide useful data for the safety profile of the combination of palbociclib and letrozole, suggesting that adverse events are predictable and manageable. Neutropenia, although common, was not accompanied by serious clinical outcomes and is likely to be the result of an on-target effect of CDK4/6 inhibition on marrow progenitor cells. The absence of serious complications resulting from palbociclib-associated neutropenia probably reflects a cytostatic rather than cytotoxic effect of the drug on bone marrow progenitor cells,23 different from what is seen with typical cytotoxic drugs. Additional analyses of the effect of palbociclib on quality-of-life measures are ongoing in the context of phase 3 studies. Taken together, the data from this study provides a proof of concept for the activity and safety of CDK4/6 inhibition in advanced, oestrogen receptor-positive, 34 HER2-negative breast cancer. The improvement in progression-free survival is substantial in this population and is accompanied by manageable toxic effects. These data clearly warrant further investigation of the efficacy and safety of palbociclib in combination with hormonal blockade, both in patients with this subtype of breast cancer and in other cancer settings. A phase 3, doubleblind, placebo-controlled study (NCT01740427) in a similar patient population (n=650) with the aim of confirming the present phase 2 findings is now fully enrolled and ongoing. Additionally, other phase 3 studies of palbociclib in combination with various anti-hormonal drugs in additional breast cancer settings are now ongoing (NCT01942135 and NCT01864746). Contributors RSF, SOK, CF, XH, STK, SR, and DJS contributed to the design of the study. RSF, DJS, and JPC were members of the steering committee that oversaw the conduct of the study. RSF, JPC, IMB, SOK, RP, ART, and NLV contributed substantially to patient recruitment. RSF, JPC, IL, KB, IMB, SOK, JE, RP, TP, MS, YS, NLV, DJS, ART, and STK contributed to data collection. JPC, RP, and SR contributed to data review. RSF, JPC, IMB, SOK, RP, CF, XH, STK, SR, and DJS contributed to data analysis. RSF, IL, KB, IMB, SOK, JE, RP, TP, MS, YS, CF, XH, STK, SR, and DJS contributed to data interpretation. RSF, JE, MS, CF, XH, STK, SR, and DJS contributed to the writing and revision of the report. DJS searched the scientific literature. DJS and XH contributed to production of the figures. All authors contributed to review of the report and approved the final submitted version. Declaration of interests RSF and DJS received research support from Pfizer and have served as consultants to the same company. DJS has common stock in Pfizer. JPC has received conference support from Pfizer. JE has received honoraria from Pfizer for attending advisory board meetings. MS has received research support, served as a consultant, and received honoraria from Pfizer. He has also received personal fees from Novartis, AstraZeneca, Roche, GlaxoSmithKline, Sanofi-Aventis, Sividon, PierreFabre, Celgene, and Eisai. CF, XH, STK, and SR are employees of Pfizer. KB, SOK, TP, IL, IMB, YS, RP, ART, and NLV declare no competing interests. Acknowledgments The study was funded by Pfizer. DJS has received funding for the preclinical work on cyclin-dependent kinase 4/6 inhibition that generated translational concepts for this clinical study from the US Department of Defense Breast Cancer Innovator Award, the Revlon/ UCLA Women’s Cancer Research Program, and the Peter and Denise Wittich Breast Cancer Program. We thank all the study participants and their families, as well as the staff of Translational Research in Oncology for their support. We also thank Nicholas J Robert (Virginia Cancer Specialists, US Oncology, Fairfax, VA, USA) and Marc Buyse (IDDI, Louvain-la-Neuve, Belgium) for their contributions. Karen Miller and Cynthia Gobbel from Complete Healthcare Communications (Chadds Ford, PA, USA) provided administrative support for the report submission (funded by Pfizer). References 1 Perou CM, Sorlie T, Eisen MB, et al. Molecular portraits of human breast tumours. Nature 2000; 406: 747–52. 2 Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin 2011; 61: 69–90. 3 Slamon D, Leyland-Jones B, Shak S, et al. Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpresses HER2. N Engl J Med 2001; 344: 783–92. 4 Baselga J, Campone M, Piccart M, et al. Everolimus in postmenopausal hormone-receptor-positive advanced breast cancer. N Engl J Med 2012; 366: 520–29. 5 Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell 2011; 144: 646–74. www.thelancet.com/oncology Vol 16 January 2015 Articles 6 7 8 9 10 11 12 13 14 15 Cancer Genome Atlas Network. Comprehensive molecular portraits of human breast tumours. Nature 2012; 490: 61–70. Malumbres M, Barbacid M. Cell cycle, CDKs and cancer: a changing paradigm. Nat Rev Cancer 2009; 9: 153–66. Stone A, Sutherland RL, Musgrove EA. Inhibitors of cell cycle kinases: recent advances and future prospects as cancer therapeutics. Crit Rev Oncog 2012; 17: 175–98. Fry DW, Harvey PJ, Keller PR, et al. Specific inhibition of cyclin-dependent kinase 4/6 by PD 0332991 and associated antitumor activity in human tumor xenografts. Mol Cancer Ther 2004; 3: 1427–38. Choi YJ, Anders L. Signaling through cyclin D-dependent kinases. Oncogene 2014; 33: 1890–903. Finn RS, Dering J, Conklin D, et al. PD 0332991, a selective cyclin D kinase 4/6 inhibitor, preferentially inhibits proliferation of luminal estrogen receptor-positive human breast cancer cell lines in vitro. Breast Cancer Res 2009; 11: R77. Finn R, Hurvitz S, Allison M, et al. Phase I study of PD 0332991, a novel, oral, cyclin-D kinase (CDK) 4/6 inhibitor in combination with letrozole, for first-line treatment of metastatic post-menopausal, estrogen receptor-positive (ER+), human epidermal growth factor receptor 2 (HER2)-negative breast cancer. Cancer Res 2009; 69 (24 suppl): abstr 5069. Mouridsen H, Gershanovich M, Sun Y, et al. Phase III study of letrozole versus tamoxifen as first-line therapy of advanced breast cancer in postmenopausal women: analysis of survival and update of efficacy from the International Letrozole Breast Cancer Group. J Clin Oncol 2003; 21: 2101–09. Grambsch PM, Therneau TM. Proportional hazards tests and diagnostics based on weighted residuals. Biometrika 1994; 81: 515–26. Chlebowski RT. Changing concepts of hormone receptor-positive advanced breast cancer therapy. Clin Breast Cancer 2013; 13: 159–66. www.thelancet.com/oncology Vol 16 January 2015 16 17 18 19 20 21 22 23 Johnston S, Pippen J Jr, Pivot X, et al. Lapatinib combined with letrozole versus letrozole and placebo as first-line therapy for postmenopausal hormone receptor-positive metastatic breast cancer. J Clin Oncol 2009; 27: 5538–46. Martin M, Loibl S, von Minckwitz G, et al. Phase III trial evaluating the addition of bevacizumab to endocrine therapy as first-line treatment for advanced breast cancer—first efficacy results from the LEA study. Cancer Res 2012; 72: abstr S1-7. Robertson JF, Ferrero JM, Bourgeois H, et al. Ganitumab with either exemestane or fulvestrant for postmenopausal women with advanced, hormone-receptor-positive breast cancer: a randomised, controlled, double-blind, phase 2 trial. Lancet Oncol 2013; 14: 228–35. Piccart M, Hortobagyi GN, Campone M, et al. Everolimus plus exemestane for hormone receptor-positive (HR+), human epidermal growth factor receptor-2-negative (HER2–) advanced breast cancer (BC): overall survival results from BOLERO-2. Eur J Cancer 2014; 50 (suppl 3): S1. Dean JL, McClendon AK, Hickey TE, et al. Therapeutic response to CDK4/6 inhibition in breast cancer defined by ex vivo analyses of human tumors. Cell Cycle 2012; 11: 2756–61. Miller TW, Balko JM, Fox EM, et al. ERα-dependent E2F transcription can mediate resistance to estrogen deprivation in human breast cancer. Cancer Discov 2011; 1: 338–51. DeMichele A, Clark AS, Heitjan D, et al. A phase II trial of an oral CDK 4/6 inhibitor, PD0332991, in advanced breast cancer. 2013 ASCO Annual Meeting; Chicago, IL, USA; May 31–June 4, 2013. Abstr 519. Roberts PJ, Bisi JE, Strum JC, et al. Multiple roles of cyclindependent kinase 4/6 inhibitors in cancer therapy. J Natl Cancer Inst 2012; 104: 476–87. 35