Chemical synthesis of the 5'-terminal structure of U1 RNA

A new method for the synthesis of N2,N2-dimethylguanosine (DMG) is developed by reductive C-S bond cleavage by use of tributyltin hydride. An improved method for the synthesis of the key intermediate (1) for construction of the 5'-terminal structure of U1 RNA, which has a trimethylated cap (TMG) structure at its 5' end, is also described. By the use of 1, several TMG-capped ribonucleosides and oligonucleotides were synthesized.     

An efficient synthesis of N3,4-diphenyl-5-(4-fluorophenyl)-2-isopropyl-1H-3-pyrrolecarboxamide, a key intermediate for atorvastatin synthesis

An efficient synthesis of N3,4-diphenyl-5-(4-fluorophenyl)-2-isopropyl-1H-3-pyrrolecarboxamide, a key intermediate for the synthesis of an effective HMG-CoA reductase inhibitor atorvastatin, is described. The synthesis is based on the 1,3-dipolar cycloaddition reaction of mesoionic munchnone (1,3-oxazolium-5-olate) with N1,3-diphenyl-2-propynamide leading to N-benzyl pyrrole, and N-debenzylation using sodium in liquid ammonia as key steps.     

Stereodivergent synthesis of diastereoisomeric carba analogs of glycal-derived vinyl epoxides: a new access to carbasugars

A convenient method for the stereoselective synthesis of diasteroisomeric vinyl epoxides (-)-2α and (-)-2β, the carba analogs of D-galactal and D-allal-derived vinyl epoxides 1α and 1β, has been elaborated starting from tri-O-acetyl-D-glucal. The key step of this synthesis is an application of the known Claisen thermal rearrangement of a glucal derivative, the vinyl allyl ether (+)-3b, which allows to switch the glycal structure into the corresponding carba analog scaffold. Epoxides (-)-2α and (-)-2β derive from the same synthetic intermediate, the trans diol (+)-5.     

14Beta-hydroxy-10-deacetylbaccatin III as a convenient, alternative substrate for the improved synthesis of methoxylated second-generation taxanes

This article describes a new, convenient, improved synthesis of the 2-debenzoyl-2-m-methoxybenzoyl-7-triethylsilyl-13-oxo-14beta-hydroxybaccatin III 1,14-carbonate, the key intermediate in the synthesis of two new second-generation antitumor taxanes.     

3-N,N-Dimethylamino-3-deoxy lincomycin: a structure-based hybrid between lincomycin and the desosamine unit of erythromycin

The observation that the desosamine sugar unit in erythromycin and the methyl thiolincosaminide portion of lincomycin occupy virtually identical sites on the 23S rRNA according to X-ray crystallograpaic data, instigated the synthesis of 3-N,N-dimethylamino-3-deoxy lincomycin as a hybrid structure. The synthesis in eight steps from lincomycin, involving a trans-diequatorial opening of an intermediate epoxide as the key step, is described.     

Synthesis of L-2'-deoxypentofuranonucleoside derivatives of thymine from D-glucose

Convergent synthesis of L-2'-deoxypentofuranonucleoside derivatives of thymine was carried out from D-glucose via 6-O-toluoyl-3-deoxy-1,2-O-isopropylidene-beta-L-lyxo-hexofuranose as a key intermediate.     

Chemical synthesis of branched RNAs: a new method for construction of synthetic units for branched RNAs by use of 2'-phosphorylation on the basis of steric control.

A 3',5'-unprotected 2'-O-bis(tert-butoxy)-phosphinyl-6-N-benzoyladenosine derivative was prepared as a key intermediate for the synthesis of branched RNAs, and used for construction of building units from which chain elongation in any of 2'-, 3'-, and 5'-directions would be possible. From this synthetic intermediate, 2'-phosphorylated ApC dimer was also synthesized.     

1-O-benzyl-2-O-methyl-rac-glycerol: a key intermediate for synthesis of ether glycerolipids.

A new scheme for synthesis of saturated 1-O-alkyl-2-O-methyl-rac-glycerolipids is described. The reductive cleavage of 2-O-methyl-1,3-O,O-benzylideneglycerol with BH3.THF complex leads to 1-O-benzyl-2-O-methyl-rac-glycerol, which is a key in intermediate for the facile preparation of 1-O-alkyl-2-O-methyl-rac-glycerol and derivatives.     

Glycoprotein inhibitors and iodophilic polysaccharide storage in group A Streptococcus pyogenes

Inhibitors of glycoprotein synthesis in eucaryotic cells also inhibited iodophilic polysaccharide (IPS) storage in group A streptococcus pyogenes. Addition of bacitracin or amphomycin, known inhibitors of polyisoprenol phosphate metabolism or lipid-linked oligosaccharide synthesis, indicated that a key intermediate must be synthesized before IPS storage could be detected. Based on inhibitor action and energy requirements the intermediate was most likely an undecaprenol pyrophosphoryl maltosaccharide. Biosynthesis of the maltosaccharide had an ATP requirement as shown by arsenate action but IPS synthesis via amylomaltase was energy independent if the lipid-linked saccharides were preformed. Maltosaccharide acceptor site blockade with 2-deoxy-d-glucose immediately inhibited IPS storage, which demonstrated the need of acceptor glucose residues for transglycosylation activity of amylomaltase. Tunicamycin failed to inhibit IPS synthesis although it was added in lethal concentrations.     

Synthesis of 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-β-d-glucopyranosylamine and dimer formation

the synthesis of 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-β-d-glucopyranosylamine (2), the key intermediate in glycopeptide synthesis, has been improved. The dimerization of 2 has been studied as a model for its activity in biological systems. The formation of β,? and α,β dimers from 2 and their interconversions could be readily followed by ₁₃C-n.m.r. spectroscopy, and a probable mechanism of their formation involving an acyclic immonium ion intermediate has been proposed.     

Novel practical synthesis of Kdn2en and its C-4 nitrogen-modified derivatives

A practical synthesis of Kdn2en and 4-amino-4-deoxy-Kdn2en has been achieved via a key intermediate, methyl 4,5,7,8,9-penta-O-acetyl-2,6-anhydro-3-deoxy-D-glycero-D-galacto-non-2- enonate, which has been prepared from Kdn in three steps in 91% overall yield.     

Synthesis of enantiopure non-natural alpha-amino acids using tert-butyl (2S)-2-[bis-(tert-butoxycarbonyl)amino]-5-oxopentanoate as key-intermediate:the first synthesis of(S)-2-amino-oleic acid.

A general method for the synthesis of enantiopure non-natural alpha-amino acids is described. The key intermediate tert-butyl (2S)-2-[bis(tert-butoxycarbonyl)amino]-5-oxopentanoate was obtained from l-glutamic acid after suitable protection and selective reduction of the gamma-methyl ester group by DIBALH. Wittig reaction of this chiral aldehyde with various ylides led to a variety of delta,epsilon-unsaturated alpha-amino acids. This methodology was applied to the synthesis of (S)-2-amino-oleic acid.     

Microbial synthesis of S (+)-1-(2-furyl)-3-pentanol: A pheromone intermediate of bark beetle

Summary The synthesis of S (+)-1-(2-furyl)-3-pentanol (3), a key intermediate in the synthesis of chalcogran (1) is described via microbial reduction of 1-(2-furyl)-1-pentene-3-one (2) using fungusRhizopus arrhizus.     

Rapid kinetic studies of acetyl-CoA synthesis: evidence supporting the catalytic intermediacy of a paramagnetic NiFeC species in the autotrophic Wood-Ljungdahl pathway

CO dehydrogenase/acetyl-CoA synthase (CODH/ACS), a key enzyme in the Wood-Ljungdahl pathway of anaerobic CO(2) fixation, is a bifunctional enzyme containing CODH, which catalyzes the reversible two-electron oxidation of CO to CO(2), and ACS, which catalyzes acetyl-CoA synthesis from CoA, CO, and a methylated corrinoid iron-sulfur protein (CFeSP). ACS contains an active site nickel iron-sulfur cluster that forms a paramagnetic adduct with CO, called the nickel iron carbon (NiFeC) species, which we have hypothesized to be a key intermediate in acetyl-CoA synthesis. This hypothesis has been controversial. Here we report the results of steady-state kinetic experiments; stopped-flow and rapid freeze-quench transient kinetic studies; and kinetic simulations that directly test this hypothesis. Our results show that formation of the NiFeC intermediate occurs at approximately the same rate as, and its decay occurs 6-fold faster than, the rate of acetyl-CoA synthesis. Kinetic simulations of the steady-state and transient kinetic results accommodate the NiFeC species in the mechanism and define the rate constants for the elementary steps in acetyl-CoA synthesis. The combined results strongly support the kinetic competence of the NiFeC species in the Wood-Ljungdahl pathway. The results also imply that the methylation of ACS occurs by attack of the Ni(1+) site in the NiFeC intermediate on the methyl group of the methylated CFeSP. Our results indicate that CO inhibits acetyl-CoA synthesis by inhibiting this methyl transfer reaction. Under noninhibitory CO concentrations (below 100 microM), formation of the NiFeC species is rate-limiting, while at higher inhibitory CO concentrations, methyl transfer to ACS becomes rate-limiting.     

Chimeric RNA with modified backbones: alternating methylene(methylimino) linked phosphodiester backbone oligonucleotides with 2'-OH and 2'-OMe groups

Synthesis of a novel ribo-MMI dimer with 2'-OH and 2'-OMe in 5'- and 3'-nucleosides, respectively is presented. The synthesis was accomplished by reductive coupling of 3'-deoxy-3'-C-formyluridine and 2'-O-methyl-5'-O-methylaminouridine via a thioacetal as the key intermediate for the top part of the dimer. Incorporation of ribo-MMI dimers into oligonucleotides increased binding affinity for target RNA.     

NMR‐based characterization of a refolding intermediate of β2‐microglobulin labeled using a wheat germ cell‐free system

In patients with dialysis‐related amyloidosis, β2‐microglobulin (β2‐m) is a major structural component of amyloid fibrils. It has been suggested that the partial unfolding of β2‐m is a prerequisite to the formation of amyloid fibrils, and that the folding intermediate trapped by the non‐native trans‐Pro32 isomer leads to the formation of amyloid fibrils. Although clarifying the structure of this refolding intermediate by high resolution NMR spectroscopy is important, this has been made difficult by the limited lifetime of the intermediate. Here, we studied the structure of the refolding intermediate using a combination of amino acid selective labeling with wheat germ cell‐free protein synthesis and NMR techniques. The HSQC spectra of β2‐ms labeled selectively at either phenylalanine, leucine, or valine enabled us to monitor the structures of the refolding intermediate. The results suggested that the refolding intermediate has an overall fold and cores similar to the native structure, but contains disordered structures around Pro32. The fluctuation of the β‐sheet regions especially the last half of the βB strand and the first half of the βE strand, both suggested to be important for amyloidogenicity, may transform β2‐m into an amyloidogenic structure.     

New Amylase Inhibitor (S-AI) from Streptomyces diastaticus var. amylostaticus No. 2476

A one-step synthesis of dimethyl 3-substituted pyrrole-2, 4-dicarboxylates by the reaction of methyl isocyanoacetate with various aldehydes in the presence of DBU was carried out. Furthermore, the reaction mechanism was investigated; it was found that the methyl α-isocyanoacrylate compound is a key intermediate of the pyrrole compound.     

Structure and function of SirC from Bacillus megaterium: a metal-binding precorrin-2 dehydrogenase

In Bacillus megaterium, the synthesis of vitamin B(12) (cobalamin) and sirohaem diverges at sirohydrochlorin along the branched modified tetrapyrrole biosynthetic pathway. This key intermediate is made by the action of SirC, a precorrin-2 dehydrogenase that requires NAD(+) as a cofactor. The structure of SirC has now been solved by X-ray crystallography to 2.8 A (1 A = 0.1 nm) resolution. The protein is shown to consist of three domains and has a similar topology to the multifunctional sirohaem synthases Met8p and the N-terminal region of CysG, both of which catalyse not only the dehydrogenation of precorrin-2 but also the ferrochelation of sirohydrochlorin to give sirohaem. Guided by the structure, in the present study a number of active-site residues within SirC were investigated by site-directed mutagenesis. No active-site general base was identified, although surprisingly some of the resulting protein variants were found to have significantly enhanced catalytic activity. Unexpectedly, SirC was found to bind metal ions such as cobalt and copper, and to bind them in an identical fashion with that observed in Met8p. It is suggested that SirC may have evolved from a Met8p-like protein by loss of its chelatase activity. It is proposed that the ability of SirC to act as a single monofunctional enzyme, in conjunction with an independent chelatase, may provide greater control over the intermediate at this branchpoint in the synthesis of sirohaem and cobalamin.     

Synthetic studies of polyene macrolide antibiotics. 3. Hydrogenation of methyl-6-deoxy-2,3-O-isopropylidene-4-methylene- alpha-L-lyxo-hexopyranoside

Experimental conditions of hydrogenation of the title compound are found (excess of Raney catalyst in benzene solution, 5 degrees C, 2h) to afford methyl 4,6-dideoxy-2,3-O-isopropylidene-4-C-methyl-alpha-L-mannopyranoside, the key intermediate in the synthesis of C33-C38 fragment of amphotericin B, with the yield as high as 57%.     

Synthesis of New Thiazolidinone Nucleoside Analogues

Abstract

The synthesis of new thiazolididone nucleoside analogues is described. Among the different proposed synthetic pathways, the condensation of various nucleic bases using TMSOTf and Et₃N as coupling reagents on a key sulfoxide thiazolidinone intermediate led to the desired compounds in a one-pot procedure. Analytical data and NMR studies confirmed the proposed structure assignment for these compounds.