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Alkaloids, the largest group among the nitrogen-containing secondary metabolites of plants, usually interact with several molecular targets. In this study, we provide evidence that six cytotoxic alkaloids (sanguinarine, chelerythrine, chelidonine, noscapine, protopine, homoharringtonine), which are known to affect neuroreceptors, protein biosynthesis and nucleic acids, also interact with the cellular cytoskeleton, such as microtubules and actin filaments, as well. Sanguinarine, chelerythrine and chelidonine depolymerized the microtubule network in living cancer cells (Hela cells and human osteosarcoma U2OS cells) and inhibited tubulin polymerization in vitro with IC50 values of 48.41 ± 3.73, 206.39 ± 4.20 and 34.51 ± 9.47 μM, respectively. However, sanguinarine and chelerythrine did not arrest the cell cycle while 2.5 μM chelidonine arrested the cell cycle in the G2/M phase with 88.27% ± 0.99% of the cells in this phase. Noscapine and protopine apparently affected microtubule structures in living cells without affecting tubulin polymerization in vitro, which led to cell cycle arrest in the G2/M phase, promoting this cell population to 73.42% ± 8.31% and 54.35% ± 11.26% at a concentration of 80 μM and 250.9 μM, respectively. Homoharringtonine did not show any effects on microtubules and cell cycle, while the known microtubule-stabilizing agent paclitaxel was found to inhibit tubulin polymerization in the presence of MAPs in vitro with an IC50 value of 38.19 ± 3.33 μM. Concerning actin filaments, sanguinarine, chelerythrine and chelidonine exhibited a certain effect on the cellular actin filament network by reducing the mass of actin filaments. The interactions of these cytotoxic alkaloids with microtubules and actin filaments present new insights into their molecular modes of action.

Three step bivariant multiple development thin-layer chromatography (BMD-TLC) technique was used in the quantitative analysis of chelerithrine, berberine, and chelidonine in extracts of greater celandine herb, roots, and herbal drugs. The separation was carried out on 10 cm × 10 cm TLC Si60 F254s glass plates in the horizontal DS-Chamber. First, the alkaloids were separated at a distance of 6 cm into two zones using toluene-ethyl acetate-methanol (1:1:1 ν/ν) solvent mixture as mobile phase. The plate was cut at 4.5 cm into two parts, lower part containing berberine and chelerithrine and upper part with chelidonine zone. The separation of berberine and chelerithrine was continued using n-propanol-formic acid-water (84:8:8 ν/ν) as mobile phase at a distance of 4.5 cm. The separation of chelidonine from accompanied compounds was achieved using the ethyl acetate-formic acid-water (7:1:1 ν/ν) solvent mixture at a distance of 5 cm. The densitometric quantification of Chelidonium alkaloids was performed at λ = 2...

Objective To evaluate the role of chelidonine isolated from ethanolic extract of Chelidonium majus in inducing apoptosis in HeLa cells and to assess the main signalling pathways involved. Methods Cells were initially treated with different concentrations of chelidonine for 48 h and the median lethal dose (LD50) value was selected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Morphological analysis of nuclear condensation and DNA damage and fragmentation were measured by 4',6-diamidino-2-phenylindole staining and comet assay. Further, reactive oxygen species (ROS) generation, cell cycle arrest and change in mitochondrial membrane potential were also examined and analyzed by flow cytometry. Evaluation of interaction of drug with CT DNA was investigated by circular dichroism (CD) spectral analysis to find any possible drug-CT DNA interaction. The mRNA and protein expressions of major signal proteins like p38, p53, protein kinase B (AKT), phosphatidylinositol 3-kinases (PI3K), Janus kinase 3 (JAK3), signal transducer and activator of transcription 3 (STAT3) and E6 and E7 oncoproteins as well as the pro-apoptotic genes and antiapoptotic genes were also estimated by reverse transcriptase-polymerase chain reaction and Western blotting. Results Based on LD(50) value (30 μg/mL) of chelidonine, three doses were selected, namely, 22.5 μg/mL (D1), 30.0 μg/mL (D2) and 37.5 μg/mL (D3). Results showed that chelidonine inhibited proliferation and induced apoptosis in HeLa cells through generation of ROS, cell cycle arrest at sub-G1 and G0/G1 stage, change in mitochondrial membrane potential and fragmentation of DNA. Results of CD spectra showed effective interaction between chelidonine and calf thymus DNA. Studies of signalling pathway revealed that chelidonine could efficiently induce apoptosis through up-regulation of expressions of p38, p53 and other pro-apoptotic genes and down-regulation of expressions of AKT, PI3K, JAK3, STAT3, E6, E7 and other antiapoptotic genes. Conclusion Chelidonine isolated from Chelidonium majus efficiently induced apoptosis in HeLa cells through possible alteration of p38-p53 and AKT/PI3 kinase signalling pathways.

Benzo[c]phenanthridine alkaloids are distributed in Papaveraceous and Rutaceous plants and classified into two categories of fully aromatized bases and partially hydrogenated bases(B/C-cis-11-hydroxyhexahydrobenzo[c]phenanthridine skeleton). They have long history, in which the first isolation of chelidonine, a partially hydrogenated base, from Papaveraceous plant, was reported in 1839. Over a century later nitidine and fagaronine, O4-type fully aromatized bases isolated from Rutaeceous plant, attracted much attention because of their antitumor activity. At present, a variety of pharmacological effects are recognized in natural and unnatural species. We, about forty years ago, started synthetic studies on fully aromatized bases focusing on structure-activity relationship of antitumor activity and succeeded in establishing widely-applicable synthetic methods. Recently the enantioselective synthesis of chelidonine was completed. Thus, in this review, we present our chemical history for the synthesis of benzo[c]phenanthridine bases.

Both enantiomers of chelidonine, a B/C-cis-11-hydroxyhexahydrobenzo[c]phenanthridine alkaloid, were synthesized by manipulation of the B/C-dehydro ring juncture of benzo[c]phenanthridine skeleton using Sharpless asymmetric dihydroxylation and stereospecific catalytic hydrogenation after introduction of oxygen functions on the C ring as key reaction steps for the construction of stereogenic centers.

In this study, we introduced a simple and sensitive method of capillary electrophoresis with ultraviolet light-emitting diode-induced native fluorescence (UV-LEDIF) detection for the determination of isoquinoline alkaloids in extracts of Chelidonium majus L. Samples were extracted with acidic methanol and the extracts were directly analysed by CE. Simultaneous determination of protopine, chelidonine, coptisine, sanguinarine, allocryptopine, chelerythrine and stylopine was performed in 20 mM phosphate buffer (pH 3.1). The baseline separation of these alkaloids was finished within 20 min. As these alkaloids have native fluorescence, they were directly detected using the commercially available UV light emitting diode without troublesome fluorescent derivatisation. Satisfactory LOD values were obtained for the studied compounds considering their appearance in natural extracts. Lower limits of detection were 0.05 μg/mL for protopine, 0.06 μg/mL for stylopine and allocryptopine, 0.07 μg/mL for chelidonine, 0.22 μg/mL for sanguinarine, 1.7 μg/mL for chelerythrine and 5.5 μg/mL for coptisine. The developed method was successfully applied to determine the contents of seven alkaloids in the aerial parts of Chelidonium majus L, which varied from 0.025 to 0.763% (w/w). Also, to demonstrate the potential of the proposed CE method, an estimation of the cytotoxic properties of selected Celandine alkaloids in a natural extract was carried out.

Trichodinids are the most common ciliates parasite present on the skin of pond-reared fish and can cause severe economic losses in aquaculture and ornamental fish breeding. The present study aims to evaluate the antiparasitic activity of the active components from Chelidonium majus L. against Trichodina sp. Bioassay-guided fractionation and isolation of the compounds with antiparasitic activity were performed on the ethanolic extract of C. majus yielding three bioactive alkaloids namely: chelidonine (1), chelerythrine (2) and sanguinarine (3). Results from in vivo antiparasitic assays revealed that these compounds when isolated could be 100% effective for the elimination of Trichodina sp. at the concentrations of 1.0, 0.8, and 0.7 mg L − 1 , with the median effective concentration (EC 50 ) values of 0.6, 0.33, and 0.32 mg L − 1 , respectively. Furthermore, the promising chelidonine, chelerythrine and sanguinarine were subjected to acute toxicity tests for the evaluation of their safety to the host ( Parabramis pekinensis ). It was found that the 48 h median lethal concentration (48 h-LC 50 ) values determined by the acute toxicity tests on P. pekinensis were 2.5, 3.8 and 1.5 mg L − 1 respectively. These results provided evidence that the isolated compounds, especially chelerythrine, can be exploited as novel antiparasitic agents for the control of Trichodina sp.

0009-3130/11/4703-0477 2011 Springer Science+Business Media, Inc. 1) Department of Pharmaceutical Technology, Faculty of Pharmacy, Tbilisi State Medical University, 22 G. Akhvlediani, Tbilisi, Georgia 0108, e-mail: annabozhadze@yahoo.com; 2) Laboratory of Alkaloids, Institute of Pharmacochemisty, 36 Sarajishvili street, Tbilisi, Georgia 0159; 3) Institute of Physical and Organic Chemistry, #5 Jikia str., Tbilisi, Georgia 0186. Published in Khimiya Prirodnykh Soedinenii, No. 3, pp. 424–426, May–June, 2011. Original article submitted March 5, 2010. Chemistry of Natural Compounds, Vol. 47, No. 3, July, 2011 [Russian original No. 3, May–June, 2011]

Dactylogyrus intermedius is one of the most common and serious cause of parasitic diseases of freshwater fish in aquaculture, and can cause morbidity and high mortality in most species of freshwater fish worldwide. To attempt controlling this parasite and explore novel potential antiparasitic agents, the present study was designed to ascertain the anthelmintic activity of Chelidonium majus L. whole plant and to isolate and characterize the active constituents against D. intermedius. The ethanol extract from C. majus whole plant showed significant anthelmintic activity against D. intermedius [EC(50) (median effective concentration) value = 71.5 mg L(-1)] and therefore subjected to further isolation and purification using various chromatographic techniques. A quaternary benzo[c]phenanthridine alkaloid exhibited significant activity against D. intermedius was obtained and identified as chelidonine. In vivo anthelmintic efficacy tests exhibited that chelidonine was 100% effective against D. intermedius at a concentration of 0.9 mg L(-1), with EC(50) value of 0.48 mg L(-1) after 48 h of exposure, which is more effective than the positive control, mebendazole (EC(50) value = 1.3 mg L(-1)). In addition, the 48-h median lethal concentration (LC(50)) for chelidonine against the host (Carassius auratus) was 4.54 mg L(-1). The resulting therapeutic index for chelidonine was 9.46. These results provided evidence that chelidonine might be potential sources of new antiparasitic drugs for the control of Dactylogyrus.

Chelidonine (CHE) is a major bioactive constituent of greater celandine, a plant used in traditional herbal medicines. CHE has widely been used as an analgesic in clinical settings. We evaluated the inhibitory effects of CHE on human cytochrome P450 enzymes. CHE produced time-, concentration-, and NADPH-dependent inhibition of CYP2D6, with K I and k inact values of 20.49 μM and 11.05 min -1 , respectively. Approximately 76% of CYP2D6 activity was suppressed after 9 minute incubation with CHE (50 μM). The loss of enzyme activity was not restored following dialysis. The estimated partition ratio of the inactivation was about 156. Quinidine, a competitive inhibitor of CYP2D6, attenuated the CHE-mediated enzyme inactivation, while glutathione and catalase/superoxide dismutase did not markedly ameliorate the inhibitory effect. Upon oxidation using potassium ferricyanide, the 15.1% activity of CYP2D6 was restored. These findings indicate that CHE acted as a mechanism-based inactivator of CYP2D6 and the observed effects may induce potential drug-drug interactions.

Comparative substrate-inhibitor analysis of catalytic properties of liver monoamine oxidases (MAO) was performed in the mature males of the American mink Mustela vison and the European mink Mustela lutreola. The action on the MAO activity was studied of alkaloids of the benzo[c]phenanthridine group: sanguinarine and chelidonine, diisoquinoline alkaloid berberine, medicinal agents “Ukrain” and “Sanguirythrin” as well as derivatives of 2-propylamine: deprenyl and chlorgylin. The latter turned out to be irreversible inhibitor of the MAO A form, whereas deprenyl-irreversible inhibitor of the MAO B form in both studied mink species. The selectivity of action of each inhibitor on the corresponding liver MAO form for the species M. vison was one order of magnitude stronger than for the species M. lutreola. All studied alkaloids as well medicinal agents on their basis have been shown to be specific irreversible inhibitors of the intermediate strength of the liver MAO A form of both mink species. They inhibit the enzymatic deamination of serotonin, tyramine, and tryptamine without affecting the deamination reaction of benzylamine and β-phenylethylamine (at concentrations of 10 mM and lower). Out of five studied isoquinoline agents, the medication “Ukrain” and alkaloid chelidonine have the highest inhibitory action; the agent “Sanguirythrin” and alkaloids berberine and sanguinarine produce the weaker monoamine oxidase effect. The revealed specificity of action of the studied inhibitors is an indirect evidence for the presence in the liver enzymes of both mink species, like in the rat liver enzyme, of several molecular forms.

A simple and rapid method for the simultaneous determination of seven isoquinoline alkaloids, protopine, chelidonine, coptisine, stylopine, sanguinarine, berberine, and chelerythrine, in Chelidonium majus L. (Ch. majus) samples by ultra-performance LC method with photodiode array detection is described. The baseline separation of these compounds was performed with (A) acetonitrile-(B) ammonium acetate (10 mM, adjusted to pH 3.0 with acetic acid) as the mobile phase using a C18 RP column (2.1x100 mm, 1.7 microm). Optimized conditions resulted in excellent peak shapes. The seven alkaloids were completely separated within 20 min. Good linear behaviors (r > or = 0.9992) over the investigated concentration ranges were observed for all the analytes. Validation proved the repeatability of the method was good and recovery was satisfactory. The validated method was successfully applied for 20 batches of Ch. majus. These results demonstrated that the ultra-performance LC photodiode array method proposed was very useful in the analysis and quality control of Ch. majus.

kelidoninin analjezik aktivitesi kuyruk kaldirma testi kullanilarak arastirilmistir. Kelidonin, C. majus’un toprak ustu kisimlarinda bulunan major alkaloitlerden birisidir. Kelidonin izolasyonu kromatografik teknikler kullanilarak gerceklestirilmistir. Izole edilen bilesigin yapisi spektroskopik (1H-NMR, 13C-NMR) ve fiziksel (erime noktasi) yontemler kullanilarak aydinlatilmistir. Bilesigin analjezik aktivitesi 5 mg/kg dozda fareler uzerinde kuyruk kaldirma testi kullanilarak degerlendirilmistir. Kelidoninin bu dozdaki analjezik aktivitesinin 90. dakikada aspirinden daha yuksek oldugu, 150. dakikada ise aspirine esit bir analjezik aktivite gosterdigi tespit edilmistir

Apoptogenic and DNA damaging effects of chelidonine (CHE) and sanguinarine (SAN), two structurally related benzophenanthridine alkaloids isolated from Chelidonium majus L. (Papaveraceae), were compared. Both alkaloids induced apoptosis in human acute T-lymphoblastic leukaemia MT-4 cells. Apoptosis induction by CHE and SAN in these cells was accompanied by caspase-9 and -3 activation and an increase in the pro-apoptotic Bax protein. An elevation in the percentage of MT-4 cells possessing caspase-3 in active form after their treatment with CHE or SAN was in parallel to a corresponding increase in the fraction of apoptotic cells. The involvement of mitochondria in apoptosis induction by both alkaloids was supported by cytochrome C elevation in cytosol, with an accompanying decrease in cytochrome C content in the mitochondrial fraction. At the same time, two alkaloids under study differed drastically in their cell cycle phase-specific effects, since only CHE arrested MT-4 cells in G(2)/M phase. It was shown earlier, that CHE, in contrast to SAN, does not interact directly with DNA. This fact is in line with DNA damaging effects of the alkaloids detected in the COMET assay. Nevertheless, apoptosis-inducing activity of CHE even slightly exceeded that of SAN.

Sanguinarine, chelerythrine and chelidonine are isoquinoline alkaloids derived from the greater celandine. They possess a broad spectrum of pharmacological activities. It has been shown that their anti-tumor activity is mediated via different mechanisms, which can be promising targets for anti-cancer therapy. We focused our study on the differential effects of these alkaloids upon cell viability, DNA damage effect and nucleus integrity in mouse primary spleen cells and mouse lymphocytic leukemic cells, L1210. Sanguinarine and chelerythrine produce a dose-dependent increase in DNA damage and cytotoxicity in both primary mouse spleen cells and L1210 cells. Chelidonine did not show a significant cytotoxicity or damage DNA in both cell types, but completely arrested growth of L1210 cells. Examination of nuclear morphology revealed more cells with apoptotic features upon treatment with chelerythrine and sanguinarine, but not chelidonine. In contrast to primary mouse spleen cells, L1210 cells showed slightly higher sensitivity to sanguinarine and chelerythrine treatment. This suggests that cytotoxic and DNA damaging effects of chelerythrine and sanguinarine are more selective against mouse leukemic cells and primary mouse spleen cells, whereas chelidonine blocks proliferation of L1210 cells. The action of chelidonine on normal and tumor cells requires further investigation.

Glioblastoma is a highly malignant tumor, characterized by an unfavorable prognosis even in response to multidisciplinary treatment strategies, owing to its high-invasive phenotype. Ukrain, a semisynthetic thiophosphoric acid derivative of the purified alkaloid chelidonine, has been used in the therapy of several solid tumors, but little is known about its effect on glioblastoma and, in general, about the molecular mechanisms responsible for its effects. In particular, we previously demonstrated that Ukrain modulates the expression of genes and proteins involved in tumor invasion, and here we investigate some unreported effects of Ukrain on human cultured glioblastoma cells. We used morphological and molecular biology methods to analyze the expression and the intracellular distribution pattern of glial fibrillary acidic protein, the expression of the gap junction protein connexin 43 and the apoptotic effect in human glioblastoma cells treated with 0.1, 1 and 10 micromol/l Ukrain for 72 h. After treatment with 10 micromol/l Ukrain, glial fibrillary acidic protein fluorescence increased and a higher number of cells displayed glial fibrillary acidic protein organized into a filamentous state. Western blot analysis of glial fibrillary acidic protein confirmed that Ukrain tended to upregulate the protein. Connexin 43 was not modulated by Ukrain both at the mRNA and at the protein level. Ukrain-induced apoptotic rate was 4.63, 10.9 and 28.9% after 0.1, 1 and 10 micromol/l Ukrain, respectively, likely mediated by cytochrome c release in the cytoplasm. Considered as a whole, these findings provide new information to complete the understanding of the mechanisms of Ukrain antitumor and chemopreventive effect, and support the possible potential of Ukrain for the therapy of brain tumors.

Background Chelidonium majus L. (Papaveraceae) (greater celandine) is a medicinal herb that is widely spread in Europe. Antitumoural activity has been reported for C. majus extracts. Hypothesis/Purpose To investigate the antitumour activity of a C. majus extract in vitro and in vivo. Study Design Cytotoxic effects of C. majus extracts were evaluated on human cancer cell lines, i.e. PANC-1 (pancreas cancer), HT-29 (colon cancer), MDA-MB-231 (breast cancer), PC-EM005 and PC-EM002 (primary endometrium cancer cells), and PANC02 (murine pancreatic adenocarcinoma cells). A preliminary in vivo study was performed to evaluate the effect of a defatted C. majus extract and UkrainTM in a highly metastatic murine pancreatic model. Methods Chelidonium majus L. herb containing 1.26% (dry weight) of total alkaloids expressed as chelidonine was used to prepare an 80% ethanolic extract (CM2). This crude extract was then defatted with n-hexane, resulting in a defatted C. majus extract (CM2B). Cytotoxic effects of the two extracts (CM2 and CM2B) were evaluated on human and murine cell lines in vitro. CM2B and UkrainTM were evaluated in a highly metastatic murine pancreatic model. Results Four main benzylisoquinoline alkaloids were identified in CM2B, i.e. chelidonine, sanguinarine, chelerythrine and protopine, using HPLC-UV. CM2 showed a high cytotoxic activity against PANC-1 (IC50, 20.7 µg/ml) and HT-29 (IC50, 20.6 µg/ml), and a moderate cytotoxic activity against MDA-MB-231 (IC50, 73.9 µg/ml). CM2 as well as CM2B showed a moderate to high cytotoxic activity against the PANC02 cell line (IC50, 34.4 and 36.0 µg/ml). Low to almost no cytotoxic effect was observed on primary endometrium cancer cells PC-EM005, PC-EM002 and on normal fibroblast cells 3T3, when treated with CM2B. Significantly less metastases were counted in mice treated with 1.2 mg/kg CM2B, but not with 3.6 mg/kg UkrainTM, compared to the control group. The extract, however, did not affect the weight of the primary tumours.

Functional gastro-intestinal diseases as the irritable bowel syndrome are very common in the population and are characterized by a broad spectrum of symptoms which mostly are related to spastic or paralytic intestinal function without defined histopathological changes of the tissue. Due to the multifactorial pathogenesis a multifactorial therapy with multi-target action seems to be reasonable. STW 5 (Iberogast), its constituent herbal extracts and some isolated compounds were used in an in vitro model provided by intestinal samples from guinea pig in order to test their activity on histamine-induced contractions and spontaneous motility, respectively. For comparison the known spasmolyticum papaverine was used. The results show that the lytic effect of the phytotherapeuticum on histamine-induced contraction represents additively the actions of the different components and corresponds to approx. 10 microM of papaverine. Spontaneous peristaltic motion was differently modulated by the various constituent extracts. The experiments with silibinin, glycyrrhicine, chelidonine, and protopine showed that the effects of the extracts were not comparable to those of the respective chemical constituents.

A new high-performance liquid chromatographic method has been developed for analysis of isoquinoline alkaloids in extracts of Chelidonium majus L. Samples were extracted with acidic methanol and the extracts were purified by solid-phase extraction on Supelclean LC-18 cartridges. Optimized conditions resulted in high recovery and reproducibility. Simultaneous determination of protopine, chelidonine, coptisine, sanguinarine, and berberine was performed by HPLC on a C18 reversed-phase column. Use of the Luna C18(2) new-generation silica-based stationary phase and 14.7:18:67.3 (v/v) acetonitrile-methanol-30 mM ammonium formate, pH 2.80, as mobile phase resulted in excellent peak shapes. Validation proved the repeatability of the method was good and recovery was satisfactory. Lower limits of detection were 0.2 ng for coptisine, 0.4 ng for sanguinarine, and 0.5 ng, for protopine, chelidonine, and berberine.

The greater celandine (Chelidonium majus L.) is a well known source of isoquinoline alkaloids with therapeutic value. The aim of our work was to develop a simple and fast method to determine the alkaloid composition in Chelidonium plant organs. TLC-densitometry seemed to be the most convenient analytical technique for routine, fast investigations and its precision was controlled by using HPLC. A densitometric method using Silica gel 60 F254 with chloroform-methanol 60:30 (v/v) and methylene chloride-methanol 97:3 (v/v) as mobile phases has been developed to quantify the main alkaloids (chelidonine, chelerythrine, sanguinarine, coptisine and berberine). The application of TLC permitted utilizing the fluorescence of alkaloids without purification and made the detection extremely sensitive. The samples were further investigated by our reliable HPLC method. Analysis was performed on a C18 column with acetonitrile-methanol-30 mM ammonium formate (pH = 2.8) 14.7:18.0:67.3 (v/v/v) as mobile phase and alkaloids were determined at 280 nm by using external standards. Our TLC-densitometric method elaborated is a simple technique for accurate and reproducible determination of Chelidonium alkaloids. The results of the two chromatographic methods showed good agreement.

The purpose of this study was to examine the relationship between the DNA intercalating characteristics and the DNA damaging capacity of four alkaloids extracted from Chelidonium majus L, as well as their toxicity towards murine NK/Ly lymphoma cells. Chelerythrine, sanguinarine and coptisine were found to be intercalated into the DNA isolated from NK/Ly cells, meanwhile, chelidonine exhibited no affinity to DNA. Sanguinarine exhibited the greatest toxicity toward NK/Ly cells, and the toxicity of the other three decreased in descending order: chelerythrine, coptisine and chelidonine. Chelerythrine and sanguinarine caused DNA damage, illustrated by the formation of comets of the third class. Coptisine was less toxic than chelerythrine and sanguinarine, and affected the formation the same class of comets in higher concentration. The quantity of comets induced by chelidonine were negligible, a finding consistent with its inability to intercalate into DNA structure. The ability of four main alkaloids of Chelidonium majus L., to intercalate into DNA isolated from murine NK/Ly lymphoma cells, correlated with their ability to induce breaks in cellular DNA and with their toxic effect towards those cells.

Chelidonine, a tertiary hexahydro-benzophenanthridine alkaloid is an inhibitor of tubulin polymerisation and has been revealed to arrest cells in G2/M. Since enhanced tyrosine kinase (TK) activity is linked to the transition from normal to the immortal malignant phenotype, the effect of 10 mM chelidonine was evaluated on TK activity in normal, transformed and malignant cell lines after 2 hours of exposure. Chelidonine caused a stimulation of TK activity in two normal cell lines (human foreskin fibroblast (Hs27) and normal monkey kidney (NMK)). In contrast, an inhibition of TK activity was observed in transformed human embryonic kidney (Graham 293) and transformed African green monkey kidney (Vero), as well as in human cervical carcinoma (HeLa) and squamous oesophageal carcinoma (WHCO5) cells. Hs27 cells exposed to chelidonine, revealed an increase in TK activity of 1.27-fold (P < 0.05). NMK cells showed a 1.15-fold increase in TK activity. A decrease in TK activity was observed in Graham 293 (0.91-fold) and Vero (0.45-fold) (P < 0.005) cells. In both HeLa and WHCO5 cells, the TK activity was reduced to 0.68-fold (P < 0.05) and 0.56-fold (P < 0.005) respectively. These data, including results from our previous studies, suggest a potential cross talk between the SAPK/JNK and TK signal transduction pathways and a possible differential effect of chelidonine on the phosphorylation status of role players involved in determining the length of G2 arrest in normal versus transformed and malignant cells.

By the end of the 18th and the beginning of the 19th century a new era began in medicine, pharmaceutics and chemistry that was strongly connected with alkaloids and alkaloid drugs. Even before that it was known that certain drugs administered in limited doses were medicines, and toxic if taken in larger doses (opium, coke leaves, belladonna roots, monkshood tubers crocus or hemlock seeds). However, the identification, isolation and structural characterization of the active ingredients of the alkaloid drugs was only possible in the mid 20th century by the use of modern extraction equipment and instrumental methods (NMR, X-ray diffraction and others).In spite of continuing use over a long time, there is still great interest in investigating new drugs, potential raw materials for the pharmaceutical industry, as well as the more detailed investigation and definition of bio-active components and the indication of their activity range, and the partial synthesis of new alkaloid molecules based on natural alkaloids. The scope of these investigations, especially in the field of semi-synthesis is to make better use of the bio-active ingredients of alkaloid drugs, i.e. to improve the pharmacological effect (stronger and prolonged effect of the medicine, decreased toxicity and side effects), or to extend or change the applications. A combined classification of alkaloids was used, based on the chemical structure and origin, i.e. the source of their isolation to study alkaloid structure. For practical reasons, the following classification of alkaloids was used: ergot alkaloids, poppy alkaloids, tropanic alkaloids purine derivative alkaloids, carbon-cyclic alkaloids, and other alkaloids. The second part of this report presents a table of general procedures for alkaloid isolation from plant drugs (extraction by water non-miscible solvents, extraction by water-miscible solvents and extraction by diluted acid solutions). Also, methods for obtaining chelidonine and glaucine as hydrochloride bases and salts were presented in more details. Data from leading world pharmacopoeias (Ph. Eur. Ill/s 2000, DAB 1996, USP 23, JP XIII, BP 1993, Ph. Jug. IV) were used in the study of application of the pure alkaloids in pharmaceutical forms with predetermined doses. A comparative study of these data shows that a great number of preparations are produced worldwide based on alkaloids and alkaloids with modified structure. These medicines have found use in modern therapeutic practice in many countries. Most products are produced on the basis of caffeine, theophylline, ephedrine, atropine, scopolamine, reserpine and pilocarpine.

A comparative study has been carried out on effects of berberine (diisoquinoline alkaloid) and sanguinarine and chelidonine (benzophenanthridine alkaloids( on erythrocyte acetylcholinesterase and serum butyrylcholinesterase from human blood. The studied alkaloids have been shown to be strong reversible inhibitors of the cholinesterase activity. Acetylcholinesterase is more sensitive to their action, than butyrylcholinesterase. The type of reversible inhibition was determined, and inhibitor constants were calculated. It is revealed that the character of inhibition is identical for the both cholinesterases. Berberine and sanguinarine are competitive-noncompetitive inhibitors, whereas chelidonine, a competitive inhibitor.

Background Chelidonine, a major bioactive, isoquinoline alkaloid ingredient in Chelidonium majus , exhibits anti-inflammatory and other pharmacological properties. However, its molecular mechanisms in asthma remain unclear. In this work we investigated chelidonine's effect and mechanism in airway inflammation in a mouse model of allergic asthma. Methods The mice were sensitized to ovalbumin followed by aerosol allergen challenges and determination of chelidonine's effect on enhanced pause (Penh), pulmonary eosinophilic infiltration, eotaxin-2, interleukin-4 (IL-4), IL-13, OVA-specific IgE production, and several transcription factors. Result Chelidonine strongly suppressed airway eosinophilia, expression of eotaxin-2, IL-4, and IL-13 cytokine production in bronchoalveolar lavage fluid (BALF). It also attenuated lung IL-17, and eotaxin-2 mRNA expression levels. Moreover, it suppressed eotaxin-2 and IL-17 production in accordance with up- and downregulation of forkhead box p3 (Foxp3), and signal transducer and activator of transcription (STAT6) expression, respectively. Conclusions Chelidonine has profound inhibitory effects on airway inflammation and this effect is caused by suppression of IL-4, eotaxin-2, and OVA-specific IgE production through the STAT6 and Foxp3 pathways. So chelidonine can improve allergic asthma in mice and be a novel anti-asthma therapeutic.

Alkaloids constitute a huge group of natural products, and some of them have been extensively studied for their activity on targets related to the central nervous system (CNS), especially neurodegenerative diseases and depression. In this context, the inhibition of monoamine oxidases (MAOs) was considered in the search for natural products with promising scaffolds for drug development. Among various classes, indole alkaloids possess biological and pharmacological properties in the CNS, mainly related to the serotonergic and glutamatergic transmission. Those containing a β-carboline moiety, such as harmane, are especially known for their inhibition of MAOs. Another class of alkaloids with a number of studies on the inhibition of MAOs is that of the isoquinoline alkaloids. Chelidonine, a representative of this group, shows specific and irreversible inhibition of MAO-A. The studies conducted so far on the selectivity between MAO-A and -B inhibitors contributed to a better understanding of structure–activity relationship (SAR) for the various classes of alkaloids. One example is desoxypeganine, a quinazoline alkaloid that presents selective inhibition on MAO-A. Due to its specific inhibition, this naturally occurring alkaloid has been clinically investigated for its potential application as an alcohol and smoking cessation aid. On the other hand, quinolone alkaloids presented specific in vitro MAO-B inhibition and are mainly considered for age-related neurodegenerative diseases. Several other examples are given in this chapter that consider selectivity on MAO isoforms and SAR issues of various classes of alkaloids.

A group of 11 derivatives of chelidonine was obtained by acylations and/or alkylations of the secondary hydroxyl group with the aim of testing their biological activity. This paper focuses on the new derivatives influence on CNS in mice. The highest activity was observed for compounds 2c, 3c and 4, which produced antinociceptive and antiserotoninergic effects, not recorded for the parent alkaloid 1.

[(+/-)-[5bRS-(5b alpha, 6beta, 12b alpha)]-5b,6,7,12b,13,14-hexahydro-13-methyl[1,3]benzodioxolo[5,6-c]-1,3-dioxolo[4,5-1] phenanthridin-6-yl]-4-[18F]fluorobenzoate (6b) an aromatic ester of chelidonine was prepared as a possible PET radiotracer. Compound (6b) was prepared in no-carrier-added (n.c.a) form from the 4-N,N,N-trimethylanilinium triflate derivative (5) in one step. The best results were obtained using Kryptofix(2.2.2)/[18F] and DMSO as the solvent at 90 degrees C. Column chromatography afforded the desired compound in overall radiochemical yield of 65-70% (EOS) with a specific activity of about 3000 Ci/mmole and radiochemical purity greater than 95% in 15 min.

Ukrain has been described as a semi-synthetic Chelidonium majus alkaloid derivative, consisting of three chelidonine alkaloids combined to triaziridide. We found the actions of Ukrain to be similar to the Chelidonium alkaloids it is prepared from, and therefore became concerned about its chemical integrity. Chemical analyses of Ukrain by thin layer chromatography, high-performance liquid chromatography and liquid chromatography-mass spectrometry was inconsistent with the proposed trimeric structure and demonstrated that at least some commercial preparations of Ukrain consist of a mixture of C. majus alkaloids (including chelidonine).

The solution conformation of the alkaloid chelidonine in CDCl3 was studied by proton steady-state NOE and DPFGSE NOE techniques. The experiments showed that the molecule adopts in solution essentially the same conformation as in the crystal state. Upon protonation with trifluoroacetic acid in CDCl3 chelidonine preserves its conformation, although a strong intramolecular N+HO hydrogen bond is formed. Conformational searches in vacuo performed with the use of GMMX software strongly suggest that the molecular geometry in the crystal state of chelidonine is also the global energetic minimum of the molecule. Copyright © 2000 John Wiley & Sons, Ltd.

A simple and effective method for the sensitive detection of alkaloids in crude plant extracts applying capillary electrophoresis with field amplified sample injection (FASI) is described. This method was compared with normal pressure injection for the determination of alkaloids in methanolic extracts from roots of Berberis vulgaris L. (Berberidaceae) and Hydrastis canadensis L. (Ranunculaceae) using a 1:1 mixture of 200 mM ammonium acetate at pH 3.1 and methanol. By introducing a short plug of 70% methanol (v/v) before electrokinetic injection with 16 kV for 8 s the concentration sensitivity was 1000-times higher compared to hydrodynamic injection for 1 s. No difference between both injection methods for selectivity and resolution of the obtained electropherograms was found. The influence of voltage and injection time on the introduced sample amount was investigated using a mixture of berberine and chelidonine as model substances.

This study evaluates the possible protective potentials of chelidonine and its poly lactide-co-glycolide (PLGA) encapsulated nano-form against cadmium chloride (CdCl₂) induced oxidative stress and hepatotoxicity in mice, ex vivo and in vivo. Acute exposure to CdCl₂ (1.0 mg/kg b.w; i.p., twice a week for 30 days) generated oxidative stress in mice through accumulation of reactive oxygen species and increased lipid peroxidation, and levels of certain liver marker enzymes (ALT, AST, ALP) with decrease in levels of GSH and certain other antioxidant enzymes (SOD, CAT, GR) in liver. Treatment with nano-chelidonine for 30 days after CdCl₂ intoxication significantly reduced oxidative stress and lipid peroxidation and restored levels of GSH, cholesterol, triglyceride and antioxidant enzymes, showing ameliorative changes in histopathology of liver. Expression pattern of certain inflammatory and apoptotic signal proteins also indicated better hepato-protective abilities of nano-chelidonine, making it a more suitable protective drug than chelidonine against cadmium toxicity in mice.

Chelidonium majus L. (Papaveraceae) has a long history as being useful for the treatment of many diseases in European countries. This plant is of great interest for its use also in Chinese herbal medicine. The plant contains, as major secondary metabolites, isoquinoline alkaloids, such as sanguinarine, chelidonine, chelerythrine, berberine and coptisine. Other compounds structurally unrelated to the alkaloids have been isolated from the aerial parts: several flavonoids and phenolic acids. C. majus extracts and its purified compounds exhibit interesting antiviral, antitumour and antimicrobial properties both in vitro and in vivo.

The determination of six different isoquinoline alkaloids from Chelidonium majus (leaves and roots) using capillary electrophoresis is described. Analyses were performed on fused silica capillaries with 66 mM phosphate buffer at pH 2.4 containing 40% methanol. The analytes were monitored by on-line ultraviolet (UV) detection to identify sanguinarine, chelerythrine, berberine and chelidonine. Both coptisine and DL-stylopine were isolated by semi-preparative high performance liquid chromatography (HPLC), and their structure assessed by UV and mass spectrometry. Capillary electrophoresis is confirmed as a useful complementary technique to HPLC.

In this study, the synergistic biological action of five celandine alkaloids in normal and cancer cells was investigated by capillary electrophoresis with light-emitting diode-induced native fluorescence detection. The specific capacity of each alkaloid to penetrate into the cells was estimated by monitoring alkaloid concentration decreases in the cell medium during incubation with murine fibroblast NIH/3T3, mouse melanoma B16F10, and human breast cancer MCF7 cell lines. Mixtures of isoquinoline alkaloids containing protopine, chelidonine, sanguinarine, allocryptopine, and stylopine were applied to cell cultures for 20 and 40 min, and the content of alkaloids in the cell media was measured by capillary electrophoresis (CE). CE separation of isoquinoline alkaloids was performed in 30 mM phosphate buffer (pH 2.5). As these alkaloids have native fluorescence, they were directly detected using the commercially available UV light-emitting diode without troublesome fluorescent derivatization. The results showed a differential ability of celandine alkaloids to penetrate into the normal and cancer cell interior, which was inversely proportional to their cytotoxic activity. While the most effective transport of celandine alkaloids from the cell medium to the cell interior was observed for normal murine fibroblast NIH/3T3 cells (about 55% of total content), cytotoxicity tests demonstrated selective and profound apoptotic effects of a five-alkaloid combination in the mouse melanoma B16F10 cell line.

Cancer cells often develop multidrug resistance (MDR) which is a multidimensional problem involving several mechanisms and targets. This study demonstrates that chelidonine and an alkaloid extract from Chelidonium majus, which contains protoberberine and benzo[c]phenanthridine alkaloids, has the ability to overcome MDR of different cancer cell lines through interaction with ABC-transporters, CYP3A4 and GST, by induction of apoptosis, and cytotoxic effects. Chelidonine and the alkaloid extract inhibited P-gp/MDR1 activity in a concentration-dependent manner in Caco-2 and CEM/ADR5000 and reversed their doxorubicin resistance. In addition, chelidonine and the alkaloid extract inhibited the activity of the drug modifying enzymes CYP3A4 and GST in a dose-dependent manner. The alkaloids induced apoptosis in MDR cells which was accompanied by an activation of caspase-3, -8,-6/9, and phosphatidyl serine (PS) exposure. cDNA arrays were applied to identify differentially expressed genes after treatment with chelidonine and the alkaloid extract. The expression analysis identified a common set of regulated genes related to apoptosis, cell cycle, and drug metabolism. Treatment of Caco-2 cells with 50 μg/ml alkaloid extract and 50 μM chelidonine for up to 48 h resulted in a significant decrease in mRNA levels of P-gp/MDR1, MRP1, BCRP, CYP3A4, GST, and hPXR and in a significant increase in caspase-3 and caspase-8 mRNA. Thus, chelidonine is a promising model compound for overcoming MDR and for enhancing cytotoxicity of chemotherapeutics, especially against leukaemia cells. Its efficacy needs to be confirmed in animal models.

Total syntheses of chelidonine and norchelidonine featuring an enamide-benzyne-[2 + 2] cycloaddition initiated cascade is described. The cascade includes a pericyclic ring-opening and intramolecular Diels-Alder reaction.

In recent years, there has been a significant expansion of the nomenclature of medicinal preparations of plant origin. To a considerable extent, this is due to the creation of complex pharmaceuticals based on several medicinal plants or, sometimes, on a rather large number of such plants. Besides the popular herbal mixtures and solid dosed medicinal forms prepared from raw plant materials, there is growing interest in liquid preparations of complex composition (extracts, elixirs, etc.). Recently, a new complex preparation for the treatment of disorders of the oral mucous membrane and periodontium was created at the State Research Institute of Medicinal and Aromatic Plants (Moscow). This liquid preparation represents a mixed aqueous-ethanol extract from seven official medicinal plants, including peppermint leaves, macleya herbs, garden sage herbs, common yarrow herbs, camomile flowers, purple coneflower herbs, and pot marigold flowers. The extract possesses antiinflammatory, wound-healing, regenerating, and antimicrobial properties. This curative effect is due to various groups of biological substances present in the composition. Garden sage, camomile, peppermint, and common yarrow contain essential oil; macleya contributes alkaloids of the chelidonine group; purple coneflower yields a complex of phenolcarboxylic acids; and pot marigold gives flavonoids, which are also present in all other plants except for macleya. The final preparation also contains carbohydrates, carotenoids, trace elements, organic and fatty acids, steroids, saponins, and other components [1 – 9]. Obviously, standardization of such a muilticomponent preparation poses a relatively complicated problem, which requires an individual approach. The general quality of the preparation is evaluated according to the State Standard OST No. 91500.05.001-00 [10], stipulating tests for the identity, quantitative content of the total of biologically active substances, dry residue (moisture content), alcohol concentration, heavy metals content, and microbiological purity. Since the preparation includes components obtained from various plants containing specific combinations of biologically active substances, the test for identity involves general qualitative group reactions for terpenoids, alkaloids, and phenolic compounds (flavonoids, phenolcarboxylic acids, etc.) and a TLC check. Flavonoids are detected using the cyanidin reaction. The presence of tannins is checked by a reaction with iron hydroxychloride. The total terpenoid content is detected by the reaction with vanillin solution in concentrated sulfuric acid and by TLC [11]. Prior to this, the terpenoids are isolated by double extraction with an organic solvent and the solvent is distilled off in vacuum at a temperature not exceeding 40°C. A qualitative analysis for terpenoids and alkaloids is performed by TLC.

The presence of adult pluripotent stem cells and the amazing regenerative capabilities make planarian flatworms an extraordinary experimental model to assess in vivo the effects of substances of both natural and synthetic origin on stem cell dynamics. This study focuses on the effects of chelidonine, an alkaloid obtained from Chelidonium majus. The expression levels of molecular markers specific for stem or differentiated cells were compared in chelidonine-treated and control planarians. The use of these markers demonstrates that chelidonine produces in vivo a significant anti-proliferative effect on planarian stem cells in a dose-dependent fashion. In response to chelidonine treatment mitotic abnormalities were also observed and the number of cells able to proceed to anaphase/telophase appeared significantly reduced with respect to the controls. Our results support the possibility that chelidonine acts on cell cycle progression by inhibition of tubulin polymerization. These studies provide a basis for preclinical evaluation in vivo of the effects of chelidonine on physiologically proliferating stem cells.

A new alkaloid, methyl 2'-(7,8-dihydrosanguinarine-8-yl)acetate (1), together with six known alkaloids, stylopine (2), protopine (3), norchelidonine (4), chelidonine (5), berberine (6), and 8-hydroxydihydrosanguinarine (7), were isolated from Chelidonium majus. Their chemical structures were primarily established using 1D and 2D NMR techniques and mass spectrometry. The anti-inflammatory activity of the isolates was examined for their inhibitory effects on LPS-induced NO production in macrophage RAW264.7 cells. Among them, compounds 5 and 7 showed strong inhibitory activities toward the LPS-induced NO production in macrophage RAW264.7 cells with IC(50) values of 7.3 and 4.5 μM, respectively. In addition, compounds 5 and 7 inhibited the inductions of COX-2 and iNOS mRNA in dose-dependent manners, indicating that these compounds attenuated the syntheses of these transcripts at the transcriptional level.

Chelidonine, sanguinarine, and chelerythrine are natural benzophenanthridine alkaloids that inhibit taxol-mediated polymerization of rat brain tubulin in the micromolar range. Chelidonine is a weak, competitive inhibitor of colchicine binding to tubulin but does not inhibit podophyllotoxin binding. On the other hand, sanguinarine inhibits both colchicine and podophyllotoxin binding to tubulin with I50 values of 32 and 46 microM, respectively, and chelerythrine inhibits with I50 values of 55 and 60 microM, respectively. The inhibition by these two agents is of the mixed type. Tubulin forms an acid-reversible pseudobase with the imminium ion of sanguinarine, probably through several of its sulfhydryl groups, as shown by the loss of the yellow color of sanguinarine and its 596-nm fluorescence emission peak. Chelidonine, on the other hand, cannot undergo such pseudobase formation, and we conclude that it acts by a different mechanism. A number of previously described pharmacologic effects of these agents may be due to their inhibition of microtubule function.

The effects of benzophenanthridine alkaloids, such as sanguinarine and chelidonine, on monoamine ­oxidase (MAO) activity in mouse brain were investigated. Sanguinarine showed an inhibitory effect on MAO activity in a concentration dependent manner (53.4% inhibition at 25 µM). However, chelidonine did not inhibit MAO activity. The IC50 value of sanguinarine was 24.5 µM. Sanguinarine inhibited non-competitively MAO activity using kynuramine as a substrate. The Ki value for sanguinarine was 22.1 µM. These results suggest that sanguinarine partially contributes to the regulation of catecholamine content. Copyright © 2001 John Wiley & Sons, Ltd.

The solid-state and solution conformations of (+)-chelidonine (1), a biologically active alkaloid, were determined by X-ray diffraction and 1H-NMR spectroscopy, X-Ray diffraction analysis revealed a conformer with B/C ‘anti-type’ cis conjunction, a half-chair of ring B, and a twist half-chair of ring C. One H2O molecule per one alkaloid molecule was cocrystallized and stabilized by H-bonding with OHC(11). Analysis of the thermal behavior of the crystal showed more thermal stability in the monohydrate than the anhydrate. The NMR measurement of concentration and temperature dependences in CDCl3 and in (CD3)2SO suggested that the OH group of 1 was intramolecularly H-bonded to N(5) in (CD3)2SO and intermolecularly H-bonded to the solvent in CDCl3. Conformational-energy calculations by the MNDO method showed that the intramolecular H-bond was little affected by the conformational stabilization of 1.

A group of 11 derivatives of chelidonine was obtained by acylations and/or alkylations of the secondary hydroxyl group with the aim of testing their biological activity. This paper focuses on the new derivatives influence on CNS in mice. The highest activity was observed for compounds 2c, 3c and 4, which produced antinociceptive and antiserotoninergic effects, not recorded for the parent alkaloid 1.

AIM: To investigate the potential effects of chelidonine, the main alkaloid of Chelidonium majus, on telomerase activity and its regulation in HepG2 cells. METHODS: Cytotoxicity of chelidonine for HepG2 cells was determined by neutral red assay. A modified polymerase chain reaction (PCR)-based telomerase repeat amplification protocol was used to estimate relative telomerase activity in chelidonine-treated cells in comparison with the untreated control cells. Relative expression level of the catalytic subunit of telomerase (hTERT) gene and P-glycoprotein (pgp) were estimated using semi-quantitative real-time reverse transcription-PCR (RT-PCR). Cell senescence in treated cells was demonstrated using a β-galactosidase test. RESULTS: Cytotoxicity of chelidonine in HepG2 cells was not dose-dependent and tended to reach plateau immediately after the living cells were reduced in number to slightly higher than 50%. However, 12 μmol/L concentration of chelidonine was considered as LD50, where the maximal attainable effects were realized. Real-time RT-PCR data showed that the expression of pgp increased three-fold in chelidonine treated HepG2 cells in comparison with the untreated controls. Morphologically, treated HepG2 cells showed apoptotic features after 24 h and a small fraction of cells appeared with single blister cell death. The relative expression level of Bcl-2 dropped to less than 50% of control cells at a sub-apoptotic concentration of chelidonine and subsequently increased to higher than 120% at LD50. Telomerase activity was reduced considerably after administration of very low doses of chelidonine, whereas higher concentrations of chelidonine did not remarkably enhance the effect. Real-time RT-PCR experiments indicated a drastic decrease in expression level of hTERT subunit of telomerase under treatment with chelidonine. Repeated treatment of cells with very low doses of chelidonine caused a decline in growth rate by 4 wk and many of the cells appeared to be aged with large volume and dark staining in the β-galactosidase assay. CONCLUSION: Chelidonine reduces telomerase activity through down-regulation of hTERT expression. Senescence induction might not be directly caused by reducing telomerase activity as it occurs after a few population doublings.

A simple and precise method using ion-pair high-performance liquid chromatography was developed for the simultaneous determination of eight isoquinoline alkaloids, namely chelidonine, berberine, protopine, coptisine, tetrahydrocoptisine, 6-methoxydihydrochelerythrine, 6-methoxydihydrosanguinarine and dihydrosanguinarine, in Chelidonium majus L.. A reversed-phase system consisting of a chemically bonded ODS silica gel column and 0.05 M tartaric acid—methanol—acetonitrile (44:10:46) containing 0.5% sodium dodecyl sulphate as the mobile phase was used. The eight alkaloids were completely separated within 40 min. The analytical results for various samples are presented.

From the underground part of Stylophorum lasiocarpum (OLIV.) FEDDE (0.58% alkaloids) (-)-stylopine and coptisine (as chloride IIa) were isolated as the principal alkaloids. Smaller quantities of sanguinarine, macarpine, chelerythrine, chelirubine, chelilutine (as the respective chlorides IIIa, IIIb, IIIc, IIId, IIIe) protopine, corysamine (as chloride IIb), berberine (as chloride IIc) and (-)-trans-N-methylstylopinium hydroxide (as iodide IVa) were also isolated. Chromatographically were detected small amounts of chelidonine, allocryptopine, cryptopine, scoulerine, isoboldine, magnoflorine and salt of cis-N-methylstylopinium. The aerial part (0.16% alkaloids) afforded (-)-stylopine, coptisine and (+)-chelidonine as dominant alkaloids and small amounts of protopine and (-)-cis-N-methylstylopinium hydroxide (as iodide IVb).Chromatography proved negligible amounts of sanguinarine, chelirubine, cryptopine, scoulerine, isoboldine, corytuberine, corysamine, berberine, magnoflorine and trans-N-methylstylopinium iodide. The so far undescribed 8-methoxydihydromacarpine (VIb) was prepared.

Projekat ministarstva br. 143026B i 143041B