Measuring nonlinear signal combination using EEG.

Relatively little is known about the processes, both linear and nonlinear, by which signals are combined beyond V1. By presenting two stimulus components simultaneously, flickering at different temporal frequencies (frequency tagging) while measuring steady-state visual evoked potentials, we can assess responses to the individual components, including direct measurements of suppression on each other, and various nonlinear responses to their combination found at intermodulation frequencies. The result is a rather rich dataset of frequencies at which responses can be found. We presented pairs of sinusoidal gratings at different temporal frequencies, forming plaid patterns that were "coherent" (looking like a checkerboard) and "noncoherent" (looking like a pair of transparently overlaid gratings), and found clear intermodulation responses to compound stimuli, indicating nonlinear summation. This might have been attributed to cross-orientation suppression except that the pattern of intermodulation responses differed for coherent and noncoherent patterns, whereas the effects of suppression (measured at the component frequencies) did not. A two-stage model of nonlinear summation involving conjunction detection with a logical AND gate described the data well, capturing the difference between coherent and noncoherent plaids over a wide array of possible response frequencies. Multistimulus frequency-tagged EEG in combination with computational modeling may be a very valuable tool in studying the conjunction of these signals. In the current study the results suggest a second-order mechanism responding selectively to coherent plaid patterns.

Macrocyclization as a Source of Desired Polypharmacology. Discovery of Triple PI3K/mTOR/PIM Inhibitors.

The PI3K/AKT/mTOR and PIM kinase pathways contribute to the development of several hallmarks of cancer. Cotargeting of these pathways has exhibited promising synergistic therapeutic effects in liquid and solid tumor types. To identify molecules with combined activities, we cross-screened our collection of PI3K/(±mTOR) macrocycles (MCXs) and identified the MCX thieno[3,2-d]pyrimidine derivative 2 as a moderate dual PI3K/PIM-1 inhibitor. We report the medicinal chemistry exploration and biological characterization of a series of thieno[3,2-d]pyrimidine MCXs, which led to the discovery of IBL-302 (31), a potent, selective, and orally bioavailable triple PI3K/mTOR/PIM inhibitor. IBL-302, currently in late preclinical development (AUM302), has recently demonstrated efficacy in neuroblastoma and breast cancer xenografts. Additionally, during the course of our experiments, we observed that macrocyclization was essential to obtain the desired multitarget profile. As a matter of example, the open precursors 35-37 were inactive against PIM whereas MCX 28 displayed low nanomolar activity.

Preclinical evaluation of a novel triple-acting PIM/PI3K/mTOR inhibitor, IBL-302, in breast cancer.

The proviral integration of Moloney virus (PIM) family of protein kinases are overexpressed in many haematological and solid tumours. PIM kinase expression is elevated in PI3K inhibitor-treated breast cancer samples, suggesting a major resistance pathway for PI3K inhibitors in breast cancer, potentially limiting their clinical utility. IBL-302 is a novel molecule that inhibits both PIM and PI3K/AKT/mTOR signalling. We thus evaluated the preclinical activity of IBL-302, in a range of breast cancer models. Our results demonstrate in vitro efficacy of IBL-302 in a range of breast cancer cell lines, including lines with acquired resistance to trastuzumab and lapatinib. IBL-302 demonstrated single-agent, anti-tumour efficacy in suppression of pAKT, pmTOR and pBAD in the SKBR-3, BT-474 and HCC-1954 HER2+/PIK3CA-mutated cell lines. We have also shown the in vivo single-agent efficacy of IBL-302 in the subcutaneous BT-474 and HCC-1954 xenograft model in BALB/c nude mice. The combination of trastuzumab and IBL-302 significantly increased the anti-proliferative effect in HER2+ breast cancer cell line, and matched trastuzumab-resistant line, relative to testing either drug alone. We thus believe that the novel PIM and PI3K/mTOR inhibitor, IBL-302, represents an exciting new potential treatment option for breast cancer, and that it should be considered for clinical investigation.