Next‐Generation o‐Nitrobenzyl Photolabile Groups for Light‐Directed Chemistry and Microarray Synthesis

Light as an external trigger is a valuable and easily controllable tool for directing chemical reactions with high spatial and temporal accuracy. Two o‐nitrobenzyl derivatives, benzoyl‐ and thiophenyl‐NPPOC, undergo photo‐deprotection with significantly improved efficiency over that of the commonly used NPPOC group. The two‐ and twelvefold increase in photo‐deprotection efficiency was proven using photolithograph synthesis of microarrays.     

Influence of free convection on the heat transfer from hot-wire probes

The present paper is concerned with the experimental determination of the influence of free convection on the heat transfer from horizontal hot-wire probes in cross flow. Free convective flow can be minimized under microgravity conditions and its quantity can be determined by comparisons with terrestrial investigations. Using the developod experimental setups it is possible to investigate all three flow regimes, i.e. pure free, mixed, and pure forced convection (o Re 0.1). It was found that the influence of free convection is limited to Reynolds numberRe 0.0045, independent of the overheat ratio. Based on the findings of Collis and Williams [12], the influence of free convection can be neglected forRe > 0.58 ·Gr ^1/3. The fluid properties are computed at the film temperature. Two correlations are established for the regime of pure forced convection. End losses to the supports were determined in a vacuum experiment, thus allowing comparison with theoretical investigations. The difference between the analytical computations and the measurement data in the range 0 Re 0.02 is caused by the three-dimensional heat transfer occuring in the experimental investigation.Die vorliegende Untersuchung beschäftigt sich mit der experimentellen Bestimmung des Einflusses der freien Konvektion auf den Wärmeübergang bei quergeströmten Hitzdrahtsonden. Die freie Konvektionsströmung kann unter Schwerelosigkeit minimiert und ihre Größe durch den Vergleich mit terrestrischen Untersuchungen ermittelt werden. Mit den erstellten Experimentaufbauten ist es möglich, alle drei Strömungsbereiche von der rein freien über die gemischte bis zur rein erzwungenen Konvektion zu untersuchen (0 <>Re 0,1). Dabei wurde festgestellt, daß der Einfluß der freien Konvektion sich unabhängig von Überhitzungsverhältnis nur bis zu einer ReynoldszahlRe 0,0045 erstreckt. Zur Vernachlässigung der freien Konvektion kann in Anlehnung an Collins und Williams folgende Beziehung angegeben werden:Re > 0,58 ·Gr ^1/3. Die Stoffwerte werden bei der Filmtemperatur berechnet. Für den Bernich der rein erzwungenen Konvektion wurden zwei Korrelationer aufgestellt. Durch einen Vakuumversuch wurden die Endverluste in die Haltespitzen ermittelt und somit der Vergleich mit theoretischen Arbeiten ermöglicht. Der Unterschied zwischen den analytischen Berechnungen und den Meßdaten im Bereich 0 Re 0,02 ergibt sich durch den dreidimensionalen Wärmeübergang bei der experimentellen Untersuchung.The authors acknowledge the support of the Deutsche Agentur für Raumfahrtangelegenheiten (DARA) under grant number 50-QV 8898-1.     

19F-und 1H-NMR-spektroskopische Untersuchungen an Hexafluoraceton-halbacetalen von Pyrimidin-nucleosiden

The reaction of OH groups of pyrimidine nucleosides with hexafluoroacetone yields hemiacetals quantitatively. The ¹⁹F NMR spectra allow unambiguous characterisation of the starting compounds and also aid in determining the selectivity of position-specific nucleoside blocking groups. On account of their greater solubility in many organic solvents, the hemiacetals offer advantages over their parent compounds in ¹H NMR spectroscopy.     

New Types of Very Efficient Photolabile Protecting Groups Based upon the [2‐(2‐Nitrophenyl)propoxy]carbonyl (NPPOC) Moiety

Based upon the photolabile [2‐(2‐nitrophenyl)propoxy]carbonyl group (NPPOC), a large number of modified 2‐(2‐nitrophenyl)propanol derivatives substituted at the phenyl ring (see 23 – 34 and 57 – 76 ) as well as at the side‐chain (see 85 – 92 and 95 – 98 ) were synthesized to improve the photoreactivity of this new type of photolabile entity. The phenyl moiety was also exchanged by the naphthalenyl group (see 102, 103, 105, 108, 110, 113 , and 114 ), the thienyl substituent (see 115, 117, 118 , and 120 ), and the benzothienyl substituent (see 121 ). The 2‐(2‐nitroaryl‐ and heteroaryl)propanols were converted with diphosgene into the corresponding carbonochloridates, which reacted subsequently with thymidine to the thymidine 5′‐(protected carbonates) 123 – 178 as the main reaction products. In several cases, the corresponding 3′‐carbonates and 3′,5′‐dicarbonates 179 – 212 were also isolated and characterized. Photolysis studies under standardized conditions (see Table) indicated that the rate of photocleavage varies in a broad range depending on the substituents. So far, the thymidine 5′‐[2‐(5‐halo‐2‐nitrophenyl)propyl carbonates] 127 – 129 , 5′‐[2‐(nitro[1,1′‐biphenyl]3‐yl)propyl carbonates] 136 – 139 , 5′‐{2‐[2‐nitro‐5‐(thianthren‐1‐yl)phenyl]propyl carbonate} ( 140 ), 5′‐[2‐(5‐naphthalenyl‐2‐nitrophenyl)propyl carbonates] 141 and 142 , and 5′‐[2‐(2‐nitro‐5‐thienylphenyl)propyl carbonates] 143 and 144 showed the best properties regarding fast and uniform deprotection. Since the nucleobases of 213 – 215 do not influence the photocleavage features, in general, the new type of photolabile building blocks allows in form of their 3′‐phosphoramidites the photolithographic formation of high‐quality biochips.     

Differential thermal study of Mg-bearing clays from saline lakes of Southern Tunisia

Clays high in Mg content occur frequently in the high saline environment of salt lakes in southern Tunisia. The DTA curves of these clays show a striking endothermic-exothermic reaction in the temperature range of 800–820°C. A strong correlation is observed between the intensity of these coupled reactions and the Mg content of the initial clay sample. The initial endothermic reaction is interpreted as the melting/dehydroxylation of the Mg-bearing smectites. The subsequent exothermic peak is interpreted as caused by the crystallisation of the new Mg-silicate phase enstatite. Therefore, the DTA is considered as a suitable method for the identification and relative quantification of high Mg clay minerals (e.g. trioctahedral smectites). Variations of the Mg content of the studied samples were well detectable by means of DTA, disclosing a distinct distribution pattern of the salt lake clays. Clues to bulk chemical composition of the initial clay assemblage can also be found in the results of the X-ray analysis of the firing products.     

Triplet‐Sensitized Photodeprotection of Oligonucleotides in Solution and on Microarray Chips

Conditions and kinetics of triplet sensitization as a method for increasing the light sensitivity of photolabile protecting groups used for the photolithographic synthesis of oligonucleotide microarrays were quantitatively studied with the photolabile 2‐(2‐nitrophenyl)propyl protecting group in homogeneous solutions and on glass substrates by using laser flash photolysis, continuous illumination with HPLC analysis, fluorescence dye labelling, and hybridization. In terms of efficiency and avoidance of chemical side reactions, 9H‐thioxanthen‐9‐one was the most‐suitable sensitizer. Both in solution and on a glass substrate, the photostationary kinetics were quantitatively modelled and the relevant kinetic parameters determined. While the sensitization kinetics was diffusion‐controlled both in solution and on the chip, the photostationary kinetics was essentially of zero order only on the chip because here the triplet‐quenching effect of the released photoproduct 2‐(2‐nitrophenyl)propene was suppressed as a consequence of the inhomogeneous reaction that took place in a narrow diffusion zone above the surface from where the photoproducts could quickly escape. The kinetic simulation allowed quantitative estimate of the density of reactive groups on the surface. It was further demonstrated that, with 9H‐thioxanthen‐9‐one as a sensitizer, high‐density oligonucleotide microarrays of high quality can be produced with one‐third of the normal exposure time.     

Intramolecular sensitization of photocleavage of the photolabile 2-(2-nitrophenyl)propoxycarbonyl (NPPOC) protecting group: photoproducts and photokinetics of the release of nucleosides

Novel photolabile protecting groups based on the 2-(2-nitrophenyl)propoxycarbonyl (NPPOC) group with a covalently linked thioxanthone as an intramolecular triplet sensitizer exhibit significantly enhanced light sensitivity under continuous illumination. Herein we present a detailed study of the photokinetics and photoproducts of nucleosides caged with these new protecting groups. Relative to the parent NPPOC group, the light sensitivity of the new photolabile protecting groups is enhanced by up to a factor of 21 at 366 nm and is still quite high at 405 nm, the wavelength at which the sensitivity of the parent compound is practically zero. A new pathway for deprotection of the NPPOC group proceeding through a nitroso benzylalcohol intermediate has been discovered to complement the main mechanism, which involves beta elimination. Under standard conditions of lithographic DNA-chip synthesis, some of the new compounds, while maintaining the same chip quality, react ten times faster than the unmodified NPPOC-protected nucleosides.     

Nucleosides. Part LIX. The 2-(4-nitrophenyl)ethylsulfonyl (Npes) Group: A New Type of Protection in Nucleoside Chemistry

The 2-(4-nitrophenyl)ethylsulfonyl (npes) group is developed as a new sugar OH-blocking group in the ribonucleoside series. Its cleavage can be performed in a β-eliminating process under aprotic conditions using 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) as the most effective base. Since sulfonates do not show acyl migration, partial protection of 1,2-cis-diol moieties is possible leading to new types of oligonucleotide building blocks. A series of Markiewicz-protected ribonucleosides 1–10 is converted into their 2′-O-[2-(4-nitrophenyl)ethylsulfonyl] derivatives 29–38 in which the 5′-O? Si bond can be cleaved by acid hydrolysis forming 39–45 . Subsequent monomethoxytritylation leads to 46–50 , and desilylation affords the 5′-O-(monomethoxytrityl)-2′-O-[2-(4-nitrophenyl)ethylsulfonyl]ribonucleosides 51–55 . Acid treatment to remove trityl groups do also not harm the npes group (→ 56–58 ). Unambiguous syntheses of fully blocked 2′-O-[2-(4-nitrophenyl)ethylsulfonyl]ribonucleosides 96–102 are achieved from the corresponding 3′-O-(tert-butyl)dimethylsilyl derivatives. Furthermore, various base-protected 5′-O-(monomethoxytrityl)- and 5′-O-(dimethoxytrityl)ribonucleosides, i.e. 59–77 , are treated directly with 2-(4-nitrophenyl)ethylsulfonyl chloride forming in all cases a mixture of the 2′,3′-di-O- and the two possible 2′- and 3′-O-monosulfonates 107–148 which can be separated into the pure components by chromatographic methods. The npes group is more labile towards DBU cleavage than the corresponding base-protecting 2-(4-nitrophenyl)ethyl (npe) and 2-(4-nitrophenyl)ethoxycarbonyl (npeoc) groups allowing selective deblocking which is of great synthetic potential.