Synthesis and physicochemical characteristics of a liver-targeted conjugate of fluorodeoxyuridine monophosphate with lactosaminated human albumin

In previous experiments fluorodeoxyuridine monophosphate (FUdRMP) was conjugated with lactosaminated human albumin (L-HSA). Fluorodeoxyuridine (FUdR) is an anticancer agent and L-HSA is a hepatotropic carrier of drugs obtained by the covalent linkage of lactose residues to the albumin molecule. The conjugate was synthesised via the imidazolide of FUdRMP at alkaline pH. Peripheral venous administration of L-HSA-FUdRMP produced enhanced FUdR levels in hepatic blood and might accomplish a non-invasive loco-regional chemotherapy of liver micrometastases. In the present paper some physicochemical characteristics of L-HSA-FUdRMP are reported. Polyacrylamide gel electrophoresis indicated that the coupling reaction did not cause covalent aggregation of the L-HSA molecules. 31P NMR spectra of the conjugate showed that FUdRMP was linked to L-HSA by phosphoamide bonds to lysine and histidine residues, and the area of the peak due to the lysine bond represented more than 80% of the spectrum of L-HSA-FUdRMP. MALDI analysis revealed a partial degradation of the peptide backbone of the conjugate which could not be detected using other methods of analysis. The degradation was not caused by the coupling of lactose molecules to albumin, but rather a consequence of FUdRMP conjugation with L-HSA. This fragmentation was dependent on the pH of the medium used for the FUdRMP coupling reaction. By decreasing the pH to 7.5, conjugates were obtained with a lower drug load but with a substantially reduced fragmentation, which should be preferred for a clinical use of L-HSA-FUdRMP.     

Electrospray ionization mass spectrometry in studies of aluminium(III)-ligand solution equilibria

Electrospray ionization mass spectrometry (ESI-MS) has been applied to the study of solution equilibria between Al(III) and the two ligands 4-hydroxy-3-pyridinecarboxylic acid (4H3P) and 3-hydroxy-4-pyridinecarboxylic acid (3H4P). The results compare well with the speciation data obtained from potentiometric, UV-visible spectroscopy, and NMR measurements. This agreement suggests the applicability of ES-MS to the study of more complicated aluminium-ligand systems.     

Is the atmospheric pressure chemical ionisation and collisionally induced methanol loss from protonated dehydroamino acids a retrosynthetic process?

Unsaturated 5(4H)-oxazolones lead, by methanolysis, to the corresponding dehydroamino acid derivatives. Interestingly, under atmospheric pressure chemical ionisation (APCI) conditions, the latter give rise, aside from abundant [M+H](+) ions, to [MHbondCH(3)OH](+) species, formally corresponding to the protonated oxazolones employed for their synthesis. Retrosynthetic processes have often been described as energetically favoured decompositions of odd-electron molecular ions but never invoked in APCI-activated fragmentations. To investigate this possible retrosynthetic process, occurring also under collisional conditions, some experiments on the deuterated analogues have been undertaken. The breakdown curves of [M+H](+) of oxazolones and [MHbondCH(3)OH](+) of the dehydroamino acid derivatives are superimposable, proving their structural identity and giving experimental evidence of the occurrence of a real retrosynthetic process from even-electron protonated molecules.     

A new program based on stochastic Liouville equation for the analysis of superhyperfine interaction in CW-ESR spectroscopy

In the slow-motion region, ESR spectra cannot be expressed as a sum of simple Lorentzian lines. Studies of Freed and co-workers, on nitroxides in liquids gained information on the microscopic models of rotational dynamics, relying much on computer programs for simulation of ESR spectra based on the stochastic Liouville equation (SLE). However, application of Freed's method to copper system of biological interest has been for a long time precluded by lack of a full program able to simulate ESR spectra containing more than one hyperfine interaction. Direct extension of the Freed's approach in order to include superhyperfine interaction is not difficult from a theoretical point of view but the resulting algorithm is problematical because it leads to substantial increase in the dimensions of the matrix related to the spin-hamiltonian operator. In this paper preliminary results of a new program, written in C, which includes the superhyperfine interactions are presented. This preliminary version of the program does not take into account a restoring potential, so it can be used only in isotropic diffusion conditions. A comparison with an approximate method previously developed in our laboratory, based on a post-convolution approach, is discussed.     

New reactivity of hydroxyallylpyridyl derivatives as C-3 carbon nucleophiles

Hydroxyallylpyridyl derivatives exhibited a peculiar thermal behavior likely ascribed to the weak acidity of the ‘picoline-type’ hydrogen atom on the C-1 carbon of the allyl residue, leading to allyl inversion products. The unprotected alcohol reacted as ‘vinylogous picoline’ carbon nucleophile with strongly activated heterocyclic counterparts. A mechanistic rationale for this unprecedented reactivity was proposed.     

Organocatalyzed enantioselective fluorocyclizations

Enantioenriched fluorinated heterocycles can be prepared through fluorocyclizations of prochiral indoles (see scheme; Ts=tosyl, Bn=benzyl, Boc=tert-butoxycarbonyl). More than twenty examples for this cascade fluorination-cyclization, which is catalyzed by cinchona alkaloids and employs N-fluorobenzenesulfonimide as the electrophilic fluorine source have been explored, and an unprecedented catalytic asymmetric difluorocyclization has also been identified.     

An integrated approach for the systematic evaluation of polymeric nanoparticles in healthy and diseased organisms

Innovative strategies that utilize nanoparticles (NPs) for a better delivery of drugs and to improve their therapeutic efficacy have been widely studied in many clinical fields, including oncology. To develop safe and reliable devices able to reach their therapeutic target, a hierarchical characterization of NP interactions with biological fluids, cells, and whole organisms is fundamental. Unfortunately, this aspect is often neglected and the development of standardized characterization methods would be of fundamental help to better elucidate the potentials of nanomaterials, even before the loading of the drugs. Here, we propose a multimodal in vitro/in vivo/ex vivo platform aimed at evaluating these interactions for the selection of the most promising NPs among a wide series of materials. To set the system, we used non-degradable fluorescent poly(methyl-methacrylate) NPs of different sizes (50, 100, and 200 nm) and surface charges (positive and negative). First we studied NP stability in biological fluids. Then, we evaluated NP interaction with two cell lines of triple-negative breast cancer (TNBC), 4T1, and MDA-MB231.1833, respectively. We found that NPs internalize in TNBC cells depending on their physico-chemical properties without toxic effects. Finally, we studied NP biodistribution in terms of tissue migration and progressive clearance in breast-cancer bearing mice. The use of highly stable poly(methyl-methacrylate) NPs enabled us to track them for a long time in cells and animals. The application of this platform to other nanomaterials could provide innovative suggestions for the development of a systematic method of characterization to select the most reliable nanodrug candidates for biomedical applications.     

Oxygen acidity of ring methoxylated 1,1-diarylalkanol radical cations bearing alpha-cyclopropyl groups. The competition between O-neophyl shift and C-cyclopropyl beta-scission in the intermediate 1,1-diarylalkoxyl radicals

A product and time-resolved kinetic study on the reactivity of the radical cations generated from cyclopropyl(4-methoxyphenyl)phenylmethanol (1) and cyclopropyl[bis(4-methoxyphenyl)]methanol (2) has been carried out in aqueous solution. In acidic solution, 1*+ and 2*+ display very low reactivities toward fragmentation, consistent with the presence of groups at Calpha (aryl and cyclopropyl) that after Calpha-Cbeta bond cleavage would produce relatively unstable carbon-centered radicals. In basic solution, 1*+ and 2*+ display oxygen acidity, undergoing -OH-induced deprotonation from the alpha-OH group, leading to the corresponding 1,1-diarylalkoxyl radicals 1r* and 2r*, respectively, as directly observed by time-resolved spectroscopy. The product distributions observed in the reactions of 1*+ and 2*+ under these conditions (cyclopropyl phenyl ketone, cyclopropyl(4-methoxyphenyl) ketone, and 4-methoxybenzophenone from 1*+; cyclopropyl(4-methoxyphenyl) ketone and 4,4'-dimethoxybenzophenone from 2*+) have been rationalized in terms of a water-induced competition between O-neophyl shift and C-cyclopropyl beta-scission in the intermediate 1,1-diarylalkoxyl radicals 1r* and 2r*.     

Robust Chemical Preservation of Digital Information on DNA in Silica with Error‐Correcting Codes

Information, such as text printed on paper or images projected onto microfilm, can survive for over 500 years. However, the storage of digital information for time frames exceeding 50 years is challenging. Here we show that digital information can be stored on DNA and recovered without errors for considerably longer time frames. To allow for the perfect recovery of the information, we encapsulate the DNA in an inorganic matrix, and employ error‐correcting codes to correct storage‐related errors. Specifically, we translated 83 kB of information to 4991 DNA segments, each 158 nucleotides long, which were encapsulated in silica. Accelerated aging experiments were performed to measure DNA decay kinetics, which show that data can be archived on DNA for millennia under a wide range of conditions. The original information could be recovered error free, even after treating the DNA in silica at 70 °C for one week. This is thermally equivalent to storing information on DNA in central Europe for 2000 years.