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Phosphatidylinositol 3-kinases (PI3Ks) regulate intracellular signaling events for multiple cell surface receptors. Phosphatidylinositol 3-kinase δ, 1 of 4 class I PI3K isoforms, is primarily found in leukocytes and regulates immune cell functions. Here, we report changes in the immune and digestive systems that were associated with AMG2519493, a highly selective small-molecule PI3Kδ inhibitor. Following 1- or 3-month oral repeat dosing in the cynomolgus monkey, changes were observed in circulating B cells, lymphoid tissues (spleen, lymph nodes, gut-associated lymphoid tissue, tonsil), and the digestive tract. Decreased circulating B cells and lymphoid cellularity in B cell-rich zones in lymphoid tissues were attributed to the intended pharmacologic activity of AMG2519493. Dose- and duration-dependent digestive system toxicity was characterized by inflammation in the large intestine and secondary opportunistic infections restricted to the digestive tract. Digestive tract changes were associated with moribundity and mortality at high-dose levels, and the effect level decreased with increased duration of exposure. These observations demonstrate the role of PI3Kδ in regulation of the immune system and of host resistance to opportunistic infections of the digestive tract.

Sotorasib (Lumakras™) is a first-in-class, non-genotoxic, small molecule inhibitor of KRAS G12C developed as an anticancer therapeutic for treatment of patients that have a high unmet medical need. Anticancer therapeutics are considered out of scope of ICH M7 guidance for control of mutagenic impurities; however, based on ICH S9 QA, mutagenicity assessments are needed for impurities that exceed the qualification threshold, consistent with ICH Q3A/B, and non-mutagenic drugs. Here, we carried out hybrid-based mutagenicity assessment of sotorasib drug substance (DS) impurities using in silico quantitative structure-activity relationship (QSAR) modelling and Ames tests (for in silico positive mutagens). We encountered contradictive mutagenicity results for 2 impurities (Beta-Chloride and PAC). PAC was negative initially by QSAR but positive in a GLP full plate Ames test and Beta-Chloride was positive by QSAR, negative in a non-GLP micro-Ames but positive in a GLP full plate Ames assay. Root cause analyses identified and characterized mutagenic contaminants, 3-chloropropionic acid in batches of Beta-Chloride and 3-chloropropionic acid and Chloro-PAC in batches of PAC, used in initial GLP full-plate Ames tests. Significant reduction of these contaminants in re-purified batches resulted in no induction of mutagenicity in follow-up GLP micro-Ames tests. In summary, root-cause analyses led to accurate mutagenicity assessment for sotorasib DS-associated impurities.

Sotorasib is a first-in-class KRAS

The implementation in the RIKEN-RAL negative muons facility of a new muon beamline monitoring and novel digital data acquisition system for gamma and X-ray spectroscopy are presented. This work also shows the high potential of the muonic atoms X-ray spectroscopy technique in non-destructive elemental characterization of archaeological samples.

Phosphatidylinositol 3-kinase (PI3K) δ is a lipid kinase primarily found in leukocytes, which regulates important cell functions. AMG2519493 was a PI3K δ-specific inhibitor in development for treatment of various inflammatory diseases. AMG2519493-related changes in the male and/or female reproductive organs were observed in the 1- and 3-month oral repeat dose toxicology studies in the rat and cynomolgus monkey. Hemorrhagic corpora lutea cysts and increased incidence of corpora lutea cysts without hemorrhage were observed in the ovaries at supra pharmacological doses in the rat. A decrease in seminiferous germ cells in the testis, indicative of spermatogenesis maturation arrest, was observed in both the rat and cynomolgus monkey. Although the characteristics were comparable, the drug systemic exposures associated with the testicular changes were very different between the 2 species. In the rat, the testicular change was only observed at supra pharmacologic exposure. Isotype assessment of PI3K signaling in rat spermatogonia in vitro indicated a role for PI3K β, but not δ, in the c Kit/PI3K/protein kinase B signaling pathway. Therefore, changes in both the ovary and testis of the rat were considered due to off target effect as they only occurred at suprapharmacologic exposure. In contrast, the testicular changes in the cynomolgus monkey (decrease in seminiferous germ cells) occurred at very low doses associated with PI3K δ-specific inhibition, indicating that the PI3K δ isoform may be important in spermatogenesis maturation in the cynomolgus monkey. Our results suggest species-related differences in PI3K isoform-specific control on reproductive organs.

Sotorasib is a first-in class KRAS

Phosphatidylinositol 3-kinases (PI3Ks) regulate intracellular signaling events for multiple cell surface receptors. Phosphatidylinositol 3-kinase δ, 1 of 4 class I PI3K isoforms, is primarily found in leukocytes and regulates immune cell functions. Here, we report changes in the immune and digestive systems that were associated with AMG2519493, a highly selective small-molecule PI3Kδ inhibitor. Following 1- or 3-month oral repeat dosing in the cynomolgus monkey, changes were observed in circulating B cells, lymphoid tissues (spleen, lymph nodes, gut-associated lymphoid tissue, tonsil), and the digestive tract. Decreased circulating B cells and lymphoid cellularity in B cell-rich zones in lymphoid tissues were attributed to the intended pharmacologic activity of AMG2519493. Dose- and duration-dependent digestive system toxicity was characterized by inflammation in the large intestine and secondary opportunistic infections restricted to the digestive tract. Digestive tract changes were associated with moribundity and mortality at high-dose levels, and the effect level decreased with increased duration of exposure. These observations demonstrate the role of PI3Kδ in regulation of the immune system and of host resistance to opportunistic infections of the digestive tract.

Supplemental Material, sj-docx-1-tpx-10.1177_0192623320966238 for Phosphatidylinositol 3-Kinase δ Inhibitor-Induced Immunomodulation and Secondary Opportunistic Infection in the Cynomolgus Monkey (Macaca fascicularis) by Jonathan A. Werner, Katsu Ishida, John Wisler, Christine Karbowski, Jackson Kalanzi, Jeanine Bussiere and Thomas M. Monticello in Toxicologic Pathology

The true purity of Roman silver coinage was hidden by enriching the surfaces of the coins. The question investigated here is whether Roman gold coins were also surfaced enriched. Two non-destructive techniques were employed to do this: X-ray fluorescence (XRF) and a ‘newer’ technique, muonic X-ray emission spectroscopy (μXES). For the latter, the momentum of the muons is controlled, allowing for the composition of the coin to be determined at various depths. Here we show that there is no surface enrichment of the Roman gold coins analysed. Furthermore, we show that XRF and μXES return congruent results at the near surface. This all supports the integrity of surface level analyses of Roman gold coins. We then discuss the broader applicability of our muon technique to the further study of Roman gold coinage, to the cultural heritage sector and to archaeological scientists more generally.

The development of elemental analysis at the ISIS pulsed neutron and muon facility, STFC Rutherford Appleton Laboratory, UK, is described. Presented are recent results from the calibration of the new setup and this shows the technique can be a powerful tool in determining the composition of materials in bulk, not just at the surface. Moreover, this technique is non-destructive and should be sensitive to all elements (perhaps with the exception of H and He).

Systemic lupus erythematous (SLE) is a heterogeneous disease lacking highly effective treatment options. Here we tested if targeting both BAFF and ICOSL has superior efficacy than single target inhibition in the mouse arthritis and lupus models. We also generated AMG 570, an ICOSL and BAFF bispecific inhibitory molecule, for potential treatment of autoimmune diseases such as SLE.Murine BAFF/ICOSL bispecific, combination of BAFF and ICOSL inhibitors or single inhibitor was evaluated in the sheep red blood cell (SRBC) challenge model, mouse collagen induced arthritis (CIA) model, or NZB/NZW lupus models. AMG 570 was tested for human and cyno BAFF and ICOSL binding affinities by Kinexa A. AMG 570 dual target blocking activities was evaluated in human and cyno BAFF and ICOSL mediated B cell and T cell assay, respectively. Pharmacodynamics effect of AMG 570 was evaluated in cynomolgus monkey.Treatment with murine ICOSL/BAFF bispecific or combination therapy was more efficacious than single ICOSL or BAFF inhibitor in mouse NZB/NZW lupus model. Dual ICOSL and BAFF inhibition was also more effective in the mouse collagen induced arthritis (CIA) model. AMG 570 was developed as the clinical bispecific lead. AMG 570 inhibits human and cynomolgus monkey ICOSL and BAFF. B cell reduction was observed after AMG 570 treatment in cynomolgus monkeys, consistent with the pharmacological effect of BAFF inhibition.By targeting both ICOSL and BAFF, AMG 570 has the potential to achieve superior efficacy in treatment of autoimmune diseases such as SLE and rheumatoid arthritis.

The idea of the CHNET-TANDEM experiment is to develop and optimize a non-destructive technique, which allows analysis deep inside the sample with a good spatial resolution, using a negative muon beam. By selecting the primary muon energies appropriately, bulk analysis can be performed without destructing the sample. The experimental setup used for this experiment, made by 2 hodoscopes and 5 HpGe, allowed us to collect very interesting preliminary data concerning scan momentum, positioning and centering of the samples by means of two hodoscopes, analysis of standard material targets and elemental characterization of Nuragic “Bronze Age” votive ship fragments.

The main features of the Data AcQuisition systems for the FAMU (on muonic atom physics) and CHNET _ TANDEM (on the development of nondestructive techniques to archaeometry) INFN projects will be described. Both the experiments exploit the RIKEN-RAL Muon Facility beam of ( 20 , 120 ) MeV ∕ c muons and the same experimental setup, which includes a wide range of detectors: HPGe detectors for high resolution spectroscopy ; LaBr 3 scintillators , with both PMT and SiPM readout, for fast and high time resolution spectroscopy; several layers of 32 SiPM readout scintillating fibers for beam monitoring with good spatial and time resolution. The performance of the DAQ in terms of conditioning and processing of such a large number of different detector signals, data storage and analysis and a few examples of the results will be presented.