Új kéntartalmú szénhidrát-származékok trypanosoma- ill. amöba-ellenes hatásának vizsgálata: szerkezet-hatás összefüggések = Testing the trypanocidal, amoebicidal activity of novel S-containing sugar derivatives: structure – activity investigations.

A Trypanosoma cruzi protozoa, a Chagas betegség kórokozója, Latin Amerikában évente 12-14 millió ember fertőzéséért felelős. A betegség kezelésére jelenleg alkalmazott szerek hatásossága nem kielégítő, különösen a betegség gyakori, krónikus formája ellen. Kísérleteket folytattunk potenciálisan tripanoszóma-ellenes vegyületek szintézisére; ennek során új, multivalens aromás glikozil-diszulfidokat és néhány diglikozil-diszulfidot állítottunk elő T.cruzi elleni tesztelés céljára. Néhány származék meglepően hatásosnak mutatkozott sejttenyészetből származó tripomasztigoták ellen: az eddig talált legaktívabb származék, az 1,4-bis(ß-D-galaktopiranozil-dithiometilén)-benzol gátlási állandója (IC50) 8.7 ±1.21 µM. Több, hasonló kémiai szerkezettel jellemzett származék is hatásosnak mutatkozott ~ 15 µM-os szinten. A tesztvegyületek citotoxikus hatását konfluens HeLa sejttenyészeteken tanulmányoztuk 18 órás inkubálást követően. Az aktív vegyületek mindegyike csekély citotoxicitást mutatott. További, igen jelentős megfigyelés, hogy az aktív származékok képesek a parazita intracelluláris proliferációját (amasztigota forma) is gátolni, amint ez a T.cruzi-val fertőzött Vero-sejtekből kibocsátott paraziták számlálása bizonyította. Véleményünk szerint ez igen figyelemre méltó eredmény, u.i. a jelenleg alkalmazott T.cruzi elleni szerek (beznidazol, nifurtimox) nem képesek a parazita intracelluláris fázisát gátolni. | Trypanosoma cruzi is the etiologic agent of Chagas disease, an endemic parasitosis in Latin America with 12–14 million people infected. Currently available drugs for treatment of this disease are unsatisfactory, particularly in its prevalent chronic form. In a search for novel chemical structures with potential trypanocidal activity we have synthesized novel aromatic multivalent glycosyl disulfides and some diglycosyl disulfides for testing against T.cruzi. Some of the tested compounds were found remarkably efficient to kill cell culture-derived trypomastigotes; 1,4-bis(ß-D-galactopyranosyl-dithiomethylene) benzene proved to be the most active derivative with an IC50 of 8.7 ±1.21 µM. Other derivatives, of similar chemical structures, were further promising candidates with low IC50 values of ~ 15 µM. The cytotoxic effects of the active and inactive compounds were evaluated after 18 h incubation with confluent HeLa cell cultures. Pleasingly, all of the active trypanocidal agents showed low cytotoxicites. Further, we observed that several of the compounds tested were also able to inhibit intracellular proliferation or parasite differentiation (amastigote form) when the number of parasites released by T.cruzi infected Vero cells was evaluated. These observations, of outstanding interest, show a remarkable effect of these compounds against the intracellular stage of T.cruzi, an activity that is missing in the currently available drugs like beznidazol and nifurtimox

Új, potenciálisan bioaktív szénhidrátok: szintetikus és NMR kutatások = Novel carbohydrates with potential biological activity: synthetic and NMR investigations

Új típusú, 3-kötéses glikozidos hidat (3KGH: -S-S-, -S-C- ) tartalmazó glikomimetikumokat szintetizáltunk. Monoszacharidokat egy központi aromás gyűrűre SS-kötéssel kapcsolva szimmetrikus oligovalens struktúrákat nyertünk. Tioéter (4->6', -S-C-) interglikozidos kiépítésével új típusú pszeudo-diszacharidokat konstruáltunk. A nem hidrolizálható kötés miatt utóbbiak a glikozilhidroláz enzimek potenciális inhibitorai. Új NMR módszereket fejlesztettünk ki a kémiai szerkezet és dinamika hatékonyabb vizsgálatára. Izotópszelektív (15N) telítés alkalmazása az STD NMR kísérletben (15N GS-STD) teljes jelátfedés esetén is lehetővé teszik fehérje-ligandum kölcsönhatások vizsgálatát. Többkötéses heteronukleáris (H,X)-, valamint 13C-13C skaláris csatolási állandók meghatározására új, hatékony 2D NMR módszereket, ill. érzékeny, 1H-detektáláson alapuló kísérleteket (CPMG-HSQMBC, ill. IPAP-DEPT-INADEQUATE és RINEPT-INADEQUATE) javasoltunk. A mért csatolási állandók és egyéb NMR adatok (NOESY/ROESY) felhasználásával elméleti molekulamodellezési módszerekkel határoztuk meg diglikozil-diszulfidok és egyéb szénhidrát-származékok konformációs preferenciáit oldatban. Szilárd fázisban röntgendiffrakciós és kiroptikai (CD) módszereket alkalmaztunk. Az aromás, oligovalens mannozil-diszulfid származékok specifikusan kötődnek a Concanavalin-A lektin-fehérjéhez. A fehérje-ligandum komplexek termodinamikai paramétereit mikrokalorimetriai (ITC), szerkezetüket STD-NMR mérésekkel határoztuk meg. | Novel glycomimetics were synthesized containing three-bond interglycosidic linkages (3BIGL, -S-S-, -S-C-). Glycopyranosyl units were attached to an aromatic scaffold through SS-linkages to obtain symmetric oligovalent structures. Non-glycosidic, 4,6'-thioether (-S-C-) -linked pseudodisaccharides were constructed via highly diastereoselective synthesis; these are potential inhibitors of glycosylhydrolases. Efficient NMR techniques were developed for in-depth studies of molecular structures and dynamics. Extension of the STD NMR experiment with an isotope selective (15N) saturation sequence allows exploration of protein-ligand interactions even in case of complete signal overlap. Novel 2D NMR methods were devised for more accurate measurement of long-range heteronuclear couplings (CPMG-HSQMBC), including sensitive 1H-detection schemes for 13C-13C scalar couplings at natural abundance (DEPT-INADEQUATE, RINEPT-INADEQUATE). Complete sets of experimental coupling constants and NOESY/ROESY data supplemented with molecular modelling allowed determination of conformational preferences of diglycosyl disulfides and other carbohydrate derivatives in solution. X-ray and chiroptical (CD) methods were applied for solid state studies. Aromatic, oligovalent mannosyl disulfides were shown to bind specifically to Concanavalin A, a plant lectin. The thermodynamic parameters and structures of the protein-ligand complexes were determined by microcalorimetry (ITC) and STD-NMR, respectively

Tri- and tetravalent mannoclusters cross-link and aggregate BC2L-A lectin from Burkholderia cenocepacia

K

Heck-oxyarylation of 2-phenyl-2H-chromenes and 1,2-dihydronaphthalenes

The Heck-oxyarylation of racemic 2-phenyl-2H-chromene [(±)-4b] and 1,2-dihydronaphthalenes (14a,b) has been studied with 2-chloromercuriphenols (5a–d) in the presence of Li2[PdCl4] catalyst. The reactions resulted in the diastereoselective formation of racemic 6- phenylpterocarpans of (6R, 6aR,11aR) relative configuration [(±)-8a–d] and their dibenzo[1,3]dioxocine analogues [(±)-12a–d] as main products, respectively. The ratio of products and the lack of regioisomeric products (13a–d) corroborated the cationic mechanism of the oxyarylation of 2H-chromenes, which has been also supported by the transformation of 14a,b under similar conditions.(doi: 10.5562/cca2103

Exploring the Syntheses of Novel Glycomimetics: Carbohydrate Derivatives with Se-S- or Se-Se- Glycosidic Linkages

K

Preparation of neuroprotective condensed 1,4-benzoxazepines by regio- and diastereoselective domino Knoevenagel–[1,5]-hydride shift cyclization reaction

Condensed O,N-heterocycles containing tetrahydro-1,4-benzoxazepine and tetrahydroquinoline moieties were prepared by a regio- and diastereoselective domino Knoevenagel–[1,5]-hydride shift cyclization reaction of a 4-aryl-2-phenyl-1,4-benzoxazepine derivative obtained from flavanone. The relative configuration of products were determined by the correlation of 3JH,H coupling data with the geometry of major conformers accessed by DFT conformational analysis. Separated enantiomers of the products were characterized by HPLC-ECD data, which allowed their configurational assignment on the basis of TDDFT-ECD calculation of the solution conformers. Two compounds showed neuroprotective activities against hydrogen peroxide (H2O2) or β-amyloid25–35 (Aβ25–35)-induced cellular injuries in human neuroblastoma SH-SY5Y cells in the range of those of positive controls

Probing pattern and dynamics of disulfide bridges using synthesis and NMR of an ion channel blocker peptide toxin with multiple diselenide bonds

Anuroctoxin (AnTx), a 35-amino-acid scorpion toxin contg. four disulfide bridges, is a high affinity blocker of the voltage-gated potassium channel Kv1.3, but also blocks Kv1.2. To improve the potential therapeutic use of the toxin, we have designed a double substituted analog, [N17A/F32T]-AnTx, which showed comparable Kv1.3 affinity to the wild-type peptide, but 2500-fold increase in the selectivity for Kv1.3 over Kv1.2. In the present study we have achieved the chem. synthesis of a Sec-analog in which all cysteine (Cys) residues have been replaced by selenocysteine (Sec) forming four diselenide bonds. To the best of our knowledge this is the first time to replace, by chem. synthesis, all disulfide bonds with isosteric diselenides in a peptide/protein. Gratifyingly, the key pharmacol. properties of the Sec-[N17A/F32T]-AnTx are retained since the peptide is functionally active. We also propose here a combined exptl. and theor. approach including NOE- and 77Se-based NMR supplemented by MD simulations for conformational and dynamic characterization of the Sec-[N17A/F32T]-AnTx. The use of such combined approach allowed us to attain unequivocal assignment of all four diselenide bonds and supplemental MD simulations allowed to characterize the conformational dynamics around each disulfide/diselenide bridge. [on SciFinder(R)

Cryptocapsinepoxide-type Carotenoids from Red Mamey, Pouteria sapota

Three new carotenoids, cryptocapsin-5,6-epoxide, 3ʹ-deoxycapsanthin-5,6-epoxide, and cryptocapsin-5,8-epoxides, have been isolated from the ripe fruits of red mamey (Pouteria sapota). Cryptocapsin-5,6-epoxide was prepared by partial synthesis via epoxidation of cryptocapsin and the (5R,6S)- and (5S,6R)-stereoisomers were identified by HPLC-ECD analysis. Spectroscopic data of the natural (anti) and semisynthetic (syn) derivatives obtained by acid-catalyzed rearrangement of cryptocapsin-5,8-epoxide stereoisomers were compared for structural elucidation. Chiral HPLC separation of natural and semisynthetic samples of cryptocapsin-5,8-epoxides was performed and HPLC-ECD analysis allowed configurational assignment of the separated stereoisomers