Squaric esters as peptide stapling reagents.

We report that squaric esters can serve as bifunctional reagents for selective peptide stapling reactions. Formation of the squaric amide staple occurs under mild conditions with amine-containing side chains. We show that short resin-bound peptides are readily stapled on solid phase and that stapling can occur at various relative positions along the peptide and with various amine tether lengths (e.g. Lysine, ornithine, etc). The squaric amide staples are stable to strong acid conditions used to cleave the stapled peptide from the resin and the stapled peptides show an increase in helicity as analyzed through circular dichroism.

Slow Release of GA3 Hormone from Polymer Coating Overcomes Seed Dormancy and Improves Germination.

Seed dormancy often hinders direct seeding efforts that are attempting to restore degraded landscapes. Gibberellic acid (GA3) can be applied to physiologically dormant seeds to induce germination, but this hormone is rarely effective, as it can degrade or be leached from the seed. We tested different polymer matrixes (polylactic acid, polyvinylpyrrolidone, and ethylcellulose) to apply and slowly release GA3 to the seed. These polymers were tested as seed coatings in either a powder, liquid, or a combination of powder and liquid forms. We found that a liquid ethylcellulose/GA3 coating generally outperformed the other polymers and applications methods using our test species Penstemon palmeri. With this top-performing treatment, seed germination was 3.0- and 3.9-fold higher at 15 °C and 25 °C, respectively. We also evaluated the liquid ethylcellulose/GA3 coating on P. comharrenus, P. strictus, P. pachyphyllus, and P. eatonii. Again, the coating had a strong treatment response, with the degree of difference related to the relative level of dormancy of the species. Growth studies were also performed in pots to ensure that the side effects of GA3 overdosing were not present. Here, we found minimal differences in root length, shoot length, or biomass between plants grown from untreated and GA3-coated seeds.

Characterization of organic crystals for second-harmonic generation.

Second-harmonic generation (SHG) is a common technique with many applications. Common inorganic single-crystalline materials used to produce SHG light are effective using short IR/visible wavelengths but generally do not perform well at longer, technologically relevant IR wavelengths such as 1300, 1550, and 2000 nm. Efficient SHG materials possess many of the same key material properties as terahertz (THz) generators, and certain single-crystalline organic THz generation materials have been reported to perform at longer IR wavelengths. Consequently, this work focuses on characterizing three efficient organic THz generators for SHG, namely, DAST (trans-4-[4-(dimethylamino)-N-methylstilbazolium] p-tosylate), DSTMS (4-N,N-dimethylamino-4'-N'-methylstilbazolium 2,4,6-trimethylbenzenesulfonate), and the recently discovered generator PNPA ((E)-4-((4-nitrobenzylidene)amino)-N-phenylaniline). All three of these crystals outperform the beta-barium borate (BBO), an inorganic material commonly used for SHG, using IR pump wavelengths (1200-2000 nm).

Structural Analysis of Non-native Peptide-Based Catalysts Using 2D NMR-Guided MD Simulations.

Proteins and enzymes generally achieve their functions by creating well-defined 3D architectures that pre-organize reactive functionalities. Mimicking this approach to supramolecular pre-organization is leading to the development of highly versatile artificial chemical environments, including new biomaterials, medicines, artificial enzymes, and enzyme-like catalysts. The use of ß-turn and α-helical motifs is one approach that enables the precise placement of reactive functional groups to enable selective substrate activation and reactivity/selectivity that approaches natural enzymes. Our recent work has demonstrated that helical peptides can serve as scaffolds for pre-organizing two reactive groups to achieve enzyme-like catalysis. In this study, we used CYANA and AmberTools to develop a computational approach for determining how the structure of our peptide catalysts can lead to enhancements in reactivity. These results support our hypothesis that the bifunctional nature of the peptide enables catalysis by pre-organizing the two catalysts in reactive conformations that accelerate catalysis by proximity. We also present evidence that the low reactivity of monofunctional peptides can be attributed to interactions between the peptide-bound catalyst and the helical backbone, which are not observed in the bifunctional peptide.

Optimizing the Local Chemical Environment on a Bifunctional Helical Peptide Scaffold Enables Enhanced Enantioselectivity and Cooperative Catalysis.

We describe a proof-of-concept study in which peptide-bound enamine and thiourea catalysts are used to facilitate the conjugate addition of cyclohexanone to nitroolefins. Our bifunctional peptide scaffold is modified to optimize the local environment around both catalysts to enhance both reactivity and enantioselectivity, affording selectivities of ≤95% ee. Circular dichroism, nuclear magnetic resonance nuclear Overhauser effect studies, and molecular dynamics simulations verify the helical structure of our catalyst in solution and the importance of the secondary structure in catalysis.

Data Mining for Terahertz Generation Crystals.

A data mining approach to discover and develop new organic nonlinear optical crystals that produce intense pulses of terahertz radiation is demonstrated. The Cambridge Structural Database is mined for non-centrosymmetric materials and these structural data are used in tandem with density functional theory calculations to predict new materials that efficiently generate terahertz radiation. This enables us to (in a relatively short time) discover, synthesize, and grow large, high-quality crystals of four promising materials and characterize them for intense terahertz generation. In a direct comparison to the current state-of-the-art organic terahertz generation crystals, these new materials excel. The discovery and characterization of these novel terahertz generators validate the approach of combining data mining with density functional theory calculations to predict properties of high-performance organic materials, potentially for a host of exciting applications.

Medium and Large N-Heterocycle Formation via Allene Hydroamination with a Bimetallic Rh(II) Catalyst.

We report the synthesis of a 2-phosphinoimidazole-derived bimetallic Rh(II) complex that enables intramolecular allene hydroamination to form 7- to 10-member rings in high yield. Monometallic Rh complexes, in contrast, fail to achieve any product formation. We demonstrate a broad substrate scope for formation of various N-heterocycles. Macrocyclizations that form 11- to 15-member N-heterocycles are also demonstrated. Mechanistic studies suggest that the reaction proceeds via reversible allene insertion with a Rh-hydride followed by C-N bond-forming reductive elimination. We hypothesize that the reactivity observed with our catalyst vs monometallic Rh complexes is derived from the bimetallic nature of our complex.

Synthesis of ɑ,ß-Unsaturated Benzotriazolyl-1,3,4-Oxadiazole Derivatives: Anticancer Activity, Cytotoxicity, and Cell Imaging.

A series of ten α , ß -unsaturated benzotriazolyl-1,3,4-oxadiazole derivatives was synthesized and all compounds were evaluated in vitro against three breast cancer cell lines (MCF-7, MDA-MB-231 and 4T1) at different concentrations (0.1, 0.5, 1, 2, 3, 4 and 5 mg/mL). The results showed that compounds 6a, 6c, 6d, 6f, 6g, and 6i displayed acceptable anticancer activity, where compound 6f was the most active on the three cell lines (IC50 = 0.80, 0.07, and 0.30 mg/mL, respectively). Regarding the cytotoxicity assay, the compounds exhibited modest toxicity on murine splenocytes and peripheral human blood cells at the highest concentration tested (5 mg/mL). Compound 6f was further evaluated at different concentrations showing moderate cytotoxicity at the 5 mg/mL concentration and negligible cytotoxicity at the minimum concentration evaluated (0.05 mg/mL). Finally, the compounds 6a, 6c, 6d, 6f, 6g, 6i, and 6j were evaluated as fluorescence markers due to their ability to be internalized into MCF-7 cells.

Custom terahertz waveforms using complementary organic nonlinear optical crystals.

Organic nonlinear optical (NLO) crystals are among the most efficient (>1%) terahertz (THz) radiation generators. However, one of the limitations of using organic NLO crystals is that the unique THz absorptions in each crystal make it difficult to obtain a strong, smooth, and broad emission spectrum. In this work, we combine THz pulses from two complementary crystals, DAST and PNPA, to effectively fill in spectral gaps, creating a smooth spectrum with frequencies out to 5 THz. The combination of pulses also increases the peak-to-peak field strength from 1 MV/cm to 1.9 MV/cm.

Enabling high-power, broadband THz generation with 800-nm pump wavelength.

The organic terahertz (THz) generation crystal BNA has recently gained traction as a source for producing broadband THz pulses. When pumped with 100 fs pulses, the thin BNA crystals can produce relatively high electric fields with frequency components out to 5 THz. However, the THz output with 800-nm pump wavelength is limited by the damage threshold of the material, particularly when using a 1 kHz or higher repetition rate laser. Here, we report that the damage threshold of BNA THz generation crystals can be significantly improved by bonding BNA to a high-thermal conductivity sapphire window. When pumped with 800-nm light from an amplified Ti:sapphire laser system, this higher damage threshold enables generation of 2.5× higher electric field strengths compared to bare BNA crystals. We characterize the average damage threshold for bare BNA and BNA-sapphire, measure peak-to-peak electric field strengths and THz waveforms, and determine the nonlinear transmission in BNA. Pumping BNA bonded to sapphire with 3 mJ 800-nm pulses results in peak-to-peak electric fields exceeding 1 MV/cm, with broadband frequency components >3 THz. This high-field, broadband THz source is a promising alternative to tilted pulse front LiNbO3 THz sources, enabling many research groups without optical parametric amplifiers to perform high-field, broadband THz spectroscopy.

CAN: A new program to streamline preparation of molecular coordinate files for molecular dynamics simulations.

Preparing molecular coordinate files for molecular dynamics (MD) simulations can be a very time-consuming process. Herein we present the development of a user-friendly program that drastically reduces the time required to prepare these molecular coordinate files for MD software packages such as AmberTools. Our program, known as charge atomtype naming (CAN), creates and uses a library of structures such as amino acid monomers to update the charge, atom type, and name of atoms in any molecular structure (mol2) file. We demonstrate the utility of this new program by rapidly preparing structural files for MD simulations for polypeptides ranging from small molecules to large protein structures. Both native and non-native amino acid residues are easily handled by this new program.

Heterogeneous layered structures for improved terahertz generation.

One of the most effective ways of generating terahertz (THz) radiation involves the conversion of short-pulsed IR or visible laser light into THz pulses at significantly lower frequencies. This conversion can be accomplished using organic crystals with nonlinear optical crystal (NLO) properties for IR to THz conversion through optical rectification. Due to the high refractive indices of organic crystals, pump laser light as well as generated THz radiation is lost from reflections at crystal surfaces. Here we report a structure composed of a layered series of materials with intermediate refractive indices designed to reduce reflective losses and improve the THz generation from organic crystals. This structure increases the transmission coefficients for both infrared pump input and THz output. We combine simple theoretical calculations with experimental data to show that a structure composed of materials with intermediate refractive indices can be used to increase generated THz intensity by nearly 50%.

γ-Tocotrienol and α-Tocopherol Ether Acetate Enhance Docetaxel Activity in Drug-Resistant Prostate Cancer Cells.

Prostate cancer is the second most commonly diagnosed cancer in men, and metastatic prostate cancer is currently incurable. Prostate cancer frequently becomes resistant to standard of care treatments, and the administration of chemotherapeutic drugs is often accompanied by toxic side effects. Combination therapy is one tool that can be used to combat therapeutic resistance and drug toxicity. Vitamin E (VE) compounds and analogs have been proposed as potential non-toxic chemotherapeutics. Here we modeled combination therapy using mixture design response surface methodology (MDRSM), a statistical technique designed to optimize mixture compositions, to determine whether combinations of three chemotherapeutic agents: γ-tocotrienol (γ-T3), α-tocopherol ether acetate (α-TEA), and docetaxel (DOC), would prove more effective than docetaxel alone in the treatment of human prostate cancer cells. Response surfaces were generated for cell viability, and the optimal treatment combination for reducing cell viability was calculated. We found that a combination of 20 µM γ-T3, 30 µM α-TEA, and 25 nm DOC was most effective in the treatment of PC-3 cells. We also found that the combination of γ-T3 and α-TEA with DOC decreased the amount of DOC required to reduce cell viability in PC-3 cells and ameliorated therapeutic resistance in DOC-resistant PC-3 cells.

Boron-Templated Dimerization of Allylic Alcohols To Form Protected 1,3-Diols via Acid Catalysis.

We report an unprecedented boron-templated dimerization of allylic alcohols that generates a 1,3-diol product with two stereogenic centers in high yield and diastereoselectivity. This acid-catalyzed reaction is achieved via in situ formation of a boronic ester intermediate that facilitates selective cyclization and formation of a cyclic boronic ester product. High yields are observed with a variety of allylic alcohols, and mechanistic studies confirm the role of boron as a template for the reaction.

Terahertz generation and optical characteristics of P-BI.

We present the structural and THz generation characteristics of the molecular salt crystal (E)-2-(4-(dimethylamino)styryl)-1,1,3-trimethyl-1H-benzo[e]indol-3-ium iodide (P-BI) using optical rectification with IR pump wavelengths. P-BI shows a peak-to-peak field ∼6 times greater than inorganic crystal GaP, and a broader THz spectrum. Data were obtained from 0-6 THz showing a significant dip in generation at 1.8 THz, similar to what has been observed with the THz generation crystal DAST at 1 THz. We characterized the power dependence of P-BI at different IR wavelengths, with optimal THz generation at the 1550-nm pump wavelength. To model THz generation as a function of P-BI crystal thickness, we measured the THz complex refractive index and the IR group index; modeling shows that imperfect phase matching leads to spectral narrowing centered at ∼2.5 THz as the crystal thickness is increased. P-BI could provide a useful alternative to inorganic THz generation crystals such a GaP.

Novel influenza inhibitors designed to target PB1 interactions with host importin RanBP5.

In search of novel targets for influenza inhibitors, a site on PB1 was selected for its high conservation and probable interaction with a host protein, RanBP5, that is key to nuclear import of PB1, where it complexes with PB2, PA, and NP to transcribe viral RNA. Docking with libraries of drug-like compounds led to a selection of five candidates that bound tightly and with a pose likely to inhibit protein binding. These were purchased and tested in vitro, found to be active, and then one was synthetically expanded to explore the structure-activity relationship. The top candidates had a carboxylic acid converted to an ester and electron-withdrawing substituents added to a phenyl group in the original structure. Resistance was slow to develop, but cytotoxicity was moderately high. Nuclear localization of PB1 and in vitro polymerase activity were both strongly inhibited.

Synthesis and characterization of ethyl benzotriazolyl acrylate-based D-π-A fluorophores for live cell-based imaging applications.

A series of eight new ethyl (Z)-benzotriazolyl acrylates 6a-d and 7a-d have been synthesized by conventional heating and microwave irradiation from ethyl benzotriazolyl acetates 3 and 4 with the corresponding aromatic aldehydes. This work reports the synthetic approach and spectroscopic characterization (1H, 13C-NMR, HRMS) of all the synthesized compounds. X-ray diffraction analyses were performed for molecules 6a, 7a and 7d. Photophysical properties of compounds were evaluated. Finally, compound 6a was tested in a human cell line and showed low to no cytotoxicity at relevant concentrations. Initial testing demonstrates its potential use as a fluid-phase fluorescent marker for live cell imaging.

C-N Bond Formation from Allylic Alcohols via Cooperative Nickel and Titanium Catalysis.

Amination of allylic alcohols is facilitated via cooperative catalysis. Catalytic Ti(O- i-Pr)4 is shown to dramatically increase the rate of nickel-catalyzed allylic amination, and mechanistic experiments confirm activation of the allylic alcohol by titanium. Aminations of primary and secondary allylic alcohols are demonstrated with a variety of amine nucleophiles. Diene-containing substrates also cyclize onto the nickel allyl intermediate prior to amination, generating carbocyclic amine products. This tandem process is only achieved under our cooperative catalytic system.

Electrophilic activation of alkynes for enyne cycloisomerization reactions with in situ generated early/late heterobimetallic Pt-Ti catalysts.

In situ formation of heterobimetallic Pt-Ti catalysts enables rapid room temperature catalysis in enyne cycloisomerization reactions. The Lewis acidic titanium atom in the ligand framework is shown to be essential for fast catalysis. A range of enyne substrates are efficiently cyclized to carbocycles and heterocycles in high yield.