Norbornene Copolymerization with Polar Monomers Catalyzed by Palladium Catalysts Containing Imidazolidin-2-imine/Guanidine Ligands.

The development of a palladium catalyst that has enhanced catalytic performance, such as low aluminum cocatalyst loading, good copolymerization ability, high molecular weight, and excellent solubility of the (co)polymers, is still a challenge in norbornene copolymerizations. Here, a series of PdCl2 and PdMeCl complexes containing differently substituted anilines and imidazolidin-2-imine/guanidine ligands was successfully synthesized and characterized. X-ray diffraction analysis results revealed that these Pd complexes adopted an almost square-planar geometry, and the six-membered chelate ring showed structural distinctions as compared to traditional N^N-based α-diimine and ß-diimine Pd complexes. These Pd complexes were activated by EtAlCl2 and then exhibited moderate activity (104-105 g mol-1 h-1) and good thermal stability (up to 90 °C) for norbornene polymerization to produce high-molecular-weight PNBs (Mn up to 96.4 kg mol-1) with narrow polydispersities (PDI as low as 1.39). These Pd complexes also exhibited good polar group tolerance in the copolymerization of norbornene with methyl 5-norbornene-2-carboxylate and methyl 10-undecenoate, in which the activity was achieved up to 7.04 × 104 g mol-1 h-1. It furnished polar functionalized norbornene-based copolymers with high molecular weight (Mn up to 63.1 kg mol-1), narrow PDI, reasonable polar monomer incorporation, and good solubility. These Pd catalysts exhibited an enhanced copolymerization ability to produce PNB or NB-based copolymers, representing significant progress in this field.

Biophysical Analysis of Small Molecule Binding to Viral RNA Structures.

RNA molecules are essential for carrying genetic information and regulating gene expression in most organisms including human pathogenic RNA and relate retro viruses. Targeting viral RNA (vRNA) structures provide broad opportunities to develop chemical tools to probe molecular virology and to discover novel targets for therapeutic intervention. An increasing number of RNA binding small molecules are being identified, stimulating increased interests in small molecule drug discovery for RNA targets. In this chapter, we describe protocols to characterize and robustly validate vRNA-small molecule (vRNA-sm) interactions starting from vRNA sample preparation, followed by small molecule screening against vRNA targets and finally to validating the vRNA-sm interactions via NMR spectroscopy and calorimetric titrations.

Multiassay Profiling of a Focused Small Molecule Library Reveals Predictive Bidirectional Modulation of the lncRNA MALAT1 Triplex Stability In Vitro.

The rapidly accelerating characterization of RNA tertiary structures has revealed their pervasiveness and active roles in human diseases. Small molecule-mediated modulation of RNA tertiary structures constitutes an attractive avenue for the development of tools for therapeutically targeting and/or uncovering the pathways associated with these RNA motifs. This potential has been highlighted by targeting of the triple helix present at the 3'-end of the noncoding RNA MALAT1, a transcript implicated in several human diseases. This triplex has been reported to decrease the susceptibility of the transcript to degradation and promote its cellular accumulation. While small molecules have been shown to bind to and impact the stability of the MALAT1 triple helix, the small molecule properties that lead to these structural modulations are not well understood. We designed a library utilizing the diminazene scaffold, which is underexplored but precedented for nucleic acid binding, to target the MALAT1 triple helix. We employed multiple assays to holistically assess what parameters, if any, could predict the small molecule affinity and effect on triplex stability. We designed and/or optimized competition, calorimetry, and thermal shift assays as well as an enzymatic degradation assay, the latter of which led to the discovery of bidirectional modulators of triple helix stability within the scaffold-centric library. Determination of quantitative structure-activity relationships afforded predictive models for both affinity- and stability-based assays. This work establishes a suite of powerful orthogonal biophysical tools for the evaluation of small molecule:RNA triplex interactions that generate predictive models and will allow small molecule interrogation of the growing body of disease-associated RNA triple helices.

Quantitative Structure-Activity Relationship (QSAR) Study Predicts Small-Molecule Binding to RNA Structure.

The diversity of RNA structural elements and their documented role in human diseases make RNA an attractive therapeutic target. However, progress in drug discovery and development has been hindered by challenges in the determination of high-resolution RNA structures and a limited understanding of the parameters that drive RNA recognition by small molecules, including a lack of validated quantitative structure-activity relationships (QSARs). Herein, we develop QSAR models that quantitatively predict both thermodynamic- and kinetic-based binding parameters of small molecules and the HIV-1 transactivation response (TAR) RNA model system. Small molecules bearing diverse scaffolds were screened against TAR using surface plasmon resonance. Multiple linear regression (MLR) combined with feature selection afforded robust models that allowed direct interpretation of the properties critical for both binding strength and kinetic rate constants. These models were validated with new molecules, and their accurate performance was confirmed via comparison to ensemble tree methods, supporting the general applicability of this platform.

R-BIND 2.0: An Updated Database of Bioactive RNA-Targeting Small Molecules and Associated RNA Secondary Structures.

Discoveries of RNA roles in cellular physiology and pathology are increasing the need for new tools that modulate the structure and function of these biomolecules, and small molecules are proving useful. In 2017, we curated the RNA-targeted BIoactive ligaNd Database (R-BIND) and discovered distinguishing physicochemical properties of RNA-targeting ligands, leading us to propose the existence of an "RNA-privileged" chemical space. Biennial updates of the database and the establishment of a website platform (rbind.chem.duke.edu) have provided new insights and tools to design small molecules based on the analyzed physicochemical and spatial properties. In this report and R-BIND 2.0 update, we refined the curation approach and ligand classification system as well as conducted analyses of RNA structure elements for the first time to identify new targeting strategies. Specifically, we curated and analyzed RNA target structural motifs to determine the properties of small molecules that may confer selectivity for distinct RNA secondary and tertiary structures. Additionally, we collected sequences of target structures and incorporated an RNA structure search algorithm into the website that outputs small molecules targeting similar motifs without a priori secondary structure knowledge. Cheminformatic analyses revealed that, despite the 50% increase in small molecule library size, the distinguishing properties of R-BIND ligands remained significantly different from that of proteins and are therefore still relevant to RNA-targeted probe discovery. Combined, we expect these novel insights and website features to enable the rational design of RNA-targeted ligands and to serve as a resource and inspiration for a variety of scientists interested in RNA targeting.

Hydrogen-Bonding-Induced Heterogenization of Nickel and Palladium Catalysts for Copolymerization of Ethylene with Polar Monomers.

The practical synthesis of polar-functionalized polyolefins using transition-metal-catalyzed copolymerization of olefins with polar monomers is a challenge; the use of heterogeneous catalysts is little explored. Herein, we report the synthesis of heterogeneous naphthoquinone-based nickel (Ni/SiO2 ) and palladium (Pd/SiO2 ) catalysts through hydrogen bonding interactions of the ligands with the silica surface. Ni/SiO2 exhibits high activities (up to 2.65×106  g mol-1 h-1 ) during the copolymerization of ethylene with 5-hexene-1-yl-acetate, affording high-molecular-weight (Mn up to 630 000) polar-functionalized semicrystalline polyethylene (comonomer incorporation up to 2.8 mol %), along with great morphology control. The resulting copolymers possess improved surface properties and great mechanical properties. Pd/SiO2 can mediate ethylene copolymerization with polar monomers with moderate activity to produce high-molecular-weight copolymers with tunable comonomer incorporation.

IRES-targeting small molecule inhibits enterovirus 71 replication via allosteric stabilization of a ternary complex.

Enterovirus 71 (EV71) poses serious threats to human health, particularly in Southeast Asia, and no drugs or vaccines are available. Previous work identified the stem loop II structure of the EV71 internal ribosomal entry site as vital to viral translation and a potential target. After screening an RNA-biased library using a peptide-displacement assay, we identify DMA-135 as a dose-dependent inhibitor of viral translation and replication with no significant toxicity in cell-based studies. Structural, biophysical, and biochemical characterization support an allosteric mechanism in which DMA-135 induces a conformational change in the RNA structure that stabilizes a ternary complex with the AUF1 protein, thus repressing translation. This mechanism is supported by pull-down experiments in cell culture. These detailed studies establish enterovirus RNA structures as promising drug targets while revealing an approach and mechanism of action that should be broadly applicable to functional RNA targeting.

Template-guided selection of RNA ligands using imine-based dynamic combinatorial chemistry.

This study establishes the applicability of imine-based dynamic combinatorial chemistry to discover non-covalent ligands for RNA targets. We elucidate properties underlying the reactivity of arylamines and demonstrate target-guided amplification of tight binders in an amiloride-based dynamic library.

Driving factors in amiloride recognition of HIV RNA targets.

Noncoding RNAs are increasingly promising drug targets yet ligand design is hindered by a paucity of methods that reveal driving factors in selective small molecule : RNA interactions, particularly given the difficulties of high-resolution structural characterization. HIV RNAs are excellent model systems for method development given their targeting history, known structure-function relationships, and the unmet need for more effective treatments. Herein we report a strategy combining synthetic diversification, profiling against multiple RNA targets, and predictive cheminformatic analysis to identify driving factors for selectivity and affinity of small molecules for distinct HIV RNA targets. Using this strategy, we discovered improved ligands for multiple targets and the first ligands for ESSV, an exonic splicing silencer critical to replication. Computational analysis revealed guiding principles for future designs and a predictive cheminformatics model of small molecule : RNA binding. These methods are expected to facilitate progress toward selective targeting of disease-causing RNAs.

Optically Transparent Functional Polyolefin Elastomer with Excellent Mechanical and Thermal Properties.

Synthesis of a variety of optically transparent polyolefin elastomers consisting of hard segment of polynorbornene and several kinds of soft segments such as atactic polypropylene, poly(ethylene-co-propylene), and poly(ethylene-co-1-hexene) was achieved. The block copolymers exhibited excellent toughness and thermal property with efficient elastic recovery. Most importantly, the introduction of hydroxyl group into polynorbornene segment not only modulated surface property of block copolymers, but also improved their mechanical properties.

Fluorescent peptide displacement as a general assay for screening small molecule libraries against RNA.

A prominent hurdle in developing small molecule probes against RNA is the relative scarcity of general screening methods. In this study, we demonstrate the application of a fluorescent peptide displacement assay to screen small molecule probes against four different RNA targets. The designed experimental protocol combined with statistical analysis provides a fast and convenient method to simultaneously evaluate small molecule libraries against different RNA targets and classify them based on affinity and selectivity patterns.

Ethylene Polymerization and Copolymerization with Polar Monomers Using Nickel Complexes Bearing Anilinobenzoic Acid Methyl Ester Ligand.

Neutral nickel complexes containing an anilinobenzoic acid methyl ester ligand are prepared and applied for the ethylene polymerization and copolymerization with polar monomers. The complex C2 containing isopropyl substituent on the aniline ligand conducts ethylene polymerization with high activity and good thermal stability. Most importantly, the catalyst promotes the copolymerization of ethylene and polar monomers with high activity (up to 277 kg·mol-1·h-1), affording ester-functionalized semicrystalline polyethylene with reasonable polar monomer content (up to 3.20 mol %).

Substituent Effects of Phenyl Group on Silylene Bridge in Stereospecific Polymerization of Propylene with C1-Symmetric Ansa-Silylene(fluorenyl)(amido) Dimethyl Titanium Complexes.

A C1-symmetric (methylphenyl)silylene-bridged (fluorenyl)(naphthylamido) titanium complex (1) and (diphenyl)silylene-bridged (fluorenyl)(naphthylamido) titanium complex (2) were synthesized and characterized by ¹H NMR, element analysis, and X-ray crystal analysis. The coordination mode of the fluorenyl ligand to the titanium metal is an η³ manner in each complex. These complexes were applied for propylene polymerization using dried modified methyaluminoxane (dMMAO) as a cocatalyst under different propylene pressures in a semi batch-type method. The catalytic activity was strongly dependent on the structure of the complex and the propylene pressure, where complex 1 exhibited the highest activity (600 kg mol-1·h-1) under a propylene pressure of 8.0 atm to produce high molecular weight polypropylene. The polypropylenes obtained were syndiotactic-rich with an rr value of 0.50, indicating that the silylene bridge was not efficient for the isospecificity of a constrained geometry catalyst (CGC). The mechanical properties of the resulting polymers depended on their microstructure.

Synthesis and Biodegradation of Poly(l-lactide-co-ß-propiolactone).

Although the copolymerizations of l-lactide (LA) with seven- or six-membered ring lactones have been extensively studied, the copolymerizations of LA with four-membered ring lactones have scarcely been reported. In this work, we studied the copolymerization of LA with ß-propiolactone (PL) and the properties of the obtained copolymers. The copolymerization of LA with PL was carried out using trifluoromethanesulfonic acid as a catalyst and methanol as an initiator to produce poly(LA-co-PL) with Mn of ~50,000 and PL-content of 6-67 mol %. The Tg values of the copolymers were rapidly lowered with increasing PL-contents. The Tm and ΔHm of the copolymers gradually decreased with increasing PL-contents, indicating their decreased crystallinity. Biodegradation test of the copolymers in compost demonstrated their improved biodegradability in comparison with the homopolymer of LA.

Synthesis of Hydroxy-Functionalized Cyclic Olefin Copolymer and Its Block Copolymers with Semicrystalline Polyolefin Segments.

Synthesis of hydroxy-functionalized cyclic olefin copolymer (COC) is achieved with remarkably high activity (up to 5.96 × 107 g-polymer mol-Ti-1 h-1 ) and controlled hydroxy group in a wide range (≈17.1 mol%) by using ansa-dimethylsilylene (fluorenyl)(amido)titanium complex. The catalyst also promotes living/controlled copolymerization to afford novel diblock copolymers consisting of hydroxy-functionalized COC and semicrystalline polyolefin sequence such as polyethylene and syndiotactic polypropylene, where the glass transition temperature of the norbornene/10-undecen-1-ol segment and each block length are controlled by comonomer composition and copolymerization time, respectively.

Supramolecular catalyst functions in catalytic amount: cucurbit[8]uril accelerates the photodimerization of Brooker's merocyanine.

Supramolecular catalysis aims to modulate chemical reactions on both selectivity and rate by taking advantage of supramolecular chemistry. However, due to the effect of product inhibition, supramolecular catalysts are usually added in stoichiometric amounts. Herein, we report a supramolecular catalysis system in which 1% of the supramolecular catalyst, cucurbit[8]uril, is able to significantly accelerate the photodimerization of Brooker's merocyanine. This catalytic process is realized in a cyclic manner because the photodimerized product can be spontaneously replaced by monomeric reactants via competitive host-guest complexation. Thus, a catalytic amount of cucurbit[8]uril is sufficient to accomplish photodimerization within 10 min. This line of research will enrich the field of supramolecular catalysis and allow the development of more efficient catalytic systems.

Substituent Effects of Adamantyl Group on Amido Ligand in Syndiospecific Polymerization of Propylene with Ansa-Dimethylsilylene(Fluorenyl)(Amido) Zirconium Complex.

A series of new fluorenylamido-ligated zirconium complexes bearing an electron-donating adamantyl group on the amido ligand were synthesized and characterized by elemental analysis, ¹H NMR, and single crystal X-ray analysis. The coordination mode of the fluorenyl ligand to the zirconium metal was η³ manner, and all the complexes were Cs-symmetric in solution. The complexes showed moderate activity (1.0 × 105 g-polymer mol-Zr-1·h-1), even at a low Al/Zr ratio of 50. The increase of propylene pressure improved the activity by one order of magnitude (up to 1.0 × 106 g-polymer mol-Ti-1·h-1). All catalyst systems gave syndiotactic polypropylene, where the complex containing the 3,6-di-t-butyl fluorenyl ligand was more effective for the enhancement of the syndiospecificity. The increase of propylene pressure also improved the syndiospecificity with the syndiotactic pentad of 0.96 and the melting point of 159 °C.

Living Polymerization of Propylene with ansa-Dimethylsilylene(fluorenyl)(cumylamido) Titanium Complexes.

A series of ansa-silylene(fluorenyl)(amido) titanium complexes (1a⁻1c, 2a, and 2b) bearing various substituents on the amido and fluorenyl ligands are synthesized and characterized by elemental analysis, ¹H NMR, and single crystal X-ray analysis. The coordination mode of the fluorenyl ligand to the titanium metal is η³ manner in each complex. The propylene polymerization is conducted with these complexes at 0 and 25 °C in a semi batch-type method, respectively. The catalytic activity of 1a⁻1c bearing cumyl-amido ligand is much higher than that of 2a and 2b bearing naphthyl group in amido ligand. High molecular weight polypropylenes are obtained with narrow molecular weight distribution, suggesting a living nature of these catalytic systems at 0 °C. The polymers produced are statistically atactic, regardless of the structure of the complex and the polymerization temperature.

Precision Chain-Walking Polymerization of trans-4-Octene Catalyzed by α-Diimine Nickel(II) Catalysts Bearing ortho-sec-Phenethyl Groups.

α-Diimine nickel complexes bearing bulky ortho-sec-phenethyl groups (bis{[N,N'-(4-methyl-2,6-di-sec-phenethylphenyl)imino]-1,2-dimethylethane}dibromonickel (1), bis{[N,N'-(4,6-dimethyl-2-sec-phenethylphenyl)imino]-1,2-dimethylethane}dibromonickel (2), bis{[N,N'-(4-methyl-2-sec-phenethylphenyl)imino]-1,2-dimethylethane}dibromonickel (3)) and {bis[N,N'-(2,4,6-trimethylphenyl)imino]-1,2-dimethylethane}dibromidonickel (4) are used as a precatalyst for the polymerization of trans-4-octene upon activation with modified methylaluminoxane. These catalysts conduct chain-walking polymerization of trans-4-octene to give polymers possessing propyl and butyl branches with high molecular weight and narrow molecular weight distribution. The branching structure depends on the nickel complex as well as the polymerization temperature, and the ratio of propyl branch was increased with increasing the bulkiness of the ligand and decreasing the polymerization temperature. Consequently, the most bulky 1 among the complexes used is found to polymerize trans-4-octene with high 1,5-regioselectivity at -20 °C to give poly(1-propylpentan-1,5-diyl).

An Amylase-Responsive Bolaform Supra-Amphiphile.

An amylase-responsive bolaform supra-amphiphile was constructed by the complexation between ß-cyclodextrin and a bolaform covalent amphiphile on the basis of host-guest interaction. The bolaform covalent amphiphile could self-assemble in solution, forming sheet-like aggregates and displaying weak fluorescence because of aggregation-induced quenching. The addition of ß-cyclodextrin led to the formation of the bolaform supra-amphiphile, prohibiting the aggregation of the bolaform covalent amphiphile and accompanying with the significant recovery of fluorescence. Upon the addition of α-amylase, with the degradation ß-cyclodextrin, the fluorescence of the supra-amphiphile would quench gradually and significantly, and the quenching rate linearly correlated to the concentration of α-amylase. This study enriches the field of supra-amphiphiles on the basis of noncovalent interactions, and moreover, it may provide a facile way to estimate the activity of α-amylase.