Second-Generation Atroposelective Synthesis of KRAS G12C Covalent Inhibitor GDC-6036.

A chromatography-free asymmetric synthesis of GDC-6036 (1) was achieved via a highly atroposelective Negishi coupling of aminopyridine 5 and quinazoline 6b catalyzed by 0.5 mol % [Pd(cin)Cl]2 and 1 mol % (R,R)-Chiraphite to afford the key intermediate (Ra)-3. An alkoxylation of (Ra)-3 with (S)-N-methylprolinol (4) and a global deprotection generates the penultimate heterobiaryl intermediate 2. A controlled acrylamide installation by stepwise acylation/sulfone elimination and final adipate salt formation and crystallization delivered high-purity GDC-6036 (1).

Atroposelective Negishi Coupling Optimization Guided by Multivariate Linear Regression Analysis: Asymmetric Synthesis of KRAS G12C Covalent Inhibitor GDC-6036.

An efficient asymmetric synthesis of a potent KRAS G12C covalent inhibitor, GDC-6036 (1), is reported. The synthesis features a highly atroposelective Negishi coupling to construct the key C-C bond between two highly functionalized pyridine and quinazoline moieties by employing a Pd/Walphos catalytic system. Statistical modeling by comparing computational descriptors of a range of Walphos chiral bisphosphine ligands to a training set of experimental results was used to inform the selection of the best ligand, W057-2, which afforded the desired Negishi coupling product (Ra)-3 in excellent selectivity. A subsequent telescoped reaction sequence of alkoxylation, global deprotection, and acrylamide formation, followed by a final adipate salt formation, furnished GDC-6036 (1) in 40% overall yield from starting materials pyridine 5 and quinazoline 6.

Bone marrow adipocytes drive the development of tissue invasive Ly6Chigh monocytes during obesity.

During obesity and high fat-diet (HFD) feeding in mice, sustained low-grade inflammation includes not only increased pro-inflammatory macrophages in the expanding adipose tissue, but also bone marrow (BM) production of invasive Ly6Chigh monocytes. As BM adiposity also accrues with HFD, we explored the relationship between the gains in BM white adipocytes and invasive Ly6Chigh monocytes by in vivo and ex vivo paradigms. We find a temporal and causal link between BM adipocyte whitening and the Ly6Chigh monocyte surge, preceding the adipose tissue macrophage rise during HFD in mice. Phenocopying this, ex vivo treatment of BM cells with conditioned media from BM adipocytes or bona fide white adipocytes favoured Ly6Chigh monocyte preponderance. Notably, Ly6Chigh skewing was preceded by monocyte metabolic reprogramming towards glycolysis, reduced oxidative potential and increased mitochondrial fission. In sum, short-term HFD changes BM cellularity, resulting in local adipocyte whitening driving a gradual increase and activation of invasive Ly6Chigh monocytes.

Asymmetric Hydrogenation of Unfunctionalized Tetrasubstituted Acyclic Olefins.

Asymmetric hydrogenation has evolved as one of the most powerful tools to construct stereocenters. However, the asymmetric hydrogenation of unfunctionalized tetrasubstituted acyclic olefins remains the pinnacle of asymmetric synthesis and an unsolved challenge. We report herein the discovery of an iridium catalyst for the first, generally applicable, highly enantio- and diastereoselective hydrogenation of such olefins and the mechanistic insights of the reaction. The power of this chemistry is demonstrated by the successful hydrogenation of a wide variety of electronically and sterically diverse olefins in excellent yield and high enantio- and diastereoselectivity.

Analysis and Control of Chain Mobility in Protein Hydrogels.

Coiled-coil domains can direct the assembly of protein block copolymers into physically cross-linked, viscoelastic hydrogels. Here, we describe the use of fluorescence recovery after photobleaching (FRAP) to probe chain mobility in reversible hydrogels assembled from engineered proteins bearing terminal coiled-coil domains. We show that chain mobility can be related to the underlying dynamics of the coiled-coil domains by application of a three-state "hopping" model of chain migration. We further show that genetic programming allows the effective mobility of network chains to be varied 500-fold through modest changes in protein sequence. Destabilization of the coiled-coil domains by site-directed mutagenesis increases the effective diffusivity of probe chains. Conversely, probe mobility is reduced by expanding the hydrophobic surface area of the coiled-coil domains through introduction of the bulky leucine surrogate homoisoleucine. Predictions from the three-state model imply asymmetric sequential binding of the terminal domains. Brownian Dynamics simulations suggest that binding asymmetry is a general feature of reversible gels, arising from a loss in entropy as chains transition to a conformationally restricted bridged state.

Glycocalyx Engineering with a Recycling Glycopolymer that Increases Cell Survival In Vivo.

Synthetic glycopolymers that emulate cell-surface mucins have been used to elucidate the role of mucin overexpression in cancer. However, because they are internalized within hours, these glycopolymers could not be employed to probe processes that occur on longer time scales. In this work, we tested a panel of glycopolymers bearing a variety of lipids to identify those that persist on cell membranes. Strikingly, we found that cholesterylamine (CholA) anchored glycopolymers are internalized into vesicles that serve as depots for delivery back to the cell surface, allowing for the display of cell-surface glycopolymers for at least ten days, even while the cells are dividing. As with native mucins, the cell-surface display of CholA-anchored glycopolymers influenced the focal adhesion distribution. Furthermore, we show that these mimetics enhance the survival of nonmalignant cells in a zebrafish model of metastasis. CholA-anchored glycopolymers therefore expand the application of glycocalyx engineering in glycobiology.

Negishi Approach to 1,5-Disubstituted 3-Amino-1H-1,2,4-triazoles.

An efficient synthesis of 1,5-disubstituted 3-amino-1H-1,2,4-triazoles has been achieved via a Negishi coupling of aryl or vinyl bromides and 1-substituted 3-amino-1H-1,2,4-triazoles in the presence of Knochel's base tetramethylpiperidinylzinc chloride lithium chloride (TMPZnCl·LiCl) and catalytic bis(di-tert-butylphenylphosphine)palladium chloride. This chemistry tolerates a variety of electronically diverse aryl or vinyl bromides and 1-substituted 3-amino-1H-1,2,4-triazoles.

Treating obesity with a novel hand-held device, computer software program, and Internet technology in primary care: the SMART motivational trial.

OBJECTIVE: The purpose of this study was to evaluate the short-term motivational effect of a technology-based weight reduction program for obese adults. METHODS: One hundred and eleven obese (37.0+/-5.8 kg/m(2)) middle aged (45.5+/-10.8 years) adults (62% female) were randomly assigned to a usual care or experimental (SMART: self-monitoring and resting metabolic rate technology) group. The usual care group received a standard nutritional program in accordance to national guidelines. All participants received a comprehensive weight management program consisting of motivational interviewing (MI) sessions and automated e-mail behavioral newsletters. Bodyweight, arterial blood pressure, and psychobehavioral constructs were assessed over 12 weeks. RESULTS: Completer analysis (n=80) indicated a significant improvement in bodyweight (-3.9%), systolic arterial pressure (-4 mmHg), and all motivational constructs following the 12-week study (p

The wheat PKABA1-interacting factor TaABF1 mediates both abscisic acid-suppressed and abscisic acid-induced gene expression in bombarded aleurone cells.

To investigate the crosstalk of abscisic acid (ABA) and gibberellin (GA) signaling in wheat (Triticum aestivum), we have focused on the transcription factor TaABF1. TaABF1 (a member of the ABA response element binding factor family) physically interacts with PKABA1, a signaling component in the ABA-suppression of GA-induced gene expression in cereal grains. Constitutive expression of TaABF1 in aleurone cells of imbibing grains completely eliminated GA-induced expression from the Amy32b promoter. In addition to its effect on Amy32b, TaABF1 strongly stimulated expression from the ABA-inducible HVA1 and HVA22 promoters. Overexpression of TaABF1 fully substituted for exogenous ABA in the induction of these two promoters. The introduction of a construct directing RNA inhibition (RNAi) of TaABF1 did not prevent either ABA-mediated or PKABA1-mediated suppression of Amy32b expression. Similarly, the RNAi construct did not prevent ABA-induction of HVA1. These results suggest that another protein may act redundantly with TaABF1 during cereal imbibition. Although TaABF1 mRNA was downregulated during imbibition of afterripened grains, transcript levels were not significantly altered by exogenous GA or ABA, suggesting that upregulation of TaABF1 at the mRNA level is not required for its role in ABA signaling. We propose a model in which TaABF1 is involved in two separate branches of ABA signaling. In this model, TaABF1 acts downstream of PKABA1 in ABA-suppression of GA-induced gene expression, and participates (independently of PKABA1) in the stimulation of ABA-induced genes.