Predictive modelling of response to neoadjuvant therapy in HER2+ breast cancer.

HER2-positive (HER2+) breast cancer accounts for 20-25% of all breast cancers. Predictive biomarkers of neoadjuvant therapy response are needed to better identify patients with early stage disease who may benefit from tailored treatments in the adjuvant setting. As part of the TCHL phase-II clinical trial (ICORG10-05/NCT01485926) whole exome DNA sequencing was carried out on normal-tumour pairs collected from 22 patients. Here we report predictive modelling of neoadjuvant therapy response using clinicopathological and genomic features of pre-treatment tumour biopsies identified age, estrogen receptor (ER) status and level of immune cell infiltration may together be important for predicting response. Clonal evolution analysis of longitudinally collected tumour samples show subclonal diversity and dynamics are evident with potential therapy resistant subclones detected. The sources of greater pre-treatment immunogenicity associated with a pathological complete response is largely unexplored in HER2+ tumours. However, here we point to the possibility of APOBEC associated mutagenesis, specifically in the ER-neg/HER2+ subtype as a potential mediator of this immunogenic phenotype.

Pilot study of bevacizumab in combination with docetaxel and cyclophosphamide as adjuvant treatment for patients with early stage HER-2 negative breast cancer, including analysis of candidate circulating markers of cardiac toxicity: ICORG 08-10 trial.

BACKGROUND: Combining bevacizumab and chemotherapy produced superior response rates compared with chemotherapy alone in metastatic breast cancer. As bevacizumab may cause hypertension (HTN) and increase the risk of cardiac failure, we performed a pilot study to evaluate the feasibility and toxicity of a non-anthracycline-containing combination of docetaxel with cyclophosphamide and bevacizumab in early stage breast cancer patients. METHODS: Treatment consisted of four 3-weekly cycles of docetaxel and cyclophosphamide (75/600 mg/m2). Bevacizumab was administered 15 mg/kg intravenously on day 1, and then every 3 weeks to a total of 18 cycles of treatment. Serum biomarker concentrations of vascular endothelial growth factor (VEGF), cardiac troponin-I (cTnI), myeloperoxidase (MPO), and placental growth factor (PlGF) were quantified using enzyme-linked immunosorbent assay (ELISA) in 62 patients at baseline and whilst on treatment to determine their utility as biomarkers of cardiotoxicity, indicated by left ventricular ejection fraction (LVEF). RESULTS: A total of 106 patients were accrued in nine sites. Median follow up was 65 months (1-72 months). Seventeen protocol-defined relapse events were observed, accounting for an overall disease-free survival (DFS) rate of 84%. The DFS rates for hormone receptor positive (HR+) and triple-negative (TN) patients were 95% versus 43%, respectively. The median time to relapse was 25 (12-54) months in TN patients versus 38 (22-71) months in HR+ patients. There have been 13 deaths related to breast cancer . The overall survival (OS) rate was 88%. The 5-year OS rate in HR+ versus TN was 95% versus 57%. None of the measured biomarkers predicted the development of cardiotoxicity. CONCLUSIONS: We observed a low relapse rate in node-positive, HR+ patients; however, results in TN breast cancer were less encouraging. Given the negative results of three large phase III trials, it is unlikely that this approach will be investigated further. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT00911716.

Impact of somatic PI3K pathway and ERBB family mutations on pathological complete response (pCR) in HER2-positive breast cancer patients who received neoadjuvant HER2-targeted therapies.

BACKGROUND: The Cancer Genome Atlas analysis revealed that somatic EGFR, receptor tyrosine-protein kinase erbB-2 (ERBB2), Erb-B2 receptor tyrosine kinase 3 (ERBB3) and Erb-B2 receptor tyrosine kinase 4 (ERBB4) gene mutations (ERBB family mutations) occur alone or co-occur with somatic mutations in the gene encoding the phosphatidylinositol 3-kinase (PI3K) catalytic subunit (PIK3CA) in 19% of human epidermal growth factor receptor 2 (HER2)-positive breast cancers. Because ERBB family mutations can activate the PI3K/AKT pathway and likely have similar canonical signalling effects to PI3K pathway mutations, we investigated their combined impact on response to neoadjuvant HER2-targeted therapies. METHODS: Baseline tumour biopsies were available from 74 patients with HER2-positive breast cancer who were enrolled in the phase II TCHL neoadjuvant study (ICORG 10-05) assessing TCH (docetaxel, carboplatin, trastuzumab) (n = 38) versus TCL (docetaxel, carboplatin, lapatinib) (n = 10) versus TCHL (docetaxel, carboplatin, trastuzumab, lapatinib) (n = 40), each for six cycles. Activating mutations in PIK3CA and ERBB family genes were identified using mass spectrometry-based genotyping. Phosphatase and tensin homolog (PTEN) expression was assessed by immunohistochemistry. RESULTS: PIK3CA and/or ERBB family mutations were detected in 23 (31.1%) tumour samples tested, whereas PTEN expression was low in 31.1% of cases tested. Mutation frequency was similar in each treatment arm (31.3% in TCH arm, 30% in TCL arm and 31.3% in TCHL arm) and was not influenced by oestrogen receptor (ER) status (27.6% in ER-negative patients, 33.3% in ER-positive patients) or progesterone receptor (PR) status (32.6% in PR-negative patients, 29% in PR-positive patients). There was no significant difference in pathological complete response (pCR) rates between 47 patients with wild-type (WT) tumours and 22 patients whose tumours carried mutations (in either PIK3CA or ERBB family genes) (42.5% vs. 54.5%; p = 0.439). Similarly, there was no significant difference in pCR rates between patients with PIK3CA/ERBB family mutated/PTEN-low (i.e., PI3K-activated) tumours and patients without PI3K activation (50% vs. 44%; p = 0.769). However, in the TCHL (but not the TCH) group, the pCR rate was higher for 9 patients with PIK3CA/ERBB family mutated tumours than for 20 patients with PIK3CA/ERBB family WT tumours (77.8% vs. 35%; p = 0.05). CONCLUSIONS: Our results indicate that patients who receive neoadjuvant TCHL and have PIK3CA/ERBB family mutated tumours may be more likely to have a pCR than patients with WT tumours. TRIAL REGISTRATION: ClinicalTrials.gov, NCT01485926 . Registered on 2 December 2011.

Temporal patterns of late bowel and bladder radiotherapy toxicity in a randomised controlled trial assessing duration of neo-adjuvant hormones in prostate cancer.

BACKGROUND: To assess the temporal patterns of late gastrointestinal (GI) and genitourinary (GU) radiotherapy toxicity and resolution rates in a randomised controlled trial (All-Ireland Cooperative Oncology Research Group 97-01) assessing duration of neo-adjuvant (NA) hormone therapy for localised prostate cancer. MATERIAL AND METHODS: Node negative patients with > 1 of: PSA > 20 ng/mL, Gleason score ≥ 7, and stage T3 or more, were included. Follow-up, including toxicity assessment, was three-monthly in the early stages and yearly thereafter. RESULTS: Median follow-up from the end of RT was 6.8 years. In the interval between 90 days following the end of RT and the last toxicity assessment, GI and GU toxicity (any grade) was found in 50% and 51% of 240 and 241 patients, respectively. For those who did develop toxicity, the median time from end of RT until the first development of any grade GI or GU toxicity was 1.2 years and 1.6 years, respectively, whilst median time to final resolution was 1.6 years and 2.2 years, respectively. Grade 2 (G2) or greater GI and GU toxicity occurred in 29 (12.1%) and 40 (16.6%) patients, respectively. The proportion with unresolved G2 + GI and GU toxicity was 89% and 79%, respectively, in year 1, 69% and 65% in year 2, 59% and 52% in year 3 and 27% and 32% in year 5. CONCLUSION: Long-term toxicities continue to occur many years after NA hormone therapy and RT. The rate of occurrence does not appear to reduce within the time frame during which our patients were followed. The percentage of patients suffering from G2 + toxicity at any time is however low. Resolution of these toxicities continues for the duration of the follow-up.

Phase II clinical trial of first or second-line treatment with bortezomib in patients with malignant pleural mesothelioma.

Based on promising preclinical efficacy of bortezomib in mesothelioma, a single-arm phase II trial (Ireland Cooperative Oncology Research Group 05-10 study), with Simon's two-stage design, was undertaken to assess efficacy of bortezomib monotherapy in the first-line (poor performance status) and second-line settings. The Bcl-2 homology domain 3-only protein Noxa has been implicated as a key inducer of apoptosis by bortezomib. Thus, in a biomarker research substudy, we hypothesized that deficiency in Noxa expression might correlate with resistance. In the second-line setting, 23 patients were enrolled. Partial response was confirmed in one patient (4.8%) who received four cycles of bortezomib. One patient had stable disease; however, progression occurred in the majority of patients within the first two cycles. Median progression-free survival and overall survival were 2.1 and 5.8 months, respectively. In the first-line setting, ten patients were accrued, and there was no evidence of objective response. In the tumor analysis, expression of Noxa was seen in all biopsies. Bortezomib monotherapy exhibits insufficient activity to warrant further investigation in unselected patients with mesothelioma.

Milling-induced disorder of pharmaceuticals: one-phase or two-phase system?

During milling, components are subjected to shear and tensile stresses, which can result in physical phase transformations. The purpose of the work described in this report is to understand the pathway by which two test compounds, d-salicin and γ-indomethacin, undergo a crystalline to amorphous transformation during cryomilling. The results show that the transformation cannot be described by a standard one-phase or two-phase disordering mechanism. In the one-phase model, a continuous set of states exist, linking perfect crystalline with completely amorphous material, whereas the two-phase model of disorder depicts the material as a binary mixture of crystalline and amorphous fractions. Instead, a model is proposed where two one-phase regions, defected crystalline and amorphous regions, are separated by a distinct transition.

Single-molecule magnets: a family of MnIII/CeIV complexes with a [Mn8CeO8]12+ core.

Four heterometallic, enneanuclear Mn8Ce clusters [Mn8CeO8(O2CMe)12(H2O)4] (4), [Mn8CeO8(O2CMe)12(py)4] (5), [Mn8CeO8(O2CPh)12(MeCN)4] [Mn8CeO8(O2CPh)12(dioxane)4] (6), and [Mn8CeO8(O2CCHPh2)12(H2O)4] (7) have been prepared by various methods. Their cores are essentially isostructural and comprise a nonplanar, saddlelike [MnIII8O8]8+ loop containing a central CeIV ion attached to the eight micro3-O2- ions. Peripheral ligation around the [Mn8CeO8]12+ core is provided by eight micro- and four micro3-O2CR- groups. Terminal ligation on four MnIII atoms is provided by H2O in 4 and 7, pyridine in 5, and MeCN/dioxane in 6. Solid-state magnetic susceptibility studies, fits of dc magnetization vs field and temperature data, and in-phase ac susceptibility studies in a zero dc field have established that complexes 4, 5, and 7 possess S=16, S=4 or 5, and S=6+/-1 spin ground states, respectively, but in all cases there are very low-lying excited states. The large variation in the ground-state spins for this isostructural family is rationalized as due to a combination of weak exchange interactions between the constituent MnIII atoms, and the presence of both nearest-neighbor and next-nearest-interactions of comparable magnitudes. Magnetization vs applied dc field sweeps on single crystals of 4.4H2O and 7.4H2O.3MeCN.2CH2Cl2 down to 0.04 K have established that these two complexes are new single-molecule magnets (SMMs). The former also shows an exchange-bias, a perturbation of its single-molecule properties from very weak intermolecular interactions mediated by hydrogen-bonding interactions with lattice-water molecules of crystallization.

Employment of 2,6-diacetylpyridine dioxime as a new route to high nuclearity metal clusters: Mn6 and Mn8 complexes.

The employment of the anion of 2,6-diacetylpyridine dioxime (dapdoH2) as a pentadentate chelate in transition metal cluster chemistry is reported. The syntheses, crystal structures, and magnetochemical characterization are described for [Mn6O2(OMe)2(dapdo)2(dapdoH)4](ClO4)2 (1), [Mn6O2(OMe)2(dapdo)2(dapdoH)4][Ca(NO3)4] (2), and [Mn8O4(OH)4(OMe)2(N3)2(dapdo)2(dapdoH)2(H2O)2] (3). The reaction of [Mn3O(O2CMe)6(py)3](ClO4) with 3 equiv of dapdoH2 (with or without 2 equiv of NEt3) in MeOH gave 1. The same cation, but with a [Ca(NO3)4]2- anion, was found in complex 2, which was obtained from the reaction in MeOH between Mn(NO3)2, Ca(NO3)2, and dapdoH2 in the presence of NEt3. In contrast, addition of NaN3 to several reactions comprising MnCl2, dapdoH2, and NEt3 in MeOH gave the octanuclear complex 3. Complexes 1-3 all possess rare topologies and are mixed-valence: 2MnII, 4MnIII for 1 and 2, and 2MnII, 6MnIII for 3. The core of the cation of 1 and 2 consists of two edge-sharing Mn4 tetrahedra at the center of each of which is a micro4-O2- ion. Peripheral ligation is provided by two micro-OMe-, four micro-dapdoH-, and two micro3-dapdo2- groups. The core of 3 consists of two [MnIIMnIII3(micro3-O)2]7+ "butterfly" units linked together by one of the micro3-O2- ions, which thus becomes micro4. Peripheral ligation is provided by four micro-OMe-, two micro-OH-, two micro-dapdoH-, and two micro4-dapdo2- groups. Variable-temperature, solid-state dc and ac magnetization studies were carried out on complexes 1-3 in the 5.0-300 K range; the data for 1 and 2 are identical. Fitting of the obtained magnetization versus field (H) and temperature (T) data by matrix diagonalization and including only axial anisotropy (zero-field splitting, D) established that 1 possesses an S=5 ground state with D=-0.24 cm(-1). For 3, low-lying excited states precluded obtaining a good fit from the magnetization data, and the ground state was instead determined from the ac data, which indicated an S=1 ground state for 3. The combined work demonstrates the ligating flexibility of pyridyl-dioxime chelates and their usefulness in the synthesis of new polynuclear Mnx clusters without requiring the co-presence of carboxylate ligands.

Coordination polymer gels: synthesis, structure and mechanical properties of amorphous coordination polymers.

The synthesis and mechanical properties of a new class of coordination polymer based materials, amorphous coordination polymer sol-gels and xerogels, is presented.

Supramolecular medicinal chemistry: mixed-ligand coordination complexes.

We report on the supramolecular synthesis and characterization of examples from three series of mixed-ligand coordination complexes composed of copper(II), a drug, and an ancillary ligand (AL). Particularly, we demonstrate how the judicious choice of an ancillary ligand affords a large degree of control over the relative lipophilicity/hydrophilicity of the complex in relation to the uncomplexed drug molecule. Furthermore, we demonstrate several important factors to consider in the design of such complexes such as the additive-constitutive nature of the partition coefficient of the ancillary ligand and the relative size of the two types of ligands.

Template synthesis and single-molecule magnetism properties of a complex with spin S = 16 and a [Mn8O8]8+ saddle-like core.

The compound [CeIVMnIII8O8(O2CMe)12(H2O)4].4H2O (1.4H2O) has been obtained from a template synthesis involving the reaction of the chain polymer {[MnIII(OH)(O2CMe)2] .(MeCO2H).(H2O)}n (3) with Ce(IV). Compound 1 contains a MnIII8 loop inside which is held the Ce(IV) ion by the bridging oxide ions. Magnetization and magnetic susceptibility studies establish that 1 has an S = 16 spin ground state, the largest yet for a Mn cluster, and displays the slow magnetization relaxation and hysteresis behavior of a single-molecule magnet (SMM). It is thus the highest spin Mn SMM discovered to date.

A new 6(5).8 topology and a distorted 6(5).8 CdSO4 topology: two new supramolecular isomers of [M2(bdc)2(L)2]n coordination polymers.

We report the crystal structures of two new coordination polymers that have similar composition but, despite having the same circuit symbol and Schläfli notation, different topologies: a novel 3D topology (USF-1) and a CdSO4-like topology.

Crystal engineering of novel cocrystals of a triazole drug with 1,4-dicarboxylic acids.

Cocrystals of the poorly soluble antifungal drug cis-itraconazole (1) with 1,4-dicarboxylic acids have been prepared. The crystal structure of the succinic acid cocrystal with 1 was determined to be a trimer by single-crystal X-ray. The trimer is comprised of two molecules of 1 oriented in antiparallel fashion to form a pocket with a triazole at either end. The extended succinic acid molecule fills the pocket, bridging the triazole groups through hydrogen-bonding interactions rather than interacting with the more basic piperazine nitrogens. The solubility and dissolution rate of some of the cocrystals are approximately the same as those of the amorphous drug in the commercial formulation and are much higher than those for the crystalline free base. The results suggest that cocrystals of drug molecules have the possibility of achieving the higher oral bioavailability common for amorphous forms of water-insoluble drugs while maintaining the long-term chemical and physical stability that crystal forms provide.

Exciplex fluorescence of ([Zn(bipy)1.5(NO3)2)].CH3OH.0.5pyrene)n: a coordination polymer containing intercalated pyrene molecules (bipy = 4,4'-bipyridine).

We report the first use of fluorescence spectroscopy to probe the environment of the cavities that are present in open framework coordination polymers.

Supramolecular isomerism in coordination compounds: nanoscale molecular hexagons and chains.

A nanoscale supramolecular hexagon, 1, and its supramolecular isomeric chain structure, 2, have been prepared from self-assembly of 5-NO2-bdc and Cu(II) cations. The hexagon is neutral and soluble and has outer and inner diameters of 3.1 and 0.8 nm, respectively.

A new supramolecular isomer of [Zn(nicotinate)2]n: a novel 4(2).8(4) network that is the result of self-assembly of 4-connected nodes.

Self-assembly of Zn(NO3)2 with m-pyridinecarboxylate (nicotinate) under mild conditions affords several products including a novel three-dimensional network [Zn(nicotinate)2]n that has connectivity defined by the circuit symbol 4(2).8(4).