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Background: Somatic ERBB2 (HER2) mutations occur in approximately 2% of patients with breast cancer and are found in a predominantly mutually exclusive manner with ERBB2 amplification. These mutations result in increased signaling and oncogenic transformation. Neratinib, a pan-ERBB irreversible tyrosine kinase inhibitor, potently inhibits growth of ERBB2 mutant tumor cell lines and xenografts. An ongoing signal-seeking phase II 'basket' study is evaluating neratinib in patients with multiple histologies harboring ERBB2 mutations (NCT01953926). Novel mutations identified in enrolled patients were characterized for biologic activity in a variety of in vitro model systems. A preliminary analysis of the HER2 non-amplified metastatic breast cancer cohort is presented. Methods: Patients with ERBB2 mutant metastatic breast cancer documented by local testing methods received single-agent oral neratinib 240 mg once daily until progression or intolerable toxicity. High-dose loperamide prophylaxis was mandatory during cycle 1. The primary endpoint was the objective response rate at 8 weeks, defined using anatomic (RECIST 1.1) and/or metabolic (PET Response Criteria) assessments. Secondary endpoints were best overall response rate, clinical benefit rate, progression-free survival, duration of response, and safety. Results: 17 patients with metastatic breast cancer were enrolled and received neratinib (13 patients are evaluable for efficacy to date). Patients had a median of 3 prior anticancer regimens. Other baseline characteristics were: median age 59 years; bone involvement 71%; visceral disease 82%. Tumor characteristics were: ductal/lobular 76%/24%; ERBB2 mutation single nucleotide variants/indels 82%/18%; HER2 amplified/non-amplified 0%/100%; hormone receptor positive/negative 82%/18%. Five patients (39%) had an objective response at 8 weeks (95% CI 14–68%). In the patients who responded, ERBB2 mutations were: 1 complete response (L755S); 4 partial responses (L755S, V777L, V777L, and L869R). The most common all-grade adverse events (in ≥15% of patients) across all cohorts (n=93) were: diarrhea (62%), fatigue (28%), nausea (36%), vomiting (30%), anemia (15%), and constipation (29%). The most common grade 3/4 adverse event was diarrhea (14%, all grade 3). Updated efficacy results, centralized genomic analyses on archival tumor samples, and in vitro characterization of novel ERBB2 mutants will be presented. Conclusions: Single-agent neratinib shows encouraging signs of clinical activity in patients with heavily pretreated, ERBB2 mutant, HER2 non-amplified metastatic breast cancer. The breast cancer cohort demonstrated sufficient activity to meet the study's pre-specified efficacy requirements according to a Simon's two-stage design, and suggests that a confirmatory trial of neratinib for targeting ERBB2 driver mutations in metastatic breast cancer is warranted. Safety was acceptable and diarrhea was manageable with loperamide prophylaxis. Citation Format: Hyman DM, Piha-Paul SA, Rodón J, Saura C, Puzanov I, Shapiro GI, Loi S, Joensuu H, Hanrahan AJ, Modi S, Lalani AS, Xu F, Garza SJ, Cutler RE, Bryce R, Meric-Bernstam F, Baselga J, Solit DB. Neratinib for ERBB2 mutant, HER2 non-amplified, metastatic breast cancer: Preliminary analysis from a multicenter, open-label, multi-histology phase II basket trial. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr PD5-05.

Background: We examined if alterations in nucleic acid variants, genes, pathways, and overall mutational load and clonal entropy are associated with pathologic complete response (pCR) and survival after neoadjuvant anti-HER2 therapies in the NeoALTTO trial. Methods: Whole exome sequencing was performed of 203 baseline biopsies with outcome information. The mean nucleotide coverage was 150x with >90% of target bases showing > 30x coverage in > 99% of samples. Somatic mutations were called by MuTect and indels by Strelka, using pooled reference normal DNA. Significantly mutated genes (FDR Results: Only 12 genes had mutation rates significantly above background and among these only PI3KCA was associated with lower pCR rate (OR=0.42, p=0.019). Genes with somatic mutations in more than 10 patients were also assessed, but none were associated with pCR or survival. Clonal entropy or adjusted mutation load also did not correlate with response. Mutations in 33 pathways showed significant association with response in the entire cohort. In the trastuzumab arm, 23 of the 33 pathways showed an association with response but none was independent of PIK3CA mutation. We constructed "PIK3CA-gene network" that included all unique genes (n=439) from theese 23 pathways. Of the 66 patients in the trastuzumab arm, 50 carried at least one mutation in one of the 439 genes and among these only 2 achieved pCR (4%) compared to 9 of 16 pCR (56%) among the wild type (OR=0.035; p < 0.001). The same genes/mutations had little impact on pCR in the lapatinib arm (pCR 20%). In the lapatinib arm, mutations in 3 pathways conferred higher probability of pCR. The "Regulation of RhoA activity" pathway, had the most significant association with pCR in the entire cohort (OR=3.77, p=0.0009) and in the lapatininb (pCR 67% vs 17%, OR=14.8, p=0.008) and lapatinib + trastuzumab (OR=3.0, p=0.06) arms, but not in the trastuzumab arm (OR=1.4, p=0.7). Event free and overall survival were also significantly higher in patients who had mutations in this pathway. Twenty seven of the 48 genes in this pathway had mutations affecting 33 patients but different genes were affected in different individuals. Conclusions: There are no high frequency recurrent single mutations associated with response to HER2-targeted therapies, other than PIK3CA. We identified several biological pathways, including RhoA activity, and a network of PIK3CA associated genes that are significantly associated with response when affected by mutations, however, different genes are mutated in different individuals. Citation Format: Pusztai L, Shi W, Jiang T, Nuciforo P, Holmes E, Harbeck N, Sotiriou C, Rimm D, Hatzis C, de la Peña L, Armour A, Piccart-Gebhart M, Baselga J. Whole exome sequencing of pre-treatment biopsies from the neoALTTO trial to identify DNA aberrations associated with response to HER2-targeted therapies. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr S5-01.

Background: PI3K pathway activation is a hallmark of hormone receptor-positive (HR+) BC cells resistant to endocrine therapy (ET). Preclinical and early clinical data suggest that combining the pan-PI3K inhibitor BUP (BKM120) with ET may provide clinical benefits in this setting. BELLE-2 (NCT01610284) is the first randomized Phase III trial to assess the efficacy and safety of a PI3K inhibitor combined with FULV in HR+ advanced BC, including a prospective analysis of whether PI3K pathway activation status measured in archival tumor tissue and ctDNA is predictive of clinical benefit. Methods: Postmenopausal women with HR+/HER2– locally advanced or metastatic BC refractory to aromatase inhibitor therapy were enrolled. After a 14-day run-in with FULV (500 mg), patients (pts) were randomized (1:1) to receive oral BUP (100 mg/day) or placebo (PBO) with FULV (500 mg per standard of care). Randomization of all pts was stratified by PI3K pathway status measured in archival tumor tissue (PIK3CA mutation or PTEN loss; activated, non-activated, unknown) and visceral disease status (present, absent). Baseline PIK3CA mutation status in ctDNA was assessed in a subset of 587 pts at trial entry. The primary endpoint was locally-assessed progression-free survival (PFS; RECIST v1.1) in the full population and PI3K pathway-activated group. Secondary endpoints included overall survival, safety, overall response rate (ORR), and clinical benefit rate (CBR). Results: 1147 pts received FULV with BUP or PBO, with 187 (16%) ongoing at data cut-off. Baseline characteristics were well balanced between the two arms: median age was 62 years, 98% had ECOG performance status of 0/1, 61% had visceral disease, 69% were sensitive to prior ET, 28% had received prior chemotherapy for metastatic BC. BELLE-2 met its primary endpoint in the full population (Table). In pts with PI3K pathway-activated tumor tissue, PFS increase for BUP+FULV vs PBO+FULV did not meet the planned endpoint. Among pts with PIK3CA status measured in ctDNA, median PFS, ORR, and CBR were significantly improved for BUP+FULV vs PBO+FULV in pts with PIK3CA-mutant tumors but not pts without. Median PFS, monthsHR95% CI; P valueORRCBR BUP+FULVPBO+FULV Full population N=11476.95.00.780.67–0.89;P The most common Grade 3/4 adverse events (AEs; ≥5% of pts; BUP+FULV vs PBO+FULV) in all pts were increased alanine aminotransferase (26 vs 1%), increased aspartate aminotransferase (18 vs 3%), hyperglycemia (15 vs 0.2%), and rash (8 vs 0%). The most common reasons for treatment discontinuation were disease progression (54 vs 73%) and AEs (13 vs 2%). Conclusions: BELLE-2 met its primary endpoint in the full population. Prespecified analyses showed that characterizing PIK3CA mutation in ctDNA at trial entry identifies pts with endocrine- resistant HR+/HER2–advanced BC, for whom BUP+FULV results in meaningful clinical benefits. Citation Format: Baselga J, Im S-A, Iwata H, Clemons M, Ito Y, Awada A, Chia S, Jagiello-Gruszfeld A, Pistilli B, Tseng L-M, Hurvitz S, Masuda N, Cortés J, De Laurentiis M, Arteaga CL, Jiang Z, Jonat W, Hachemi S, Le Mouhaër S, Di Tomaso E, Urban P, Massacesi C, Campone M. PIK3CA status in circulating tumor DNA (ctDNA) predicts efficacy of buparlisib (BUP) plus fulvestrant (FULV) in postmenopausal women with endocrine-resistant HR+/HER2– advanced breast cancer (BC): First results from the randomized, phase III BELLE-2 trial. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr S6-01.

Mammography (Mx) and breast ultrasound (US) are the most commonly used diagnostic imaging modalities to estimate primary tumor size at the time of diagnosis. However, there are uncertainties regarding their use in the context of neoadjuvant therapy to predict pathologic complete response (pCR) or event-free survival (EFS). In this study, we sought to determine the value of Mx and US in predicting outcomes in women with HER2-positive breast cancer treated within the Neoadjuvant Lapatinib and/or Trastuzumab Treatment Optimisation (Neo-ALTTO) trial. METHODS Neo-ALTTO enrolled 455 women with invasive HER2-positive breast cancer and compared rates of pCR to neoadjuvant lapatinib, trastuzumab, and their combination. Each anti-HER2 therapy was given alone for 6 weeks, followed by 12 weeks of the same therapy plus weekly paclitaxel prior to surgery. Mx and US were requested at baseline, week 6 and before surgery. Central imaging review was not pre-planned and two independent investigators (SDC and HAA Jr), blinded to assigned treatment and clinical outcomes, reviewed the measurements reported for each imaging modality and assigned the corresponding RECIST category of response. Responders were defined as patients who had either a partial or complete response (CR + PR). We evaluated the association between radiological response at week 6/surgery with both pCR and EFS. RESULTS A total of 340 (77%) and 267 (61%) pts had an evaluable US and Mx at weeks 6; and 309 (70%) and 248 (56%) pts had an evaluable US and Mx at the time of surgery. Early response (CR + PR) in the primary tumors was observed after 6 weeks of treatment in 32% pts by Mx and in 43% pts by US. pCR rates were twice as high for early responders than non-responders (week 6: 46% vs 23% by US, p = CONCLUSION Our results show that both Mx and US are underused during neo-adjuvant treatment, and further recommendations regarding the use of both imaging modalities should be explored prospectively. US may be used to assess early response to preoperative treatment in patients with HER2 positive breast cancer receiving anti-HER2 therapies, whereas Mx appears to be more useful in detecting residual disease at the time of surgery. Citation Format: Di Cosimo S, Campbell C, Hazim Jr HA, Curigliano G, Criscitiello C, Crescenzo R, de la Pena L, Piccart-Gebhart MJ, Eidtmann H, Baselga J, de Azambuja E. Breast ultrasound and mammography and response to neoadjuvant lapatinib, trastuzumab and their combination in HER2 positive breast cancer: Results from Neo-ALTTO (BIG 1-06). [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P6-01-04.

Efficacy and safety if ixabepilone plus capecitabine in elderly patients with anthracycline and taxane-pretreated metastatic breast cancer

Background Many patients in Europe do not receive adequate relief of pain because of excessive regulatory restrictions on the availability and accessibility of opioids. This is a major public health problem. The aim of the study is to evaluate and report on opioid availability and the legal and regulatory barriers to accessibility across the countries of Europe. Methods European Society for Medical Oncology and European Association for Palliative Care national representatives reported data regarding survey of opioid availability and accessibility. Formulary adequacy is evaluated relative to the World Health Organization (WHO) essential drugs list and the International Association for Hospice and Palliative Care list of essential medicines for palliative care. Overregulation is evaluated according to the guidelines for assessment of national opioid regulations of the WHO. Results Data were reported on the availability and accessibility of opioids for the management of cancer pain in 21 Eastern European countries and 20 Western European countries. Results are presented describing the availability and cost of opioids for cancer pain in each surveyed country and nine forms of regulatory restrictions. Conclusions Using standards derived from the WHO and International Narcotics Control Board, this survey has exposed formulary deficiencies and excessive regulatory barriers that interfere with appropriate patient care in many European countries. There is an ethical and public health imperative to address these issues.

Background On the basis of clinical activity of capecitabine and gemcitabine for metastatic breast cancer, we carried out a multicenter phase II clinical trial on the combination of these two agents in advanced anthracycline-pretreated breast cancer patients. Main objectives were to assess its efficacy and safety profile. Patients and methods Seventy-six anthracycline-pretreated breast cancer patients were evaluated and were stratified according to previous treatment of advanced disease (group-1: not previously treated and group-2: previously treated). Study treatment consisted of gemcitabine 1000 mg/m2, i.v., as 30 min-infusion, days 1 and 8 every 21 days, plus oral capecitabine 830 mg/m2 b.i.d., days 1–14 every 21 days. Results Overall response rate was 61% for group-1, 48.5% for group-2 and 55.2% for the whole population. Clinical benefit rate was 73% for group-1, 80% for patients in group-2 and 76% for all patients. Median time to progression was 13.0 months for group-1, 8.2 months for group-2 and 11.1 months for the whole population. Most frequent grade 3–4 observed toxic effects per patient were neutropenia (60%), asymptomatic liver toxicity (13.5%), asthenia (14%) and hand–foot syndrome (16%). Only one patient presented febrile neutropenia. No treatment-related deaths occurred. Conclusion Combination of gemcitabine and capecitabine is an active and safe regimen in anthracycline-pretreated breast cancer patients.

Glass transition temperatures (Tᵍ) of epoxy resins, diglycidyl ether of bisphenol-A cured with piperidine, have been measured by differential scanning calorimetry and dynamic mechanical thermal analysis in order to follow the curing process. Optimum cure conditions have been established as 3% PP at 120°C. Tᵍs of the neat epoxy system and those modified with 5 and 16% of a carboxyl-terminated reactive rubber copolymer of butadiene and acrylonitrile (CTBN) have been determined as a function of the cure time. Addition of 5% of CTBN does not modify the process, but the higher CTBN content accelerates the initial stage. Tᵍs after long cure times (Tᵍx) are lower for the modified systems than for the neat, showing that CTBN is not fully segregated. The amounts of CTBN dissolved in the epoxy crosslinked network have been estimated. The authors thank Dr A. J. Kinloch for advice in this investigation. We are grateful to CIBAGEIGY for kindly providing the samples. Financial support by the Comisión de lnvestigación Científica y Técnica is acknowledged (89/0435).

Increasing life expectancy in the general population has led to a rise in the incidence of cancer and new challenges with regard to the diagnosis, therapy and prognosis of this disease.To assess prognostic factors in the initial work-up of patients ultimately diagnosed with cancer in an Internal Medicine Service, particularly those related with age.A prospective study was undertaken with 224 patients ultimately diagnosed with cancer, as confirmed by histological or cytological study. The neoplasms included respiratory, gastrointestinal, genitourinary, metastatic adenocarcinoma of unknown origin, gynaecological, hepatobiliary and others. Before reaching the diagnosis, the following factors were investigated in all patients: functional status [Karnofsky Performance Status (KPS)], comorbidity (Charlson scale), body mass index (BMI), serum cholesterol and albumin concentrations, cognitive level (Mini-mental test), quality of life (Short Form 36 questionnaire), and extension of the disease according to established criteria. Survival at 1 year was analysed. Statistical analyses were done with spss 11.0 for Windows, using a forward stepwise (likelihood ratio) method to construct the model and a Cox multivariate model for the survival analysis.A total of 224 patients, 167 men (74.5%) and 57 women (25.5%), with a mean age of 66.1 +/- 12.3 years were studied. KPS wasor= 70 in 84% and comorbidity was zero or one in 74%. BMI was 24.25 +/- 4.3, cholesterol 180.7 +/- 4.3, albumin 3.32 +/- 0.5 and Mini-mental score 25.4 +/- 3.7. Metastasis was seen in 131 patients (58.5%) and local disease in 93 cases (41.5%). One-year survival was 38.8% (87 patients) with a mean of 203.8 +/- 143 days. In the Cox analysis, the independent predictive factors for survival were KPS [hazard ratio (HR) = 0.951; 95% CI = 0.930-0.974; p0.01], metastatic dissemination (HR = 2.422; 95% CI = 1.643-3.571; p0.01), physical quality of life (HR = 0.978; 95% CI = 0.962-0.995; p0.01) and albumin (HR = 0.653; 95% CI = 0.455-0.936; p0.01).In the initial work-up of patients ultimately diagnosed with cancer in an Internal Medicine Service, functional status, dissemination, the physical component in the quality of life scale and serum albumin levels were independent prognostic factors for survival. Age was not an independent prognostic factor and should not be used as a basis for adopting diagnostic or therapeutic decisions in these patients.

1094 Background: Lap has single agent activity against BM in patients (pts) with HER2+ (ErbB2+) BC. Lap + Cap increases response rate, time to progression, and progression-free survival when compared with Cap alone in pts with refractory HER2+ BC. An exploratory analysis of the latter study (EGF100151) showed fewer pts on Lap + Cap developed BM at first relapse compared with Cap alone. The LEAP and ATU were designed to provide access to Lap before commercial availability and were initiated after EGF100151 enrollment was halted. Eligibility requirements for these programs were similar to EGF100151, i.e., pts had disease progression following prior taxane, anthracycline, and trastuzumab therapy. Although data were not formally collected via case report forms, we surveyed LEAP and ATU sites to ascertain the percent of pts with progressive BM at entry and response to Lap + Cap. Methods: LEAP/ATU sites that enrolled 15 or more pts were queried to provide data via worksheets or narratives regarding progressive ...

The ectodomain of different transmembrane molecules is released by a proteolytic event known as shedding. The metalloprotease disintegrin proTNF-alpha converting enzyme (TACE) is responsible for the shedding of various proteins, including protransforming growth factor-alpha (proTGF-alpha) and amyloid-beta precursor protein (APP). Inactive TACE accumulates in the early secretory pathway of cell mutants (M1 and M2) defective in proTGF-alpha and APP shedding. Although previous evidences indicated that the component mutated in M1 and M2 cells is different from TACE, recent results show the existence of two heterozygous point mutations in TACE from M2 cells. Here, we show that wild-type TACE stably transfected in M2 cells is processed, transported to the cell surface, and rescues the proTGF-alpha and APP shedding-defective phenotype. Furthermore, M1 cells also express mutant TACE and transfection with wild-type TACE restores the wild-type phenotype. Therefore, different inactivating mutations result in the accumulation of TACE in the early secretory pathway, emphasizing the importance of the initial steps in the biosynthesis of TACE.

Background: The PI3K/AKT/mTOR pathway has been reported altered in up to 40% of hormone receptor-positive (HR+) breast cancers (BC). Furthermore, pathway activation has been implicated in resistance to endocrine therapy, including aromatase inhibitors (AI). BKM120 is an oral inhibitor of all 4 isoforms of class I PI3K (α, β, γ, δ). In combination with fulvestrant, BKM120 has demonstrated pro-apoptotic effects in vitro, and achieved near-complete tumor regression in HR+ PIK3CA-mutated estrogen-independent xenografts. Preliminary clinical activity was observed with BKM120 in patients with BC, both as a single agent and in combination with letrozole (Mayer, et al. ASCO 2012: #510) or trastuzumab (Saura, et al. SABCS 2011: #PD09-03). Study design: This is a Phase III randomized, double-blind, placebo-controlled trial of BKM120 or placebo in combination with fulvestrant in postmenopausal women with HR+/HER2– locally advanced or metastatic BC that progressed on or after AI treatment (BELLE-2; CBKM120F2302; NCT01610284). After a 14-day fulvestrant run-in phase to allow determination of tumor PI3K pathway activation status, patients are randomized (1:1) to receive continuous BKM120 (100 mg/d) or placebo, and fulvestrant 500 mg at C1D15, and D1 of all cycles thereafter; 1 cycle=28 days. Randomization is stratified according to PI3K pathway activation (PIK3CA mut and/or PTEN mut and/or loss of PTEN protein expression by IHC), and visceral disease status. Study treatment is given until disease progression or until study discontinuation for any reason. Eligibility criteria include: postmenopausal women with HR+/HER2– locally advanced or metastatic BC refractory to AI (progression while on AI, or within 4 wks [metastatic] or 12 mos [adjuvant] of last AI dose); no more than 1 previous line of chemotherapy for advanced disease; and available archival tumor tissue for analysis of PI3K-related biomarkers. Co-primary endpoints: PFS in the full and PI3K pathway-activated populations based on local investigator assessment (RECIST 1.1). Key secondary endpoints: overall survival (OS) in both full and PI3K pathway-activated populations. Other secondary endpoints: PFS and OS (PI3K non-activated/unknown population), overall response rate and clinical benefit rate (both in full, PI3K-activated, PI3K non-activated/unknown populations), safety (CTCAE 4.03), pharmacokinetics of BKM120 and fulvestrant, and patient-reported quality of life (EORTC QLQ-C30 and QLQ-BR23). Statistical methods: PFS and OS will be estimated using the Kaplan–Meier method for both full and PI3K pathway activated populations. Both primary and key secondary endpoints will be tested using a stratified log-rank test at levels α governed by a graphical gate-keeping procedure. A stratified Cox regression will be used to estimate the hazard ratio, along with two-sided 95% confidence interval. Target accrual: Estimated enrollment for BELLE-2 is 842 patients. Recruitment in this Global study is ongoing with planned enrollment of patients in North America, South America, Europe, Asia and Africa. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr OT2-3-09.

Curing of an epoxy resin was studied by steady-state fluorescence and correlationed with IR methods to get more insight on the physical processes occurring during the curing of an epoxy/amine system. We have synthesized and tested a set of 9-anthroic acid (AA) derivatives, that are sensitive to changes in the medium properties. Its fluorescence response is analyzed and compared and it is concluded that N -dibutylanthroic acid amide shows the best performance.

The cure kinetics of DGEBA with aliphatic diamines was studied on the diffusion-controlled region. Two different diamines (ethylenediamine (EDA) and N -(methyl)ethylenediamine (MEDA)) were selected, to achieve different cross-linked networks. Additionally, reaction of DGEBA with butylamine was followed to compare non-cross-linked systems with the above mentioned ones. Curing temperatures were between 20 and 60 °C. Concentrations of epoxy and primary amine functionality were both followed by near FTIR, and T g was analyzed by DSC. An autocatalytic model was applied. Diffusion control kinetic in cross-linked polymers was analyzed in terms of free volume decrease. In that region, a model based on a WLF equation provides a good fit to the experimental data.

The HER2/neu gene encodes a 185 kDa transmembrane receptor (HER2) that belongs to the epidermal growth factor receptor family and has intrinsic tyrosine kinase activity. HER2 is overexpressed in 25-30% of breast cancers and is suggested to have a direct role in the pathogenesis and clinical aggressiveness of HER2 overexpressing tumors. A murine monoclonal antibody, 4D5, directed against the extracellular domain of HER2, is a potent inhibitor of growth of human breast cancer cells overexpressing HER2 in vitro and in xenograft models. To facilitate clinical investigation, 4D5 was humanized by inserting the complementary determining regions of 4D5 into the framework of a consensus human IgG1. The resulting recombinant humanized anti-HER2 MAb, trastuzumab, was found to inhibit the growth of human cancer cells and tumor xenografts overexpressing HER2. Data from phase II trials in women with breast cancer whose tumors overexpress HER2 have shown that trastuzumab has a favorable toxicity profile, is active as a single agent and induces long-lasting objective tumor responses. In combination studies, there was no evidence that trastuzumab enhanced the toxicity of cisplatin and the pharmacokinetic parameters of trastuzumab were unaltered by coadministration of cisplatin. Furthermore, clinical response rates were higher than those reported with either agent alone in a similar patient population. Results of a multicenter, phase III clinical trial of chemotherapy (doxorubicin- or paclitaxel-based) plus trastuzumab as compared to chemotherapy alone in patients with advanced breast cancers overexpressing HER2 showed a significant enhancement in the effects of chemotherapy on time to disease progression, response rates and survival with coadministration of trastuzumab, without increases in overall severe adverse events. Myocardial dysfunction syndrome, similar to that observed with anthracyclines, was reported more commonly with chemotherapy plus trastuzumab. Positive results from clinical studies led to the approval of trastuzumab in the U.S in October 1998 for the treatment of metastatic breast cancer in patients with tumors overexpressing HER2. Since then, the MAb has also been marketed in Switzerland and Canada.

The factors and mechanisms that transduce the intracellular signals sent upon activation of the receptor for the epidermal growth factor (EGFR) and related receptors are reasonably well understood and, in fact, are the targets of anti-tumor drugs. In contrast, less is known about the mechanisms implicated in sending the signals that activate these receptors. Here we show that when its proteolytic shedding is prevented, the transmembrane form of the transforming growth factor-alpha (proTGF-alpha) interacts with, but does not activate, the EGFR. Thus, shedding seems to control not only the availability of the soluble form of the growth factor (TGF-alpha) but also the activity of the transmembrane form. The activity of the protease responsible for the shedding of proTGF-alpha, tumor necrosis factor-alpha converting enzyme (TACE), is required for the activation of the EGFR in vivo and for the development of tumors in nude mice, indicating a crucial role of TACE in tumorigenesis. In agreement with this view, TACE is dramatically overexpressed in the majority of mammary tumors analyzed. Collectively, this evidence points to TACE as a promising target of anti-tumor therapy.

Publicación derivada de la comunicación presentada en: International Conference on Advanced Materials Processing Technologies [AMPT'01] y publicada en: Proceedings of the International Conference on Advanced Materials Processing Technologies [AMPT'01]: Universidad Carlos III de Madrid, Leganés, Spain, 18-21 September, 2001 Aqueous solutions of three silanes to cover silica microfibres were used, being the 3-aminopropyltriethoxysilane (APTES) and the 3-aminopropylmethyldiethoxysilane (APDES) the reagents for preparing them at the proportion of (APTES/APDES): 1/0, 1/1 and 0/1. The 1-pyrene sulphonyl chloride (PSC) fluorescent moiety was chemically attached to the silanised substrate via the sulphonamide formation. The hydrolytic degradation phenomenon of the silane coupling layer was studied as a function of: (i) temperature, (ii) coating layer type and (iii) pH (4, 7 and 10). The hydrolytic damage in the coupling region of the silica microfibres composite materials occurs under an equilibrium process. It was obtained the activation energies (Ea) for the hydrolytic damage considering the rate to reach the equilibrium. The values of Ea depended on the type of coating and on the pH. As a consequence, the rate of hydrolytic damage could be related to the proportion of Sisilane–O–Sisilane, while the OH− groups were thought to catalyse the reaction.

It is very important to control the distribution of stresses in order to prevent catastrophic failures during working life of polymer materials. At the present moment, there are several methods to test the distribution of stresses in materials. In the last years, many numerical methods have been proposed, but they have limitations. On the other hand, experimental methods are difficult to perform in all cases, for example when “in situ” determinations are required. In the present work, the use of environmentally sensitive fluorescence molecules (fluorophores) to estimate the tensile state in polymers is discussed. Pyrene, p -quaterphenyl, 1,6-diphenyl-1,3,5-hexatriene (DPhHT), 1,4-bis(5-phenyloxazol-2-yl)benzene and dansyl derivatives, whose fluorescences are sensitive to different types of changes (polarity, microviscosity, etc.), were immersed in one epoxy mixture. After that, the epoxy mixture was cured in a mould to prepare size controlled samples. The specimens were uniaxially loaded. In most of the cases the fluorescence suffers small variations with the epoxy deformation. To evaluate these changes, two photophysical parameters were used: (i) the integrated intensity and (ii) the first moment in the wavenumber of the emission band, 〈 ν 〉. Related to the fluorescence intensity, the dansyl moiety seems to be the most sensitive; however, for 〈 ν 〉 the DPhHT showed the highest changes.

Water absorption is a common problem in epoxy-based composites, especially in interfacial regions. In contrast to epoxy resins, silicones are hydrophobic materials. Modification of epoxy thermosets with silicones should improve physical properties because of decrease in water diffusion. Bisphenol-A diglycidyl ether (DGEBA) was cured with a synthesised aminopropyl perfunctionalised siloxane. Water absorption was analysed by different techniques, including near infrared (FTIR) water absorbance at 5200 cm−1 and gravimetry. Fick’s law diffusion coefficients were calculated. Swelling equilibrium of the resin was also analysed. Absorbed water catalyses the curing reaction strongly, allowing to reach high conversions.

Protein ectodomain shedding is a specialized type of regulated proteolysis that releases the extracellular domain of transmembrane proteins. The metalloprotease disintegrin tumor necrosis factor-α-converting enzyme (TACE) has been convincingly shown to play a central role in ectodomain shedding, but despite its broad interest, very little is known about the mechanisms that regulate its activity. An analysis of the biosynthesis of TACE in mutant cell lines that have a gross defect in ectodomain shedding (M1 and M2) shows a defective removal of the prodomain that keeps TACE in an inactive form. Using LoVo, a cell line that lacks of active furin, and α1-Antitrypsin Portland, a protein inhibitor of proprotein convertases, we show that TACE is normally processed by furin and other proprotein convertases. The defect in MI and M2 cells is due to a blockade of the exit of TACE from the endoplasmic reticulum. The processing of other zinc-dependent metalloproteases, previously suggested to participate in activated ectodomain shedding is normal in the mutant cells, indicating that the component mutated is highly specific for TACE. In summary, the characterization of shedding-defective somatic cell mutants unveils the existence of a specific mechanism that directs the proteolytic activation of TACE through the control of its exit from the ER., This work was supported by grants from the Spanish Comisión Interministerial de Ciencia y Tecnología (SAF2000-0203), Fundació La Marató de TV3, Fondo de Investigación Sanitaria (PI021003) and the EMBO Young Investigator Program (to J. A.), and predoctoral fellowships from the Fundació “la Caixa” and from the Fundació per a la Recerca i Docència dels Hospitals Vall d’Hebron (to M. B.-P. and S. R.-P., respectively).

In the present work, using the pyrene fluorescence response, the kinetic of the cure process at the interface of a composite material is studied. Silica microfibres were surface coated with 3-aminopropyltriethoxysilane (APTES), labelled with the pyrene group and dispersed in a mixture based on diglycidylether of bisphenol A (DGEBA) and ethylenediamine (EDA). The fluorescence of the pyrene was used to monitor, at different temperatures, the cure process at the epoxy/microfibres interface. A simple kinetic model was discussed from the fluorescence data and an apparent activation energy was obtained ( E a =33 kJ/mol). On the other hand, the cure process of the whole mixture was followed by FT-IR and by DSC and their results were compared with those obtained by fluorescence.

Background Approximately 20% of breast cancer (BC) patients (pts) have HER2−positive tumors. While the adjuvant use of the anti-HER2 humanized monoclonal antibody trastuzumab (T) has been shown to improve disease-free (DFS) and overall survival (OS), not all pts treated with this agent benefit from this therapy. Pertuzumab (P) is a humanized monoclonal antibody that inhibits HER2 dimerization and induces ADCC with a complementary mechanism of action to T. In HER2−positiveadvanced BC, T and P is active in pts who have progressed to T. In the neoadjuvant setting, T and P in combination with chemotherapy (CT) nearly doubled the pathological complete response rate compared to either T or P administered in combination with CT (45.8% vs 29% vs 24%, respectively). Therefore, comprehensive HER2 blockade with two anti-HER2 monoclonal antibodies warrants further investigation in the adjuvant setting. Trial Design: APHINITY is a prospective, randomized, multicenter, double-blind, placebo-controlled study in pts with HER2−positive primary BC who have had an excision of their tumor. Pts will be randomized to one of 2 arms (1:1 ratio). The investigational arm will comprise of a course of adjuvant CT (investigators choice) consisting of either an anthracycline-taxane or taxane-platin containing regimens and T and P for 1 year. The comparator arm will consist of the same adjuvant CT backbone with T and placebo for 1 year. Major Eligibility Criteria: 1. Non-metastatic primary BC histologically confirmed and adequately excised 2. Node-positive or node-negative: for patients with node-positive disease (pN ≥1), any pT except T0; for patients with node-negative disease (pN0), tumor size must be >1.0 cm OR for tumor size between >0.5 cm and ≤1.0 cm, at least one of the following features will be required: histologic grade 3 OR negative for ER and PgR OR age 3. The interval between definitive surgery for BC and randomization must be at least 3 weeks but no more than 7 weeks 4. Baseline LVEF ≥55% 5. HER2−positive BC confirmed by a central laboratory and defined as: IHC 3+ in >10% immunoreactive cells OR HER2 gene amplification by in situ hybridization [ISH] (ratio of HER2 gene signals to centromere 17 signals ≥2) Aims: The primary objective is to compare invasive disease-free survival (IDFS) between both treatment arms. Secondary objectives include comparing IDFS including second non-BC, DFS, OS, recurrence-free interval (RFI), distant RFI, cardiac safety, overall safety and health-related quality of life in the two treatment arms. Statistical Methods: Pts will be stratified based on nodal status, type of adjuvant CT regimen, hormone receptor status and geographical region. The study is designed to have an 80% power to test the null hypothesis of no true difference in risk of an IDFS event (HR = 1) versus the alternative hypothesis of a difference (HR = 0.75) in hazard rates with a 5%, 2-sided significance level. Target accrual: 3806; Present accrual: Start Q4 2011 Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr OT1-02-04.

Aim: In CLEOPATRA, 808 pts with HER2-positive MBC were randomized to receive 1L placebo (Pla) + T + D or Ptz + T + D. At primary analysis, pertuzumab was shown to increase progression-free survival significantly, with a strong trend to OS benefit. At a second interim analysis (May 2012), OS was improved to a degree which was both statistically significant and clinically meaningful (HR = 0.66, 95% CI 0.52–0.84; P = 0.0008) but the median OS in pts who received Ptz was not reached. Here we report results of a subsequent prespecified OS analysis. Methods: This OS analysis was planned when ≥385 deaths were reported. The log-rank test, stratified by prior treatment status and geographic region, was used to compare OS between arms, applying the Lan-DeMets a-spending function with an O'Brien-Fleming threshold of p ≤ 0.0456. The Kaplan–Meier approach was used to estimate median OS in both arms; a stratified Cox proportional hazard model was used to estimate HR and 95% CIs. Subgroup analyses of OS were performed for stratification factors and other key baseline characteristics. Results: Median follow-up time was 50 months (mos) and the statistically significant improvement in OS in favor of Ptz + T + D was maintained (HR = 0.68, 95% CI 0.56–0.84; p = 0.0002). Median OS was 40.8 mos in the Pla arm and 56.5 mos in the Ptz arm, the difference at the medians being 15.7 mos. The OS benefit in predefined subgroups was consistent with previous observations. It is to be noted that following the previous report of OS benefit, cross-over therapy was allowed and 48 pts in the Pla arm crossed over to the Ptz arm. The safety profile of Ptz + T + D in the overall population and in pts who crossed over to the Ptz arm was consistent with the known safety profile of Ptz and the long-term cardiac safety profile was maintained. Conclusions: 1L treatment with Ptz + T + D significantly improved OS for pts with HER2-positive MBC compared with Pla + T + D, providing a 15.7 mo increase in the median values. The 56.5 mo median OS is unprecedented in 1L MBC and this substantial improvement confirms the Ptz regimen as 1L standard of care for pts with HER2-positive MBC. Disclosure: S. Swain: acts as an uncompensated consultant for Genentech/Roche. Her institution has received research funding from Genentech/Roche, Pfizer, Puma, Sanofi-Aventis and Bristol-Myers Squibb; S. Kim: has received research funding from Novartis; J. Cortes: has provided consultancy for Novartis, Celgene and Roche and has received honoraria from Novartis, Celgene, Eisai and Roche; M. Campone: has provided consultancy for Novartis/Servier and has received honoraria from Novartis; J. Ferrero: has provided consultancy for Sanofi and has received honoraria from Novartis and Roche; A. Schneeweiss: has provided consultancy for Medac, Celgene and Roche, has received honoraria from Novartis, Celgene, Eisai, Astrazeneca, GlaxoSmithKline, Pfizer and Roche and has received research funding from Roche and Celgene; S. Heeson and G. Ross: is an employee of and owns shares in Roche; E. Clark: is an employee of Roche and owns shares in AstraZeneca; M.C. Benyunes: is an employee of Genentech/Roche; J. Baselga: has provided consultancy for Roche. All other authors have declared no conflicts of interest.

Aim: A phase III randomized, double-blinded, placebo (PLC)-controlled study of sorafenib (S) in combination with capecitabine (C) was conducted in patients with HER2-negative, locally advanced or metastatic breast cancer. The primary endpoint was progression-free survival (PFS) per independent review. Secondary endpoints included overall survival (OS), time to progression (TTP), objective response rate (ORR), disease control rate (DCR) and safety. Methods: Patients resistant to prior taxane and resistant to prior anthracycline or for whom further anthracycline was not indicated, and having received no more than one prior regimen for advanced disease were randomized 1:1 to PLC or S (600 mg daily, po, 21-day schedule) in combination with C (1000 mg/m2, bid, po, 14/7 days on/off). Results: A total of 537 patients were randomized: 266 to S + C and 271 to PLC + C. The two arms were generally well balanced; median age 54 years; 69% hormone receptor positive and 31% triple negative; and 43%/57% having received 0/1 prior lines of chemotherapy for metastatic disease. Median PFS was 5.5 mo for S + C and 5.4 mo for PLC + C; HR 0.973 (95% CI: 0.779, 1.217; one-sided p-value = 0.406). Median OS was 18.9 mo for S + C and 20.3 mo for PLC + C; HR 1.195 (95% CI: 0.943, 1.513; one-sided p-value = 0.930). There were no differences in TTP, ORR or DCR. The incidence of grade 3 AEs was 58.5% vs 39.3%, grade 4 AEs 5.8% vs 4.5%, and grade 5 AEs 6.2% vs 4.5%, S + C vs PLC + C arms, respectively. The most commonly reported grade 3 AEs included palmar-plantar erythrodysesthesia syndrome (16% vs 8%), hypertension (14% vs 2%), fatigue (7% vs 3%) diarrhea (4% vs 6%), neutropenia (4% vs 5%), and anemia (5% vs 2%). The average daily dose for C was 1734 mg/m2 vs 1914 mg/m2 with a median duration of 18 wks vs 23 wks for C in S + C and PLC + C arms, respectively. There was one drug-related grade 5 AE (hepatic failure) reported in the S + C arm. Conclusions: S + C did not improve PFS compared to PLC + C for patients with HER2-negative, locally advanced or metastatic breast cancer. The types of AEs observed were consistent with the known safety profiles of S and C. There was a trend toward worse OS in the S + C arm that is not readily explained by study treatment-related toxicity. Disclosure: P. Gomez: Honoraria: Bayer; B. Melichar: Honoraria: Bayer, Roche; J. Bergh: Clinical research: Bayer, Amgen, Astra-Zeneca, Merck, Pfizer, Roche and Sanofi-Aventis; P. Maeda, L. Huang and J. Zhang: Employment: Bayer HealthCare Pharmaceuticals. All other authors have declared no conflicts of interest.

Aim: The Adjuvant Lapatinib and/or Trastuzumab Treatment Optimisation (ALTTO) Trial is a randomised phase III adjuvant breast cancer trial comparing 3 oral lapatinib (L)-containing regimens with trastuzumab (T), each given for 1 year. DFS results of L + T and T—> L compared to T, presented at ASCO 2014, showed a non-significant DFS benefit. Results of L vs T and key secondary analyses are presented for the first time. Methods: 8381 patients (pts) were randomised to receive either T (N = 2097), L (N = 2100), T —> L (N = 2091), or L + T (N = 2093) from 2007-2011. Primary comparison of L vs T evaluated non-inferiority at a margin of 1.11 and the futility boundary for this arm was crossed resulting in premature arm closure (Aug 2011). Pts in the L arm who were free of disease recurrence at that time were offered T as adjuvant treatment. Key secondary analyses including DFS by hormone receptor status and CNS endpoints are now presented. Results: In 4197 pts and at a median follow-up of 4.5 years, 366 (17%) DFS events in the L and 301 (14%) DFS events in the T arms were observed. 1090 (52%) pts in the L arm received at least one T dose. In post-hoc analysis, there was evidence that these pts benefitted from receiving T [hazard ratio (HR) = 0.67; 95% CI 0.49-0.91 from a time-dependent Cox model of DFS]. The non-inferiority HR was 1.34 (95% CI 1.15-1.56) in the ITT population; 1.45 for hormone receptor negative, 1.23 for hormone receptor positive. CNS as first site of metastases occurred in 2% of cases in all arms. AEs of any grade were more common with L than T: diarrhoea (64% vs 20%), rash or erythema (54% vs 20%) and hepatobiliary (25% vs 16%). Any cardiac events were infrequent in both arms, but more common in T compared to L (4.5% vs 1.9%; p Conclusions: Overall, L added to T did not statistically improve DFS. L did not appear to decrease the rate of CNS as first site of metastases. T confers a better outcome compared to L in pts with HER2-positive EBC and remains the standard of care. There is evidence of T benefit even when initiated later in the course of follow-up. Disclosure: E. De Azambuja: Disclosed an advisory role with GSK; honoraria from Roche; research funding from GSK; and travel grants from GSK/Roche; G. Viale: disclosed an advisory role with Roche and Dako; and honoraria from GSK, Roche, and Dako; R. Crescenzo: disclosed compensated employment with GlaxoSmithKline; and stock ownership with Glaxo SmithKline; K. Pritchard: disclosed an advisory role with Roche; honoraria from Roche; research funding from Roche; and expert testimony on behalf of Roche; C. Jackisch: disclosed honoraria from Roche and GSK; F. Boyle: disclosed honoraria from GlaxoSmithKline; and research funding from GlaxoSmithKline; R.D. Gelber: disclosed research funding from GSK and Roche; M. Piccart: disclosed consultancy with PharmaMar; honoraria from Amgen, Astellas, AstraZeneca, Bayer, Eli Lilly, Invivis, MSD, Novartis, Pfizer, Roche-Genentech, Sanofi Aventis, Symphogen, Synthon, and Verastem; research grants from most companies. All other authors have declared no conflicts of interest.

Large pivotal phase II and III clinical trials investigated the therapeutic efficacy and safety of the humanized anti-HER2 monoclonal antibody, Herceptin, alone and in combination with standard chemotherapy, respectively, in HER2-positive metastatic breast cancer. Eligible patients were HER2 2+ and 3+ overexpressors, as determined by immunohistochemistry (IHC). Herceptin was well tolerated in both trials. Single-agent second/third-line Herceptin produced durable tumor responses. First-line Herceptin in combination with chemotherapy, in particular paclitaxel, significantly improved time to disease progression, duration of response and time to treatment failure. Combination therapy also provided a significant 25% improvement in overall survival. These clinical benefits have led to the approval of Herceptin for clinical use in the USA and elsewhere. Greater efficacy was noted in IHC 3+ patients compared with the overall population in both trials. Retrospective fluorescent in situ hybridization (FISH) testing of patient-tumor HER2 status revealed similar clinical outcomes in IHC 3+ and FISH-positive patients, consistent with the reported high concordance between IHC and FISH. Responses in the single-agent Herceptin trial were seen exclusively in FISH-positive patients. Approximately a quarter of HER2 2+ patients test FISH positive and may therefore benefit from therapy. Numerous studies are underway or planned to evaluate other Herceptin combinations and regimens in the metastatic and adjuvant settings.

The Her2/neu (c-erbB-2) oncogene encodes a 185-kDa protein tyrosine kinase which is overexpressed in 20% of breast adenocarcinomas and is recognized by a humanized anti-Her2/neu monoclonal antibody (mAb) (rhu4D5 or Herceptin). Natural killer (NK) cells are capable of mediating antibody-dependent cell cytotoxicity (ADCC) against antibody-coated targets via their expression of a low-affinity receptor for IgG (FcgammaRIII or CD16). NK cells can be expanded in cancer patients via the administration of low-dose interleukin-2 (IL-2) and become potent cytotoxic effectors following exposure to high doses of IL-2. We tested IL-2-activated NK cells against Her2/neu+ (MCF-7Her2/neu) and Her2/neu- (MDA-468) breast cancer cell lines in a 4-h 51Cr-release cytotoxicity assay in the presence or absence of rhu4D5 mAb (effector : target ratio = 10 : 1). Specific lysis of rhu4D5-coated MCF-7Her2/neu and MDA-468 target cells by IL-2-activated NK cells was 35% and 3%, respectively (p0.05). Lysis was less than 5% when targets were treated with either the non-humanized mu4D5 mAb or control huIgG. Lysis of rhu4D5-coated MCF-7Her2/neu cells was inhibited by 80 % when NK cells were pre-treated with an anti-Fc receptor antibody prior to use in the cytotoxicity assay. Enhanced ADCC of MCF-7Her2/neu target cells was seen when the effector cells consisted of mononuclear cells obtained from a patient demonstrating significant expansion of NK cells secondary to therapy with low-dose IL-2. Serum from patients receiving infusions of rhu4D5 mAb could substitute for exogenous antibody in the ADCC assay. NK cells activated by rhu4D5-coated tumor cells in the presence of IL-2 also produced large amounts of IFN-gamma with concomitant up-regulation of cell-surface activation markers CD25 and CD69. These results lend support to the concurrent use of rhu4D5 mAb and IL-2 therapy in patients with cancers that express the Her2/neu oncogene.

The search for new methods of treating cancer, combined with advances in our understanding of carcinogenesis, molecular biology and technology, has resulted in the development of novel biologic agents with proven clinical efficacy. One such agent is trastuzumab (Herceptin), a humanized monoclonal antibody that targets the human epidermal growth factor receptor-2 (HER2). HER2 is a member of a family of receptors that interact with each other and various ligands to stimulate various intracellular signal transduction pathways involved in cell growth control. HER2 is overexpressed in 20%-30% of women with breast cancer and is associated with aggressive tumor characteristics and poor prognosis. Trastuzumab is the first humanized monoclonal antibody to be approved for therapeutic use and the first oncogene-targeted treatment with proven survival benefit in women with HER2-positive metastatic breast cancer. However, its mechanism of action has not been fully characterized and appears to be complex. This paper reviews current knowledge of the mechanism of action of trastuzumab, including HER2 protein downregulation, prevention of HER2-containing heterodimer formation, initiation of G1 arrest and induction of p27, prevention of HER2 cleavage, inhibition of angiogenesis, and induction of immune mechanisms. The significance of these mechanisms for selection of concomitant chemotherapy is also considered.

The expression of the activated mitogen-activated kinases/extracellular signal-regulated kinases (ERKs) ERK1 and ERK2 was characterized in 101 humanhead and neck squamous carcinoma specimens. Activated ERK1/2were detected at different levels in the majority of these tumors, as assayed by immunostaining with an antibody specific for the dually phosphorylated and activated ERK1 and ERK2. ERK1/2 activation levels were higher in tumors with advanced regional lymph node metastasis (P = 0.048) and in relapsed tumors (P = 0.021). The expression of epidermal growth factor (EGF) receptor (P = 0.037), transforming growth factor alpha (TGF-alpha; P0.001), and HER2 (P = 0.066; positive trend) correlated with activation of ERK1/2. In a multivariate analysis, both TGF-alpha (P0.0001) and HER2 (P = 0.045) were independently correlated with ERK1/2 activation. In turn, activation of ERK1/2 was associated with a higher Ki-67 proliferative index (P = 0.002). In EGF receptor-dependent model cells (A431 and DiFi), a specific EGF receptor tyrosine kinase inhibitor ("Iressa"; ZD1839) and a chimeric anti-EGF receptor antibody ("Cetuximab"; C225) inhibited ERK 1/2 activation at concentrations that inhibited autocrine cell proliferation. In patients on treatment with C225, the activation of ERK1/2 in skin, an EGF receptor-dependent tissue, was lower compared with control skin. Parallel changes were seen in keratinocyte Ki67 proliferation indexes in skin from C225-treated patients. Taken together, these studies provide support for a role of activation of ERK1/2 in head and neck squamous carcinoma and a correlation with EGF receptor/TGF-alpha expression. The inhibition of ERK1/2 activation in vitro and in vivo by compounds targeting the EGF receptor points to the interest of ERK1/2 as potential surrogate markers of EGF-receptor signaling in clinical therapeutic studies.

HER2 is a ligand-less tyrosine kinase receptor of the ErbB family that is frequently overexpressed in breast cancer. It undergoes proteolytic cleavage that results in the release of the extracellular domain and the production of a truncated membrane-bound fragment, p95. We show that HER2 shedding is activated by 4-aminophenylmercuric acetate (APMA), a well-known matrix metalloprotease activator, in HER2-overexpressing breast cancer cells. The HER2 p95 fragment, which appears after APMA-induced cleavage, is phosphorylated. We analyzed 24 human breast cancer specimens, and a phosphorylated M(r) 95,000 HER2 band could be detected in some of them, which indicated that the truncated receptor is also present in vivo. The activation of HER2 shedding by APMA in cells was blocked with batimastat, a broad-spectrum metalloprotease inhibitor. Trastuzumab (Herceptin; Genentech, San Francisco, CA), a humanized monoclonal antibody directed at the HER2 ectodomain, which has been shown to be active in patients with HER2-overexpressing breast cancer, inhibited basal and induced HER2 cleavage and, as a consequence, the generation of phosphorylated p95. This inhibitory effect of trastuzumab was not shared by 2C4, an antibody against a different epitope of the HER2 ectodomain. The inhibition of basal and APMA-induced cleavage of HER2 by trastuzumab preceded antibody-induced receptor down-modulation, which indicated that the effect of trastuzumab on cleavage was not attributable to a decrease in cell-surface HER2 induced by trastuzumab. We propose that the inhibition of HER2 cleavage and prevention of the production of an active truncated HER2 fragment represent a novel mechanism of action of trastuzumab.

Background: Modulation of estrogen synthesis or its receptor activity represents the treatment of choice for postmenopausal women with estrogen receptor-positive (ER+) breast cancer (BC). However, despite initial response to hormonal manipulation, most tumors ultimately develop endocrine resistance. The emerging evidence that ERα can signal in both ligand-dependent and independent manner supports the development of agents that are not only competitive ERα antagonists but also reduce steady state levels of the receptor and thus limit both modes of signaling. ARN-810 is a novel, non-steroidal ERα antagonist that induces proteasomal-mediated degradation of ERα in breast cancer cell lines at picomolar concentrations. This unique biological profile coupled with good oral pharmacokinetics (PK) produced tumor regression in both tamoxifen-sensitive and resistant BC models. Importantly, in the tamoxifen-resistant model, response to ER modulators like fulvestrant and tamoxifen was limited to tumor growth arrest. Trial Design: This is a safety and PK study of ARN-810 administered orally on a continuous daily dosing regimen with a PK lead-in. The incidence of dose limiting toxicity (DLT) will be assessed in the first cycle of treatment (28 days). Patients will be assigned to escalating doses of ARN-810 in cohorts of 3-6 patients per dose until determination of the maximum tolerated dose (MTD) and/or recommended Phase 2 Dose (RP2D) using standard 3+3 criteria. The MTD and/or RP2D will be defined as the dose with ≤1 out of 6 patients with DLT. Key eligibility criteria include postmenopausal women with locally advanced or metastatic ER+ (HER2-) BC, ECOG PS 0/1, and adequate organ function. Untreated or symptomatic brain metastases and endometrial disorders are exclusionary. At the MTD and/or RP2D, expansion cohorts of patients with disease progression after ≤2 prior hormonal therapies (≤1 in the metastatic setting) or prior therapy with fulvestrant will be enrolled. Plasma PK after first dose and at steady-state will be analyzed using non-compartmental methods. Pre- and post-treatment functional imaging with [18F]-fluoroestradiol (FES)-PET to assess tumor ER binding and paired tumor biopsies to assess for ER target modulation by IHC and gene expression will be performed. Objectives: The primary objective is to determine the MTD and/or RP2D of ARN-810. In addition to safety and PK, evaluation of biomarkers of pharmacodynamic (PD) response with FES-PET and ER target gene expression will be explored along with preliminary antitumor activity. Statistical Methods: Descriptive statistics will be used to summarize patient characteristics, treatment administration/compliance, safety and PK parameters, antitumor activity endpoints, and exploratory biomarkers. Status: To date, 3 patients have been enrolled in the first cohort at 100 mg/day. The target accrual for the dose escalation will depend upon the observed safety and PK/PD profile, which will determine the number of escalations. During dose expansion, 30 patients will be enrolled. More information about this study can be found at http://clinicaltrials.gov/ct2/show/NCT01823835?term = ARN-810. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr OT3-2-07.

Background: The NeoALTTO study demonstrated a significantly higher breast pathological complete response (pCR) rate (NSABP definition – ypT0/is) with dual HER2 blockade using lapatinib and trastuzumab compared with single HER2 blockade using either lapatinib or trastuzumab (51.3% vs. 24.7% vs. 29.5%, respectively; p< 0.01 for both) in HER2 positive primary breast cancer (BC). A similar pattern was seen for locoregional pCR (ypT0/is ypN0) (Baselga J et al. Lancet 2012). Material and Methods: From January, 2008, to December, 2009, 455 patients were randomized from 99 participating sites in 23 countries. Patients were randomized to receive either lapatinib 1500 mg/d (154 pts), or trastuzumab 4 mg/kg IV loading dose followed by 2 mg/kg IV weekly (149 pts), or lapatinib 1000 mg/d with trastuzumab for a total of 6 weeks (152 pts). After this biological window, patients continued on the same targeted therapy plus weekly paclitaxel 80 mg/m2 for a further 12 weeks, until definitive surgery (total neoadjuvant therapy duration of 18 weeks). After surgery, patients received 3 cycles of adjuvant FEC followed by the same targeted therapy as in the biological window of the neoadjuvant phase for a further 34 weeks (to complete 52 weeks of anti-HER2 therapy), with on-going follow-up planned until 10 years after last randomised patient. Secondary objectives included disease-free survival and overall survival (OS). Following current practice and draft FDA recommendations for neoadjuvant trial endpoints, an amendment was made to the protocol secondary objectives (released in May 2013). DFS (from surgery) was replaced by event-free survival (EFS) from randomisation; OS from surgery was correspondingly replaced with OS from randomisation, and examination of the association between these survival endpoints and locoregional pCR (ypT0/is ypN0) was added as a secondary objective. EFS was defined as the time from randomization to first EFS event (breast cancer relapse, second primary malignancy or death without recurrence). OS was defined as the time from randomization to death from any cause. Results: The clinical cut-off date for the first planned analysis of these secondary objectives was on 26 May 2013, three years after the date of last surgery. Data cleaning is ongoing and analysis will be completed by November 2013. Results for EFS, OS and their association with pCR will be presented at the meeting. Funding: GlaxoSmithKline is the sponsor of this trial. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr S1-01.

In summary, monoclonal antibodies directed at two of the class I growth factor receptors, EGFR and HER2, have shown to be active and well tolerated. These findings represent proof of concept that the use of monoclonal antibodies against growth factor receptors is a valid and promising approach in the therapy of patients with cancer. Current research goals include to define the optimal combinations with conventional chemotherapeutic agents and with radiation therapy, determine the best therapy candidates and to expand clinical trials to other tumour types.

Trastuzumab (Herceptin; Genentech, Inc, So. San Francisco, CA) is a high-affinity, humanized anti-HER2 antibody that has shown benefit in the therapy of patients with metastatic HER2-overexpressing breast cancer. Results from initial clinical trials of trastuzumab as a single agent or in combination with chemotherapy established important guidelines for the therapy of HER2-positive tumors, but represent an early phase of clinical development. Preclinical studies have shown additive and synergistic effects of trastuzumab when given in combination with several chemotherapeutic agents, and a series of clinical trials exploring these new combinations is under way or will be started shortly. Substantial effort will be directed toward promising schedules and combinations, such as weekly paclitaxel with trastuzumab or combinations of platinum, taxanes, and trastuzumab. Due to the unexpected cardiac toxicity observed with the combination of doxorubicin plus trastuzumab, new studies will be designed to prospectively assess cardiac function and will incorporate anthracyclines with less cardiotoxicity, such as liposomal doxorubicin. Combination studies are not limited to cytotoxic agents, as laboratory and clinical data have demonstrated that HER2 overexpression results in resistance to hormonal therapy. Therefore, a series of studies combining hormonal treatments with trastuzumab is being considered. Finally, the integration of trastuzumab into the adjuvant and neoadjuvant settings will be studied by United States and European cooperative groups.

The HER2 gene (also known as neu and as c-erb-B2) encodes a 185-kd transmembrane glycoprotein receptor with intrinsic tyrosine kinase activity. HER2 is overexpressed in 25% to 30% of human breast cancers, plays a role in the pathogenesis of breast cancer, and predicts for a worse prognosis in patients with metastatic disease. Trastuzumab (Herceptin; Genentech, Inc, So. San Francisco, CA), a humanized monoclonal antibody that targets the HER2 oncogene receptor, was shown to be active in preclinical models. In initial phase I clinical trials, trastuzumab was found to be safe and to exhibit dose-dependent pharmacokinetics. Three phase II studies of single-agent trastuzumab, which was administered weekly in the outpatient setting, have now been conducted in patients with HER2-overexpressing metastatic breast cancer. In the initial phase II study, the response rate was 11% in a heavily pretreated patient population. In a pivotal follow-up study of single-agent trastuzumab, more than 200 patients who had received at least one prior chemotherapeutic regimen for metastatic disease were entered. Despite a number of unfavorable baseline characteristics, the response rate reported by an independent response evaluation committee was 15%. A more recent study in previously untreated patients has shown a 23% response rate. The median duration of response in these trials has ranged from 6.6 to 9.1 months. In these three phase II studies, trastuzumab has been shown to be safe. The most clinically significant adverse event has been cardiac dysfunction syndrome, which occurred in less than 5% of patients. Trastuzumab is not associated with the other commonly observed side effects of chemotherapy, such as alopecia, mucositis, and neutropenia. The results from these studies demonstrate that trastuzumab is active and safe in patients with metastatic HER2-overexpressing breast cancer.

Systemic oncologic therapies can cause multiple emergency situations. There are, however, two unique emergencies directly related to chemotherapy administration: drug extravasation and hypersensitivity reactions (HSRs). Most drugs can cause varying degrees of local tissue injury when extravasated. The medical management of extravasation is based on proper maintenance of the intravenous line, application of local cooling or warming for certain extravasations, and the use of antidotes to prevent the local toxic action of the extravasated drug. Antidotes that appear useful include hyaluronidase (for vinca alkaloids, epipodophyllotoxins, and paclitaxel), sodium thiosulfate (for mechlorethamine and cisplatin), and dimethylsulfoxide (DMSO) (for anthracyclines and mitomycin C). HSRs, another potential adverse effect of chemotherapy administration, are a major concern for therapy with taxanes and with L-asparaginase, and their administration requires the use of premedication to prevent these reactions. Once a HSR has occurred, therapy may be continued by using an analog drug or by the administration of premedication as prophylaxis, in particular if the reaction was minor. On the other hand, it is also pertinent to become acquainted with the emergencies induced by biological agents, taking into consideration their increasing usages. In addition to interferons and interleukin-2 (IL-2), both of which have been in clinical use for several years, cytokine-toxin fusion proteins (DAB3891L-2) and two monoclonal antibodies (rituximab and trastuzumab) were recently approved for cancer therapy. The distinct toxicity profiles of these agents are reviewed.

Breast carcinomas express high levels of ErbB receptors and their ligands, and their overexpression has been associated with a more aggressive clinical behavior. For these reasons therapies directed at these receptors have the potential to be useful anti-cancer treatments. A series of monoclonal antibodies (MAbs)3 directed against the EGF (ErbB1) receptor and the closely related HER2/Neu (ErbB2) receptor are currently under evaluation. These MAbs have shown promising preclinical activity and "chimeric" and "humanized" MAbs have been produced in order to obviate the problem of host immune reactions. These antibodies are currently being tested in clinical trials either alone or in combination with chemotherapeutic agents. Clinical activity with one of these antibodies, trastuzumab, a humanized anti-ErbB2 MAb, has been documented in patients with breast cancer in a series of clinical trials and has recently been approved for the therapy of patients with metastatic ErbB2 overexpressing breast cancer. In addition to antibodies, compounds that inhibit receptor tyrosine kinases have shown significant preclinical activity and are currently being evaluated in the clinic.