Characterisation of Metallo-Cyclodextrins for Chiral Separations

Almost half of commercial pharmaceuticals are chiral and therefore effective analytical techniques to detect and quantify enantiomers are important in the pharmaceutical sector to ensure drug safety and efficacy. In this work copper(II) complexes of aminoalkane derivatives of β-cyclodextrin (CDEn, CDPn and CDBn) were investigated to determine if they provide a versatile solution for the direct separation of the enantiomers of DOPA, benserazide and carbidopa using capillary electrophoresis. Tyrosine was used as a model compound. Electronic spectroscopy was used in order to determine if copper(II) coordinates to the amino-CDs and to determine if further coordination to the guest species occurs. Decreases in λmax for the d-d transitions on coordination of the metal ion suggest the formation of a CuCDAm binary complex with CDEn and CDPn acting as bidentate ligands and CDBn acting as a monodentate ligand. A further blue shift on the introduction of a guest species suggests further coordination of copper(II) and formation of ternary complexes of tyrosine and DOPA with all three metallo-complexes of the aminoalkane derivatives. A ternary complex also formed between CuCDEn and Lcarbidopa. No coordination was evident with benserazide. These results suggest that the use of these metallo-cyclodextrin derivatives for enantioselection may be guest specific if ternary complex formation is required for separation. Cicular dichroism (c.d.) can show the differential inclusion of enantiomers in cyclodextrin cavities and therefore was used here to determine if the metallocyclodextrins can differentiate between enantiomers. Increases in intensity of the bands in the spectra indicate deeper inclusion of a guest in the CD cavity. From the results obtained in this study it can be shown that the binary complex CuCDEn is the most ii enantioselective material of those studied towards both tyrosine and DOPA. CuCDPn and CuCDBn showed no enantioselection. The stoichiometry of all complexes was determined using electronic spectroscopy and Job’s method of continuous variation. A 1:1 binary complex formed between Cu(II) and CDEn. It was also shown that CuCDEn formed a 1:1 ternary complex with tyrosine and a 2:1 complex with DOPA. Results also suggested that both 1:1 and 2:1 complexes of CuCDEn and L-carbidopa were formed. Using CE in normal polarity mode and a background electrolyte (BGE) at pH 6.8, separation of the enantiomers of DOPA was not achieved using CuCDEn as a chiral selector. The stability of the binary complex is limited by pH and therefore CDEn alone was used as the chiral selector. Separation of L-DOPA from L-carbidopa was achieved using CDEn in a BGE at pH 2.5 and a resolution of 2.36 was obtained. However it was still not possible to separate the isomers of DOPA. An alternative charged CD derivative, sulfated-β-cyclodextrin was investigated and was found to successfully separate the enantiomers of D,L-DOPA in a BGE of pH 2.5 and a resolution of 3.62 was obtained. These results suggest that amino-cyclodextrin derivatives are guest specific for chiral separations in capillary electrophoresis and do not represent a universal method for separations required for regulatory compliance. Since separations were obtained using the anionic sulfated derivatives it is suggested that host molecules with a permanent charge are more useful and therefore inclusion is not the only factor needed for separation to be successful

Poly(ethylene glycol)-Based Peptidomimetic “PEGtide” of Oligo-Arginine allows for efficient siRNA Transfection and gene inhibition

While a wide range of experimental and commercial transfection reagents are currently available, persistent problems remain regarding their suitability for continued development. These include the transfection efficiency for difficult-to-transfect cell types and the risks of decreased cell viability that may arise from any transfection that does occur. Therefore, research is now turning toward alternative molecules that improve the toxicity profile of the gene delivery vector (GDV), while maintaining the transfection efficiency. Among them, cell-penetrating peptides, such as octa-arginine, have shown significant potential as GDVs. Their pharmacokinetic and pharmacodynamic properties can be enhanced through peptidomimetic conversion, whereby a peptide is modified into a synthetic analogue that mimics its structure and/or function, but whose backbone is not solely based on α-amino acids. Using this technology, novel peptidomimetics were developed by co- and postpolymerization functionalization of substituted ethylene oxides, producing poly(ethylene glycol) (PEG)-based peptidomimetics termed “PEGtides”. Specifically, a PEGtide of the poly(α-amino acid) oligo-arginine [poly(glycidylguanidine)] was assessed for its ability to complex and deliver a small interfering ribonucleic acid (siRNA) using a range of cell assays and high-content analysis. PEGtide–siRNA demonstrated significantly increased internalization and gene inhibition over 24 h in Calu-3 pulmonary epithelial cells compared to commercial controls and octa-arginine-treated samples, with no evidence of toxicity. Furthermore, PEGtide–siRNA nanocomplexes can provide significant levels of gene inhibition in “difficult-to-transfect” mouse embryonic hypothalamic (mHypo N41) cells. Overall, the usefulness of this novel PEGtide for gene delivery was clearly demonstrated, establishing it as a promising candidate for continued translational research

Synthesis, characterisation and DNA intercalation studies of regioisomers of ruthenium (II) polypyridyl complexes

Regioisomers of the functional group of the main ligand (L) on a series of [Ru(phen)2L]2+and [Ru(bpy)2L]2+ complexes, where phen is 1,10 phenanthroline and bpy is 2,2′-bipyridine, were synthesised to investigate the interaction with deoxyribonucleic acid (DNA) as potential therapeutics. UV–Vis binding titrations, thermal denaturation and circular dichroism were used to evaluate their interaction with DNA. The conclusions indicated the significance of the auxiliary ligand; especially 1,10-phenanthroline has on the binding constants (Kb). The systematic variation of auxiliary ligand(phen or bpy), and polypyridyl ligand (4-(1H-Imidazo[4,5-f][1,10]phenanthrolin-2-yl)benzonitrile (CPIP), 2-(4-formylphenyl)imidazo[4,5-f] [1,10] phenanthroline (FPIP), 2-(4-bromophenyl)imidazo[4,5-f][1,10]phenanthroline (BPIP) and 2-(4-nitrophenyl)imidazo[4,5-f] [1,10] phenanthroline (NPIP), split in terms of functional group change were investigated for DNA interaction. The CPIP analogues in particular were investigated for the regioisomerism (ortho, meta, para) effect of the nitrile group on the ligand. It was found that both the DNA interaction could be tailored through the systematic variation of the electronic nature of the individual auxiliary ligand and to a lesser extent the functional group and regioisomeric change. Preliminary cell line studies have been carried out to determine the selectivity of the complexes against cell lines such as A375 (Skin Cancer), HeLa (Cervical Cancer), A549 (Lung Cancer), Beas2B (Lung Normal Cell) and MCF-7 (Breast Cancer). Complexes which had strong DNA interactions in the binding studies have proven to be the most efficacious against certain cell lines. Establishing well-defined structure property relationships when looking at trends in spectroscopic properties and DNA binding will aid in the intelligent design of potential therapeutic complexes

New Low Accretion-Rate Magnetic Binary Systems and their Significance for the Evolution of Cataclysmic Variables

Discoveries of two new white dwarf plus M star binaries with striking optical cyclotron emission features from the Sloan Digital Sky Survey (SDSS) brings to six the total number of X-ray faint, magnetic accretion binaries that accrete at rates < 10^{-13} Msun/yr, or <1% of the values normally encountered in cataclysmic variables. This fact, coupled with donor stars that underfill their Roche lobes and very cool white dwarfs, brand the binaries as post common-envelope systems whose orbits have not yet decayed to the point of Roche-lobe contact. They are pre-magnetic CVs, or pre-Polars. The systems exhibit spin/orbit synchronism and apparently accrete by efficient capture of the stellar wind from the secondary star, a process that has been dubbed a ``magnetic siphon''. Because of this, period evolution of the binaries will occur solely by gravitational radiation, which is very slow for periods >3 hr. Optical surveys for the cyclotron harmonics appear to be the only means of discovery, so the space density of pre-Polars could rival that of Polars, and the binaries provide an important channel of progenitors (in addition to the asynchronous Intermediate Polars). Both physical and SDSS observational selection effects are identified that may help to explain the clumping of all six systems in a narrow range of magnetic field strength around 60 MG.Comment: 25 pages, 13 figures, Accepted to Ap

Observations of the Magnetic Cataclysmic Variable VV Puppis with the Far Ultraviolet Spectroscopic Explorer

We present the first far-ultraviolet (FUV) observations of the magnetic cataclysmic variable VV Puppis, obtained with the Far Ultraviolet Spectroscopic Explorer satellite. In addition, we have obtained simultaneous ground-based optical photometric observations of VV Pup during part of the FUV observation. The shapes of the FUV and optical light curves are consistent with each other and with those of past observations at optical, extreme-ultraviolet, and X-ray wavelengths. Time-resolved FUV spectra during the portion of VV Pup's orbit when the accreting magnetic pole of the white dwarf can be seen show an increasing continuum level as the accretion spot becomes more directly visible. The most prominent features in the spectrum are the O VI 1031.9A, 1037.6A emission lines. We interpret the shape and velocity shift of these lines in the context of an origin in the accretion funnel near the white dwarf surface. A blackbody function with T > 90,000 K provides an adequate fit to the FUV spectral energy distribution of VV Pup.Comment: 18 pages, 6 figures, 1 table; to be published in the Astronomical Journa

Time-Resolved HST Spectroscopy of Four Eclipsing Magnetic Cataclysmic Variables

Time-resolved HST UV eclipse spectrophotometry is presented for the magnetic CVs V1309 Ori, MN Hya, V2301 Oph, and V1432 Aql. Separation of the light curves into wavebands allows the multiple emission components to be distinguished. Photospheric hot spots are detected in V1309 Ori and V2301 Oph. The emission- line spectra of V1309 Ori and MN Hya are unusual, with the strength of N V 1240 and N IV 1718 suggesting an overabundance of nitrogen. Three epochs of observation of the asynchronous V1432 Aql cover ~1/3 of a 50-day lap cycle between the white dwarf spin and binary orbit. The light curves vary from epoch to epoch and as a function of waveband. The dereddened UV spectrum is extremely bright and the spectral energy distribution coupled with the duration of eclipse ingress indicate that the dominant source of energy is a hot (T~35,000K) white dwarf. Undiminished line emission through eclipse indicates that the eclipse is caused by the accretion stream, not the secondary star. The hot white dwarf, combined with its current asynchronous nature and rapid timescale for relocking, suggests that V1432 Aql underwent a nova eruption in the past 75-150 yr. The reversed sense of asynchronism, with the primary star currently spinning up toward synchronism, is not necessarily at odds with this scenario, if the rotation of the magnetic white dwarf can couple to the ejecta during the wind phase of the eruption.Comment: To appear in ApJ Part 1; 25 pages, 12 figure

Synthesis, analytical characterization, and monoamine transporter activity of the new psychoactive substance 4-methylphenmetrazine (4-MPM), with differentiation from its ortho- and meta- positional isomers.

The availability of new psychoactive substances (NPS) on the recreational drug market continues to create challenges for scientists in the forensic, clinical and toxicology fields. Phenmetrazine (3-methyl-2-phenylmorpholine) and an array of its analogs form a class of psychostimulants that are well documented in the patent and scientific literature. The present study reports on two phenmetrazine analogs that have been encountered on the NPS market following the introduction of 3-fluorophenmetrazine (3-FPM), namely 4-methylphenmetrazine (4-MPM), and 3-methylphenmetrazine (3-MPM). This study describes the syntheses, analytical characterization, and pharmacological evaluation of the positional isomers of MPM. Analytical characterizations employed various chromatographic, spectroscopic, and mass spectrometric platforms. Pharmacological studies were conducted to assess whether MPM isomers might display stimulant-like effects similar to the parent compound phenmetrazine. The isomers were tested for their ability to inhibit uptake or stimulate release of tritiated substrates at dopamine, norepinephrine and serotonin transporters using in vitro transporter assays in rat brain synaptosomes. The analytical characterization of three vendor samples revealed the presence of 4-MPM in two of the samples and 3-MPM in the third sample, which agreed with the product label. The pharmacological findings suggest that 2-MPM and 3-MPM will exhibit stimulant properties similar to the parent compound phenmetrazine, whereas 4-MPM may display entactogen properties more similar to 3,4-methylenedioxymethamphetamine (MDMA). The combination of test purchases, analytical characterization, targeted organic synthesis, and pharmacological evaluation of NPS and their isomers is an effective approach for the provision of data on these substances as they emerge in the marketplace

Analytical characterization and pharmacological evaluation of the new psychoactive substance 4-fluoromethylphenidate (4F-MPH) and differentiation between the (±)-threo and (±)-erythro diastereomers.

Misuse of (±)-threo-methylphenidate (methyl-2-phenyl-2-(piperidin-2-yl)acetate; Ritalin®; MPH) has long been acknowledged, but the appearance of MPH analogs in the form of 'research chemicals' has only emerged in more recent years. 4-Fluoromethylphenidate (4F-MPH) is one of these recent examples. This study presents the identification and analytical characterization of two powdered 4F-MPH products that were obtained from an online vendor in 2015. Interestingly, the products appeared to have originated from two distinct batches given that one product consisted of (±)-threo-4F-MPH isomers whereas the second sample consisted of a mixture of (±)-threo and (±)-erythro 4F-MPH. Monoamine transporter studies using rat brain synaptosomes revealed that the biological activity of the 4F-MPH mixture resided with the (±)-threo and not the (±)-erythro isomers based on higher potencies determined for blockage of dopamine uptake (IC 4F-MPH  = 66 nM vs. IC (±)-threo = 61 nM vs. IC (±)-erythro = 8,528 nM) and norepinephrine uptake (IC 4F-MPH  = 45 nM vs. (±)-threo = 31 nM vs. IC (±)-erythro = 3,779 nM). In comparison, MPH was three times less potent than (±)-threo-4F-MPH at the dopamine transporter (IC  = 131 nM) and around 2.5 times less potent at the norepinephrine transporter (IC  = 83 nM). Both substances were catecholamine selective with IC values of 8,805 nM and >10,000 nM for (±)-threo-4F-MPH and MPH at the serotonin transporter. These findings suggest that the psychostimulant properties of (±)-threo-4F-MPH might be more potent in humans than MPH. Copyright © 2017 John Wiley & Sons, Ltd