High-throughput purification of combinatorial libraries I: a high-throughput purification system using an accelerated retention window approach

We have developed a high-throughput purification system to purify combinatorial libraries at a 50-100-mg scale with a throughput of 250 samples/instrument/day. We applied an accelerated retention window method to shorten the purification time and targeted one fraction per injection to simplify data tracking, lower QC workload, and simplify the postpurification processing. First, we determined the accurate retention time and peak height for all compounds using an eight-channel parallel LC/UV/MS system, and calculated the specific preparative HPLC conditions for individual compounds. The preparative HPLC conditions include the compound-specific gradient segment for individual compounds with a fixed gradient slope and the compound-specific UV or ELSD threshold for triggering a fraction collection device. A unique solvent composition or solvent strength was programmed for each compound in the preparative HPLC in order to elute all compounds at the same target time. Considering the possible deviation of the predicted retention time, a 1-min window around the target time was set to collect peaks above a threshold based on UV or ELSD detection. Dual column preparative instruments were used to maximize throughput. We have purified more than 500 000 druglike compounds using this system in the past 3 years. We report various components of this high-throughput purification system and some of our purification results.     

Migratory-insertion reactions involving the thiocarbonyl ligand. dihapto-thioacyl complexes from the rearrangement of arylthiocarbonyl complexes of osmium(II). Structure of Os(η2-CSR)(η1-O2CCF3)(CO)(PPh3)2

Reaction of HgR₂ with OsHCl(CS)(PPh₃)₃ yields red, five-coordinate, OsRCl-(CS)(PPh₃)₂ (R = p-tolyl). From this have been derived the compounds OsRX(CS)(PPh₃)₂ with X = Br, I, S₂CNEt₂, O₂CMe, O₂CCF₃. These compounds add an additional ligand, MeCN, CO or CNR to form colourless, six coordinate arylthiocarbonyl complexes, which undergo migratory-insertion reactions to form red, dihapto-thioacyl complexes. The crystal structure of a representative example, Os(η²-CSR)(η¹-O₂CCF₃)(CO)PPh₃)₂ has been determined. The red equant crystals are orthorhombic, space group P2₁2₁2₁, a 11.584(1), b 19.184(2), c 18.90(1) Å, V 4199 ų, Z  4. The structure was solved by conventional heavy-atom methods and refined by full-matrix least-squares employing anisotropic thermal parameters for all non-hydrogen atoms except the carbon atoms of the triphenylphosphines. The final R factor is 0.057 for 2868 observed reflections.The coordination geometry in the monomeric complex is that of an octahedron distorted by the constraints of the ligands. The triphenyl phosphine ligands are mutually trans; the equatorial plane contains carbonyl, monohapto-trifluoroacetate, and dihapto-thioacyl ligands. Bond distances and angles are OsP 2.405, 2.407(4) Å; POsP 173.9(1)°; OsCO 1.83(2) Å; Os-O (trifluoroacetate) 2.206(11) Å; OsC (thioacyl) 1.91(2); OsS 2.513(6); CS 1.72 Å. The CS bond length implies a reduction in bond order from 2.0 to approx. 1.5 upon coordination to the metal.The η²-thioacyl ligand in Os(η²-CSR)Cl(CNR)(PPh³)₂ is methylated with methyl triflate and further reaction with LiCl produces the thiocarbene complex OsCl₂(C[SMe]R)(CNR)(PPh₃)₂.     

Generation and reactivity of bis(2,6-diethylphenyl)germanium(II)

The title intermediate ($\text{3a}$) is produced on photolysis of hexakis(2,6-diethyl-phenyl)cyclotrigermane ($\text{1}$) or bis(2,6-diethylphenyl)bis(trimethylsilyl)germane ($\text{4}$) as evidenced by trapping experiments, and thermally dimerizes to tetrakis(2,6-diethyl-phenyl)digermene ($\text{2a}$). Diarylgermylenes such as $\text{3a}$ do not form stable triethylamine adducts (e.g. $\text{5a}$) as has been previously reported.     

Thermal analysis of Vectran® fibers and films

Vectra^® liquid crystalline polymers (LCP's) were introduced as commercial products in the mid-1980's. The first of these (Vectra A130) was a wholly aromatic thermotropic copolyester ofp-hydroxybenzoic acid and 6-hydroxy-2-naphthoic acid. Vectra A130 is a thermotropic LCP that can be melt spun into filaments that on heat treatment are characterized by high strength and high modulus. Vectra resin can also be extruded into films. In the fiber or film form this material is commercially known as Vectran^®. Heat treatment enhances the tensile strength of Vectran fiber variants. Because of this, the elucidation of the physical transformations taking place in the internal structure of the material during heating has always been an important subject. Several thermal techniques are used to indicate clearly that what is observed as a glass transition is unlike the conventional glass transition in typical semicrystalline polymers. There is also an indication of the presence of multiple states of mesophase aggregation that collapse into a single state when taken to high enough temperatures.     

Molecular weights of backbones in graft copolymers

Copolymerization of a monomer X with an existing polymer yields a copolymer contaminated by poly-X homopolymer and ungrafted polymer. Measurements on the pure copolymer can yield only the molecular weights of the cumulative backbone portion and of the cumulative graft portion. However, if the grafts can be isolated, then the molecular weights of their individual chains may be determined also. Since it is not isolatable, the backbone is not amenable to similar treatment. Equations are derived for the molecular weights (of any average) of individual backbone chains; they are expressed in terms of the following experimentally accessible quantities: (i) molecular weight of original polymer; (ii) molecular weight of ungrafted portion: and (iii) fraction by weight of the original polymer which remains ungrafted. The procedure is thus the analogue, applied to the backbone, of chemical isolation applied to the grafts. The treatment has been examined for copolymers prepared by grafting acrylic acid to nylon-6 by mutual irradiation.     

Determination of in-core power in low energy research reactors by measurement of16N and18F in the primary coolant

The measurement of¹⁶N and¹⁸F activity in the primary coolant of the JASON Argonaut reactor has been used to monitor in-core reactor power. The¹⁶N is produced by the¹⁶O(n, p)¹⁶N reaction and the 6.1 MeV photopeak was measured on-line using a BGO detector adjacent to the primary coolant circuit. These data provided a relative measure of power stability during steady state operation and a measure of linearity at different power levels. The¹⁸F is produced in the primary coolant by the¹⁸O(p, n)¹⁸F reaction and aliquots of primary coolant were sampled from the reactor dump tank for off-line radiochemical analysis. The¹⁸F was separated as trimethylfluorosilane and the activity was determined by measurement of the 0.511 MeV annihilation photopeak using a NaI(TI) detector. The measured¹⁸F activity was used to determine actual in-core reactor power using both ab-initio calculations and by comparison of results with a calibrated power reactor. The¹⁸F data also provided a method of nomalising the¹⁶N data for direct monitoring of in-core reactor power in JASON.     

Multifunctional redox catalysts as selective enhancers of oxidative stress

Certain cancer cells proliferate under conditions of oxidative stress (OS) and might therefore be selectively targeted by redox catalysts. Among these catalysts, compounds containing a chalcogen and a quinone redox centre are particularly well suited to respond to the presence of OS. These catalysts combine the specific electrochemical features of quinones and chalcogens. They exhibit high selectivity and efficiency against oxidatively stressed rat PC12, human Jurkat and human Daudi cells in cell culture, where their mode of action most likely involves the catalytic activation of existent and the generation of new reactive oxygen species. The high efficiency and selectivity shown by these catalysts makes them interesting for the development of anti-cancer drugs.     

Molecular potential-energy surfaces for chemical reaction dynamics

This paper reviews the construction of molecular potential-energy surfaces by an interpolation method which has been developed over the last several years. The method uses ab initio quantum chemistry calculations of the molecular electronic energy in an automated procedure to construct global potential- energy surfaces which can be used to simulate chemical reactions with either classical or quantum dynamics. The methodology is explained and several applications are presented to illustrate the approach. Received: 22 February 2002 / Accepted: 2 May 2002 / Published online: 6 November 2002 Correspondence to: M. A. Collins e-mail: collins@rsc.anu.edu.au Acknowledgements. The methods described in this overview are the result of collaborations with former members of my group, in particular with Josef Ischtwan, Meredith Jordon, Keiran Thompson and Ryan Bettens. I am also indebted for inspiration gained from many discussions with my colleagues Leo Radom and Donghui Zhang (National University of Singapore). This work has been supported by the Supercomputer Facility of the Australian National University and the Australian Partnership for Advanced Computing.