N-Acylamino Saccharin as an Emerging Cysteine-Directed Covalent Warhead and Its Application in the Identification of Novel FBPase Inhibitors toward Glucose Reduction.

With a resurgence of covalent drugs, there is an urgent need for the identification of new moieties capable of cysteine bond formation. Herein, we report on the N-acylamino saccharin moieties capable of novel covalent reactions with cysteine. Their utility as alternative electrophilic warheads was demonstrated through the covalent modification of fructose-1,6-bisphosphatase (FBPase), a promising target associated with cancer and type 2 diabetes. The cocrystal structure of title compound W8 bound with FBPase unexpectedly revealed that the N-acylamino saccharin moiety worked as an electrophile warhead that covalently modified the noncatalytic C128 site in FBPase while releasing saccharin, suggesting a previously undiscovered covalent reaction mechanism of saccharin derivatives with cysteine. Treatment of title compound W8 displayed potent inhibition of glucose production in vitro and in vivo. This newly discovered reactive warhead supplements the current repertoire of cysteine covalent modifiers while avoiding some of the limitations generally associated with established moieties.

Analysis and comparison of seed protein, oil, and sugars in edamame dried using two oven-drying methods and mature soybeans.

BACKGROUND: Edamame, a vegetable soybean (Glycine max) grown mainly in Asia, has high nutritional and market value and is a relatively new crop to North America. By 2 years of field trials, we evaluated the seed composition traits in 54 genotypes to analyze the differences and relationship between edamame seeds dried by two oven-drying methods and mature soybeans. RESULTS: The genotypic differences were significant for all the traits investigated. Significant differences also existed between the two sets of dried edamame and mature seeds. Protein content in mature soybean averaged 426.8 g kg-1 , and 432.8 g kg-1 and 405.6 g kg-1 for shelled-dried and unshelled-dried edamame respectively. Oil content in shelled-dried and unshelled-dried edamame averaged 206.3 g kg-1 and 212.6 g kg-1 respectively, and 195.8 g kg-1 for mature soybean. Sucrose content in mature soybean (60.2 g kg-1 ) was approximately 1.5 and 3 times that of unshelled-dried and shelled-dried edamame respectively. Mature soybean also exhibited the highest concentrations of stachyose and total sugars, followed by unshelled-dried and shelled-dried edamame. The broad-sense heritability estimates of traits in mature soybean (49.41-89.16%) were higher than those of edamame (10.26-78.96%). Higher broad-sense heritability was uncovered for protein and oil, but lower estimates for sugars, fiber, and ash. Positive correlations were detected between the two sets of edamame seeds and mature soybean for protein and oil (r = 0.63-0.88). CONCLUSION: The results suggest that indirect selection through mature seeds is helpful for the improvement of protein and oil in edamame, whereas the improvement of seed sugars in edamame is more challenging. © 2020 Society of Chemical Industry.

Design, Synthesis, and Biological Evaluation of Orally Available First-Generation Dual-Target Selective Inhibitors of Acetylcholinesterase (AChE) and Phosphodiesterase 5 (PDE5) for the Treatment of Alzheimer's Disease.

Through drug discovery strategies of repurposing and redeveloping existing drugs, a series of novel tadalafil derivatives were rationally designed, synthesized, and evaluated to seek dual-target AChE/PDE5 inhibitors as good candidate drugs for Alzheimer's disease (AD). Among these derivatives, 1p and 1w exhibited excellent selective dual-target AChE/PDE5 inhibitory activities and improved blood-brain barrier (BBB) penetrability. Importantly, 1w·Cit (citrate of 1w) could reverse the cognitive dysfunction of scopolamine-induced AD mice and exhibited an excellent effect on enhancing cAMP response element-binding protein (CREB) phosphorylation in vivo, a crucial factor in memory formation and synaptic plasticity. Moreover, the molecular docking simulations of 1w with hAChE and hPDE5A confirmed that our design strategy was rational. In summary, our research provides a potential selective dual-target AChE/PDE5 inhibitor as a good candidate drug for the treatment of AD, and it could also be regarded as a small molecule probe to validate the novel AD therapeutic approach in vivo.

Novel Vilazodone-Tacrine Hybrids as Potential Multitarget-Directed Ligands for the Treatment of Alzheimer's Disease Accompanied with Depression: Design, Synthesis, and Biological Evaluation.

Depression is one of the most frequent psychiatric complications of Alzheimer's disease (AD), affecting up to 50% of the patients. A novel series of hybrid molecules were designed and synthesized by combining the pharmacophoric features of vilazodone and tacrine as potential multitarget-directed ligands for the treatment of AD with depression. In vitro biological assays were conducted to evaluate the compounds; among the 30 hybrids, compound 1e showed relatively balanced profiles between acetylcholinesterase inhibition (IC50 = 3.319 ± 0.708 µM), 5-HT1A agonist (EC50 = 107 ± 37 nM), and 5-HT reuptake inhibition (IC50 = 76.3 ± 33 nM). Compound 1e displayed tolerable hepatotoxicity and moderate hERG inhibition activity, and could penetrate the blood-brain barrier in vivo. Furthermore, an oral intake of 30 mg/kg 1e·HCl could significantly improve the cognitive function of scopolamine-induced amnesia mice and alleviate the depressive symptom in tail suspension test. The effectivity of 1e validates the rationality of our design strategy.

Functional characterization of Corynebacterium glutamicum mycothiol S-conjugate amidase.

The present study focuses on the genetic and biochemical characterization of mycothiol S-conjugate amidase (Mca) of Corynebacterium glutamicum. Recombinant C. glutamicum Mca was heterologously expressed in Escherichia coli and purified to apparent homogeneity. The molecular weight of native Mca protein determined by gel filtration chromatography was 35 kDa, indicating that Mca exists as monomers in the purification condition. Mca showed amidase activity with mycothiol S-conjugate of monobromobimane (MSmB) in vivo while mca mutant lost the ability to cleave MSmB. In addition, Mca showed limited deacetylase activity with N-acetyl-D-glucosamine (GlcNAc) as substrate. Optimum pH for amidase activity was between 7.5 and 8.5, while the highest activity in the presence of Zn2+ confirmed Mca as a zinc metalloprotein. Amino acid residues conserved among Mca family members were located in C. glutamicum Mca and site-directed mutagenesis of these residues indicated that Asp14, Tyr137, His139 and Asp141 were important for activity. The mca deletion mutant showed decreased resistance to antibiotics, alkylating agents, oxidants and heavy metals, and these sensitive phenotypes were recovered in the complementary strain to a great extent. The physiological roles of Mca in resistance to various toxins were further supported by the induced expression of Mca in C. glutamicum under various stress conditions, directly under the control of the stress-responsive extracytoplasmic function-sigma (ECF-σ) factor SigH.

Stacking of a stearoyl-ACP thioesterase with a dual-silenced palmitoyl-ACP thioesterase and ∆12 fatty acid desaturase in transgenic soybean.

Soybean (Glycine max (L.) Merr) is valued for both its protein and oil, whose seed is composed of 40% and 20% of each component, respectively. Given its high percentage of polyunsaturated fatty acids, linoleic acid and linolenic acid, soybean oil oxidative stability is relatively poor. Historically food processors have employed a partial hydrogenation process to soybean oil as a means to improve both the oxidative stability and functionality in end-use applications. However, the hydrogenation process leads to the formation of trans-fats, which are associated with negative cardiovascular health. As a means to circumvent the need for the hydrogenation process, genetic approaches are being pursued to improve oil quality in oilseeds. In this regard, we report here on the introduction of the mangosteen (Garcinia mangostana) stearoyl-ACP thioesterase into soybean and the subsequent stacking with an event that is dual-silenced in palmitoyl-ACP thioesterase and ∆12 fatty acid desaturase expression in a seed-specific fashion. Phenotypic analyses on transgenic soybean expressing the mangosteen stearoyl-ACP thioesterase revealed increases in seed stearic acid levels up to 17%. The subsequent stacked with a soybean event silenced in both palmitoyl-ACP thioesterase and ∆12 fatty acid desaturase activity, resulted in a seed lipid phenotype of approximately 11%-19% stearate and approximately 70% oleate. The oil profile created by the stack was maintained for four generations under greenhouse conditions and a fifth generation under a field environment. However, in generation six and seven under field conditions, the oleate levels decreased to 30%-40%, while the stearic level remained elevated.

Structure and mechanical properties of new biomass-based nanocomposite: castor oil-based polyurethane reinforced with acetylated cellulose nanocrystal.

New nanocomposites consisting of a castor oil-based polyurethane matrix filled with acetylated cellulose nanocrystals (ACNs) were developed. The ACN exhibited improved dispersion in tetrahydrofuran as a blending medium, and reduced polarity as compared with unmodified cellulose nanocrystals, resulting in a high loading level of 25 wt% in the nanocomposite. As the ACN loading-level increased from 0% to 25%, the tensile strength and Young's modulus of the nanocomposites increased from 2.79 MPa to 10.41 MPa and from 0.98 MPa to 42.61 MPa, respectively. When the ACN loading-level was 10 wt%, the breaking elongation of the nanocomposites reached the maximum value of more than twice that of the polyurethane. The enhanced mechanical performance was primarily attributed to the formation of a three-dimensional ACN network and strong interfacial interactions between filler and matrix. This work produced new polyurethane-based nanocomposites containing modified cellulose nanocrystal with a high biomass content. Its high performance could contribute to potential applications.