A novel series of aromatic esters (1a-1m) related to the Amaryllidaceae alkaloid (AA) haemanthamine were designed, synthesized and tested in vitro with particular emphasis on the treatment of neurodegenerative diseases. Some of the synthesized compounds revealed promising acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibitory profile. Significant human AChE (hAChE) inhibition was demonstrated by 11-O-(3-nitrobenzoyl)haemanthamine (1j) with IC50value of 4.0 ± 0.3 µM. The strongest human BuChE (hBuChE) inhibition generated 1-O-(2-methoxybenzoyl)haemanthamine (1g) with IC50 value 3.3 ± 0.4 µM. Moreover, 11-O-(2-chlorbenzoyl)haemanthamine (1m) was able to inhibit both enzymes in dose-dependent manner. The mode of hAChE and hBuChE inhibition was minutely inspected using enzyme kinetic analysis in tandem with in silico experiments, the latter elucidating crucial interaction in 1j-, 1m-hAChE and 1g-, 1m-hBuChE complexes. The blood-brain barrier (BBB) permeability was investigated applying the parallel artificial membrane permeation assay (PAMPA) to predict the CNS availability of the compounds.

• Klíčová slova
• Acetylcholinesterase, Alzheimer’s disease, Amaryllidaceae, Butyrylcholinesterase, Docking studies, Haemanthamine,
• MeSH
• acetylcholinesterasa chemie   metabolismus   MeSH
• alkaloidy amarylkovitých chemie   metabolismus   terapeutické užití   MeSH
• Alzheimerova nemoc farmakoterapie   patologie   MeSH
• Amaryllidaceae chemie   metabolismus   MeSH
• butyrylcholinesterasa chemie   metabolismus   MeSH
• cholinesterasové inhibitory chemická syntéza   metabolismus   terapeutické užití   MeSH
• estery chemie   MeSH
• fenantridiny chemie   metabolismus   terapeutické užití   MeSH
• hematoencefalická bariéra účinky léků   metabolismus   MeSH
• kinetika MeSH
• lidé MeSH
• simulace molekulového dockingu MeSH
• vazebná místa MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• acetylcholinesterasa MeSH
• alkaloidy amarylkovitých MeSH
• butyrylcholinesterasa MeSH
• cholinesterasové inhibitory MeSH
• estery MeSH
• fenantridiny MeSH
• hemanthamine MeSH Prohlížeč

Twelve derivatives 1a-1m of the β-crinane-type alkaloid haemanthamine were developed. All the semisynthetic derivatives were studied for their inhibitory potential against both acetylcholinesterase and butyrylcholinesterase. In addition, glycogen synthase kinase 3β (GSK-3β) inhibition potency was evaluated in the active derivatives. In order to reveal the availability of the drugs to the CNS, we elucidated the potential of selected derivatives to penetrate through the blood-brain barrier (BBB). Two compounds, namely 11-O-(2-methylbenzoyl)-haemanthamine (1j) and 11-O-(4-nitrobenzoyl)-haemanthamine (1m), revealed the most intriguing profile, both being acetylcholinesterase (hAChE) inhibitors on a micromolar scale, with GSK-3β inhibition properties, and predicted permeation through the BBB. In vitro data were further corroborated by detailed inspection of the compounds' plausible binding modes in the active sites of hAChE and hBuChE, which led us to provide the structural determinants responsible for the activity towards these enzymes.

• Klíčová slova
• Alzheimer’s disease, Amaryllidaceae, acetylcholinesterase, butyrylcholinesterase, docking studies, glycogen synthase kinase-3β inhibition, haemanthamine,
• MeSH
• alkaloidy amarylkovitých chemie   metabolismus   MeSH
• Alzheimerova nemoc metabolismus   MeSH
• Amaryllidaceae chemie   metabolismus   MeSH
• fenantridiny chemie   metabolismus   MeSH
• hematoencefalická bariéra metabolismus   MeSH
• kinasa glykogensynthasy 3beta metabolismus   MeSH
• lidé MeSH
• ligandy MeSH
• molekulární konformace MeSH
• molekulární modely MeSH
• molekulární struktura MeSH
• permeabilita MeSH
• simulace molekulového dockingu MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• alkaloidy amarylkovitých MeSH
• fenantridiny MeSH
• hemanthamine MeSH Prohlížeč
• kinasa glykogensynthasy 3beta MeSH
• ligandy MeSH

Scadoxus puniceus (Amaryllidaceae), a medicinal plant of high value in South Africa, is used as a component of a traditional herbal tonic prescribed to treat several ailments. Ultra-high performance liquid chromatography-tandem mass spectrometry quantified the phenolic compounds in different organs of S. puniceus. Gravity column chromatography was used to separate fractions and active compounds. The structure of these compounds was determined using 1D and 2D nuclear magnetic resonance and mass spectroscopic techniques. A microplate technique was used to determine the acetylcholinesterase inhibitory activity of the pure compounds. Metabolite profiling revealed a greater profusion of hydroxycinnamic acids (69.5%), as opposed to hydroxybenzoic acids (30.5%). Chlorogenic acid was the most abundant (49.6% of hydroxycinnamic acids) compound. In addition to chlorogenic acid, the study is the first to report the presence of sinapic, gallic, and m-hydroxybenzoic acids in the Amaryllidaceae. Chromatographic separation of S. puniceus led to the isolation of haemanthamine (1), haemanthidine (2), and a rare chlorinated amide, metolachlor (3), the natural occurrence of which is described for the first time. Haemanthamine, haemanthidine, and metolachlor displayed strong acetylcholinesterase inhibitory activity (IC50 ; 23.1, 23.7, and 11.5 μM, respectively). These results substantiate the frequent use of S. puniceus as a medicinal plant and hold much promise for further pharmaceutical development.

• Klíčová slova
• Amaryllidaceae, NMR, alkaloids, chromatography, paintbrush lily, phenolics,
• MeSH
• acetamidy chemie   izolace a purifikace   metabolismus   farmakologie   MeSH
• alkaloidy amarylkovitých chemie   izolace a purifikace   metabolismus   farmakologie   MeSH
• Amaryllidaceae chemie   metabolismus   MeSH
• cholinesterasové inhibitory chemie   izolace a purifikace   farmakologie   MeSH
• fenantridiny chemie   izolace a purifikace   metabolismus   farmakologie   MeSH
• kyseliny kumarové chemie   izolace a purifikace   metabolismus   farmakologie   MeSH
• léčivé rostliny chemie   MeSH
• rostlinné extrakty chemie   izolace a purifikace   metabolismus   farmakologie   MeSH
• tandemová hmotnostní spektrometrie MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Jihoafrická republika MeSH
• Názvy látek
• acetamidy MeSH
• alkaloidy amarylkovitých MeSH
• cholinesterasové inhibitory MeSH
• fenantridiny MeSH
• hemanthamine MeSH Prohlížeč
• hemanthidine MeSH Prohlížeč
• kyseliny kumarové MeSH
• metolachlor MeSH Prohlížeč
• rostlinné extrakty MeSH

Adult rats were orally administered with a single dose of sanguinarine (10 mg SA per 1 kg body weight) in 1.0 ml water. In the plasma and the liver, dihydrosanguinarine (DHSA) was identified as a SA metabolite by high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC/ESI-MS). Significantly higher levels of DHSA were found in both the plasma and the liver in comparison with those of SA. SA and DHSA were not detected in the urine. The formation of DHSA might be the first step of SA detoxification in the organism and its subsequent elimination in phase II reactions. Benz[c]acridine (BCA), in the literature cited SA metabolite, was found neither in urine nor in plasma and liver.

• MeSH
• alkaloidy farmakokinetika   MeSH
• benzofenantridiny MeSH
• biotransformace MeSH
• fenantridiny metabolismus   MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací metody   MeSH
• isochinoliny MeSH
• krysa rodu Rattus MeSH
• potkani Wistar MeSH
• reprodukovatelnost výsledků MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• alkaloidy MeSH
• benzofenantridiny MeSH
• dihydrosanguinarine MeSH Prohlížeč
• fenantridiny MeSH
• isochinoliny MeSH
• sanguinarine MeSH Prohlížeč

Correlated ab initio as well as semiempirical quantum chemical calculations and molecular dynamics simulations were used to study the intercalation of cationic ethidium, cationic 5-ethyl-6-phenylphenanthridinium and uncharged 3,8-diamino-6-phenylphenanthridine to DNA. The stabilization energy of the cationic intercalators is considerably larger than that of the uncharged one. The dominant energy contribution with all intercalators is represented by dispersion energy. In the case of the cationic intercalators, the electrostatic and charge-transfer terms are also important. The DeltaG of ethidium intercalation to DNA was estimated at -4.5 kcal mol(-1) and this value agrees well with the experimental result. Of six contributions to the final free energy, the interaction energy value is crucial. The intercalation process is governed by the non-covalent stacking (including charge-transfer) interaction while the hydrogen bonding between the ethidium amino groups and the DNA backbone is less important. This is confirmed by the evaluation of the interaction energy as well as by the calculation of the free energy change. The intercalation affects the macroscopic properties of DNA in terms of its flexibility. This explains the easier entry of another intercalator molecule in the vicinity of an existing intercalation site.

• MeSH
• chemické modely * MeSH
• DNA chemie   MeSH
• ethidium chemie   MeSH
• fenantridiny chemie   metabolismus   MeSH
• interkalátory chemie   MeSH
• kationty chemie   MeSH
• konformace nukleové kyseliny MeSH
• kvantová teorie MeSH
• molekulární konformace MeSH
• molekulární modely MeSH
• molekulární struktura MeSH
• počítačová simulace MeSH
• termodynamika MeSH
• vodíková vazba MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA MeSH
• ethidium MeSH
• fenantridiny MeSH
• interkalátory MeSH
• kationty MeSH

Inhibition of porcine pancreas and human saliva alpha-amylase (EC 3.2.1.1) by sanguinarine and chelerythrine was studied. The inhibition of alpha-amylase was assayed using a biosensor method which utilises a flow system equipped with a peroxide electrode. 250 microM sanguinarine and 250 microM chelerythrine cause complete inhibition of 1.9 nkat alpha-amylase from porcine pancreas. The same concentration of sanguinarine and chelerythrine caused 23.9% and 7.5% inhibition, respectively, of 1.9 nkat alpha-amylase from human saliva. Mixed type and partially reversible inhibition was found for both alpha-amylases treated with either alkaloid.

• MeSH
• alfa-amylasy antagonisté a inhibitory   metabolismus   MeSH
• alkaloidy metabolismus   farmakologie   MeSH
• benzofenantridiny MeSH
• biosenzitivní techniky MeSH
• časové faktory MeSH
• fenantridiny metabolismus   farmakologie   MeSH
• inhibitory enzymů farmakologie   MeSH
• isochinoliny MeSH
• kinetika MeSH
• lidé MeSH
• prasata MeSH
• stabilita enzymů MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• alfa-amylasy MeSH
• alkaloidy MeSH
• benzofenantridiny MeSH
• chelerythrine MeSH Prohlížeč
• fenantridiny MeSH
• inhibitory enzymů MeSH
• isochinoliny MeSH
• sanguinarine MeSH Prohlížeč

Using the 32P-postlabeling assay, we investigated the ability of quaternary benzo[c]phenanthridine alkaloids, sanguinarine, chelerythrine and fagaronine, to form DNA adducts in vitro. Two enhanced versions of the assay (enrichment by nuclease P1 and 1-butanol extraction) were utilized in the study. Hepatic microsomes of rats pre-treated with beta-naphthoflavone or those of uninduced rats, used as metabolic activators, were incubated in the presence of calf thymus DNA and the alkaloids, with NADPH used as a cofactor. Under these conditions sanguinarine and chelerythrine, but not fagaronine, formed DNA adducts detectable by 32P-postlabeling. DNA adduct formation by both alkaloids was found to be concentration dependent. When analyzing different atomic and bond indices of the C11-C12 bond (ring B) in alkaloid molecules we found that fagaronine behaved differently from sanguinarine and chelerythrine. While sanguinarine and chelerythrine showed a preference for electrophilic attack indicating higher potential to be activated by cytochrome P450, fagaronine exhibited a tendency for nucleophilic attack. Our results demonstrate that sanguinarine and chelerythrine are metabolized by hepatic microsomes to species, which generate DNA adducts.

• MeSH
• adukty DNA biosyntéza   MeSH
• alkaloidy metabolismus   farmakologie   MeSH
• benzofenantridiny MeSH
• fenantridiny metabolismus   farmakologie   MeSH
• fytogenní protinádorové látky metabolismus   farmakologie   MeSH
• interkalátory metabolismus   farmakologie   MeSH
• isochinoliny MeSH
• jaterní mikrozomy účinky léků   metabolismus   MeSH
• krysa rodu Rattus MeSH
• kvantová teorie MeSH
• potkani Sprague-Dawley MeSH
• radioizotopy fosforu analýza   MeSH
• systém (enzymů) cytochromů P-450 metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adukty DNA MeSH
• alkaloidy MeSH
• benzofenantridiny MeSH
• chelerythrine MeSH Prohlížeč
• fagaronine MeSH Prohlížeč
• fenantridiny MeSH
• fytogenní protinádorové látky MeSH
• interkalátory MeSH
• isochinoliny MeSH
• radioizotopy fosforu MeSH
• sanguinarine MeSH Prohlížeč
• systém (enzymů) cytochromů P-450 MeSH