Multivalent Small-Molecule Pan-RAS Inhibitors.

Design of small molecules that disrupt protein-protein interactions, including the interaction of RAS proteins and their effectors, may provide chemical probes and therapeutic agents. We describe here the synthesis and testing of potential small-molecule pan-RAS ligands, which were designed to interact with adjacent sites on the surface of oncogenic KRAS. One compound, termed 3144, was found to bind to RAS proteins using microscale thermophoresis, nuclear magnetic resonance spectroscopy, and isothermal titration calorimetry and to exhibit lethality in cells partially dependent on expression of RAS proteins. This compound was metabolically stable in liver microsomes and displayed anti-tumor activity in xenograft mouse cancer models. These findings suggest that pan-RAS inhibition may be an effective therapeutic strategy for some cancers and that structure-based design of small molecules targeting multiple adjacent sites to create multivalent inhibitors may be effective for some proteins.

Regulation of ferroptotic cancer cell death by GPX4.

Ferroptosis is a form of nonapoptotic cell death for which key regulators remain unknown. We sought a common mediator for the lethality of 12 ferroptosis-inducing small molecules. We used targeted metabolomic profiling to discover that depletion of glutathione causes inactivation of glutathione peroxidases (GPXs) in response to one class of compounds and a chemoproteomics strategy to discover that GPX4 is directly inhibited by a second class of compounds. GPX4 overexpression and knockdown modulated the lethality of 12 ferroptosis inducers, but not of 11 compounds with other lethal mechanisms. In addition, two representative ferroptosis inducers prevented tumor growth in xenograft mouse tumor models. Sensitivity profiling in 177 cancer cell lines revealed that diffuse large B cell lymphomas and renal cell carcinomas are particularly susceptible to GPX4-regulated ferroptosis. Thus, GPX4 is an essential regulator of ferroptotic cancer cell death.

Distal Biceps Brachii Tendon Transfer for Re-establishing Extrinsic Finger Function: Feasibility Study in Cadavers.

PURPOSE: To determine the anatomic feasibility of transferring the biceps brachii tendon into either the extensor digitorum communis (EDC) or flexor digitorum profundus (FDP), determine the excursion imparted to EDC and FDP tendons after transfer, and compare the work capacity of the cadaver biceps to previously published data on the biceps as well as the recipient muscles by calculating the physiologic cross-sectional area (PCSA). METHODS: Four fresh-frozen cadaver shoulder-elbow-wrist specimens were used to measure tendon excursion that can be obtained with transfer of the distal biceps tendon into either the EDC or FDP. Two cadavers had distal biceps-to-EDC transfer performed, and the other 2 had distal biceps-to-FDP performed. Passive ranging of each elbow from flexion to extension and active loading at 90° of elbow flexion were then performed on each specimen to determine tendon excursion. An analysis of the PCSA of the biceps muscle was performed on each specimen. RESULTS: Distal biceps-to-EDC transfer resulted in an average of 24 mm of tendon excursion with passive loading, and 24 mm of tendon excursion with active loading. Distal biceps-to-FDP transfer resulted in an average of 24 mm of tendon excursion with passive loading, and 24 mm of tendon excursion with active loading. The average PCSA was 3.6 cm2. CONCLUSIONS: Transfer of the distal biceps tendon into the EDC or FDP is anatomically feasible and provides roughly 24 mm of tendon excursion to the tendon units. The PCSA in the specimens used is slightly lower than other published data; it closely approximates the PCSA of the EDC, but is only half of the PCSA of the FDP in previously published data. CLINICAL RELEVANCE: The findings suggest potentially novel transfer options for restoring finger flexion and extension in patients lacking FDP or EDC function.

Modulatory profiling identifies mechanisms of small molecule-induced cell death.

Cell death is a complex process that plays a vital role in development, homeostasis, and disease. Our understanding of and ability to control cell death is impeded by an incomplete characterization of the full range of cell death processes that occur in mammalian systems, especially in response to exogenous perturbations. We present here a general approach to address this problem, which we call modulatory profiling. Modulatory profiles are composed of the changes in potency and efficacy of lethal compounds produced by a second cell death-modulating agent in human cell lines. We show that compounds with the same characterized mechanism of action have similar modulatory profiles. Furthermore, clustering of modulatory profiles revealed relationships not evident when clustering lethal compounds based on gene expression profiles alone. Finally, modulatory profiling of compounds correctly predicted three previously uncharacterized compounds to be microtubule-destabilizing agents, classified numerous compounds that act nonspecifically, and identified compounds that cause cell death through a mechanism that is morphologically and biochemically distinct from previously established ones.

Comparison of radiation exposure in lumbar pedicle screw placement with fluoroscopy vs computer-assisted image guidance with intraoperative three-dimensional imaging.

BACKGROUND/OBJECTIVE: Little is known about the long-term effects of chronic exposure to ionizing radiation. Studies have shown that spine surgeons may be exposed to significantly more radiation than that observed in surgery on the appendicular skeleton. Computer-assisted image guidance systems have been shown in preliminary studies to enable accurate instrumentation of the spine. Computer-assisted image guidance systems may have significant application to the surgical management of spinal trauma and deformity. The objective of this study was to compare C-arm fluoroscopy and computer-assisted image guidance in terms of radiation exposure to the operative surgeon when placing pedicle screw-rod constructs in cadaver specimens. METHODS: Twelve single-level (2 contiguous vertebral bodies) lumbar pedicle screw-rod constructs (48 screws) in 4 fresh cadavers were placed using standard C-arm fluoroscopy and computer-assisted image guidance (Stealth Station with Iso-C(3D)). Pedicle screw-rod constructs were placed at L1-L2, L3-L4, and L5-S1 in 4 fresh cadaver specimens. Imaging was alternated between C-arm fluoroscopy and computer-assisted image guidance with StealthStation Iso-C(3D). Radiation exposure was measured using ring and badge dosimeters to monitor the thyroid, torso, and index finger. Postprocedure CT scans were obtained to judge accuracy of screw placement. RESULTS: Mean radiation exposure to the torso was 4.33 +/- 2.66 mRem for procedures performed with standard fluoroscopy and 0.33 +/- 0.82 mRem for procedures performed with computer-assisted image guidance. This difference was statistically significant (P= 0.012). Radiation exposure to the index finger and thyroid was negligible for all procedures. The accuracy of screw placement was similar for both techniques. CONCLUSIONS: Computer-assisted image guidance systems allow for the safe and accurate placement of pedicle screw-rod constructs with a significant reduction in exposure to ionizing radiation to the torso of the operating surgeon.

Discovery of Potent and Selective Leads against Toxoplasma gondii Dihydrofolate Reductase via Structure-Based Design.

Current treatment of toxoplasmosis targets the parasite's folate metabolism through inhibition of dihydrofolate reductase (DHFR). The most widely used DHFR antagonist, pyrimethamine, was introduced over 60 years ago and is associated with toxicity that can be largely attributed to a similar affinity for parasite and human DHFR. Computational analysis of biochemical differences between Toxoplasma gondii and human DHFR enabled the design of inhibitors with both improved potency and selectivity. The approach described herein yielded TRC-19, a promising lead with an IC50 of 9 nM and 89-fold selectivity in favor of Toxoplasma gondii DHFR, as well as crystallographic data to substantiate in silico methodology. Overall, 50% of synthesized in silico designs met hit threshold criteria of IC50 < 10 µM and >2-fold selectivity favoring Toxoplasma gondii, further demonstrating the efficiency of our structure-based drug design approach.

Small Molecule that Reverses Dexamethasone Resistance in T-cell Acute Lymphoblastic Leukemia (T-ALL).

Glucocorticoids are one of the most utilized and effective therapies in treating T-cell acute lymphoblastic leukemia. However, patients often develop resistance to glucocorticoids, rendering these therapies ineffective. We screened 9517 compounds, selected for their lead-like properties, chosen from among 3 372 615 compounds, against a dexamethasone-resistant T-ALL cell line to identify small molecules that reverse glucocorticoid resistance. We synthesized analogues of the most effective compound, termed J9, from the screen in order to define the scaffold's structure-activity relationship. Active compounds restored sensitivity to glucocorticoids through upregulation of the glucocorticoid receptor. This compound and mechanism may provide a strategy for overcoming glucocorticoid resistance in patients with T-ALL.

Pharmacological inhibition of cystine-glutamate exchange induces endoplasmic reticulum stress and ferroptosis.

Exchange of extracellular cystine for intracellular glutamate by the antiporter system xc (-) is implicated in numerous pathologies. Pharmacological agents that inhibit system xc (-) activity with high potency have long been sought, but have remained elusive. In this study, we report that the small molecule erastin is a potent, selective inhibitor of system xc (-). RNA sequencing revealed that inhibition of cystine-glutamate exchange leads to activation of an ER stress response and upregulation of CHAC1, providing a pharmacodynamic marker for system xc (-) inhibition. We also found that the clinically approved anti-cancer drug sorafenib, but not other kinase inhibitors, inhibits system xc (-) function and can trigger ER stress and ferroptosis. In an analysis of hospital records and adverse event reports, we found that patients treated with sorafenib exhibited unique metabolic and phenotypic alterations compared to patients treated with other kinase-inhibiting drugs. Finally, using a genetic approach, we identified new genes dramatically upregulated in cells resistant to ferroptosis.DOI: http://dx.doi.org/10.7554/eLife.02523.001.

Privileged scaffolds for library design and drug discovery.

This review explores the concept of using privileged scaffolds to identify biologically active compounds through building chemical libraries. We hope to accomplish three main objectives: to provide one of the most comprehensive listings of privileged scaffolds; to reveal through four selected examples the present state of the art in privileged scaffold library synthesis (in hopes of inspiring new and even more creative approaches); and also to offer some thoughts on how new privileged scaffolds might be identified and exploited.

Anatomic reconstruction of thumb metacarpophalangeal joint ulnar collateral ligament using an interference screw docking technique.

Complete rupture of the ulnar collateral ligament injury of the thumb metacarpophalangeal (MCP) joint can be a debilitating injury resulting in decreased grip and pinch strength. The spectrum of injury to this ligament varies from simple strain to complete rupture of both proper and accessory ligaments. Generally, this is a result of an abduction force transmitted across the thumb MCP joint.Physical examination assists in identification of these injuries and the extent of ligamentous injury. Ecchymoses and pain over the ligament are usually present in the acute injury. Chronic injuries may not be painful. A palpable mass may be present. The valgus stress test across this joint in 30 degrees of flexion provides information on the competency of the proper ligaments, whereas valgus stress in extension assesses the accessory collateral ligaments. This stress test is useful in the acute or chronic setting.Stabilization of the thumb MCP joint in chronic injuries can require reconstruction. We describe an anatomic reconstruction using a tendon graft and a bioabsorbable interference screw.