Designed Small-Molecule Inhibitors of the Anthranilyl-CoA Synthetase PqsA Block Quinolone Biosynthesis in Pseudomonas aeruginosa.

The Gram-negative bacterial pathogen Pseudomonas aeruginosa uses three interconnected intercellular signaling systems regulated by the transcription factors LasR, RhlR, and MvfR (PqsR), which mediate bacterial cell-cell communication via small-molecule natural products and control the production of a variety of virulence factors. The MvfR system is activated by and controls the biosynthesis of the quinolone quorum sensing factors HHQ and PQS. A key step in the biosynthesis of these quinolones is catalyzed by the anthranilyl-CoA synthetase PqsA. To develop inhibitors of PqsA as novel potential antivirulence antibiotics, we report herein the design and synthesis of sulfonyladeonsine-based mimics of the anthranilyl-AMP reaction intermediate that is bound tightly by PqsA. Biochemical, microbiological, and pharmacological studies identified two potent PqsA inhibitors, anthranilyl-AMS (1) and anthranilyl-AMSN (2), that decreased HHQ and PQS production in P. aeruginosa strain PA14. However, these compounds did not inhibit production of the virulence factor pyocyanin. Moreover, they exhibited limited bacterial penetration in compound accumulation studies. This work provides the most potent PqsA inhibitors reported to date and sets the stage for future efforts to develop analogues with improved cellular activity to investigate further the complex relationships between quinolone biosynthesis and virulence factor production in P. aeruginosa and the therapeutic potential of targeting PqsA.

Tailored α-methylene-γ-butyrolactones and their effects on growth suppression in pancreatic carcinoma cells.

A selected series of racemic α-methylene-γ-butyrolactones (AMGBL) were synthesized via allylboration and screened against three human pancreatic cancer cell lines (Panc-1, MIA PaCa-2, and BxPC-3). This systematic study established a discernible relationship between the substitution pattern of AMGBL and their anti-proliferative activity. ß,γ-diaryl-AMGBLs, particularly those with a trans-relationship exhibited higher potency than parthenolide and LC-1 against all three cell lines.

Asymmetric aldol reaction with diisopinocampheyl enolborinates of propionates.

A convenient and general, reagent-controlled, diastereo- and enantioselective aldol reaction of diisopinocampheylboron enolates of esters, followed by reduction, has been developed as an alternative to crotylboration-ozonolysis. This protocol was then exploited for the double diastereoselective synthesis of the C11-C17 subunit of (-)-dictyostatin.

Effects of gabapentin on cocaine self-administration, cocaine-triggered relapse and cocaine-enhanced nucleus accumbens dopamine in rats.

Gabapentin is a gamma-aminobutyric acid (GABA) analogue, with GABAmimetic pharmacological properties. Gabapentin is used for the treatment of seizures, anxiety and neuropathic pain. It has been proposed that gabapentin may be useful in the treatment of cocaine dependence. However, clinical trials with gabapentin have shown conflicting results, while preclinical studies are sparse. In the present study, we investigated the effects of gabapentin on intravenous cocaine self-administration and cocaine-triggered reinstatement of drug-seeking behavior, as well as on cocaine-enhanced dopamine (DA) in the nucleus accumbens (NAc). We found that gabapentin (25-200 mg/kg, i.p., 30 min or 2 h prior to cocaine) failed to inhibit intravenous cocaine (0.5 mg/kg/infusion) self-administration under a fixed-ratio reinforcement schedule or cocaine-triggered reinstatement of cocaine-seeking behavior. In vivo microdialysis showed that the same doses of gabapentin produced a modest increase (approximately 50%, p<0.05) in extracellular NAc GABA levels, but failed to alter either basal or cocaine-enhanced NAc DA. These data suggest that gabapentin is a weak GABA-mimic drug. At the doses tested, it has no effect in the addiction-related animal behavioral models here tested. This is in striking contrast to positive findings in the same animal models shown by another GABAmimetic--gamma-vinyl GABA (see companion piece to present article).

Synthesis of (E)- and (Z)-alpha-alkylidene-gamma-aryl-gamma-butyrolactones via alkenylalumination of oxiranes.

Alkenylalumination of substituted styrene oxides with [alpha-(ethoxycarbonyl)alkenyl]diisobutylaluminum, in the presence of BF(3).Et(2)O, affords the corresponding (Z)-alpha-alkylidene-gamma-aryl-gamma-hydroxy esters in 81-100% Z-selectivity. Chromatographic separation of isomers, followed by lactonization with trifluoroacetic acid, provides isomerically pure (Z)-alpha-alkylidene-gamma-aryl-gamma-butyrolactones in 53-78% overall yield. Isomerization of the (Z)-alkylidene hydroxyl esters using LDA, followed by protonation using a bulky proton source, such as BHT, provides a simple route to the corresponding alpha-(E)-alkylidene-gamma-phenyl-gamma-hydroxy esters in 72-78% yield, which were cyclized to obtain the corresponding (E)-butyrolactones in 78-85% yield.

Acidity-directed synthesis of substituted gamma-butyrolactones from aliphatic aldehydes.

The strength of the Lewis or Brønsted acids controls the formation of either beta,gamma-disubstituted-alpha-methylene-gamma-butyrolactones or gamma-substituted-alpha-alkylidene-gamma-butyrolactones via the lactonization or oxonia cope rearrangement-lactonization, respectively, of the borate intermediates resulting from the crotylboration of aliphatic aldehydes with ester-containing crotylboronates, such as (E)-methyl 2-boramethyl-2-butenoates.

Control of stereochemistry: a general synthesis of cis- or trans-beta,gamma-disubstituted-gamma-butyrolactones following Z-crotylboration.

[reaction; see text] A general and practical procedure for the highly diastereoselective preparation of either the cis- or trans-beta,gamma-disubstituted-gamma-butyrolactones by appropriate choice of Lewis or Bronsted acid catalysts during crotylboration or lactonization is reported. The cis-stereochemistry of the Z-crotylboration product can be inverted with strong acids during lactonization. A carbocation mechanism and catalytic cycle has been proposed.

Studies towards the synthesis of epothilone A via organoboranes.

Studies towards the synthesis of epothilone A via organoboranes have been described. A modified procedure for the large-scale preparation of B-gamma,gamma-dimethylallyldiisopinocampheylborane from prenyl alcohol has been developed. This reagent, upon reaction with various aldehydes, provides the corresponding alpha,alpha-dimethylhomoallylic alcohols in high enantioselectivities. The application of this reagent for the synthesis of the C1-C6 subunit of epothilone has been demonstrated. Alternatively, inter- and intramolecular asymmetric reduction protocols have also been utilized for the synthesis of the C1-C6 subunit of epothilone A. The synthesis of the C7-C21 fragment of epothilone A involving asymmetric alkoxyallyl- and crotylboration using alpha-pinene-derived reagents has also been described.

Synthesis of 4-substituted homoallylic alcohols via a one-pot tandem Lewis-acid catalyzed crotylboration-[3,3]-sigmatropic rearrangement.

Crotylboration of aldehydes with E- or Z-crotylboronates in the presence of catalytic amounts of indium triflate provides the corresponding 4-substituted homoallylic alcohols.

Novel functionalized trisubstituted allylboronates via Hosomi-Miyaura borylation of functionalized allyl acetates.

[reaction: see text] A series of novel functionalized achiral and chiral allylboronates have been synthesized via the nucleophilic addition of boronates on allyl acetates derived via vinylalumination or Baylis-Hillman reaction of aldehydes. These reagents, upon allylboration with aldehydes, furnish beta-substituted-alpha-methylene-gamma-butyrolactones stereoselectively.