- Characterization of the first naturally thermostable terpene synthases and development of strategies to improve thermostability in this family of enzymes
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The terpenoid family of natural products is being targeted for heterologous microbial production as a cheaper and more reliable alternative to extraction from plants. The key enzyme responsible for diversification of terpene structure is the class-I terpene synthase (TS), and these often require engineering to improve properties such as thermostability, robustness and catalytic activity before they are suitable for industrial use. Improving thermostability typically relies on screening a large number of mutants, as there are no naturally thermostable TSs described upon which to base rational design decisions. We have characterized the first examples of natural TSs exhibiting thermostability, which catalyse the formation of the sesquiterpene τ-muurolol at temperatures up to 78 °C. We also report an enzyme with a kcat value of 0.95 s?1 at 65 °C, the highest kcat recorded for a bacterial sesquiterpene synthase. In turn, these thermostable enzymes were used as a model to inform the rational engineering of another TS, with the same specificity but low sequence identity to the model. The newly engineered variant displayed increased thermostability and turnover. Given the high structural homology of the class-I TS domain, this approach could be generally applicable to improving the properties of other enzymes in this class. Database: Model data are available in the PMDB database under the accession number PM0080780.
- Styles, Matthew Q.,Nesbitt, Edward A.,Marr, Scott,Hutchby, Marc,Leak, David J.
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Read Online
- Concise synthesis of the tetracyclic framework of azadiradione: Tandem radical cyclization route
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Azadiradione, a limonoid isolated from the neem plant, possesses a unique stereostructure distinct from that of the steroids. We describe a concise synthesis of the tetracyclic framework of azadiradione by a tandem radical cyclization process.
- Yamashita, Shuji,Naruko, Akito,Yamada, Takahiro,Hayashi, Yujiro,Hirama, Masahiro
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Read Online
- A mild method for the replacement of a hydroxyl group by halogen: 3. the dichotomous behavior of α-haloenamines towards allylic and propargylic alcohols
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A study of the deoxyhalogenation of allylic and propargylic alcohols with tetramethyl-α-halo-enamines is reported. Primary allylic and primary and secondary propargylic alcohols gave the corresponding halides in high yields. Secondary allylic and propargylic alcohols yielded the corresponding secondary halides but the reaction also produced some rearranged primary halides (I > Br > Cl). The reactions with tertiary allylic and tertiary propargylic alcohols gave several products and was therefore of little synthetic value. However, the addition of triethylamine to the reaction mixture or the use of lithium alkoxide instead of alcohol brought about a major change of the course of the reaction which led to amides carrying an allyl or an allenyl group at C2. This was shown to result from a Claisen-Eschenmoser rearrangement of an intermediate α-allyloxy- or propargyloxy-enamine.
- Munyemana, Fran?ois,Patiny, Luc,Ghosez, Léon
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- Characterization of a Sesquiterpene Synthase Catalyzing Formation of Cedrol and Two Diastereoisomers of Tricho-Acorenol from Euphorbia fischeriana
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A sesquiterpene synthase gene was identified from the transcriptome of Euphorbia fischeriana Steud, and the function of its product EfTPS12 was characterized by in vitro biochemical experiments and synthetic biology approaches. EfTPS12 catalyzed conversion of farnesyl diphosphate into three products, including cedrol (1) and eupho-Acorenols A (2) and B (3) (two diastereoisomers of tricho-Acorenol), thereby being named EfCAS herein. The structures of 2 and 3 were determined by spectroscopic methods and comparison of experimental and calculated electronic circular dichroism spectra. EfCAS is the first example of a plant-derived sesquiterpene synthase that is capable of synthesizing acorane-Type alcohols. This study also documents that synthetic biology approaches enable large-scale preparation of volatile terpenes and thereby substantially facilitate characterization of corresponding terpene synthases and elucidation of the structures of their products.
- Zhu, Jianxun,Liu, Lihong,Wu, Maobo,Xia, Guiyang,Lin, Pengcheng,Zi, Jiachen
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p. 1780 - 1786
(2021/06/21)
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- SYNTHESIS OF E,E-FARNESOL, FARNESYL ACETATE AND SQUALENE FROM FARNESENE VIA FARNESYL CHLORIDE
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The present disclosure provides methods for preparing polyunsaturated hydrocarbons, such as E,E-farnesol, farnesyl acetate and squalene, by base catalyzed addition of a dialkylamine to a 3-methylene-1-alkene, such as farnesene. The present disclosure also provides compositions including one more farnesene derivatives prepared using the disclosed methods.
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Paragraph 0081
(2019/12/28)
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- Repositioning Salirasib as a new antimalarial agent
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Malaria is a serious tropical disease that kills thousands of people every year, mainly in Africa, due to Plasmodium falciparum infections. Salirasib is a promising cancer drug candidate that interferes with the post-translational modification of Ras. This S-farnesyl thiosalicylate inhibits isoprenylcysteine carboxyl methyltransferase (ICMT), a validated target for cancer drug development. There is a high homology between the human and the parasite enzyme isoforms, in addition to being a druggable target. Looking to repurpose its structure as an antimalarial drug, a collection of S-substituted derivatives of thiosalicylic acid were prepared by introducing 1,2,3-triazole as a diversity entry point or by direct alkylation of the thiol. We further investigated the in vitro toxicity of FTS analogues to Plasmodium falciparum in the asexual stages and in Vero cells. An antiplasmodial activity assay was performed using a simple, high-sensitivity methodology based on nanoluciferase (NLuc)-transfected P. falciparum parasites. The results showed that some of the analogs were active at low micromolar concentration, including Salirasib. The most potent member of the series has S-farnesyl and the 1,2,3-triazole moiety substituted with phytyl. However, the compound substituted with methyl-naphthyl shows promising physicochemical and activity values. The low cytotoxicity in eukaryotic cells of the most active analogs provided good therapeutic indices, being starting-point candidates for future antimalarial drug development.
- Porta, Exequiel O. J.,Bofill Verdaguer, Ignasi,Perez, Consuelo,Banchio, Claudia,Ferreira De Azevedo, Mauro,Katzin, Alejandro M.,Labadie, Guillermo R.
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p. 1599 - 1605
(2019/09/30)
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- Concise synthesis of artemisinin from a farnesyl diphosphate analogue
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Artemisinin is one of the most potent anti-malaria drugs and many often-lengthy routes have been developed for its synthesis. Amorphadiene synthase, a key enzyme in the biosynthetic pathway of artemisinin, is able to convert an oxygenated farnesyl diphosphate analogue directly to dihydroartemisinic aldehyde, which can be converted to artemisinin in only four chemical steps, resulting in an efficient synthetic route to the anti-malaria drug.
- Tang, Xiaoping,Demiray, Melodi,Wirth, Thomas,Allemann, Rudolf K.
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p. 1314 - 1319
(2017/09/30)
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- Optimising Terpene Synthesis with Flow Biocatalysis
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Sesquiterpenes are an important family of natural products, many of which exhibit important pharmaceutical and agricultural properties. They are biosynthesised from farnesyl diphosphate in sesquiterpene synthase catalysed reactions. Here, we report the development of a highly efficient segmented flow system for the enzyme-catalysed continuous flow production of sesquiterpenes. Design of experiment (DoE) methods were used to optimise the performance of the flow biocatalysis, and quantitative yields were achieved by using an operationally simple but highly effective segmented flow system.
- Tang, Xiaoping,Allemann, Rudolf K.,Wirth, Thomas
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supporting information
p. 414 - 418
(2017/01/24)
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- ADHESION PREVENTING AGENT
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The present invention relates to an adhesion preventing agent comprising an amphipathic compound having the following general formula (I): wherein X and Y each denotes a hydrogen atom or together denote an oxygen atom, n denotes an integer from 0 to 2, m denotes the integer 1 or 2, AA: AA denotes a single bond or double bond, and R denotes a hydrophilic group having two or more hydroxyl groups.
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Paragraph 0372; 0373
(2016/04/20)
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- ADHESION PREVENTING AGENT
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PROBLEM TO BE SOLVED: To provide a compound available as an easily applicable adhesion preventing agent. SOLUTION: The invention provides an amphipathic compound represented by the general formula (II) in the figure. (In the formula, X and Y either each denote a hydrogen atom or together denote an oxygen atom, n denotes an integer from 0 to 2, m denotes the integer 1 or 2, and R denotes a hydrophilic group obtained by removal of one hydroxyl group from any one selected from the group consisting of glycerol, erythritol, pentaerythritol, diglycerol, and glyceric acid.) SELECTED DRAWING: None COPYRIGHT: (C)2016,JPOandINPIT
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Paragraph 0211-0212
(2016/12/01)
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- ISOPRENE OLIGOMER, POLYISOPRENE, PROCESSES FOR PRODUCING THESE MATERIALS, RUBBER COMPOSITION, AND PNEUMATIC TIRE
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The invention relates to an isoprene oligomer that contains a trans structural moiety and a cis structural moiety, which can be represented by the following formula (1), wherein at least 1 atom or group in the trans structural moiety is replaced by another atom or group. The invention also relates to a polyisoprene, which is biosynthesized using the isoprene oligomer and isopentenyl diphosphate. Further, this invention provides a rubber composition comprising the isoprene oligomer and/or the polyisoprene, and a pneumatic tire, including tire components (e.g., treads and sidewalls) formed from the rubber composition. wherein n represents an integer from 1 to 10; m represents an integer from 1 to 30; and Y represents a hydroxy group, a formyl group, a carboxy group, an ester group, a carbonyl group, or a group represented by the following formula (2):
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Paragraph 268
(2014/06/25)
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- Staphylococcus aureus penicillin-binding protein 2 can use depsi-lipid ii derived from vancomycin-resistant strains for cell wall synthesis
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Vancomycin-resistant Staphylococcus aureus (S. aureus) (VRSA) uses depsipeptide-containing modified cell-wall precursors for the biosynthesis of peptidoglycan. Transglycosylase is responsible for the polymerization of the peptidoglycan, and the penicillin-binding protein 2 (PBP2) plays a major role in the polymerization among several transglycosylases of wild-type S. aureus. However, it is unclear whether VRSA processes the depsipeptide-containing peptidoglycan precursor by using PBP2. Here, we describe the total synthesis of depsi-lipid I, a cell-wall precursor of VRSA. By using this chemistry, we prepared a depsi-lipid II analogue as substrate for a cell-free transglycosylation system. The reconstituted system revealed that the PBP2 of S. aureus is able to process a depsi-lipid II intermediate as efficiently as its normal substrate. Moreover, the system was successfully used to demonstrate the difference in the mode of action of the two antibiotics moenomycin and vancomycin. Copyright
- Nakamura, Jun,Yamashiro, Hidenori,Miya, Hiroto,Nishiguchi, Kenzo,Maki, Hideki,Arimoto, Hirokazu
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supporting information
p. 12104 - 12112
(2013/09/23)
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- Kinetic studies of Micrococcus luteus B-P 26 undecaprenyl diphosphate synthase reaction using 3-desmethyl allylic substrate analogs
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In order to investigate the substrate binding feature of undecaprenyl diphosphate synthase from Micrococcus luteus B-P 26 with respect to farnesyl diphosphate and a reaction intermediate, (Z,E,E)-geranylgeranyl diphosphate, we examined the reactivity of artificial substrate analogs, 3-desmethyl farnesyl diphosphate and 3-desmethyl Z-geranylgeranyl diphosphate, which lack the methyl group at the 3-position of farnesyl diphosphate and Z-geranylgeranyl diphosphate, respectively. Undecaprenyl diphosphate synthase did not accept either of the 3-desmethyl analogs as the allylic substrate, indicating that the methyl group at the 3-position of the allylic substrate is important in the undecaprenyl diphosphate synthase reaction. These analogs showed different inhibition patterns in the cis-prenyl chain elongation reaction with respect to the reactions of farnesyl diphosphate and Z-geranylgeranyl diphosphate as allylic substrate. These results suggest that the binding site for the natural substrate farnesyl diphosphate and those for the intermediate allylic diphosphate, which contains the cis-prenyl unit, are different during the cis-prenyl chain elongation reaction.
- Fujikura, Keitaro,Maki, Yuji,Ohya, Norimasa,Satoh, Mikiya,Koyama, Tanetoshi
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p. 851 - 855
(2008/09/18)
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- Solid-phase organic synthesis of polyisoprenoid alcohols with traceless sulfone linker
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(Chemical Equation Presented) Solid-phase organic synthesis of polyprenols with a traceless sulfone linker is described. The polymerbound benezenesulfinate is first linked with the "tail" building blocks of isoprenyl chlorides via S-alkylation. With use of dimsyl anion as an appropriate base, the polymer-bound α-sulfonyl carbanion is generated and coupled with other "body" building blocks in an efficient manner. After repeated processes and a global palladium-catalyzed desulfonation with LiEt3BH as the reducing agent, the desired polyprenols with various chain lengths and geometrical configurations are obtained in 32-59% overall yields. The solid-phase synthesis offers the advantage in facile isolation of polyprenols without tedious operation or time-consuming purification.
- Chang, Yi-Fan,Liu, Chen-Yu,Guo, Chih-Wei,Wang, Yen-Chih,Fang, Jim-Min,Cheng, Wei-Chieh
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supporting information; experimental part
p. 7197 - 7203
(2009/05/09)
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- Synthesis and structure-activity relationships of novel warfarin derivatives
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4-Hydroxycoumarins such as warfarin 1 have been the mainstay of oral anticoagulation therapy for over 20 years. Yet little detail is known about the molecular interactions between 4-hydroxycoumarins with vitamin K epoxide reductase (VKER), inhibition of w
- Gebauer, Markus
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p. 2414 - 2420
(2007/10/03)
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- Redox-active therapeutics for treatment of mitochondrial diseases and other conditions and modulation of energy biomarkers
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Methods of treating or suppressing mitochondrial diseases, such as Friedreich's ataxia (FRDA), Leber's Hereditary Optic Neuropathy (LHON), mitochondrial myopathy, encephalopathy, lactacidosis, stroke (MELAS), or Kearns-Sayre Syndrome (KSS) are disclosed,
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Page/Page column 37; 38
(2010/11/25)
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- Mycobacterium tuberculosis H37Rv3377c encodes the diterpene cyclase for producing the halimane skeleton
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The cloning and functional expression of Mycobacterium tuberculosis Rv3377c in Escherichia coli revealed that this gene encodes the diterpene cyclase for producing (+)-5(6),13-halimadiene-15-ol, which accepts geranylgeranyldiphosphate as the intrinsic substrate. The Royal Society of Chemistry 2005.
- Nakano, Chiaki,Okamura, Tomoo,Sato, Tsutomu,Dairi, Tohru,Hoshino, Tsutomu
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p. 1016 - 1018
(2007/10/03)
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- Pheromone synthesis, CXCVI. Synthesis of germacrene-B and its extension to the synthesis of (±)-9-methylgermacrene-B, the racemate of the male- produced sex pheromone of the sandfly Lutzomyia longipalpis from Lapinha, Brazil
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Both germacrene-B (2) and 9-methylgermacrene-B [(±)-1] were synthesized by employing cyclization reactions [8 → 9 and (±)-20 → (±)-21] as the key steps. The latter [(±)-1] was shown to be the racemate of the male-produced sex pheromone of the sandfly Lutz
- Muto, Shin-Etsu,Nishimura, Yutaka,Mori, Kenji
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p. 2159 - 2165
(2007/10/03)
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- Incorporation of farnesyl pyrophosphate derivertives into abscisic acid and its biosynthetic intermediates in Cercospora cruenta
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To investigate the transformation from (2E,6E)-farnesyl pyrophosphate to (2Z,4E)-γ-ionylideneethanol in the abscisic acid-producing fungi, Cercospora cruenta, plausible [2-14C]-C15 intermediates were prepared and fed. Substrates such as (2E,6E)-farnesyl pyrophosphate, (2Z,4E)-γ-ionylideneethanol and its pyrophosphate were incorporated into ABA and its known biosynthetic precursors. It is suggested that (2E,6E)-farnesyl pyrophosphate is converted to (2Z,4E)-γ-ionylideneethanol in four consecutive steps: dehydrogenation, isomerization, cyclization and hydrolysis.
- Yamamoto, Hirotaka,Oritani, Takayuki
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p. 821 - 824
(2007/10/03)
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- Method for lowering cholesterol employing a phosphonomethylphosphinate squalene synthetase inhibitor
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A method is provided for inhibiting cholesterol biosynthesis by inhibiting de novo squalene production employing methylene phosphonoalkylphosphinate compounds.
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- A novel approach towards the synthesis of pyrophosphate analogues of farnesyl pyrophosphate
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The synthesis of two new analogues (i.e. 3 and 4) of farnesyl pyrophosphate (FPP) containing a phosphonophosphate and a phosphonophosphate moiety was accomplished via the phosphonomorpholidate.The latter was easily obtained by treatment of a phosphonic chloride with morpholine.
- Valentijn, A. R. P. M.,Berg, O. van den,Marel, G. A. van der,Cohen, L. H.,Boom, J. H. van
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p. 563 - 566
(2007/10/02)
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- Method for preparing a phosphonic acid ester
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A method is provided for preparing a phosphonic acid ester having the structure STR1 wherein R1 is farnesyl or a derivative or analog thereof, and R2 is lower alkyl, by treating a farnesyl halide R1 Hal(Hal=Cl,Br,I) with an alkoxide of the structure STR2 wherein M is an alkali metal and R2c is lower alkyl. The resulting phosphonic acid ester is an intermediate in preparing a squalene synthetase inhibitor which is used for inhibiting cholesterol biosynthesis.
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- Phosphorus-containing squalene synthetase inhibitors
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Compounds which are inhibitors of cholesterol biosynthesis (by inhibiting de novo squalene biosynthesis), and thus are useful as hypocholesterolemic agents and antiatherosclerotic agents are provided which have the structure STR1 wherein m is 0, 1, 2 or 3; n is 0, 1, 2, 3 or 4; Y1 and Y2 are H or halogen; R2, R3 and R4 may be the same or different and are independently H, metal ion, C1 to C8 alkyl or C3 to C12 alkenyl; X is O, S, NH or NCH2 R15 wherein R15 is H or C1 to C5 alkyl; and R1 is R5 --Q1 --Q2 --Q3 -- wherein R5, Q1, Q2 and Q3 are as defined herein; and when m is o, X is other than S; and if m is o and X is 0, then n is 1, 2, 3 or 4; including all stereoisomers thereof.
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- A simple synthetic process for the elaboration of oligoprenols by stereospecific coupling of di-, tri-, or oligoisoprenoid units
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An effective method of allyl-allyl coupling which is position and stereospecific leads in a simple way to oligoprenols from E-allylic halides.
- Corey,Shieh
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p. 6435 - 6438
(2007/10/02)
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- SYNTHESIS OF ALKYL HALIDES UNDER NEUTRAL CONDITIONS
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Primary and secondary alcohols are efficiently converted to the corresponding alkyl halides under neutral conditions.
- Munyemana, Francois,Frisque-Hesbain, Anne-Marie,Devos, Alain,Ghosez, Leon
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p. 3077 - 3080
(2007/10/02)
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- INVESTIGATION OF ION-CATALYZED TELOMERIZATION. XXIV. THE COMPOSITION OF SESQUITERPENE CHLORIDES - THE DIADDUCTS OF ISOPRENE WITH ITS MONOHYDROCHLORIDES
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The isomeric composition of primary allylic sesquiterpene chlorides, i.e., the diadducts of the isoprene telomer with its monohydrochlorides, was interpreted by 13C NMR spectroscopy.The diadducts are formed predominantly by telomerization chain growth, and the chemical structure and composition are determined by the chemical structure of the taxogen.The possibility of increasing the yield with increase in the excess of the taxogen is suggested.
- Siirde, K. E.,Erm, A. Yu.,Muks, E. A.,Vyalimyae, T. K.,Krumm, L. L.,Leets, K. V.
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p. 832 - 836
(2007/10/02)
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- Phosphorylation of Isoprenoid Alcohols
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Procedures for the synthesis and purification of 20 isoprenoid diphosphates and methanediphosphonate analogues from the corresponding alcohols are described.The alcohols are activated for phosphorylation by conversion of homoallylic systems to tosylates and allylic systems to halides.The activated intermediates are treated with tris(tetra-n-butylammonium) salts of pyrophosphoric, methanediphosphonic, or difluoromethanediphosphonic acid to obtain the corresponding esters in yields 34-80percent.Chromatography on cellulose is a general method for purification of isoprenoid diphosphates, and procedures are decribed for compounds with C5 to C20 hydrocarbon moieties.The displacement by pyrophosphate occurs with inversion of configuration, and the procedure can be used to prepare isoprenoid diphosphates with chiral C1 methylene groups in high optical purity from the corresponding alcohols.
- Davisson, V. Jo,Woodside, Andrew B.,Neal, Timothy R.,Stremler, Kay E.,Muehlbacher, Manfred,Poulter, C. Dale
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p. 4768 - 4779
(2007/10/02)
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- Complexes η3-allylpalladium : isomerisation et photolyse ; synthese du squalene
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Complexation of primary allylic chlorides in the presence of Pdo may lead to a mixture of syn and anti η3-allylpalladium complexes.Photolysis or thermolysis also causes syn-anti isomerisation.A mixture of squalene stereoisomers is obtained by photolysis of the η3-farnesylpalladium complexes.
- Muzart, J.,Pete, J. P.
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- PHOTOCHEMICAL TRANSFORMATIONS-III; ORGANIC IODIDES (Part 3): GERANYL AND NERYL IODIDES AND 2(E),6(E)- AND 2(Z),6(E)-FARNESYL IODIDES
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Solution photolysis of geranyl and neryl iodides, and 2(E),6(E)- and 2(Z),6(E)-farnesyl iodides has been carried out.Products arising from simple elimination as well as ?-participation are formed.Thus, both geranyl and neryl iodides furnished, besides some unidentified compounds, myrcene, cis-ocimene, limonene and terpinolene, though in different proportions.Likewise, the sesquiterpene analogues yielded different amounts of trans-β-farnesene, β-bisabolene, trans-α-bisabolene and ar-curcumene.Results have been discussed in terms of ionic intermediates.
- Saplay, K. M.,Damodaran, N. P.,Dev, Sukh
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p. 2999 - 3004
(2007/10/02)
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- STEREOCONTROLLED SYNTHESIS OF CONJUGATED POLYENE ISOPRENOIDS USING PHOSPHINE OXIDE ANION INTERMEDIATES
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The potential for the Horner-Wittig reaction in the stereocontrolled synthesis of conjugated polyene isoprenoids is investigated.It is shown that metallations of the Z- and E-phosphine oxides (4 and 5), followed by reaction with Z- and E-citrals lead, in single steps, to the corresponding isomeric pentaenes (6) showing complete preservation of the Z- and E-geometries in the starting materials.Although the single step reactions leading to 6 proceed in low overall yields (10-20percent), the olefinations occur with >98percent E-stereoselectivity.Isolation of the intermediate β-hydroxyphosphine oxides (e.g. 14), followed by subsequent treatment with s odium hydride in dimethylformamide, demonstrate that these features of the reaction are associated largely with the remarkably slow rates of elimination from the erythro-intermediates in comparison with the threo-intermediates.Condensation between 4 and the ester-aldehyde (20) leads to the Z-6 ester 21 (34percent).After conversion of 21 into the tetraenal (25), reaction with the phosphine oxide (26), followed by separation of the erythro-β-hydroxy-phosphine oxide intermediate 27 and treatment with NaH in DMF, leads to the di-Z-octaene (28).The octaene 28 shows closely similar spectral data to those found in natural di-Z-phytofluene (2) isolated from fruit of the Tangerine tomato, Lycopersicon escalentum var.In a similar manner, reaction between the Z-phosphine oxide (4) and the dialdehyde (29) produces the di-Z-nonaene (33) which has the same chromophore as that found in di-Z-ζ-carotene (3) from Tangerine tomato fruits.
- Clough, John M.,Pattenden, Gerald
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p. 3911 - 3920
(2007/10/02)
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