1,4-Dihydropyridine Derivatives: Dihydronicotinamide Analogues—Model Compounds Targeting Oxidative Stress

Many 1,4-dihydropyridines (DHPs) possess redox properties. In this review DHPs are surveyed as protectors against oxidative stress (OS) and related disorders, considering the DHPs as specific group of potential antioxidants with bioprotective capacities. They have several peculiarities related to antioxidant activity (AOA). Several commercially available calcium antagonist, 1,4-DHP drugs, their metabolites, and calcium agonists were shown to express AOA. Synthesis, hydrogen donor properties, AOA, and methods and approaches used to reveal biological activities of various groups of 1,4-DHPs are presented. Examples of DHPs antioxidant activities and protective effects of DHPs against OS induced damage in low density lipoproteins (LDL), mitochondria, microsomes, isolated cells, and cell cultures are highlighted. Comparison of the AOA of different DHPs and other antioxidants is also given. According to the data presented, the DHPs might be considered as bellwether among synthetic compounds targeting OS and potential pharmacological model compounds targeting oxidative stress important for medicinal chemistry

1,4 dihidropiridinski derivati povećavaju ekspresiju gena Psma3, Psmb5 i Psmc6 u glasničkoj RNA štakora

The ubiquitin-proteasome system modifies different cellular and protein functions. Its dysregulation may lead to disrupted proteostasis associated with multiple pathologies and aging. Pharmacological regulation of proteasome functions is already an important part of the treatment of several diseases. 1,4-dihydropyridine (1,4-DHP) derivatives possess different pharmacological activities, including antiaging and neuroprotective. The aim of this study was to investigate the effects of several 1,4-DHP derivatives on mRNA expression levels of proteasomal genes Psma3, Psmb5, and Psmc6 in several organs of rats. Rats were treated with metcarbatone, etcarbatone, glutapyrone, styrylcarbatone, AV-153-Na, or AV-153-Ca per os for three days. mRNA expression levels were determined with real-time polymerase chain reaction (PCR). For AV-153-Na and AV-153-Ca, we also determined the expression of the Psma6 gene. In the kidney, metcarbatone, etcarbatone, styrylcarbatone, and AV-153-Na increased the expression of all analysed genes. Glutapyrone increased the expression of Psmb5 and Psmc6 but did not affect the expression of Psma3. In the blood, glutapyrone increased Psmb5 expression. In the liver, AV-153-Na increased the expression of Psma6 and Psmc6 but lowered the expression of Psmb5, while AV-153-Ca only increased Psma6 expression. The ability of 1,4-DHP derivatives to increase the expression of proteasome subunit genes might hold a therapeutic potential in conditions associated with impaired proteasomal functions, but further research is needed.Ubikvitin-proteasomski sustav utječe na različite funkcije bjelančevina i stanica. Poremećaji u njegovoj regulaciji mogu dovesti do poremećaja u proteostazi koji su povezani s nastankom različitih bolesti i sa starenjem, ali se sustav može regulirati u liječenju pojedinih bolesti lijekovima poput 1,4 dihidropiridinskih (1,4 DHP) derivata, koji štite živčani sustav i usporavaju starenje. Cilj ovoga istraživanja bio je utvrditi djelovanje nekoliko 1,4 DHP derivata na ekspresiju glasničke RNA (mRNA) u proteasomskim genima Psma3, Psmb5 i Psmc6 u više štakorskih organa. Štakori su tri dana dobivali oralne doze metkarbatona, etkarbatona, glutapirona, stirilkarbatona, AV-153-Na ili AV-153-Ca, a genska se ekspresija u mRNA utvrdila polimeraznom lančanom reakcijom u stvarnom vremenu (engl. real-time polymerase chain reaction, krat. PCR). Osim toga, utvrdili smo djelovanje derivata AV‑153‑Na i AV‑153‑Ca na ekspresiju gena Psma6. U bubrezima su metkarbaton, etkarbaton, stirilkarbatoni AV‑153‑Na povećali ekspresiju svih analiziranih gena. Glutapiron je povećao ekspresiju Psmb5 i Psmc6, ali ne i Psma3. U krvi je glutapiron povećao gensku ekspresiju Psmb5. U jetrima je AV‑153‑Na povećao ekspresiju Psma6 i Psmc6, istodobno smanjivši ekspresiju Psmb5. AV‑153‑Ca utjecao je samo na Psma6, povećavši mu ekspresiju. Sposobnost 1,4-DHP derivata da povećaju gensku ekspresiju proteasomskih podjediničnih proteina obećava u smislu mogućnosti liječenja bolesti povezanih s poremećenom proteasomskom funkcijom, no nužna su daljnja istraživanja

Kratki pregled genotoksičnoga i genoprotektivnoga djelovanja derivata 1,4-dihidropiridina

This review summarises current knowledge about the genotoxic and genoprotective effects of 1,4-dihydropyridines (DHP) with the main focus on the water-soluble 1,4-DHPs. Most of these water-soluble compounds manifest very low calcium channel blocking activity, which is considered “unusual” for 1,4-DHPs. Glutapyrone, diludine, and AV-153 decrease spontaneous mutagenesis and frequency of mutations induced by chemical mutagens. AV-153, glutapyrone, and carbatones protect DNA against the damage produced by hydrogen peroxide, radiation, and peroxynitrite. The ability of these molecules to bind to the DNA may not be the only mechanism of DNA protection, as other mechanisms such as radical scavenging or binding to other genotoxic compounds may take place and enhance DNA repair. These uncertainties and reports of high 1,4-DHP concentrations damaging the DNA call for further in vitro and in vivo preclinical research, pharmacokinetic in particular, as it can help pinpoint the exact mechanism(s) of the genotoxic and/or genoprotective action of 1,4-DHPs.Ovaj pregledni rad donosi sažetak onoga što smo dosad naučili o genotoksičnom i genoprotektivnom djelovanju 1,4-dihidropiridina (DHP), s posebnom pažnjom na 1,4-DHP-ove topljive u vodi. Većina tih u vodi topljivih spojeva slabo aktivira blokiranje kalcijevih kanala, što se smatra “neuobičajenim” za 1,4-DHP-ove. Glutapiron, diludin i AV-153 ublažavaju spontanu mutagenezu i učestalost mutacija prouzročene kemijskim mutagenima. AV-153, glutapiron i karbatoni štite DNA od oštećenja prouzročenih vodikovim peroksidom, zračenjem i peroksinitritom. Sposobnost tih molekula da se vežu za DNA vjerojatno nije jedini mehanizam njegove zaštite, budući da su mogući i drugi mehanizmi, poput uklanjanja radikala ili vezanja za druge genotoksične spojeve koji pospješuju popravak DNA. Zbog tih nepoznanica i izvještaja da visoke koncentracije 1,4-DHP-ova oštećuju DNA, potrebno je napraviti daljnja neklinička istraživanja in vitro i in vivo, napose ona farmakokinetička, budući da mogu pomoći razaznati točne mehanizme genotoksičnoga i/ili genoprotektivnoga djelovanja derivata 1,4-dihidropiridina

Growth Modulation of Human Cells in Vitro by Mild Oxidative Stress and 1,4-Dihydropyridine Derivative Antioxidants

Reactive oxygen species and lipid peroxidation products are not only cytotoxic but may also modulate signal transduction in cells. Accordingly, antioxidants may be considered as modifiers of cellular redox signaling. Therefore, the effects of two novel synthetic antioxidants, analogues of 1,4-dihydropyridine derivatives, cerebrocrast and Z41-74 were analysed in vitro on human osteosarcoma cell line HOS, the growth of which can be modulated by lipid peroxidation. The cells were pretreated with either cerebrocrast or Z41-74 and afterwards exposed to mild, copper induced lipid peroxidation or to 4-hydroxynonenal (HNE), the end product of lipid peroxidation. The results obtained have shown that both antioxidants exert growth modulating effects interfering with the lipid peroxidation. Namely, cells treated with antioxidants showed increased metabolic rate and cell growth, thereby attenuating the effects of lipid peroxidation. Such biomodulating effects of cerebrocrast and Z41-74 resembled growth modulating effects of HNE, suggesting that the antioxidants could eventually promote cellular adaptation to oxidative stress interacting with redox signaling and hydroxynonenal HNE-signal transduction pathways. This may be of particular relevance for better understanding the beneficial role of hydroxynonenal HNE in cell growth control. Therefore, cerebrocrast and Z41-74 could be convenient to study further oxidative homeostasis involving lipid peroxidation

Growth Modulation of Human Cells in Vitro by Mild Oxidative Stress and 1,4-Dihydropyridine Derivative Antioxidants

Reactive oxygen species and lipid peroxidation products are not only cytotoxic but may also modulate signal transduction in cells. Accordingly, antioxidants may be considered as modifiers of cellular redox signaling. Therefore, the effects of two novel synthetic antioxidants, analogues of 1,4-dihydropyridine derivatives, cerebrocrast and Z41-74 were analysed in vitro on human osteosarcoma cell line HOS, the growth of which can be modulated by lipid peroxidation. The cells were pretreated with either cerebrocrast or Z41-74 and afterwards exposed to mild, copper induced lipid peroxidation or to 4-hydroxynonenal (HNE), the end product of lipid peroxidation. The results obtained have shown that both antioxidants exert growth modulating effects interfering with the lipid peroxidation. Namely, cells treated with antioxidants showed increased metabolic rate and cell growth, thereby attenuating the effects of lipid peroxidation. Such biomodulating effects of cerebrocrast and Z41-74 resembled growth modulating effects of HNE, suggesting that the antioxidants could eventually promote cellular adaptation to oxidative stress interacting with redox signaling and hydroxynonenal HNE-signal transduction pathways. This may be of particular relevance for better understanding the beneficial role of hydroxynonenal HNE in cell growth control. Therefore, cerebrocrast and Z41-74 could be convenient to study further oxidative homeostasis involving lipid peroxidation

Modifikacije ekspresije gena i proteina koji su uključeni u popravak DNA i metabolizam dušikova oksida karbatonidima [derivati dinatrij-2,6-dimetil-1,4-dihidropiridin-3,5-bis(karboniloksiacetata)] u kontrolnih i dijabetičkih štakora

Studies on the pathogenesis of diabetes mellitus complications indicate that the compounds reducing free radicals and enhancing DNA repair could be prospective as possible remedies. Carbatonides, the disodium-2,6-dimethyl-1,4-dihydropyridine-3,5-bis(carbonyloxyacetate) derivatives, were tested for these properties. EPR spectroscopy showed that metcarbatone was an effective scavenger of hydroxyl radicals produced in the Fenton reaction, etcarbatone, and propcarbatone were less effective, styrylcarbatone was ineffective. UV/VIS spectroscopy revealed that styrylcarbatone manifested a hyperchromic effect when interacting with DNA, while all other carbatonides showeda hypochromic effect. Rats with streptozotocin induced type 1 DM were treated with metcarbatone, etcarbatone or styrylcarbatone (all compounds at doses 0.05 mg kg-1 or 0.5 mg kg-1) nine days after the DM approval. Gene expression levels in kidneys and blood were evaluated by quantitative RT-PCR; protein expression - immunohistochemically in kidneys, heart, sciatic nerve, and eyes; DNA breakage - by comet assay in nucleated blood cells. Induction of DM induced DNA breaks; metcarbatone and styrylcarbatone (low dose) alleviated this effect. Metcarbatone and etcarbatone up-regulated mRNA and protein of eNOS in kidneys of diabetic animals; etcarbatone also in myocardium. Etcarbatone reduced the expression of increased iNOS protein in myocardium, nerve, and kidneys. iNos gene expression was up-regulated in kidneys by etcarbatone and metcarbatone in diabetic animals. In blood, development of DM increased iNos gene expression; etcarbatone and metcarbatone normalised it. Etcarbatone up-regulated the expression of H2AX in kidneys of diabetic animals but decreased the production of c-PARP1. Taken together, our data indicate that carbatonides might have a potential as drugs intended to treat DM complications.Rezultati ispitivanja patogeneze komplikacija šećerne bolesti (diabetes mellitus – DM) upozoravaju na to da bi tvari koje smanjuju nastanak slobodnih radikala a pospješuju popravak DNA mogle biti obećavajuće u budućem liječenju te bolesti. U ovome su istraživanju ispitana navedena svojstva karbatonida, derivata dinatrij-2,6-dimetil-1,4-dihidropiridin-3,5-bis(karboniloksiacetata). EPR spektroskopija je pokazala da je metkarbaton učinkovit sakupljač hidroksilnih radikala koji nastaju Fentonovom reakcijom. Etkarbaton i propkarbaton su bili malo manje učinkoviti, a stirilkarbaton nije bio učinkovit. UV/VIS spektroskopija pokazala je hiperkromni učinak stirilkarbatona u interakciji s DNA, a svi ostali karbatonidi pokazali su hipokromni učinak. Štakori kod kojih je DM tipa 1 induciran streptozotocinom tretirani su metkarbatonom, etkarbatonom ili stirilkarbatonom (sve tvari su davane u dozi 0,05 mg kg-1 ili 0,5 mg kg-1) tijekom devet dana nakon što je potvrđeno da je izazvan DM. Razine ekspresije gena u bubrezima procijenjene su kvantitativnim RT-PCR-om, ekspresija proteina – imunohistokemijski u bubrezima, srcu, ishijadičnom živcu i očima, a lom DNA – komet-testom bijelim krvnim stanicama. Indukcija DM-a uzrokovala je lomove u DNA; metkarbaton i stirilkarbaton (niska doza) ublažili su ovaj učinak. Metkarbaton i etkarbaton pojačali su mRNA i ekspresiju proteina eNOS-a u bubrezima dijabetičkih životinja; etkarbaton je takav učinak pokazao i u miokardu. Etkarbaton je smanjio ekspresiju iNOS proteina u miokardu, živcu i bubrezima. Ekspresija iNOS-a bila je pojačana u bubrezima primjenom etkarbatona i metkarbatona u dijabetičkih životinja. Razvoj DM povećao je ekspresiju iNOS-a u krvi, a etkarbaton i metkarbaton vratili su ju na normalne vrijednosti. Etkarbaton je pojačao ekspresiju H2AX u bubrezima dijabetičkih životinja, ali je smanjio proizvodnju c-PARP1. Naši podaci upućuju na potencijal karbatonida kao lijekova za liječenje komplikacija uzrokovanih dijabetesom

Dihydropyridine Derivatives as Cell Growth Modulators In Vitro

The effects of eleven 1,4-dihydropyridine derivatives (DHPs) used alone or together with prooxidant anticancer drug doxorubicin were examined on two cancer (HOS, HeLa) and two nonmalignant cell lines (HMEC, L929). Their effects on the cell growth (3H-thymidine incorporation) were compared with their antiradical activities (DPPH assay), using well-known DHP antioxidant diludine as a reference. Thus, tested DHPs belong to three groups: (1) antioxidant diludine; (2) derivatives with pyridinium moieties at position 4 of the 1,4-DHP ring; (3) DHPs containing cationic methylene onium (pyridinium, trialkylammonium) moieties at positions 2 and 6 of the 1,4-DHP ring. Diludine and DHPs of group 3 exerted antiradical activities, unlike compounds of group 2. However, novel DHPs had cell type and concentration dependent effects on 3H-thymidine incorporation, while diludine did not. Hence, IB-32 (group 2) suppressed the growth of HOS and HeLa, enhancing growth of L929 cells, while K-2-11 (group 3) enhanced growth of every cell line tested, even in the presence of doxorubicin. Therefore, growth regulating and antiradical activity principles of novel DHPs should be further studied to find if DHPs of group 2 could selectively suppress cancer growth and if those of group 3 promote wound healing

Crystal structure and metabolic activity of 4-(thien-2-yl)-2-methyl-5-oxo-1,4,5,6,7,8-hexahydroquinoline-3-carboxylic acid ethoxycarbonylphenylmethylester

In the title compound, C25H25NO5S, which exhibits metabolism-regulating activity, the 1,4-dihydropyridine ring adopts a flattened boat conformation while the cyclohexenone ring is in an envelope conformation. Molecules in the crystal are assembled into C(6) chains along the a-axis direction via N—H...O hydrogen bonds. The thienyl fragment is disordered over two sets of sites in a 0.7220 (19):0.2780 (19) ratio