Evaluation of darunavir-derived HIV-1 protease inhibitors incorporating P2' amide-derivatives: Synthesis, biological evaluation and structural studies.

We report here the synthesis and biological evaluation of darunavir derived HIV-1 protease inhibitors and their functional effect on enzyme inhibition and antiviral activity in MT-2 cell lines. The P2' 4-amino functionality was modified to make a number of amide derivatives to interact with residues in the S2' subsite of the HIV-1 protease active site. Several compounds exhibited picomolar enzyme inhibitory and low nanomolar antiviral activity. The X-ray crystal structure of the chloroacetate derivative bound to HIV-1 protease was determined. Interestingly, the active chloroacetate group converted to the acetate functionality during X-ray exposure. The structure revealed that the P2' carboxamide functionality makes enhanced hydrogen bonding interactions with the backbone atoms in the S2'-subsite.

Cytotoxic Assessment of 3,3-Dichloro-ß-Lactams Prepared through Microwave-Assisted Benzylic C-H Activation from Benzyl-Tethered Trichloroacetamides Catalyzed by RuCl2(PPh3)3.

Natural and synthetic ß-lactam derivatives constitute an interesting class of compounds due to their diverse biological activity. Mostly used as antibiotics, they were also found to have antitubercular, anticancer and antidiabetic activities, among others. In this investigation, six new 3,3-dichloro-ß-lactams prepared in a previous work were evaluated for their hemolytic and cytotoxic properties. The results showed that the proposed compounds have non-hemolytic properties and exhibited an interesting cytotoxic activity toward squamous cell carcinoma (A431 cell line), which was highly dependent on the structure and concentration of these ß-lactams. Among the molecules tested, 2b was the most cytotoxic, with the lowest IC50 values (30-47 µg/mL) and a promising selectivity against the tumor cells compared with non-tumoral cells.

The potential for bacteria from carbon-limited deep terrestrial environments to participate in chlorine cycling.

Bacteria capable of dehalogenation via reductive or hydrolytic pathways are ubiquitous. Little is known, however, about the prevalence of bacterial dechlorination in deep terrestrial environments with a limited carbon supply. In this study we analyzed published genomes from three deep terrestrial subsurface sites: a deep aquifer in Western Siberia, the Sanford Underground Research Facility in South Dakota, USA, and the Soudan Underground Iron Mine (SUIM) in Minnesota, USA to determine if there was evidence to suggest that microbial dehalogenation was possible in these environments. Diverse dehalogenase genes were present in all analyzed metagenomes, with reductive dehalogenase and haloalkane dehalogenase genes the most common. Taxonomic analysis of both hydrolytic and reductive dehalogenase genes was performed to explore their affiliation; this analysis indicated that at the SUIM site, hydrolytic dehalogenase genes were taxonomically affiliated with Marinobacter species. Because of this affiliation, experiments were also performed with Marinobacter subterrani strain JG233 ('JG233'), an organism containing three predicted hydrolytic dehalogenase genes and isolated from the SUIM site, to determine whether hydrolytic dehalogenation was an active process and involved in growth on a chlorocarboxylic acid. Presence of these genes in genome appears to be functional, as JG233 was capable of chloroacetate dechlorination with simultaneous chloride release. Stable isotope experiments combined with confocal Raman microspectroscopy demonstrated that JG233 incorporated carbon from 13C-chloroacetate into its biomass. These experiments suggest that organisms present in these extreme and often low-carbon environments are capable of reductive and hydrolytic dechlorination and, based on laboratory experiments, may use this capability as a competitive advantage by utilizing chlorinated organic compounds for growth, either directly or after dechlorination.

Rh(I)-Catalyzed Allenic Pauson-Khand Reaction to Access the Thapsigargin Core: Influence of Furan and Allenyl Chloroacetate Groups on Enantioselectivity.

Thapsigargin (Tg) is a potent SERCA pump inhibitor with the potential to treat cancer and COVID-19. We have extended the scope of the asymmetric allenic Pauson-Khand reaction to furan-tethered allene-ynes, a stereoconvergent transformation affording the 5,7,5-ring system of Tg in good yields and high enantioselectivity. Computational studies of the oxidative cyclization step show that the furan and chloroacetate groups contribute to this high selectivity.

Slow glycosylation: Activation of trichloroacetimidates under mild conditions using lithium salts and the role of counterions.

Glycosylations were carried out with the two glycosyl donors 4-O-acetyl-2,3-O-isopropylidene-1-O-trichloroacetimidoyl-α-l-rhamnopyranose and 2,3,4-tri-O-benzyl-1-O-trichloro-acetimidoyl-α-l-rhamnopyranose in combination with the two alcohols 1-adamantanol and l-menthol as model glycosyl acceptors. As catalysts, the five lithium salts LiNTf2, LiI, LiClO4, LiPF6 and LiOTf were investigated. We demonstrated that both lithium and the respective counterions are playing a role in the activation of trichloroacetimidate glycosyl donors at rt. Under these very mild conditions, the glycosylations are slow and completed in two to eight days. Depending on the counterion, the rate and yield of the reaction differs; however, the selectivity of all investigated lithium salts is deficient.

Tris(pentafluorophenyl)borane-Catalyzed Stereoselective C-Glycosylation of Indoles with Glycosyl Trichloroacetimidates: Access to 3-Indolyl-C-glycosides.

An efficient and highly regioselective method for the synthesis of 3-indolyl-C-glycosides has been developed through coupling of glycosyl trichloroacetimidates with a wide range of substituted indoles in the presence of catalytic amounts of B(C6F5)3 within a few minutes. This methodology has a wide scope of substrates under mild reaction conditions and provides exclusively ß-stereoselective 3-indolyl-C-glycosides in 64-87% yields.

Ab Initio Molecular Dynamics Simulations of the SN1/SN2 Mechanistic Continuum in Glycosylation Reactions.

We report a computational approach to evaluate the reaction mechanisms of glycosylation using ab initio molecular dynamics (AIMD) simulations in explicit solvent. The reaction pathways are simulated via free energy calculations based on metadynamics and trajectory simulations using Born-Oppenheimer molecular dynamics. We applied this approach to investigate the mechanisms of the glycosylation of glucosyl α-trichloroacetimidate with three acceptors (EtOH, i-PrOH, and t-BuOH) in three solvents (ACN, DCM, and MTBE). The reactants and the solvents are treated explicitly using density functional theory. We show that the profile of the free energy surface, the synchronicity of the transition state structure, and the time gap between leaving group dissociation and nucleophile association can be used as three complementary indicators to describe the glycosylation mechanism within the SN1/SN2 continuum for a given reaction. This approach provides a reliable means to rationalize and predict reaction mechanisms and to estimate lifetimes of oxocarbenium intermediates and their dependence on the glycosyl donor, acceptor, and solvent environment.

An efficient synthetic route to O-(2-O-benzyl-3,4-di-O-acetyl-α/ß-l-fucopyranosyl)-trichloroacetimidate.

An efficient synthetic route to prepare O-(2-O-benzyl-3,4-di-O-acetyl-α/ß-l-fucopyranosyl)-trichloroacetimidate from l-fucose was developed by introducing the thiophenyl group at the anomeric center and the benzylidene functional group to protect the 3 and 4 positions. Although three approaches were considered, the best result was obtained when, after the 2-hydroxyl benzylation, both protective groups were simultaneously removed by using acetic anhydride and perchloric acid supported on silica as catalyst. Selective deacetylation of the obtained tri-O-acetate followed by the reaction of the resultant hemiacetal with trichloroacetonitrile and DBU afforded the trichloroacetimidate with an overall yield of 56% from the l-fucose.

Trichloroacetyl chloride, CCl3COCl, as an alternative Cl atom precursor for laboratory use and determination of Cl atom rate coefficients for n-CH2[double bond, length as m-dash]CH(CH2)xCN (x = 3-4).

An investigation of CCl3COCl was conducted with the purpose of using the compound as an alternative Cl atom precursor in laboratory settings. CCl3COCl can be used with or without O2 as a source of Cl atoms and photolysis studies in air and N2 diluent displayed COCl2 and CO as being the major photolysis products. Relative rate studies were performed to determine the Cl atom rate coefficients for reaction with CH3Cl and C2H2 and the results were in agreement with literature values. Cl atom rate coefficients for reaction with n-CH2[double bond, length as m-dash]CH(CH2)3CN and n-CH2[double bond, length as m-dash]CH(CH2)4CN were determined as (2.95 ± 0.58) × 10-10 and (3.73 ± 0.60) × 10-10 cm3 molecule-1 s-1, respectively. CCl3COCl requires UV-C irradiation, so not all molecules are feasible for use in e.g. relative rate studies. Furthermore, it is recommended to perform experiments with O2 present, as this minimizes IR feature disturbance from product formation.

A visible light promoted O-glycosylation with glycosyl trichloroacetimidates using eosin Y as an organo photoacid.

A photoacid catalyzed O-glycosylation of alcohols with glycosyl trichloroacetimidates in the presence of commercially available phenolic photoacids, fluorescein, 4',5'-dibromo-fluorescein, and eosin Y under visible light irradiation by blue LEDs was developed. The method is operationally simple without neutralization and proceeds at room temperature.

Dialkylation of Indoles with Trichloroacetimidates to Access 3,3-Disubstituted Indolenines.

2-Substituted indoles may be directly transformed to 3,3-dialkyl indolenines with trichloroacetimidate electrophiles and the Lewis acid TMSOTf. These reactions provide rapid access to complex indolenines which are present in a variety of complex natural products and medicinally relevant small molecule structures. This method provides an alternative to the use of transition metal catalysis. The indolenines are readily transformed into spiroindoline systems which are privileged scaffolds in medicinal chemistry.

An anticancer Os(II) bathophenanthroline complex as a human breast cancer stem cell-selective, mammosphere potent agent that kills cells by necroptosis.

Conventional chemotherapy is mostly effective in the treatment of rapidly-dividing differentiated tumor cells but has limited application toward eliminating cancer stem cell (CSC) population. The presence of a very small number of CSCs may contribute to the development of therapeutic resistance, metastases, and relapse. Thus, treatment failure by developing novel anticancer drugs capable of effective targeting of CSCs is at present a major challenge for research focused on chemotherapy of cancer. Here, we show that Os(II) complex 2 [Os(η6-pcym)(bphen)(dca)]PF6 (pcym = p-cymene, bphen = bathophenanthroline, and dca = dichloroacetate), is capable of efficient and selective killing CSCs in heterogeneous populations of human breast cancer cells MCF-7 and SKBR-3. Notably, its remarkable submicromolar potency to kill CSCs is considerably higher than that of its Ru analog, [Ru(η6-pcym)(bphen)(dca)]PF6 (complex 1) and salinomycin, one of the most selective CSC-targeting compounds hitherto identified. Furthermore, Os(II) complex 2 reduces the formation, size, and viability of three-dimensional mammospheres which more closely reflect the tumor microenvironment than cells in traditional two-dimensional cultures. The antiproliferation studies and propidium iodide staining using flow cytometry suggest that Os(II) complex 2 induces human breast cancer stem cell death predominantly by necroptosis, a programmed form of necrosis. The results of this study demonstrate the promise of Os(II) complex 2 in treating human breast tumors. They also represent the foundation for further preclinical and clinical studies and applications of Os(II) complex 2 to comply with the emergent need for human breast CSCs-specific chemotherapeutics capable to treat chemotherapy-resistant and relapsed human breast tumors.

Degradation of haloacetic acids with the Fenton-like and analysis by GC-MS: use of bioassays for monitoring of genotoxic, mutagenic and cytotoxic effects.

In this study, a method was developed to evaluate the degradation of haloacetic acids (HAAs) in water by a heterogenous Fenton-like process catalyzed by cobalt-doped magnetite nanoparticles (Fe3 - xCoxO4), extraction of the contaminants by liquid-liquid extraction (LLE), and analysis by gas chromatography-mass spectrometry (GC-MS). The developed method was efficient in the degradation of HAAs, with the following degradation values: 63%, 62%, 30%, 39%, 37%, 50%, 84%, 41%, and 79% for monochloroacetic acid, monobromoacetic acid, dichloroacetic acid, trichloroacetic acid, bromochloroacetic acid, dibromoacetic acid, bromodichloroacetic acid, dibromochloroacetic acid, and tribromoacetic acid compounds, respectively. Through the application of the Allium cepa test, the cytotoxicity, genotoxicity, and mutagenicity of HAAs were evaluated. The results confirm its genotoxic and mutagenic effects on Allium cepa meristematic cells. Through this study, it was possible to verify the effectiveness of the developed method and its potential as a proposal for environmental remediation.

Application of different advanced oxidation processes for the removal of chloroacetic acids using a planar falling film reactor.

Advanced oxidation processes (AOPs) are considered as an effective and promising method for the degradation and mineralization of aqueous recalcitrant organic pollutants. In this study, application of ozonation and various types of AOPs including photocatalysis, Fenton alone and their combinations were investigated and compared for the degradation and mineralization of chloroacetic acids (CAAs) in aqueous solutions, using a planar falling film reactor. CAAs are widely available in water treated by chlorination processes and are resistance against ozonation in the darkness. The results of the present work showed that the plain ozonation was inefficient method for the destruction of the CAAs as only about 2% degradation was observed after 90 min treatment. However, the best results were achieved by ozone in combinations with other oxidation processes. Furthermore, a synergistic effect on the removal rate was observed when these processes were exposed to the UVA light. Among the examined processes, combination of photo-Fenton with ozonation was found to be the fastest one for CAAs degradation. The effects of different parameters such as initial concentration of Fe2⁺, H2O2 and CAAs in photo-Fenton combined with ozonation were investigated. The optimum ratio of 0.12 of Fe2⁺/H2O2 concentration was found to give the best result for CAAs degradation. The degree of CAAs mineralization, measured by the total organic carbon removal, as well as the effect of falling liquid film flow rate on the removal of CAAs were also studied and discussed.

Ester Formation via Symbiotic Activation Utilizing Trichloroacetimidate Electrophiles.

Trichloroacetimidates are useful reagents for the synthesis of esters under mild conditions that do not require an exogenous promoter. These conditions avoid the undesired decomposition of substrates with sensitive functional groups that are often observed with the use of strong Lewis or Brønsted acids. With heating, these reactions have been extended to benzyl esters without electron-donating groups. These inexpensive and convenient methods should find application in the formation of esters in complex substrates.

Occurrence of disinfection by-products in swimming pools and the estimated resulting cytotoxicity.

Swimming pools are disinfected to protect against the risk of microbial disease, however, the formation of disinfection by-products (DBPs) is an unwanted consequence. While many studies have reported the occurrence of commonly investigated DBPs (trihalomethanes and haloacetic acids) in pools, few studies have investigated emerging DBP classes, such as the haloketones or haloacetaldehydes, and the nitrogenous haloacetamides, halonitromethanes, haloacetonitriles and N-nitrosamines. This study investigated the occurrence of sixty four DBPs from the eight aforementioned DBP classes in pools employing different treatment methods. Approximately 70% of the DBPs were detected in at least one of the pools, with most concentrations being equal to or greater than those previously reported. Chloral hydrate (trichloroacetaldehyde) was one of many DBPs detected in all chlorinated waters (202 to 1313 µg/L), and, on a molar basis, was the predominant DBP. Several other DBPs, namely chloroacetic acid, dichloroacetic acid, trichloroacetic acid, dichloroacetamide, dibromoacetamide, dibromochloroacetamide and trichloroacetamide, and many of the N-nitrosamines, were measured at concentrations greater than previously reported: up to 200 to 479 µg/L for the haloacetic acids, 56 to 736 µg/L for the haloacetamides and up to 1093 ng/L for some N-nitrosamines. The higher disinfectant residuals required to be employed in Australian pools, and poor pool management (e.g. of chlorine residual and pH) are likely factors contributing to these relatively high DBP concentrations. Where possible, the cytotoxicity values of the investigated DBPs were evaluated, with chloral hydrate representing over 90% of the total chronic cytotoxicity despite only representing up to 64% of the total molar DBP concentration. This study is the first report of bromodichloroacetaldehyde and bromochloroacetaldehyde in pools and is the first investigation of N-nitrosamines in a brominated pool. Furthermore, this work aids in understanding DBPs in both chlorine and bromine treated pools, the latter being the subject of only limited previous studies.

Organocatalytic Direct α-Selective N-Glycosylation of Amide with Glycosyl Trichloroacetimidate.

Through the synergistic catalytic effect of the halogen bond (XB) donor and thiourea catalyst, a direct α-selective N-glycosylation of the amide residue of asparagine derivative was achieved using readily accessible glycosyl trichloroacetimidate. n-Butyl methyl ether was found to be the most suitable solvent for the α-selectivity.

Promoter free allylation of trichloroacetimidates with allyltributylstannanes under thermal conditions to access the common 1,1'-diarylbutyl pharmacophore.

1,1'-Diarylbutyl groups are a common pharmacophore found in many biologically active small molecules. To access these systems under mild conditions, the reaction of diarylmethyl trichloroacetimidates with allyltributylstannanes was explored. Simply heating allyltributylstannane with the trichloroacetimidate resulted in substitution of the imidate with an allyl group. Unlike other methods used to access these systems, no strong base, transition metal catalyst, Brønsted acid or Lewis acid promoter was required to affect the transformation. Conversions are best with electron rich benzylic trichloroacetimidate systems, where excellent yields are achieved just by refluxing the reactants together in nitromethane.

ß-L-Arabinofuranosylation Conducted by 5-O-(2-pyridinecarbonyl)-L-arabinofuranosyl Trichloroacetimidate.

We describe a ß-L-arabinofuranosylation method by employing the 5-O-(2-pyridinecarbonyl)-L-arabinofuranosyl trichloroacetimidate 10 as a donor. This approach allows a wide range of acceptor substrates, especially amino acid acceptors, to be used. Stereoselective synthesis of ß-(1,4)-L-arabinofuranosyl-(2S, 4R)-4-hydroxy-L-proline (ß-L-Araf-L-Hyp4) and its dimer is achieved readily by this method. Both the stereoselectivities and yields of the reactions are excellent. To demonstrate the utility of this methodology, the preparation of a trisaccharide in a one-pot manner was carried out.

One-pot Microwave-assisted Conversion of Anomeric Nitrate-esters to Trichloroacetimidates.

The goal of the following procedure is to provide a demonstration of the one-pot conversion of a 2-azido-1-nitrate-ester to a trichloroacetimidate glycosyl donor. Following azido-nitration of a glycal, the product 2-azido-1-nitrate ester can be hydrolyzed under microwave-assisted irradiation. This transformation is usually achieved using strongly nucleophilic reagents and extended reaction times. Microwave irradiation induces hydrolysis, in the absence of reagents, with short reaction times. Following denitration, the intermediate anomeric alcohol is converted, in the same pot, to the corresponding 2-azido-1-trichloroacetimidate.