Supplementary data for the article: Jevtić, I. I.; Savić Vujović, K.; Srebro, D.; Vučković, S.; Ivanović, M. D.; Kostić-Rajačić, S. V. Synthesis and Pharmacological Evaluation of Novel Cis and Trans 3-Substituted Anilidopiperidines. Pharmacol. Rep 2020, 72 (4), 1069–1075. https://doi.org/10.1007/s43440-020-00121-2

Supplementary material for: [ https://doi.org/10.1007/s43440-020-00121-2]Related to published version: [https://cherry.chem.bg.ac.rs/handle/123456789/4211

μ-opioid/D2 dopamine receptor pharmacophore containing ligands: Synthesis and pharmacological evaluation

Herein, the synthesis and pharmacological evaluation of 13 novel compounds, designed as potential heterobivalent ligands for μ-opioid receptor (MOR) and dopamine D2 receptors (D2DAR), are reported. The compounds consisted of anilido piperidine and N-aryl piperazine moieties, joined by a variable-length methylene linker. The two moieties represent MOR and D2DAR pharmacophores, respectively. The synthesis encompassed four steps, securing the final products in 28–42 % overall yields. The approach has a considerable synthetic potential, providing access to various related structures. Pharmacological tests involved in vitro competitive assay for D2DAR using [3H] spiperon, as a standard radioligand, and in vivo antinociceptive tests for MOR. The measured dopamine affinities were modest to low, while antinociceptive activity was completely absent. Therefore, the compounds of the general structure prepared in this research are unlikely to be useful as opioid–dopamine receptor heterobivalent ligands

Modified Sabouraud dextrose agar for isolation and identification of dermatophytes

The most common causative agents of dermatomycoses are fungi belonging to genders Trichophyton, Microsporum and Epidermophyton. Media mainly used for isolation of dermatophytes are mycobiotic agar, dermatophyte test medium Sabouraud agar (original formula or modification by Emmons) with or without antibiotics and cycloheximide. Peptones are the most important components of the media, which enable adequate reproductivity in identification of dermatophytes. Standard medium for isolation of dermatophytes is not produced in our country. The aim of the study was to create an optimal easily accessible and economic medium which enables isolation and identification of dermatophytes according to criteria for morphological diagnosis provided by identification guides. We examined 57 strains of Trichophyton, 24 of Microsporum and 5 of Epidermophyton floccosum (E. floccosum). Each strain was seeded on Sabouraud dextrose agar (Torlak Serbia and Montenegro), Sabouraud maltose agar (Torlak), two experimental modified Sabouraud dextrose agar media marked as SA-2 and SA-3 (Torlak) Sabouraud-Chloramp- henicole agar (Biomerieux, France) Sabouraud-Chloramphenicole agar (Himedia, India), Glucose-peptone agar (Himedia, India) and Sabouraud Emmons dextrose Agar with Chloramphenicole and Cycloheximide (Biolife, Italy). Colony morphology of Trichophyton mentagrophytes (T. mentagrophytes) was uni- form on all the media, while morphology of Trichophyton rubrum (T. rubrum) and Microsporum canis (M. canis) depended more on the media type. Colonies of E. floccosum were typical and uniform on all the media, as were the control species of Trichophyton schoenleinii (T. schoenleinii) and Trichophyton soudanense (T. soudanense). Experimental modified Sabouraud dextrose agar (Torlak) marked as SA-3 demonstrated the best results in identification of dermatophytes in this study

Modifikovani Saburo dekstrozni agar za izolaciju i identifikaciju dermatofita

The most common causative agents of dermatomycoses are fungi belonging to genders Trichophyton, Microsporum and Epidermophyton. Media mainly used for isolation of dermatophytes are mycobiotic agar, dermatophyte test medium Sabouraud agar (original formula or modification by Emmons) with or without antibiotics and cycloheximide. Peptones are the most important components of the media, which enable adequate reproductivity in identification of dermatophytes. Standard medium for isolation of dermatophytes is not produced in our country. The aim of the study was to create an optimal easily accessible and economic medium which enables isolation and identification of dermatophytes according to criteria for morphological diagnosis provided by identification guides. We examined 57 strains of Trichophyton, 24 of Microsporum and 5 of Epidermophyton floccosum (E. floccosum). Each strain was seeded on Sabouraud dextrose agar (Torlak Serbia and Montenegro), Sabouraud maltose agar (Torlak), two experimental modified Sabouraud dextrose agar media marked as SA-2 and SA-3 (Torlak) Sabouraud-Chloramp- henicole agar (Biomerieux, France) Sabouraud-Chloramphenicole agar (Himedia, India), Glucose-peptone agar (Himedia, India) and Sabouraud Emmons dextrose Agar with Chloramphenicole and Cycloheximide (Biolife, Italy). Colony morphology of Trichophyton mentagrophytes (T. mentagrophytes) was uniform on all the media, while morphology of Trichophyton rubrum (T. rubrum) and Microsporum canis (M. canis) depended more on the media type. Colonies of E. floccosum were typical and uniform on all the media, as were the control species of Trichophyton schoenleinii (T. schoenleinii) and Trichophyton soudanense (T. soudanense). Experimental modified Sabouraud dextrose agar (Torlak) marked as SA-3 demonstrated the best results in identification of dermatophytes in this study.Dermatomikoze su široko rasprostranjena oboljenja svugde u svetu. Najčešći uzročnici dermatomikoza su gljive iz rodova Trichophyton, Microsporum i Epidermophyton. Za izolaciju dermatofita uglavnom se koriste mikobiotski agar, dermatofit-test agar, Saburo agar sa originalnom formulom ili Emonsova modifikacija Saburo agara sa i bez antibiotika i cikloheksimida. Peptoni su najznačajnija komponenta podloga koja omogućava adekvatnu reproduktivnost u identifikaciji dermatofita. U našoj zemlji se ne proizvodi standardna podloga za izolaciju dermatofita. Cilj ovog rada je bio da se kreira optimalna, lako dostupna i ekonomična podloga, koja omogućava izolaciju i identifikaciju dermatofita prema kriterijumima za morfološku dijagnostiku predviđenim vodičima za identifikaciju. Pregledano je 57 sojeva roda Trichophyton, 24 sojeva roda Microsporum i 5 sojeva Epidermophyton floccosum (E. floccosum). Svaki soj je zasejan na Saburo dekstrozni agar (Torlak), Saburo maltozni agar (Torlak), dve eksperimentalne podloge nastale modifikovanjem sastava standardnog Saburo dekstroznog agara pod radnim nazivom SA-2 i SA-3 (Torlak) Saburo-hloramfenikol agar (Biomerieux, Francuska), Saburo-hloramfenikol agar (Himedia, Indija), glukozo peptonski agar (sa peptonom Himedia Indija) i Saburo Emons dekstrozni agar sa hloramfenikolom i cikloheksimidom (Biolife, Italija). Kriterijumi za ocenu ispitivanih podloga ustanovljeni su na osnovu poznate morfologije koju sojevi dermatofita pokazuju na mikobiotskom agaru (Mycosel, Oxoid, Velika Britanija) i na osnovu ključa za identifikaciju gljiva. Morfologija kolonija Trichophyton mentagrophytes (T. mentagrophytes) bila je uniformna na svim podlogama, dok je morfologija Trichophyton rubrum (T. rubrum) i Microsporum canis (M. canis) više zavisila od vrste podloge nego od soja. Kolonije E. floccosum su bile tipične i uniformne na svim ispitivanim podlogama, kao i kontrolni sojevi Trichophyton schoenleinii (T. schoenleinii) i Trichophyton soudanense (T. soudanense). Eksperimentalna podloga SA-3 pokazala je najbolje rezultate u identifikaciji dermatofita u ovoj studiji. Makroskopska i mikroskopska morfologija dermatofita na ovoj podlozi najviše odgovara morfologiji ovih gljiva na standardnim podlogama u kojima se kao glavna komponenta nalazi mikološki pepton. Saburo dekstrozni agar (Torlak), koji se dosada u našoj zemlji koristio za primarnu izolaciju dermatofita, može se koristiti u identifikaciji M. canis jer potpomaže produkciju makrokonidija već u prvoj nedelji inkubacije. Na svim podlogama T. mentagrophytes se može dijagnostikovati nakon 7 dana, dok je za identifikaciju ostalih vrsta dermatofita potrebno više od 14 dana

Synthesis and pharmacological evaluation of novel cis and trans 3-substituted anilidopiperidines

Background: 4-Anilidopiperidine class of synthetic opioid analgesics, with it’s representative fentanyl, are by far the most potent and clinically significant for the treatment of the severe chronic and surgical pain. However, side effects of μ-opioids are often quite serious. In order to improve the pharmacological profile of this class of opioid analgesics, a novel fentanyl analogs were designed, synthesized and evaluated in vivo for their antinociceptive activity. Methods: The title compounds were prepared using known synthetic transformations, including N-bromoacetamide mediated Hofmann rearrangement, highly selective carbamate cleavage with trimethylsilyl iodide and dehydration of carboxamide group to nitrile in the presence of SOCl2. The antinociceptive activity of the synthesized compounds was determined by tail-immersion and formalin test. Results: The scalable synthetic route towards novel fentanyl analogs bearing nitrogen groups in position C3 of piperidine ring is designed. In addition, Hofmann rearrangement was substantially improved for the more efficient synthesis of previously published 3-substituted fentanyl analogs. The series of ten fentanyl analogs was tested in vivo for their antinociceptive activity. The most potent compound of the series was found to be cis-4, based on the determined ED50 values in tail-immersion test. Conclusion: Of ten compounds tested for their antinociceptive activity, compound cis-4 is characterized by high potency, rapid beginning and short duration of action and due to this might be incorporated in different pharmaceutical forms.Supplementary material: [https://cherry.chem.bg.ac.rs/handle/123456789/4212

3-alkil analozi fentanila - studija odnosa strukture i aktivnost

Introduction. Fentanyl belongs to 4-anilidopiperidine class of synthetic opioid analgesics. It is characterized by high potency, rapid onset and short duration of action. A large number of fentanyl analogues have been synthesized so far, both to establish the structure-activity-relationship (SAR) and to find novel, clinically useful analgesic drugs. Objective. In this study, newly synthesized 3-alkyl fentanyl analogues were examined for analgesic activity and compared with fentanyl. Methods. Analgesic activity was assessed by tail-immersion test in rats. Results. The relative potency was: (±)cis-3-methyl fentanyl (8) gt (±)trans-3-methyl fentanyl (2) $(±)cis-3-ethyl fentanyl (1.5) gt fentanyl (1)$ (±)trans-3-ethyl fentanyl (0.9) gt (±)cis-3-butyl fentanyl (0.064) $(±)trans-3-butyl fentanyl (0.035) gt (±)cis-3-benzyl fentanyl (0.008)$ (±)trans-3-benzyl fentanyl (0.0055). (±)Cis-3-iso-propyl fentanyl, and (±)cis-3-phenethyl fentanyl are inactive in doses up to 5 mg/kg. The duration of action (ED99) was: (±)cis-3-methyl fentanyl (90 min) gt (±)trans-3-methyl fentanyl (40 min) # (±)cis-3-ethyl fentanyl (60 min) (±)trans-3-ethyl fentanyl (40 min) # fentanyl (50 min) = (±)cis-3-butyl fentanyl (50 min) = (±)trans-3-butyl fentanyl (50 min) = (±)cis-3-benzyl fentanyl (50 min) = (±)trans-3-benzyl fentanyl (50 min). Symbols gt and lt denotes p lt 0.05. Conclusion. It is concluded that the analgesic potency of 3-alkyl fentanyl analogues is influenced by the steric factor (voluminosity of the group at the position 3 of the piperidine ring and the cis/trans isomerism). Otherwise, with the exception of 3-methyl fentanyl, the duration of action of 3-alkyl fentanyl analogues is not significantly affected by the stereochemistry.Uvod. Fentanil pripada grupi sintetskih opioidnih analgetika, 4-anilidopiperidina. Karakteriše ga visoka potentnost, brz početak i kratko trajanje dejstva. Do sada je sintetisan veliki broj analoga fentanila, kako u cilju određivanja odnosa strukture i farmakološke aktivnosti, tako i u cilju pronalaženja novog klinički korisnog analgetika. Cilj rada. U studiji su ispitivana analgetska dejstva novosintetisanih 3-alkil analoga fentanila i poređena sa fentanilom. Metod rada. Merenje analgetskog dejstva ispitivanih jedinjenja vršeno je uz pomoć testa potapanja repa pacova u toplu vodu. Rezultati. Relativna jačina jedinjenja iznosila je: (±)cis-3-metil fentanil (8) gt (±)trans-3-metil fentanil (2) (±)cis-3-etil fentanil (1,5) gt fentanil (1) $(±)trans-3-etil fentanil (0,9) gt (±)cis-3-butil fentanil (0,064)$ (±)trans-3-butil fentanil (0,035) gt (±)cis-3-benzil fentanil (0,008) (±)trans-3-benzil fentanil (0,0055). (±)Cis-3-izopropil fentanil i (±)cis-3-fenetil fentanil nisu ispoljili dejstvo u dozama do 5 mg/kg. Dužina dejstva (ED99) iznosila je: (±)cis-3-metil fentanil (90 min) gt (±)trans-3-metil fentanil (40 min) # (±)cis-3-etil fentanil (60 min) (±)trans-3-etil fentanil (40 min) # fentanil (50 min) = (±)cis-3-butil fentanil (50 min) = (±)trans-3-butil fentanil (50 min) = (±)cis-3-benzil fentanil (50 min) = (±)trans-3-benzil fentanil (50 min). Oznake gt i lt označavaju P lt 0.05. Zaključak Zaključeno je da na analgetsku jačinu 3-alkil analoga fentanila utiče sterni faktor (voluminoznost grupe na položaju 3 piperidinskog prstena i cis/trans izomerizam). Inače, uz izuzetak 3-metil fentanila, stereohemija ne utiče značajno na dužinu dejstva 3-alkil analoga fentanila