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A) Structure of JNJ-42541707, a structurally related compound to JNJ-42756493. B) 72h growth inhibition (IC50) of JNJ-42541707 against 236 cancer cell lines from multiple origins color coded based on FGFR1,2,4 mRNA overexpression and FGFR WT.

In vitro kinase inhibition by DiscoverX KinomeScan assay. For details see (19)

Inhibitory activity of Brivanib and JNJ-42756943 in kinase (top) and BaF3 kinase dependent proliferation (bottom) assays and ratio of FGFRs/VEGFR2 activities.

JNJ-42756493 anti-proliferative activity against cancer cells lines from multiple origins.. Detailed data supporting Figure 2.

Summary 1.Nectar is a crucial energy resource that strongly mediates the interactions between plants and animal pollinators or plant defenders. Previous research has shown that nectar is commonly colonized by microorganisms, most commonly bacteria and yeasts, which can have a strong impact on nectar chemistry. However, at present little is known about the effects of microorganisms on the fitness of animals feeding on nectar. 2.We used three nectar bacteria representing different metabolic groups (Asaia sp., Lactococcus sp. and Rosenbergiella sp.) and the common generalist aphid parasitoid Aphidius ervi (Haliday) (Hymenoptera: Braconidae) to test the hypothesis that different nectar-dwelling bacteria affect nectar consumption and insect longevity differently by altering the chemistry of nectar. 3.Bacteria significantly affected nectar chemistry by altering its acidity, sugar and amino acids composition/concentration and by adding compounds synthesized by the microbes. Although inoculation with bacteria did not affect nectar consumption, a significant difference in insect longevity was observed. The impact on longevity was species specific, with Lactococcus being beneficial and Asaia having a detrimental effect.​ 4.Bacteria have a strong impact on nectar chemistry and changes in nectar chemistry may not only influence the foraging behavior of flower-visiting animals and impact on plant fitness, but also influence the fitness of nectar-consuming organisms. As effects were species dependent, changes in nectar chemistry induced by different bacteria may have contrasting effects on the interactions between plants and insects. It is therefore essential to know how different microbes alter nectar chemistry to understand the relationships between plants, nectar-inhabiting microbes and nectar-consuming animals. This article is protected by copyright. All rights reserved.

Fibroblast growth factor (FGF) signaling plays critical roles in key biological processes ranging from embryogenesis to wound healing and has strong links to several hallmarks of cancer. Genetic alterations in FGF receptor (FGFR) family members are associated with increased tumor growth, metastasis, angiogenesis, and decreased survival. JNJ-42756493, erdafitinib, is an orally active small molecule with potent tyrosine kinase inhibitory activity against all four FGFR family members and selectivity versus other highly related kinases. JNJ-42756493 shows rapid uptake into the lysosomal compartment of cells in culture, which is associated with prolonged inhibition of FGFR signaling, possibly due to sustained release of the inhibitor. In xenografts from human tumor cell lines or patient-derived tumor tissue with activating FGFR alterations, JNJ-42756493 administration results in potent and dose-dependent antitumor activity accompanied by pharmacodynamic modulation of phospho-FGFR and phospho-ERK in tumors. The results of the current study provide a strong rationale for the clinical investigation of JNJ-42756493 in patients with tumors harboring FGFR pathway alterations. Mol Cancer Ther; 16(6); 1010–20. ©2017 AACR.

Summary Fungi of the genus Aspergillus are widespread in the environment. Some Aspergillus species, most commonly Aspergillus fumigatus, may lead to a variety of allergic reactions and life‐threatening systemic infections in humans. Invasive aspergillosis occurs primarily in patients with severe immunodeficiency, and has dramatically increased in recent years. There are several factors at play that contribute to aspergillosis, including both fungus and host‐related factors such as strain virulence and host pulmonary structure/immune status, respectively. The environmental tenacity of Aspergilllus, its dominance in diverse microbial communities/habitats, and its ability to navigate the ecophysiological and biophysical challenges of host infection are attributable, in large part, to a robust stress‐tolerance biology and exceptional capacity to generate cell‐available energy. Aspects of its stress metabolism, ecology, interactions with diverse animal hosts, clinical presentations and treatment regimens have been well‐studied over the past years. Here, we synthesize these findings in relation to the way in which some Aspergillus species have become successful opportunistic pathogens of human‐ and other animal hosts. We focus on the biophysical capabilities of Aspergillus pathogens, key aspects of their ecophysiology and the flexibility to undergo a sexual cycle or form cryptic species. Additionally, recent advances in diagnosis of the disease are discussed as well as implications in relation to questions that have yet to be resolved., Some Aspergillus species, most commonly Aspergillus fumigatus, can induce allergic reactions and life‐threatening systemic infections in humans. Here, we discuss the potency of Aspergillus as an opportunistic pathogen of humans and other animal hosts. We focus on the biophysical capabilities of pathogenic aspergilli; key aspects of their ecophysiology; and their ability to undergo a sexual cycle or form cryptic species. Recent advances in diagnosis of the disease are discussed, as well as outstanding scientific questions.

Aphid parasitoids are commonly used in the biological control of aphids. However, their success in biological control largely depends on the availability of carbohydrate-rich food as an energy source for maintenance and reproduction. Therefore, as these resources have become rare in modern agricultural systems, external sugar sources like flowering plants or artificial sugar solutions are more and more used to provide the biocontrol agents with the necessary sugars. When developing such artificial food sources, it is essential to carefully select the sugars that best support the target parasitoids without benefiting non-target insects, such as pest insects or hyperparasitoids. Here, we investigated the gustatory response and longevity of two commonly used aphid parasitoids (Aphidius colemani and Aphidius matricariae) and their hyperparasitoid Dendrocerus aphidum when provided with one of eight plant- and/or insect-derived sugars (fructose, galactose, glucose, melibiose, melezitose, rhamnose, sucrose and trehalose). Our results showed that the studied insect species consumed the largest amounts of sugars that are most commonly found in honeydew (sucrose, fructose, glucose and melezitose) and also survived best when feeding on these sugars. Both Aphidius spp. survived well on melibiose, whereas D. aphidum performed poorly on this sugar. When melibiose was offered in a mixture with glucose, a significant reduction in longevity was observed for D. aphidum when compared to glucose only, while this was less pronounced for Aphidius spp. This knowledge can be exploited in tailoring food sources to selectively support Aphidius parasitoids, enhancing the biological control of aphids.

Summary Invasive aspergillosis (IA) has become increasingly common and is characterised by high morbidity and mortality. Upcoming resistance threatens treatment with azoles and highlights the continuous need for novel therapeutics. This laboratory study investigated the in vitro and in vivo potential of the alkylphospholipid oleylphosphocholine (OlPC) against Aspergillus. In vitro activities of OlPC, miltefosine, posaconazole and voriconazole were determined for Aspergillus fumigatus, A. niger, A. terreus and A. flavus. In vivo efficacy of OlPC was evaluated in a systemic A. fumigatus mouse model, adopting a short-term and long-term oral or intraperitoneal dosing regimen. OlPC showed good in vitro activity against A. fumigatus (IC50 = 1.04 μmol l−1). Intraperitoneal administration of 50 mg kg−1 day−1 OlPC significantly reduced the fungal organ burdens at 4 days post-infection (dpi). Although 5- and 10-day OlPC treatment improved survival, organ burdens were not affected at 10 and 15 dpi. While this study showed excellent in vitro activity of OlPC against Aspergillus spp., its therapeutic efficacy in an acute mouse model for IA was less convincing. Given the limited therapeutic options in the current antifungal market for invasive infections, OlPC activity should be assessed in a less stringent in vivo model, potentially in combination treatment with other already marketed antifungal drugs.

Brettanomyces (Dekkera) bruxellensis is an ascomycetous yeast of major importance in the food, beverage and biofuel industry. It has been isolated from various man-made ecological niches that are typically characterized by harsh environmental conditions such as wine, beer, soft drink etc. Recent comparative genomics studies revealed an immense intraspecific diversity, but it is still unclear whether this genetic diversity also leads to systematic differences in fermentation performance and (off-)flavor production, and to what extent strains have evolved to match their ecological niche. Here, we present an evaluation of the fermentation properties of eight genetically diverse B. bruxellensis strains originating from beer, wine and soft drinks. We show that sugar consumption and aroma production during fermentation are determined by both the yeast strain and composition of the medium. Furthermore, our results indicate a strong niche adaptation of B. bruxellensis, most clearly for wine strains. For example, only strains originally isolated from wine were able to thrive well and produce the typical Brettanomyces-related phenolic off-flavors 4-ethylguaiacol and 4- ethylphenol when inoculated in red wine. Sulfite tolerance was found as a key factor explaining the observed differences in fermentation performance and off-flavor production. Sequence analysis of genes related to phenolic off-flavor production, however, revealed only marginal differences between the isolates tested, especially at the amino acid level. Altogether, our study provides novel insights in the Brettanomyces metabolism of flavor production, and is highly relevant for both the wine and beer industry. ispartof: FEMS Yeast Research vol:17 issue:1 ispartof: location:England status: published

To meet their carbohydrate requirements, adult parasitoids exploit a broad range of sugar resources, including floral and extrafloral nectar and honeydew. Although honeydew might be the predominant sugar source, especially in agricultural systems, it often is nutritionally inferior to sugar sources like nectar. Given its broad availability, it may be expected that sugar-feeding insects have evolved specialized adaptations to deal with this typically inferior sugar source. This would apply especially to organisms that have a close association with honeydew producers. Here, we hypothesized that parasitoids of honeydew-producing insects should show a pronounced response to sugars, such as fructose, sucrose, melezitose, and trehalose, and to a lesser extent glucose. To test this hypothesis, we investigated sugar consumption, feeding behavior and survival of the aphid parasitoid Aphidius ervi on several sugars (equiweight solutions). Our results show that A. ervi adults consumed typical honeydew sugars (sucrose, fructose, trehalose, and melezitose) the most, while consuming considerably less glucose or melibiose. Rhamnose, which does not occur in aphid honeydew, was not, or was only marginally, consumed. When different sugars were provided at the same time, A. ervi adults preferred sucrose or fructose over glucose or melezitose. Furthermore, pre-exposure to sucrose or fructose significantly reduced subsequent intake of glucose, suggesting an acquired distaste for glucose after being previously exposed to highly preferred sugars such as sucrose and fructose. Altogether, this study shows that A. ervi adults prefer sugars (fructose, melezitose, trehalose, and sucrose) that are overrepresented in aphid honeydew and show a lower preference to one (glucose) that is underrepresented in honeydew.

Although Aspergillus infections pose a growing threat to immunocompromised individuals, the limited range of existing drugs does not allow efficient management of invasive aspergillosis. Moreover, drug resistance is becoming increasingly common. Given that drug discovery relies on high-quality animal studies, careful design of in vivo models for invasive aspergillosis could facilitate the identification of novel antifungals. In this review, we discuss key aspects of animal models for invasive aspergillosis, covering laboratory animal species, immune modulation, inoculation routes, Aspergillus strains, treatment strategies and efficacy assessment, to enable the reader to tailor specific protocols for different types of preclinical antifungal evaluation study.

Background Pyrrolo[1,2-α][1,4]benzodiazepines (PBDs) have been described as a novel class of antifungal compounds with activity against dermatophytes and Aspergillus fumigatus. The initial structureactivity relationship showed that compounds with a chlorine substitution at position 7 have a higher activity compared with regioisomers or other substituents. Methods The present study evaluated more analogues with a 7-chlorine-substitution in vitro against a broad panel of clinically relevant fungal species. The Microsporum canis model in guinea pigs was used to assess the in vivo efficacy after oral and topical administration. Results IC50 values in the low micromolar range (IC50 0.68.0 μM for dihydro-PBDs; 0.10.7 μM for oxidized PBDs) confirmed the potent and selective in vitro activity of PBDs against dermatophytes, while the activity against A. fumigatus and Candida parapsilosis was slightly lower. For dihydro-PBDs, para-substitution showed superior activity, while oxidized compounds with a meta-substitution performed best. Oxidized Compound O with meta-CF2CH3-substitution showed excellent IC50 values of 0.6 μM against M. canis, 2.0 μM against Trichophyton mentagrophytes and 0.7 μM against Trichophyton rubrum, matching or outperforming the activity of itraconazole (IC50 values of 2.0, 0.4 and 0.6 μM, respectively). In vivo, topical application of a 0.25% formulation of Compound O gave a lesion reduction of >90% compared with placebo-treated animals. Oral administration of this compound at 20 mg/kg showed superior therapeutic efficacy compared with the reference drug itraconazole. Conclusions In conclusion, PBDs with a chlorine atom at position 7 are very promising antifungal candidates with convincing in vitro and in vivo activity particularly against dermatophytes and should be studied in greater detail to explore their full potential in the treatment of dermatophytoses.

This paper describes the development of poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles (NPs) and nanosuspensions with the polyene antibiotic amphotericin B (AmB). The nanoformulations were prepared using nanoprecipitation and were characterised with respect to size, zeta potential, morphology, drug crystallinity and content. Standard in vitro sensitivity tests were performed on MRC-5 cells, red blood cells, Leishmania infantum promastigotes and intracellular amastigotes and the fungal species Candida albicans, Aspergillus fumigatus and Trichophyton rubrum. The in vivo efficacy was assessed and compared to that of Fungizone® and AmBisome® in the acute A. fumigatus mouse model at a dose of 2.5 and 5.0 mg/kg AmB equivalents. The developed AmB nanoformulations were equivalently or more effective against the different Leishmania stages and axenic fungi in comparison with the free drug. The in vitro biological activity, and especially hemolytic activity, clearly depended on the preparation parameters of the different nanoformulations. Further, we demonstrated that the superior in vitro antifungal activity could be extrapolated to the in vivo situation. At equivalent dose, the optimal AmB-loaded PLGA NP was about two times and the AmB nanosuspension about four times more efficacious in reducing the total burden than AmBisome®. The developed AmB nanomedicines could represent potent and cost-effective alternatives to Fungizone® and AmBisome®.

The triazole antifungal pramiconazole (Stiefel, a GSK company) was compared with itraconazole, miconazole, and terbinafine in vitro and in vivo . Potent in vitro activities against Candida spp. (50% inhibitory concentration [IC 50 ], 0.04 to 1.83 μM) and Microsporum and Trichophyton spp. (IC 50 , 0.15 to 1.34 μM) were obtained but not, however, against other filamentous molds and zygomycetes. In the M. canis guinea pig model and C. albicans vulvovaginitis rat model, pramiconazole was superior to the reference compounds after oral and topical administration.

It is becoming increasingly clear that many solid tumors are not a homogeneous mass, but instead consist of highly heterogeneous microsystems comprised of multiple tumor subpopulations together with stromal cells. Such heterogeneity is often attributed to genetic and epigenetic alterations that promote functional specialization of different tumor subpopulations, thereby supporting the growth and differentiation of the primary tumor and subsequent invasion and metastases. This intratumoral complexity serves as a base for a range of intrinsic and acquired resistance mechanisms that are responsible for the failure of targeted therapies, especially in highly heterogeneous tumors such as lung cancer. JNJ-42756493 is a nanomolar inhibitor of fibroblast growth factor receptors (FGFRs) 1, 2, 3 and 4, that is currently being evaluated in Phase I clinical trials. As part of our ongoing preclinical program to elucidate potential mechanisms of resistance to JNJ-42756493, we have developed and characterized a range of cell-based models, representing different solid tumor histologies which show resistance to JNJ-42756493 and other FGFR inhibitors. Here, we describe the outgrowth of pre-existing subpopulations of cells that were intrinsically resistant to FGFR inhibition, from different FGFR1-amplified lung cancer subtypes: NCI-H1581, large cell (LCLC), and DMS114, small cell lung carcinoma (SCLC) cells. Following expansion of these resistant clones, a range of comparative ‘omics’ approaches (microarray analysis, RNA sequencing, shRNA knockdown screening, exome sequencing, and phospho RTK array analysis) were employed to explore the molecular mechanisms of resistance. Comprehensive profiling of these intrinsically resistant subpopulations has revealed that the LCLC and SCLC models have divergent mechanisms of resistance. Met pathway activation was found in LCLC model whilst IGF1R was found to be activated in the SCLC model resistant to FGFR inhibition. This is a novel observation of IGF1R pathway activation in FGFR1 amplified small cell lung cancer models that are resistant to FGFR inhibition. Blocking Met or IGF1R pathways with targeted therapies was observed to prevent the emergence of intrinsic resistance observed in these FGFR driven tumors. Follow-up experiments were performed in vitro and in vivo to confirm these intrinsic mechanisms of resistance and these data help build a rationale for combinations of targeted therapies in the clinic. Finally, our data indicates that pre-existing heterogeneity is likely to be a key driver for the appearance of resistance to targeted therapy in independent lung cancer models. Citation Format: Eleonora Jovcheva, Souichi Ogata, Kelly Van De Ven, Caroline Paulussen, Inez Van de Weyer, Hans De Wolf, Hugo Ceulemans, Steve McClue, Jorge Vialard, Timothy Perera. Identification of alternative mechanisms of resistance to FGFR inhibitor treatment in FGFR1-amplified large cell compared to FGFR1-amplified small cell lung cancer models. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-329. doi:10.1158/1538-7445.AM2014-LB-329

The ability of drugs to penetrate into cells and tissues is a fundamentally important property that is essential for the efficacy of drugs targeting intracellular processes. Many weakly basic small molecule drugs are known to accumulate extensively in acidic sub-cellular compartments such as lysosomes, through a process called ion trapping that is dependent on the physico-chemical properties of a compound. ‘Lysosomal uptake’ has been shown to significantly influence the activity of many agents, including anticancer drugs, both positively and negatively. Here we describe the unique physico-chemical properties of JNJ-42756493, a potent inhibitor of fibroblast growth factor receptors (FGFRs) 1, 2, 3 and 4, currently undergoing clinical testing for the treatment of malignancies with FGFR pathway activating alterations. JNJ-42756493 has weakly basic and lipophilic characteristics, is highly permeable, and is fluorescent when exposed to UV light. The fluorescent property of JNJ-42756493 enabled rapid and convenient visualization of the drug (spatial and temporal localization) within the lysosomes of cultured cells exposed to JNJ-42756493. In order to evaluate the influence of this lysosomotropic property on the activity of JNJ-42756493, FGFR phosphorylation was monitored in a series of wash-out experiments performed in various cellular models with FGFR1 or FGFR2 amplifications. This comparative washout analysis demonstrated prolonged FGFR kinase inhibition by JNJ-42756493 in comparison to other FGFR inhibitors lacking lysosomotropic properties. This qualitative fluorescence-based data was complemented by accurate quantification of the amount of compound taken up by the cells through the use of radiolabelled JNJ-42756493. Our results unequivocally demonstrate that JNJ-42756493 accumulates at high concentrations within the lysosomal compartment of cells without undergoing metabolism. The continued, long lasting effects on the target FGF receptor kinase inhibition following drug washout, suggest a novel intrinsic slow release of the unmodified drug from these intracellular stores. The lysosomotropic property of JNJ-42756493 is proposed to be a major contributor to the efficient and persistent inhibition of FGFR phosphorylation leading to potent cellular and in vivo activity. Citation Format: Eleonora Jovcheva, Caroline Paulussen, Patrick Van Bergen, Yolanda Chong, Jorge Vialard, Laurence Mevellec, Patrick Angibaud, Timothy Perera. Unique physicochemical properties of the potent FGFR 1, 2, 3 and 4 inhibitor JNJ-42756493 contribute to prolonged target shutdown. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-233. doi:10.1158/1538-7445.AM2014-LB-233