65954-93-0Relevant academic research and scientific papers
Synthesis of pppA2'p5'A2'p5'A γ-amidates by one pot procedure from A2'p5'A2'p5'A
Nyilas, Agnes
, p. 2517 - 2518 (1997)
One pot synthesis of γ-amidate of pppA2'p5'A2'p5'A described here using phosphoroxychloride and bis-tri-n-butylammonium pyrophosphate for preparing cyclic trimetaphosphate intermediate opening up with n-decylamin.
High yield synthesis, purification and characterisation of the RNase L activators 5'-triphosphate 2′-5′-oligoadenylates
Morin,Rabah,Boretto-Soler,Tolou,Alvarez,Canard
experimental part, p. 345 - 352 (2011/10/07)
Upon viral infection, double-stranded viral RNA is detected very early in the host cell by several cellular 2′-5′ oligoadenylate synthetases, which synthesize 2′-5′ adenylate oligonucleotides that activate the cellular RNase L, firing an early primary antiviral response through self and non-self RNA cleavage. Transfecting cells with synthetic 2′-5′ adenylate oligonucleotides activate RNase L, and thus provide a useful shortcut to study the early steps of cellular and viral commitments into this pathway. Defined 2′-5′ adenylate oligonucleotides can be produced in vitro, but their controlled synthesis, purification, and characterisation have not been reported in detail. Here, we report a method suitable to produce large amounts of 2-5As of defined lengths in vitro using porcine OAS1 (pOAS) and human OAS2 (hOAS). We have synthesized a broad spectrum of 2-5As at the milligram scale and report an HPLC-purification and characterisation protocol with quantified yield for 2-5A of various lengths.
Role of the stereochemistry of 3'-fluoro-3'-deoxy analogues of 2-5A in binding to and activation of mouse RNase L
Kalinichenko, Elena N.,Podkopaeva, Tatjana L.,Poopeiko, Nicolai E.,Kelve, Merike,Saarma, Mart,et al.
, p. 43 - 50 (2007/10/02)
The synthesis of two sets of analogues of 2-5A trimer containing 9-(3-fluoro-3-deoxy-β-D-xylo-furanosyl)adenine (AF) or 3'-fluoro-3'-deoxyadenosine (AF) at different positions of the chain is described, along with the preparation of the corresponding 5'-monophosphates and 5'-diphosphorylated (core) trimers.The ability of each ribo and xylo isomeric pair of fluorodeoxy analogues of 2-5A (i) to compete with p3(A2'p)3A3'pC3'p for binding to RNase L in L929 cell extracts, and (ii) to activate the partially purified RNase L from L929 cell extracts to hydrolyze poly(U), was compared to that of the related 3'-deoxy analogue and the parent trimer, p3A3, using radiobinding and RNase L-(2',5')pentaadenylate(core)-agarose assays, respectively.Evidence is presented to show that the stereochemistry of the trimers plays an important role, specifically in the second process.The most striking observation is that, compared to 2-5A, p3A(AF)A was found to be nine times more effective an activator of RNase L, whereas isomeric p3A(AF)A is 30 times less effective.
Mg(II) ION-MEDIATED CONVERSION OF MONO- AND OLIGONUCLEOTIDES TO 5'-POLYPHOSPHATES IN AQUEOUS SOLUTION
Sawai, Hiroaki,Inaba, Yoshiko,Hirano, Atsushi,Wakai, Hiromichi,Shimazu, Masamitsu
, p. 4801 - 4804 (2007/10/02)
5'-Polyphosphates of mono- and oligonucleotides were prepared from the corresponding 5'-monophosphates with phosphorotriimidazolide or phosphorotribenzimidazolide mediated by Mg(II) or Mn(II) ion in aqueous solution.
Expedient Chemical Synthesis of Sequence-Specific 2',5'-Oligonucleotides
Imai, Jiro,Torrence, Paul F.
, p. 1418 - 1426 (2007/10/02)
A rapid chemical approach to the preparation of sequence-specific 2',5'-oligonucleotides and analogues of 2-5A is described.For instance, reaction of the 5'-phosphoromorpholidate of adenosine (MopA, 14) with the 5'-phosphoroimidazolidate of inosine (ImpI, 17), under conditions of lead ion catalysis, gave MopA2'p5'I (19a) in 21percent yield.Acid hydrolysis of 19a gave pA2'p5'I, which then was converted to the corresponding 5'-phosphoroimidazolidate Imp5'A2'p5'I (19c) through redox condensation with triphenylphosphine, imidazole, and 2,2-dipyridyl disulfide.Lead ion catalyzed condensation of 19c with Mop5'A (14) gave Mop5'A2'p5'A2'p5'I (30) in 17percent yield.By acid hydrolysis, 30 could be converted to the corresponding 5'-monophosphate, or, by reaction with pyrophosphate in DMF, to the corresponding 2-5A analogue, ppp5'A2'p5'A2'p5'I (5b).The following oligonucleotides were prepared by using similar methodology: ppp5'A2'p5'A2'p5'A (1b), ppp5'A2'p5'I2'p5'A (4b), ppp5'I2'p5'A2'p5'A (3b), and ppp5'A2'p5'A2'p5'(2'dA) (2b).
Bis(2,2,2-trichloroethyl) Phosphorochloridite as a Reagent for the Phosphorylation of Oligonucleotides: Preparation of 5'-Phosphorylated 2',5'-Oligoadenylates
Imai, Jiro,Torrence, Paul F.
, p. 4015 - 4021 (2007/10/02)
Bis(2,2,2-trichloroethyl) phosphorochloridite was found to be a useful reagent for the phosphorylation of protected nucleosides and oligonucleotides especially when the phosphate blocking group was 2,2,2-trichloroethyl and the hydroxyl protecting group was tert-butyldimethylsilyl.Thus compounds 2a-c,f,g could be phosphorylated to the corresponding 5'-phosphotriesters 4a-c,f,g in yields of 75-99percent.Removal of the protecting groups (to give the 5'-monophosphates 6a-c,f,g) was achieved by zinc-copper couple/2,4-pentanedione/DMF treatment to remove the 2,2,2-trichloroethyl group and tetrabutylammonium fluoride/THF treatment to remove the tert-butyldimethylsilyl groups.A method was found that permits preparation of reproducibly active zinc-copper couple.As a hydroxyl protecting group, the isopropylidene moiety was somewhat less useful in conjunction with the use of bis(2,2,2-trichloroethyl) phosphorochloridite.Thus, upon phosphorylation of 2d and 2e, the phosphotriesters 4d and 4e were obtained in yields of 83percent and 61percent, respectively.Deblocking of the isopropylidene groups was accomplished with formic acid at room temperature to give 6a and 6b in yields of 74percent and 70percent, respectively.The 5'-phosphorylated 2'-5'-linked oligonucleotides 6f and 6g were converted to the corresponding 5'-triphosphates to give compounds 1a and 1b which are found in extracts of interferon-treated cells upon incubation with double-stranded RNA.
