Robust arterial input function surrogate measurement from the superior sagittal sinus complex signal for fast dynamic contrast-enhanced MRI in the brain.

PURPOSE: Accurately estimating the arterial input function for dynamic contrast-enhanced MRI is challenging. An arterial input function is typically determined from signal magnitude changes related to a contrast agent, often leading to underestimation of peak concentrations. Alternatively, signal phase recovers the accurate peak concentration for straight vessels but suffers from high noise. A recent method proposed to fit the signal in the complex plane by combining the advantages of the previous 2 methods. The purpose of this work is to refine this complex-based method to determine the venous output function (VOF), an arterial input function surrogate, from the superior sagittal sinus. METHODS: We propose a state-of-the-art complex-based method that includes direct compensation for blood inflow and signal phase correction accounting for the curvature of the superior sagittal sinus, generally assumed collinear with B0 . We compared the magnitude-, phase-, and complex-based VOF determination methods against various simulated biases as well as for 29 brain metastases patients. RESULTS: Angulation of the superior sagittal sinus relative to B0 varied widely within patients, and its effect on the signal phase caused an underestimation of peak concentrations of up to 65%. Correction significantly increased the VOF peak concentration for the phase- and complex-based VOFs in the cohort. The phase-based method recovered accurate peak concentrations but lacked precision in the tail of the VOF. Our complex-based VOF completely recovered the effect of inflow and resulted in a high-peak concentration with limited noise. CONCLUSION: The new complex-based method resulted in high-quality VOF robust against superior sagittal sinus curvature and variations in patient positioning.

Increased precision in the intravascular arterial input function with flow compensation.

PURPOSE: In this study, we investigate the effects of pulsatile flow and inflow on dynamic susceptibility-contrast MRI intravascular arterial input function measurement in human brain arteries and measure how they are affected by first-order flow compensation. METHODS: A dual-echo single-shot EPI sequence with alternating flow compensation gradients was used to acquire dynamic susceptibility-contrast images with electrocardiogram monitoring. The dynamic signal variations measured inside the middle cerebral and internal carotid arteries were associated to the pulsatile arterial blood velocities measured with a single-slice quantitative flow sequence throughout the cardiac cycle. RESULTS: Major inverse correlations between intravascular signal and blood velocity were found for the standard single-shot EPI sequence. Flow compensation reduces these correlated variations that contribute to signal physiological noise. This causes a significant twofold increase of intravascular SNR in the middle cerebral and the internal carotid arteries (2.3 ± 0.9, P = 0.03) and (2.0 ± 0.9, P = 0.04), respectively; and reduced phase SD for the internal carotid arteries (0.72 ± 0.14, P = 0.004). The correction proposed in this work translates into a quantitative arterial input function with reduced noise in the internal carotid arteries. CONCLUSION: The physiological noise added by pulsatile flow and inflow for intravascular arterial input function measurement in the brain arteries is significantly reduced by flow compensation.

Molecular mechanism by which a potent hepatitis C virus NS3-NS4A protease inhibitor overcomes emergence of resistance.

Although optimizing the resistance profile of an inhibitor can be challenging, it is potentially important for improving the long term effectiveness of antiviral therapy. This work describes our rational approach toward the identification of a macrocyclic acylsulfonamide that is a potent inhibitor of the NS3-NS4A proteases of all hepatitis C virus genotypes and of a panel of genotype 1-resistant variants. The enhanced potency of this compound versus variants D168V and R155K facilitated x-ray determination of the inhibitor-variant complexes. In turn, these structural studies revealed a complex molecular basis of resistance and rationalized how such compounds are able to circumvent these mechanisms.

Xanthates as synthetic equivalents of oxyacyl radicals: access to lactones under tin-free conditions.

In addition to their utility in Barton-McCombie deoxygenations, xanthates can engage in 5-exo-trig radical cyclizations to afford lactones after oxidative workup. In this paper, we describe a tin-free protocol that provides direct access to lactones via hydrolysis of labile thioketal intermediates. Analysis of several systems of varying complexity reveals that the reaction is most applicable for constrained systems in which the reacting center is prepositioned near the radical-accepting alkene.

Valorization of dairy waste and by-products through microbial bioprocesses.

Waste is an inherent and unavoidable part of any process which can be attributed to various factors such as process inefficiencies, usability of resources and discarding of not so useful parts of the feedstock. Dairy is a burgeoning industry following the global population growth, resulting in generation of waste such as wastewater (from cleaning, processing, and maintenance), whey and sludge. These components are rich in nutrients, organic and inorganic materials. Additionally, the presence of alkaline and acidic detergents along with sterilizing agents in dairy waste makes it an environmental hazard. Thus, sustainable valorization of dairy waste requires utilization of biological methods such as microbial treatment. This review brings forward the current developments in utilization and valorization of dairy waste through microbes. Aerobic and anaerobic treatment of dairy waste using microbes can be a sustainable and green method to generate biofertilizers, biofuels, power, and other biobased products.

Valorization of sugar beet pulp to value-added products: A review.

The processing of sugar beet in the sugar production industry releases huge amounts of sugar beet pulp as waste which can be considered a valuable by-product as a source of cellulose, hemicellulose, and pectin. Valorization of sugar beet pulp into value added products occurs through acid hydrolysis, hydrothermal techniques, and enzymatic hydrolysis. Biochemical conversion of beet pulp into simple fermentable sugars for producing value added products occurs through enzymatic hydrolysis is a cost effective and eco-friendly process. While beet pulp has predominantly been used as a fodder for livestock, recent developments in its biotechnological valorization have unlocked its value as a feedstock in the production of biofuels, biohydrogen, biodegradable plastics, and platform chemicals such as lactic acid, citric acid, alcohols, microbial enzymes, single cell proteins, and pectic oligosaccharides. This review brings forward recent biotechnological developments made in the valorization of sugar beet pulp into valuable products.

Novel Miniaturized Drug Conjugate Leverages HSP90-driven Tumor Accumulation to Overcome PI3K Inhibitor Delivery Challenges to Solid Tumors.

The PI3K pathway is considered a master regulator for cancer due to its frequent activation, making it an attractive target for pharmacologic intervention. While substantial efforts have been made to develop drugs targeting PI3K signaling, few drugs have been able to achieve the inhibition necessary for effective tumor control at tolerated doses. HSP90 is a chaperone protein that is overexpressed and activated in many tumors and as a consequence, small-molecule ligands of HSP90 are preferentially retained in tumors up to 20 times longer than in normal tissue. We hypothesize that the generation of conjugates that use a HSP90-targeting ligand and a payload such as copanlisib, may open the narrow therapeutic window of this and other PI3K inhibitors. In support of this hypothesis, we have generated a HSP90-PI3K drug conjugate, T-2143 and utilizing xenograft models, demonstrate rapid and sustained tumor accumulation of the conjugate, deep pathway inhibition, and superior efficacy than the PI3K inhibitor on its own. Selective delivery of T-2143 and the masking of the inhibitor active site was also able to mitigate a potentially dose-limiting side effect of copanlisib, hyperglycemia. These data demonstrate that by leveraging the preferential accumulation of HSP90-targeting ligands in tumors, we can selectively deliver a PI3K inhibitor leading to efficacy in multiple tumor models without hyperglycemia in mice. These data highlight a novel drug delivery strategy that allows for the potential opening of a narrow therapeutic window through specific tumor delivery of anticancer payloads and reduction of toxicity.

Iron-catalyzed 1,4-hydroboration of 1,3-dienes.

A chemo-, regio-, and stereoselective iron-catalyzed 1,4-hydroboration of dienes that affords gamma-disubstituted allylboranes has been developed. 1,4-Hydroboration of 2-substituted dienes forms allylborane products with (E)-trisubstituted double bonds exclusively.

Targeting the Somatostatin Receptor 2 with the Miniaturized Drug Conjugate, PEN-221: A Potent and Novel Therapeutic for the Treatment of Small Cell Lung Cancer.

Small cell lung cancer (SCLC) is an aggressive neuroendocrine carcinoma with a 95% mortality rate with no improvement to treatment in decades, and new therapies are desperately needed. PEN-221 is a miniaturized peptide-drug conjugate (∼2 kDa) designed to target SCLC via a Somatostatin Receptor 2 (SSTR2)-targeting ligand and to overcome the high proliferation rate characteristic of this disease by using the potent cytotoxic payload, DM1. SSTR2 is an ideal target for a drug conjugate, as it is overexpressed in SCLC with limited normal tissue expression. In vitro, PEN-221 treatment of SSTR2-positive cells resulted in PEN-221 internalization and receptor-dependent inhibition of cellular proliferation. In vivo, PEN-221 exhibited rapid accumulation in SSTR2-positive SCLC xenograft tumors with quick clearance from plasma. Tumor accumulation was sustained, resulting in durable pharmacodynamic changes throughout the tumor, as evidenced by increases in the mitotic marker of G2-M arrest, phosphohistone H3, and increases in the apoptotic marker, cleaved caspase-3. PEN-221 treatment resulted in significant antitumor activity, including complete regressions in SSTR2-positive SCLC xenograft mouse models. Treatment was effective using a variety of dosing schedules and at doses below the MTD, suggesting flexibility of dosing schedule and potential for a large therapeutic window in the clinic. The unique attributes of the miniaturized drug conjugate allowed for deep tumor penetration and limited plasma exposure that may enable long-term dosing, resulting in durable tumor control. Collectively, these data suggest potential for antitumor activity of PEN-221 in patients with SSTR2-positive SCLC.

Discovery of an SSTR2-Targeting Maytansinoid Conjugate (PEN-221) with Potent Activity in Vitro and in Vivo.

Somatostatin receptor 2 (SSTR2) is frequently overexpressed on several types of solid tumors, including neuroendocrine tumors and small-cell lung cancer. Peptide agonists of SSTR2 are rapidly internalized upon binding to the receptor and linking a toxic payload to an SSTR2 agonist is a potential method to kill SSTR2-expressing tumor cells. Herein, we describe our efforts towards an efficacious SSTR2-targeting cytotoxic conjugate; examination of different SSTR2-targeting ligands, conjugation sites, and payloads led to the discovery of 22 (PEN-221), a conjugate consisting of microtubule-targeting agent DM1 linked to the C-terminal side chain of Tyr3-octreotate. PEN-221 demonstrates in vitro activity which is both potent (IC50 = 10 nM) and receptor-dependent (IC50 shifts 90-fold upon receptor blockade). PEN-221 targets high levels of DM1 to SSTR2-expressing xenograft tumors, which has led to tumor regressions in several SSTR2-expressing xenograft mouse models. The safety and efficacy of PEN-221 is currently under evaluation in human clinical trials.

Utilization of methyloleate in production of microbial lipase.

In this article, we report the development and optimization of an industrial culture medium for the production of extracellular lipase in the yeast Yarrowia lipolytica. Until now olive oil in combination with glucose was used as the carbon source and inducer for the production of lipase. Our results demonstrate that methyloleate, a cheap hydrophobic compound, could efficiently substitute olive oil as the inducer and carbon source for lipase production. A new process of lipase production was developed yielding a twofold increase in the level of production compared with the levels in previous reports.

Simple method provides resolution of albumin, lipoprotein, free fraction, and chylomicron to enhance the utility of protein binding assays.

Medicinal chemists have been encouraged in recent years to embrace high speed protein binding assays. These methods employ dialysis membranes in 96-well format or spin filters. Membrane-based methods do not separate lipoprotein binding from albumin binding and introduce interference despite membrane binding controls. Ultracentrifugation methods, in contrast, do not introduce interference if density gradients can be avoided and they resolve lipoprotein from albumin. A new generation of compact, fast ultracentrifuges facilitates the rapid and fully informative separation of plasma into albumin, albumin/fatty acid complex, lipoprotein, protein-free, and chylomicron fractions with no need of salt or sugar density gradients. We present a simple and fast ultracentrifuge method here for two platinum compounds and a taxane that otherwise bound irreversibly to dialysis membranes and which exhibited distinctive lipoprotein binding behaviors. This new generation of ultracentrifugation methods underscores a need to further discuss protein binding assessments as they relate to medicinal chemistry efforts.

Discovery of hepatitis C virus NS3-4A protease inhibitors with improved barrier to resistance and favorable liver distribution.

Given the emergence of resistance observed for the current clinical-stage hepatitis C virus (HCV) NS3 protease inhibitors, there is a need for new inhibitors with a higher barrier to resistance. We recently reported our rational approach to the discovery of macrocyclic acylsulfonamides as HCV protease inhibitors addressing potency against clinically relevant resistant variants. Using X-ray crystallography of HCV protease variant/inhibitor complexes, we shed light on the complex structural mechanisms by which the D168V and R155K residue mutations confer resistance to NS3 protease inhibitors. Here, we disclose SAR investigation and ADME/PK optimization leading to the identification of inhibitors with significantly improved potency against the key resistant variants and with increased liver partitioning.

Iron-catalyzed 1,4-addition of alpha-olefins to dienes.

A new intermolecular, stereo- and regioselective iron-catalyzed 1,4-addition of alpha-olefins to 1,3-dienes using as low as 1 mol % of an iminopyridine-ferrous chloride complex was developed. Importantly, both double bonds of the linear 1,4-diene addition products are obtained with absolute stereocontrol.

Expedient stereoselective synthesis of coronafacic acid through intramolecular Diels-Alder cyclization.

A stereoselective synthesis of coronafacic acid, a natural component of the phytotoxin coronatin, was achieved using an intramolecular Diels-Alder reaction as the key step. The triene precursor bearing a substituted diene and a vinylketone as dienophile was synthesized and then tested in the thermal intramolecular cyclization. We have devised a new strategy to assemble the E,Z-diene through the stereoselective aldol reaction of an ester enolate followed by a stereoselective dehydration. Following the thermal cyclization, the corresponding hydrindanone thereby obtained with the desired relative stereochemistry could easily be converted into the natural product. The synthesis of the coronafacic acid was accomplished in six steps in 29% overall yield.

Expedient synthesis of cyclopropane alpha-amino acids by the catalytic asymmetric cyclopropanation of alkenes using iodonium ylides derived from methyl nitroacetate.

A highly enantioselective (up to 97.5% ee) and diastereoselective (95:5 dr trans/cis) Cu(I)-catalyzed cyclopropanation of alkenes using phenyliodonium ylide generated in situ from iodosobenzene and methyl nitroacetate is reported. The cyclopropanation took place with high enantioselectivity for a wide range of alkenes, and the reaction was performed at room temperature. 1-Nitrocyclopropyl esters are versatile building blocks to access the corresponding cyclopropane amino esters and aminocyclopropanes in two and three steps, respectively, from commercially available products.

Synthesis of novel D-glucuronic acid fatty esters using Candida antarctica lipase in tert-butanol.

Glucuronic acid n-alkyl esters, a novel class of promising biosurfactants and their corresponding glucose esters with the same side-chain length, were synthesized by direct esterification in a non-aqueous phase (tert-butanol) using an immobilized lipase.