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321.
U Junker D Bevec C Barske C Kalfoglou S Escaich M Dobrovnik J Hauber E B?hnlein 《Canadian Metallurgical Quarterly》1996,7(15):1861-1869
Previously, we described two mutants of the cellular Rev co-factor, eukaryotic initiation factor 5A (eIF-5A M13 and M14), which suppress human immunodeficiency virus type 1 (HIV-1) SF2 replication in clonal T cell lines. This study introduced the notion that it is possible to develop gene therapies against infectious agents on the basis of mutant host factors required for viral replication. In this report, we provide further evidence to support this new paradigm and describe murine leukemia virus (MLV)-based retroviral vectors expressing three different eIF-5A mutants from the viral long terminal repeat (LTR). HIV-1 replication (SF2, HXB-3) was reduced up to 2 orders of magnitude in transduced, polyclonal T cell populations. All eIF-5A mutants also showed antiviral activity (approximately seven-fold reduction) in a chronic HIV-1 infection model. Expression of eIF-5A mutant M13 delta in peripheral blood lymphocytes (PBLs) showed no difference in proliferation and metabolic activity as determined in a 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT)-assay, suggesting that expression of this type of mutant protein is not associated with cellular toxicity. In summary, these data suggest that gene therapy for HIV-1 infection can be developed on the basis of mutants of the Rev co-factor eIF-5A. 相似文献
322.
Sarah A. Berger Dr. Christopher Grimm Jonathan Nyenhuis Dr. Stefan E. Payer Dr. Isabel Oroz-Guinea Dr. Joerg H. Schrittwieser Prof. Dr. Wolfgang Kroutil 《Chembiochem : a European journal of chemical biology》2023,24(11):e202300170
Mass spectrometry-based high-throughput screening methods combine the advantages of photometric or fluorometric assays and analytical chromatography, as they are reasonably fast (throughput ≥1 sample/min) and broadly applicable, with no need for labelled substrates or products. However, the established MS-based screening approaches require specialised and expensive hardware, which limits their broad use throughout the research community. We show that a more common instrumental platform, a single-quadrupole HPLC-MS, can be used to rapidly analyse diverse biotransformations by flow-injection mass spectrometry (FIA-MS), that is, by automated infusion of samples to the ESI-MS detector without prior chromatographic separation. Common organic buffers can be employed as internal standard for quantification, and the method provides readily validated activity and selectivity information with an analytical run time of one minute per sample. We report four application examples that cover a broad range of analyte structures and concentrations (0.1–50 mM before dilution) and diverse biocatalyst preparations (crude cell lysates and whole microbial cells). Our results establish FIA-MS as a versatile and reliable alternative to more traditional methods for screening enzymatic reactions. 相似文献
323.
Willi Kantlehner Michael Hauber Erwin Haug Claus Schallenmüller Claudia Regele 《Advanced Synthesis \u0026amp; Catalysis》2000,342(7):682-699
Orthoamides. LIV. Contributions to the Chemistry of Azavinylogous Orthoformic Acid Amide Derivatives The azavinylogous aminalester 3 reacts with primary amines to give amidines 5 and 6 . In the reaction of 3 with aniline the azavinylogous amidine 7 is produced additionally to the amidine 5c . Ethylendiamine is formylated at both aminogroups, the bis‐amidine 8 thus formed is transformed to the salts 9a , b . Benzoxazole and benzimidazole can be prepared from 3 and o‐aminophenol and o‐phenylenediamine, resp. carboxylic acid amides, urea, thiourea, aromatic acid hydrazides 17 and the sulfonylhydrazide 19 are formylated by 3 at nitrogen to give N‐acylated formamidines 14 , 16 , 18 , 20 . From 3 and aliphatic acid hydrazides 17 and alkylhydrazines, resp., can be obtained 1,2,4‐triazole 21 and 1‐alkyl‐1,2,4‐triazoles 22a , b , resp. N,N‐dimethylcyanacetamide ( 32 ) reacts with 3 and the orthoamide 4a , resp., to give a mixture of the formylated compound 34 and the amidine 33 . The reaction conditions are of low influence on the ratio in which 33 and 34 are formed. The orthoamide 4b and 32 react to afford a mixture of the amidine 35 and the enamine 36 . Hydrogensulfide acts on 3 giving N,N‐dimethylthioformamide ( 37 ). From 3 and 1‐alkynes 41 can be prepared the amidines 42 . Hydrolysis of 42b affords phenylpropiolaldehyde ( 43 ). The alkylation of the aminalester 3 gives rise to the formation of vinylogous amidiniumsalts 1c and 1d , resp., additionally is formed the amide acetal 2a . The salt 1d can also be prepared from 3 and borontrifluoride‐ether. Iodide reacts with N,N‐dimethylformamide acetals 12a , b in an unclear, complicated manner giving orthoesters 53 , N,N‐dimethylformamide, alkyliodides, alcohols, ammoniumiodides 46 and carbondioxide. The action of halogens on 3 affords the salts 1a , b , c , e , f depending on the chosen stoichiometric ratio. Aromatic aldehydes are suited for trapping azavinylogous carbenes formed on thermolysis of 3 — 1,3‐oxazoles 69 are the reaction products. From 3 and propionaldehyde the amidine 65 can be obtained with low yield. Carbondisulfide transforms 3 to the azavinylogous salt 66 . The preparation of the azavinylogous orthoamide 4a is described. The thermolysis of 3 and 4a , resp., gives rise to the formation of the triaminopyrimidine 67 . Treatment of 1a with lithiumdiisopropylamide affords the triaminopyrazine 68 , which can also be obtained by thermolysis of 3 in the presence of sodium hydride. Azavinylogous carbenes are thought to be the intermediates. 相似文献
324.
A useful and specific method for the the title reaction ( 1 → 2 ) involves treatment with NaBH4 in CF3COOH whereas other procedures from the literature yield hard‐toseparate mixtures. 1‐Arylcyclohexanols 3 give mainly cyclohexenes with borohydride/trifluoroacetic acid. Their reduction to compounds 4 ,however, is possible with MeSO3H/NaBH4. 相似文献
325.
Willi Kantlehner Rüdiger Stieglitz Michael Hauber Erwin Haug Claudia Regele 《Advanced Synthesis \u0026amp; Catalysis》2000,342(3):256-268
The formamidinium salts 11a, c as well as the nitrile 12 react with sodiumhydride/dimethylamine in the presence of trimethylborate to give the ortho formic acid amide 3a . The orthoamides 6a and 16 can be prepared from the iminium salts 15 and 14 , resp. by the same procedure. Treatment of the azavinylogous formamidinium salt 15 with sodiumhydride and piperidine or morpholine in the presence of trime‐thylborate affords the orthoamides 6c and 6d , resp. By transamination of the azavinylogous aminalester 5a are accessible the orthoamides 6b—d . The vinylogous orthocarbonic acid derivative 17 can be obtained from the salt 14 and sodium alcoholates. The action of sodiumhydride, dimethylamine and trimethylborate on the iminium salt 18 produces a mixture of the orthocarbonic acid derivatives 7a, 8a, 9a . When the guanidinium salt 20 is treated with the same reagents the ortho‐amides 3a and 10a are obtained. The reduction of the salt 20 with sodiumhydride in the presence of several activating reagents (e.g. tetrabutyl orthotitanate, aluminiumisopropylate, trimethylborate) affords the orthoamide 3a . The reduction of the iminium salts 18 and 24 does not proceed clean, giving mixtures of various orthoformic acid derivatives. The form‐amidine 25 can be prepared by reduction of the salt 15 with sodiumhydride/trimethylborate with good yields. By the action of the corresponding carbanions on the guanidinium salt 20 can be obtained the carboxylic acid orthoamides 26—33 . By the same procedure the orthoamides of alkyne carboxylic acids 36a—h, j—n are accessible. 相似文献