Diversity-oriented synthesis: producing chemical tools for dissecting biology.

Small molecule modulators of biological function can be discovered by the screening of compound libraries. However, it became apparent that some human disease related targets could not be addressed by the libraries commonly used which typically are comprised of large numbers of structurally similar compounds. The last decade has seen a paradigm shift in library construction, with particular emphasis now being placed on increasing a library's structural, and thus functional diversity, rather than only its size. Diversity-oriented synthesis (DOS) aims to generate such structural diversity efficiently. This tutorial review has been written to introduce the subject to a broad audience and recent achievements in both the preparation and the screening of structurally diverse compound collections against so-called 'undruggable' targets are highlighted.

Palladium-catalysed cross-coupling of vinyldisiloxanes with benzylic and allylic halides and sulfonates.

The Hiyama cross-coupling reaction is a powerful method for carbon-carbon bond formation. To date, the substrate scope of this reaction has predominantly been limited to sp(2)-sp(2) coupling reactions. Herein, the palladium-catalysed Hiyama type cross-coupling of vinyldisiloxanes with benzylic and allylic bromides, chlorides, tosylates and mesylates is reported. A wide variety of functional groups were tolerated, and the synthetic utility of the methodology was exemplified through the efficient total synthesis of the cytotoxic natural product bussealin A. In addition, the antiproliferative ability of bussealin A was evaluated in two cancer-cell lines.

Synthesis and biological profiling of tellimagrandin I and analogues reveals that the medium ring can significantly modulate biological activity.

A novel synthesis of the ellagitannin natural product tellimagrandin I and a series of medium ring analogues is described. These compounds were all subsequently screened for redox activity, ability to precipitate protein and cellular phenotype in HeLa cells. From this we have shown that all properties can be modulated independently by varying ring size and by moving the ester out of conjugation with the biaryl ring system. Increasing ring size increased redox activity and cytotoxicity, leading to the identification of a compound (10) which was significantly more cytotoxic. In addition compounds identified with a redox active scaffold and low cytotoxicity may be employed as a new class of redox probes.

Chemical genetics.

Chemical genetics can be defined as the study of biological systems using small molecule tools. Cell permeable and selective small molecules modulate gene product function rapidly, reversibly and can be administered conditionally in either a cellular or organismal context. The small molecule approach provides exacting temporal and quantitative control and is therefore an extremely powerful tool for dissecting biological processes. This tutorial review has been written to introduce the subject to a broad audience and highlights recent developments within the field in four key areas of biology: modulating protein-protein interactions, malaria research, hepatitis C virus research, and disrupting RNA interference pathways.

Facile synthesis of 3-nitro-2-substituted thiophenes.

A new approach to 3-nitro-2-substituted thiophenes has been developed. Exposure of commercially available 1,4-dithane-2,5-diol to nitroalkenes in the presence of 20% triethylamine results in a tandem Michael-intramolecular Henry reaction to form the corresponding tetrahydrothiophene. Subsequent microwave irradiation on acidic alumina in the presence of chloranil effects the solvent free dehydration and aromatization to form 3-nitro-2-substituted thiophenes cleanly and rapidly. A simple workup procedure removes the requirement for purification by chromatography in most cases.

Discovery of a small-molecule binder of the oncoprotein gankyrin that modulates gankyrin activity in the cell.

Gankyrin is an ankyrin-repeat oncoprotein whose overexpression has been implicated in the development of many cancer types. Elevated gankyrin levels are linked to aberrant cellular events including enhanced degradation of tumour suppressor protein p53, and inhibition of gankyrin activity has therefore been identified as an attractive anticancer strategy. Gankyrin interacts with several partner proteins, and a number of these protein-protein interactions (PPIs) are of relevance to cancer. Thus, molecules that bind the PPI interface of gankyrin and interrupt these interactions are of considerable interest. Herein, we report the discovery of a small molecule termed cjoc42 that is capable of binding to gankyrin. Cell-based experiments demonstrate that cjoc42 can inhibit gankyrin activity in a dose-dependent manner: cjoc42 prevents the decrease in p53 protein levels normally associated with high amounts of gankyrin, and it restores p53-dependent transcription and sensitivity to DNA damage. The results represent the first evidence that gankyrin is a "druggable" target with small molecules.

Diversity-oriented synthesis as a tool for identifying new modulators of mitosis.

The synthesis of diverse three-dimensional libraries has become of paramount importance for obtaining better leads for drug discovery. Such libraries are predicted to fare better than traditional compound collections in phenotypic screens and against difficult targets. Herein we report the diversity-oriented synthesis of a compound library using rhodium carbenoid chemistry to access structurally diverse three-dimensional molecules and show that they access biologically relevant areas of chemical space using cheminformatic analysis. High-content screening of this library for antimitotic activity followed by chemical modification identified 'Dosabulin', which causes mitotic arrest and cancer cell death by apoptosis. Its mechanism of action is determined to be microtubule depolymerization, and the compound is shown to not significantly affect vinblastine binding to tubulin; however, experiments suggest binding to a site vicinal or allosteric to Colchicine. This work validates the combination of diversity-oriented synthesis and phenotypic screening as a strategy for the discovery of biologically relevant chemical entities.

Synergistic organocatalysis in the kinetic resolution of secondary thiols with concomitant desymmetrization of an anhydride.

Kinetic resolution is an important method for the separation of racemates into their component enantiomers. Thiols are precursors to a variety of organosulfur compounds, with high utility in both chemistry and chemical biology, yet there is a surprising dearth of methodologies for their direct and efficient catalytic kinetic resolution. Here, we demonstrate an organocatalytic process involving the highly enantioselective desymmetrization of an achiral electrophile with the simultaneous kinetic resolution of a racemic thiol. The preparative potential of the methodology is exemplified by the synthesis of a drug precursor antipode in excellent yield and enantioselectivity as a by-product of a process that also resolves a sec-thiol substrate with a selectivity of S = 226 (that is, both thiol antipodes produced in >95% ee at 51% conversion). In a second example a racemic sec-thiol representing the stereocentre-containing core of the anti-asthma drug (R)-Montelukast was resolved with synthetically useful selectivity under mild conditions.

Glottic insertion of the ProSeal LMA occurs in 6% of cases: a review of 627 patients.

PURPOSE: Glottic insertion of the ProSeal Laryngeal Mask Airway (PLMA) has received little attention in the anesthesiology literature. We investigated the incidence and depth of insertion associated with this important cause for a failed insertion attempt with the PLMA. METHODS: With Institutional Review Board approval, we reviewed 15-months' use of the PLMA. Diagnosis of glottic insertion involved a test with children's bubble solution placed on the drain tube port, as well as a fibreoptic examination of the airway of patients experiencing airway obstruction. Patients were anesthetized and paralyzed and the PLMA was inserted deflated with the fingertip method (women size 4, men size 5). The cuff was inflated and a soap membrane established on the drain tube port. Glottic insertion was diagnosed by applying fingertip pressure to the patient's chest wall and observing pulmonary exhalation via the drain tube and bubble formation. The PLMA was then removed and reinserted without further assessment. For all patients, we used a fibrescope to determine the cause of unexplained airway obstruction after the PLMA was considered successfully inserted. RESULTS: There were 627 patients (391 women, 236 men). We diagnosed glottic insertion in 38/627 (6.1%) patients, 37 by the soap membrane test and one with airway obstruction and direct fibreoptic visualization of malposition. Following glottic insertion, successful reinsertion of the PLMA behind the larynx was always associated with greater depth of insertion by an average 2.0 cm. CONCLUSION: Glottic insertion can be easily and quickly diagnosed and our results suggest the incidence and importance of malposition are under-reported in the literature.

Maximum minute ventilation test for the ProSeal laryngeal mask airway.

UNLABELLED: One of the distinguishing features of the ProSeal laryngeal mask airway (PLMA) is that it can cause upper airway obstruction, even when it is correctly inserted behind the cricoid cartilage. We used a hyperventilation test, the maximum minute ventilation test (MMV test), to aid in the diagnosis of upper airway obstruction after PLMA insertion. The patient was briefly hyperventilated for 15 s yielding a MMV value equal to 4 x (breaths/15 s) x (exhaled tidal volume). MMV values were collected in 317 adult women and men over 6 mo. Critical MMV values were obtained in 17 of 317 patients, 15 of 317 (4.7%) of which were due to insertion of the PLMA. The PLMA was removed in seven of 317 (2.2%) patients. The most common cause of upper airway obstruction due to the PLMA was laryngeal obstruction. This refers to compression of supraglottic and glottic structures with resulting narrowing and compromise of the airway. A second, much less common, form of airway obstruction was bilateral cuff infolding with or without downfolding of the epiglottis. Finally, we discuss the margin of safety for minute ventilation, defined as the excess of the MMV over and above basal minute ventilation requirements for the patient. With critical MMV, the margin of safety is drastically reduced or nonexistent. IMPLICATIONS: One of the distinguishing features of the ProSeal laryngeal mask airway (PLMA) is that it can cause upper airway obstruction, even when it is correctly inserted behind the cricoid cartilage. We used a hyperventilation test, the maximum minute ventilation test, to aid in the diagnosis of upper airway obstruction after PLMA insertion.

Gastric distention in a spontaneously ventilating patient with a ProSeal laryngeal mask airway.

IMPLICATIONS: Partial upper airway obstruction during spontaneous respiration with a ProSeal laryngeal mask airway can result in gastric distention.

Esophageal insufflation with normal fiberoptic positioning of the ProSeal laryngeal mask airway.

IMPLICATIONS: Esophageal insufflation can occur simultaneously with venting from the drain tube during positive pressure ventilation with the ProSeal laryngeal mask airway.