Phase I pharmacological and bioavailability study of oral diflomotecan (BN80915), a novel E-ring-modified camptothecin analogue in adults with solid tumors.

PURPOSE: Diflomotecan (BN80915) is an E-ring modified camptothecin analogue that possesses greater lactone stability in plasma compared with other topoisomerase I inhibitors, a potential advantage for antitumor activity. As with other camptothecins, oral administration has pharmacological and clinical advantages. This Phase I study was performed to assess the feasibility of the administration of oral diflomotecan, to determine the maximum-tolerated, dose its bioavailability, and to explore the pharmacokinetics. EXPERIMENTAL DESIGN: An initial i.v. bolus was administered to assess the bioavailability of diflomotecan. Fourteen days later, diflomotecan was administered p.o. once daily for 5 days to adult patients with solid malignant tumors and repeated every 3 weeks. BN80915 and its open lactone form BN80942 were measured. RESULTS: Twenty-two patients entered the study and received a total of 57 cycles of oral diflomotecan at flat dose levels of 0.10, 0.20, 0.27, and 0.35 mg. The main toxicity was hematological, but some patients experienced alopecia, mild gastrointestinal toxicity, and fatigue. At the 0.35-mg dose level, 2 of 4 patients experienced dose-limiting toxicity comprising grade 3 thrombocytopenia with epistaxis and febrile neutropenia in 1 patient and uncomplicated grade 4 neutropenia lasting for >7 days in another. Toxicity was acceptable at the 0.27-mg dose level at which dose-limiting toxicities were observed in 3 of 12 patients (grade 4 neutropenia > 7 days, complicated by fever in 1 patient but without other signs of infection). After two cycles of diflomotecan, 6 patients had disease stabilization, which was maintained in 2 patients for 9 months and >1 year, respectively. Diflomotecan pharmacokinetics were linear over the dose range studied. Systemic exposure correlated with the fall in WBC counts. The mean oral bioavailability (+/-SD) was 72.24 +/- 59.2% across all dose levels. Urinary excretion of BN80915 was very low. CONCLUSIONS: The recommended oral diflomotecan dose for Phase II studies is 0.27 mg/day x 5 every 3 weeks. This regimen is convenient and generally well tolerated with a favorable pharmacokinetic profile and high but variable bioavailability.     

INAUGURAL ARTICLE by a Recently Elected Academy Member:Sustained stoichiometric imbalance and its ecological consequences in a large oligotrophic lake

Considerable attention is given to absolute nutrient levels in lakes, rivers, and oceans, but less is paid to their relative concentrations, their nitrogen:phosphorus (N:P) stoichiometry, and the consequences of imbalanced stoichiometry. Here, we report 38 y of nutrient dynamics in Flathead Lake, a large oligotrophic lake in Montana, and its inflows. While nutrient levels were low, the lake had sustained high total N: total P ratios (TN:TP: 60 to 90:1 molar) throughout the observation period. N and P loading to the lake as well as loading N:P ratios varied considerably among years but showed no systematic long-term trend. Surprisingly, TN:TP ratios in river inflows were consistently lower than in the lake, suggesting that forms of P in riverine loading are removed preferentially to N. In-lake processes, such as differential sedimentation of P relative to N or accumulation of fixed N in excess of denitrification, likely also operate to maintain the lake’s high TN:TP ratios. Regardless of causes, the lake’s stoichiometric imbalance is manifested in P limitation of phytoplankton growth during early and midsummer, resulting in high C:P and N:P ratios in suspended particulate matter that propagate P limitation to zooplankton. Finally, the lake’s imbalanced N:P stoichiometry appears to raise the potential for aerobic methane production via metabolism of phosphonate compounds by P-limited microbes. These data highlight the importance of not only absolute N and P levels in aquatic ecosystems, but also their stoichiometric balance, and they call attention to potential management implications of high N:P ratios.

The emergence of the Anthropocene era has been marked by major changes in all of Earth’s major biogeochemical cycles (1). For example, fluxes of carbon (C) (as CO₂) to the atmosphere have increased by ∼14% during the last 120 y largely due to fossil fuel combustion. Fluxes of nitrogen (N) into the biosphere have increased by at least 100% due to application of the Haber-Bosch reaction for fertilizer production, land use change favoring N-fixing legumes, and conversion of atmospheric N₂ to available forms (NO_x) by high temperature combustion of petroleum and fossil gas (2). Finally, large-scale mining of phosphorus (P)-rich geological deposits for production of fertilizers has amplified rates of P cycling in the biosphere by ∼400% (1). Each of these perturbations has biophysical and ecological impacts at differing time and space scales. For C, its accumulation in the atmosphere has altered Earth’s radiative balance, warming the planet and perturbing precipitation patterns globally. Amplified inputs of reactive N to the Earth system enter the hydrosphere and, thus, potentially lead to overenrichment of lakes, rivers, and coastal oceans across broad regions. Amplifications of P inputs often impair water quality at watershed and local scales (3), stimulating phytoplankton production and contributing, along with N, to harmful algal blooms, fish kills, and “dead zones” (4, 5). These differential amplifications and their contrasting spatial scales indicate that ecosystems are experiencing not only absolute changes in biogeochemical cycling, but also perturbations in the relative inputs and outputs of biologically important elements (6). Studies of elemental coupling and uncoupling in ecosystems are not yet widespread, but emerging work has shown how C, N, and P are differentially processed as they pass through watersheds (7).The potential for differential alteration in supplies of N and P to aquatic ecosystems suggests that understanding the nutrient status of a water body requires knowledge of not only absolute supplies of limiting nutrients, but also their relative proportions (i.e., their N:P stoichiometry). This work has been facilitated in recent years by the emergence of the theory of ecological stoichiometry (8). For example, seminal work by Redfield (9) found that N:P ratios in marine organic matter were tightly constrained around 16:1 (molar, here and throughout), a value that may represent the central tendency for the N:P ratio of phytoplankton undergoing balanced growth in which major pools of N (protein) and P (RNA) are produced at the same rate (10). In lakes, N:P ratios show much wider variation—around a value of ∼30—perhaps reflecting the biogeochemical connections of lakes to terrestrial systems where N:P ratios have a similar value and range of variation (11). Nevertheless, this classic “Redfield ratio” of 16:1 can be thought of as representing a balanced nutrient supply for primary producers in pelagic ecosystems. When the ratios of N and P supplied deviate from this balanced ratio, primary limitation of growth by N (when N:P is low) or by P (when N:P is high) can occur. For example, phytoplankton growth in lakes with imbalanced total N: total P (TN:TP) ratios that exceed 30:1 is generally P limited (12). Disproportionate inputs of N relative to P from atmospheric deposition can increase lake TN:TP ratios and shift lake phytoplankton from N to P limitation (13), inducing P limitation in zooplankton (14). Imbalanced N:P ratios in nutrient supplies can also shift the competitive advantage among phytoplankton and enhance production of potentially toxic compounds during harmful algal blooms. For example, skewed supplies of N relative to P can increase production of N-rich secondary compounds by phytoplankton, while disproportionate inputs of P relative to N can induce production of C-based toxins (15). High N:P ratios can also enhance proliferation of fungal parasites of phytoplankton (16).Imbalanced N:P ratios can impact aquatic ecosystems in other ways. For example, they can alter the functioning of food webs. In particular, shifts in nutrient supply regimes that enhance P limitation can impede energy flow in trophic interactions because biomass of P-limited primary producers is of low quality for animals due to its low P content (8). Ecosystem shifts to high N:P ratios and more prevalent P limitation can also impact the cycling of the greenhouse-active gas methane (CH₄) because phosphate limitation can result in production of methane under aerobic conditions in both marine and freshwater phytoplankton and bacteria (17, 18). Both chemoheterotrophic and photoautotrophic bacteria (e.g., Pseudomonas, SAR11, Trichodesmium, Synechococcus) can metabolize organic P compounds, called phosphonates, to acquire P. Microbial cleavage of one type of phosphonate, methylphosphonic acid (MPn), to acquire P results in formation of methane (17). While it is likely that anaerobic methane production due to oxygen depletion in response to P-driven eutrophication is the dominant process connecting P to methane dynamics, the significance of aerobic phosphonate metabolism to global methane cycles remains to be assessed. However, contributions are potentially large, given the prevalence of P limitation in both freshwater and marine ecosystems. In light of emerging trends that suggest overall increases in ecosystem N:P ratios due to human impacts (6), these trophic and biogeochemical impacts of stoichiometric imbalance show that it is critical to consider not only absolute levels of nutrients, but also their stoichiometry. In particular, high N:P ratios can accentuate P limitation, causing a suite of ecological impacts that, currently, are poorly described.In this paper, we illustrate the utility of stoichiometric approaches by combining analyses of long-term records of nutrient supply and dynamics, together with contemporary experiments, to examine how imbalances in N:P stoichiometry (e.g., strong divergence from classic Redfield proportions) influence plankton ecology and biogeochemistry across multiple scales in Flathead Lake, a large lake in western Montana. The lake is itself relatively unperturbed by human impacts and, thus, maintains low overall nutrient levels. However, the strong stoichiometric imbalance that we describe makes Flathead Lake appropriate for assessing ecosystem consequences of what appear to be general trends of increasing N:P ratios in global ecosystems (6). Numerous limnological properties of the lake and its inflow rivers have been monitored continuously for several decades, including concentrations of various forms of N and P. Thus, these time-series data allow us not only to assess long-term variability or stability in the stoichiometry of N and P in the lake and its river inflows over decadal time scales, but also to connect its stoichiometry with potential consequences for nutrient limitation, food web dynamics, and biogeochemical cycling under low-nutrient conditions.     

Deconvoluting the Obesity and Breast Cancer Link: Secretome, Soil and Seed Interactions

Obesity is associated with increased risk of breast cancer in postmenopausal women and is linked with poor prognosis in pre- and postmenopausal breast cancer patients. The mechanisms underlying the obesity-breast cancer connection are becoming increasingly clear and provide multiple opportunities for primary to tertiary prevention. Several obesity-related host factors can influence breast tumor initiation, progression and/or response to therapy, and these have been implicated as key contributors to the complex effects of obesity on cancer incidence and outcomes. These host factors include components of the secretome, including insulin, insulin-like growth factor-1, leptin, adiponectin, steroid hormones, cytokines, vascular regulators, and inflammation-related molecules, as well as the cellular and structural components of the tumor microenvironment. These secreted and structural host factors are extrinsic to, and interact with, the intrinsic molecular characteristics of breast cancer cells (including breast cancer stem cells), and each will be considered in the context of energy balance and as potential targets for cancer prevention.