Automated and enhanced extraction of a small molecule-drug conjugate using an enzyme-inhibitor interaction based SPME tool followed by direct analysis by ESI-MS

We report a novel, fast, and automatic SPME-based method capable of extracting a small molecule-drug conjugate (SMDC) from biological matrices. Our method relies on the extraction of the drug conjugate followed by direct elution into an electrospray mass spectrometer (ESI-MS) source for qualitative and quantitative analysis. We designed a tool for extracting the targeting head of a recently synthesized SMDC, which includes acetazolamide (AAZ) as high-affinity ligand specific to carbonic anhydrase IX. Specificity of the extraction was achieved through systematic optimization. The design of the extraction tool is based on noncovalent and reversible interaction between AAZ and CAII that is immobilized on the SPME extraction phase. Using this approach, we showed a 330% rise in extracted AAZ signal intensity compared to a control, which was performed in the absence of CAII. A linear dynamic range from 1.2 to 25 μg/ml was found. The limits of detection (LOD) of extracted AAZ from phosphate-buffered saline (PBS) and human plasma were 0.4 and 1.2 μg/ml, respectively. This with a relative standard deviation of less than 14% (n = 40) covers the therapeutic range.

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The synthesis of unsymmetrically N-substituted chiral 1,4,7-triazacyclononanes

A number of chiral unsymmetrically N-substituted 1,4,7-triazacyclononane ligands have been prepared by modular methods. The key step in the synthesis centres on the macrocyclisation of three tertiary amide precursors under standard Richman-Atkins conditions which allows for subsequent N-functionalisation.     

Screening of Three Transition Metal‐Mediated Reactions Compatible with DNA‐Encoded Chemical Libraries

The construction of DNA‐encoded chemical libraries (DECLs) crucially relies on the availability of chemical reactions, which are DNA‐compatible and which exhibit high conversion rates for a large number of diverse substrates. In this work, we present our optimization and validation procedures for three copper and palladium‐catalyzed reactions (Suzuki cross‐coupling, Sonogashira cross‐coupling, and copper(I)‐catalyzed alkyne‐azide cycloaddition (CuAAC)), which have been successfully used by our group for the construction of large encoded libraries.     

Small Targeted Cytotoxics: Current State and Promises from DNA‐Encoded Chemical Libraries

The targeted delivery of potent cytotoxic agents has emerged as a promising strategy for the treatment of cancer and other serious conditions. Traditionally, antibodies against markers of disease have been used as drug‐delivery vehicles. More recently, lower molecular weight ligands have been proposed for the generation of a novel class of targeted cytotoxics with improved properties. Advances in this field crucially rely on efficient methods for the identification and optimization of organic molecules capable of high‐affinity binding and selective recognition of target proteins. The advent of DNA‐encoded chemical libraries allows the construction and screening of compound collections of unprecedented size. In this Review, we survey developments in the field of small ligand‐based targeted cytotoxics and show how innovative library technologies will help develop the drugs of the future.     

Identification of Structure–Activity Relationships from Screening a Structurally Compact DNA‐Encoded Chemical Library

Methods for the rapid and inexpensive discovery of hit compounds are essential for pharmaceutical research and DNA‐encoded chemical libraries represent promising tools for this purpose. We here report on the design and synthesis of DAL‐100K, a DNA‐encoded chemical library containing 103 200 structurally compact compounds. Affinity screening experiments and DNA‐sequencing analysis provided ligands with nanomolar affinities to several proteins, including prostate‐specific membrane antigen and tankyrase 1. Correlations of sequence counts with binding affinities and potencies of enzyme inhibition were observed and enabled the identification of structural features critical for activity. These results indicate that libraries of this type represent a useful source of small‐molecule binders for target proteins of pharmaceutical interest and information on structural features important for binding.     

Electrochemical insertion of lithium in mechanochemically synthesized Zn2SnO4

We studied the electrochemical insertion of Li in mechanochemically prepared Zn(2)SnO(4). The mechanism of the electrochemical reaction was investigated by using X-ray diffraction, nuclear magnetic resonance spectroscopy, and M?ssbauer spectroscopy. Changes in the morphology of the Zn(2)SnO(4) particles were studied by in situ scanning electron microscopy. The results were compared with mixtures of SnO(2) + ZnO and with Zn(2)SnO(4) prepared by conventional solid-state synthesis and showed that the mechanochemically prepared Zn(2)SnO(4) exhibits the best cyclic stability of these samples.     

Radio-frequency muon spin resonance (RFμSR) experiments on condensed matter

In this paper we present an overview of the radio-frequency muon spin resonance (RFμSR) technique, an analogue to continuous-wave NMR, and an introduction to time-integral (TI) and time-differential (TD) RFμSR on muons in diamagnetic or in paramagnetic environments. The general form of the resonance line for TI-RFμSR as well as the expression for the time-dependence of the longitudinal muon spin polarization at resonance are given. Since RFμSR does not require phase coherence of the muon spin ensemble, this technique allows us to investigate muon species that are generated by transitions from, or in the course of reactions of, a precursor muon species even if in transverse-field (TF) μSR measurements the signal is lost due to dephasing. This ability of RFμSR is clearly demonstrated by measurements on doped Si. In this example, at low temperatures, a very pronounced signal from a muon species in diamagnetic environment has been found in RFμSR measurements, whereas in TFμSR experiments only a very small signal from muons in diamagnetic environment could be detected and a large fraction of the implanted muons escaped detection. These findings could be interpreted in terms of the delayed formation of a diamagnetic muonium-dopant complex, and, due to the large diamagnetic RFμSR signal, the RFμSR technique is a unique tool to study how the variation of parameters and experimental conditions such as illumination affects formation and behavior of these complexes. First results obtained on illuminated boron doped Si are reported. However, as illustrated by the example of experiments on the muonated radical in solid C₆₀, results from conventional TI-RFμSR cannot always be interpreted unambiguously since different parameters, namely the fraction of muons forming the investigated muon species, the longitudinal and the transverse relaxation rates, have similar effects on height and shape of the RFμSR resonance line. These ambiguities, however, may be resolved by collecting time-differential data. With this extension RFμSR becomes a very powerful complementary method to TFμSR in the studies of dynamic effects.