Alkyl and carboxylalkyl esters of 4'-demethyl-4-deoxypodophyllotoxin: synthesis, cytotoxic, and antitumor activity.

Esters of 4'-demethyl-4-deoxypodophyllotoxin (DDPT) with alkanoic acids and alkanedioic acids were prepared and tested for cytotoxic and antitumor activity. Among 19 esters, esters of propanoic acid, tetradecanedioic acid, 13-carboxyundecanoic acid, and hexadecanedioic acid improved the antitumor activity compared with that of the starting compounds, DDPT.

Cytotoxic 2',5'-dihydroxychalcones with unexpected antiangiogenic activity.

A series of 2',5'-dihydroxychalcones were synthesized and evaluated for cytotoxicity against tumor cell lines and human umbilical venous endothelial cells (HUVEC). It was found that chalcones with electron-withdrawing substituents on the B ring exhibited potent cytotoxicity against a variety of tumor cell lines while compounds with electron-releasing groups were less potent in general. Those compounds with B ring replaced by extended or heteroaromatic rings exhibited significant bioactivity. Several compounds were shown to have marked cytotoxic selectivity towards HUVECs. Especially, among the synthesized compounds, 2-chloro-2',5'-dihydroxychalcone (2-3) showed the highest selectivity index up to 66 in comparison to HCT116 cells. This compound also exhibited strong inhibitory effects on the HUVEC tube formation in an in vitro model. When administered into BDF1 mice bearing Lewis lung carcinoma cells at 50 mg kg(-1) day(-1), 2-3 was found to inhibit the growth of tumor mass by 60.5%.

Pulsatilla saponin D: the antitumor principle from Pulsatilla koreana.

By bioassay-guided separation, an already known saponin, Pulsatilla saponin D was isolated from the root of Pulsatilla koreana Nakai as a antitumor component when evaluated by in vivo antitumor activity as well as in vitro cytotoxic activity test. It showed potent inhibition rate of tumor growth (IR, 82%) at the dose of 6.4 mg/kg on the BDF1 mice bearing LLC cells.

5-Arylidene-2(5H)-furanone derivatives: synthesis and structure-activity relationship for cytotoxicity.

Thirty-eight 5-arylidene-2(5H)-furanone derivatives possessing halo-, methoxy-, oxo-, dioxo-, and thiophenyl groups as well as anthraquinone and naphthquinone moieties were synthesized, and their cytotoxicity was evaluated against various cancer cell lines. The introduction of halogen atoms or nitro group at aromatic ring of 5-arylidene-2(5H)-furanone was shown to increase the cytotoxicity with 5-(3-nitrobenzylidene)-2(5H)-furanone (21) being the most potent. Among anthracenyl or naphthalenyl derivatives, (E)-5-[2-(1,4-dimethoxy-9,10-dioxo) anthracenyl]-2(5H)-furanone (34) showed the most potent cytotoxic activity.

2,3-dibenzylbutyrolactones and 1,2,3,4-tetrahydro-2-naphthoic acid gamma-lactones: structure and activity relationship in cytotoxic activity.

Dibenzyl-g-butyrolactone and 1,2,3,4-tetrahydro-2-naphthoic acid gamma-lactone (TNL) derivatives were synthesized and evaluated for cytotoxic activity against some cancer cell lines. It was found that TNL derivatives with a shorter distance between C-4 in ring A and C'-2 in ring C were more cytotoxic, while dibenzyl-gamma-butyrolactones with a longer one were nearly inactive. In TNL series, presence of 3,4-dioxy group in ring A and 2-methoxy group in ring C was essential for the enhancement of the activity.

Antiangiogenic activity of Bupleurum longiradiatum on human umbilical venous endothelial cells.

The ethyl acetate fraction of Bupleurum longiradiatum was found to have an inhibitory effect on the tube-like formation of human umbilical venous endothelial (HUVE) cells. The active compounds, isolated from the fraction, were identified as acetylbupleurotoxin (P1) and bupleurotoxin (P2). The compounds P1 and P2 completely inhibited the tube-like formation of HUVE cells at 30 microg/ml, below the cytotoxic concentration. But, they did not exhibit antitumor activity on BDF1 mice bearing Lewis lung carcinoma cells despite their antiangiogenic activity.

Synthesis and cytotoxicity of 2,5-dihydroxychalcones and related compounds.

A series of 2, 5-dihydroxychalcones and related compounds were synthesized, and their cytotoxicities against tumor cell lines and human umbilical venous endothelial cells (HUVEC) evaluated. It was found that chalcones, with electron-withdrawing substituents on an A ring, exhibited significant cytotoxicities. Among the synthesized compounds, 2'-chloro-2, 5-dihydroxychalcone (9) was most potent, with an IC50 value as low as 0.31 microg/mL. This compound also exhibited a significant cytotoxic selectivity toward HUVEC.

Synthesis and cytotoxicity of some rigid derivatives of methyl 2,5-dihydroxycinnamate.

Eight rigid compounds designed as esterase-stable analogues of methyl 2,5-dihydroxycinnamate (1) were synthesized. These derivatives include 2-(2',5'-dihydroxybenzylidene)cyclopentenone (3a), 2-(2',5'-dihydroxybenzylidene)cyclohexanone (3b), 2,6-bis(2',5'-dihydroxybenzylidene)cyclohexanone (4b), 2,6-bis(2',5'-dihydroxybenzylidene)cyclopentenone (4a), (E)-3-(2',5'-dihydroxybenzylidene)pyrrolidin-2-one (5), (E)-5-(2',5'-dihydroxybenzylidene)-1,2-isothiazolidine-1,1-dioxide (6), 4-(2',5'-dihydroxyphenyl)-5H-furan-2-one (7), and 3-(2',5'-dihydroxyphenyl)cyclopent-2-ene-1-one (8). Among the eight compounds, the furanone 7 and cyclopentenone 8 showed the most potent cytotoxicity with IC50 values of 0.39-0.98 microg/mL. Compound 8 was further brominated, phenylated and methylated at the alpha position to give three corresponding analogues, including 2-bromo-3-(2',5'-dihydroxyphenyl)cyclopent-2-ene-1-one (24), 3-(2',5'-dihydroxyphenyl)-2-phenylcyclopent-2-ene-1-one (27), and 3-(2',5'-dihydroxyphenyl)-2-methylcyclopent-2-ene-1-one (28). Among the three, the most enhanced activity was observed with the phenylated compound 27.

Synthesis, cytotoxicity and antitumor activity of 2,3-diarylcyclopent-2-ene-1-ones.

Two series of 2,3-diarylcyclopent-2-ene-1-ones including 2-aryl-3-(2,5-dihydroxyphenyl)cyclopent-2-ene-1-ones (2a-2f) and 3-aryl-2-3',4',5'-trimethoxyphenyl)cyclopent-2-ene-1-one (3a-3j) were synthesized and evaluated for the cytotoxicity against three tumor cell lines; B16F10, HCT116 and A431. It was found that the 3,4,5-trimethoxy substituent was optimal for the bioactivity of compounds in series 2. Meanwhile, compounds in series 3 exhibited the most potent cytotoxicity with 3-aryl ring being 4-methoxyphenyl (compound 3f), (3-hydroxy-4-methoxy)phenyl (compound 3e), or (3-amino-4-methoxy)phenyl (compound 3j).

New constituents from Crinum latifolium with inhibitory effects against tube-like formation of human umbilical venous endothelial cells.

Six compounds (1-6) were isolated from the methanol extract of Crinum latifolium by bioassay-guided separation. Among the six isolates, compounds 2 and 6 were new metabolites. Their structures were established as 4-senecioyloxymethyl-3,4-dimethoxycoumarin (2) and 5,6,3'-trihydroxy-7,8,4'-trimethoxyflavone (6) based on spectroscopic analyses. Compound 2 was found to be strongly inhibitory against the in vitro tube-like formation of human umbilical venous endothelial cells (HUVECs) while manifesting no cytotoxicity in tumor cell lines (B16F10, HCT116). Significant inhibitory activity (inhibition percentage, 53.5%) was still observed at concentrations as low as 1 microg/mL. Compound 6 showed a modest inhibitory effect on the tube-like formation of HUVECs. Other compounds, including cycloartenol (1), 4',7-dihydroxy-3'-methoxyflavan (3), 4',7-dihydroxyflavan (4), and 2',4',7-trihydroxydihydrochalcone (5) were found to be nearly inactive.

Triterpenoid saponins from the roots of Pulsatilla koreana.

Six new saponins, five lupanes (1-5) and one oleanane (6), along with 11 known saponins, were isolated from the roots of Pulsatilla koreana. The structures of the new saponins were found to be 23-hydroxy-3beta-[(O-alpha-L-rhamnopyranosyl-(1-->2)-O-[O-beta-D-glucopyranosyl-(1-->4)]-alpha-L-arabinopyranosyl)oxy]lup-20(29)-en-28-oic acid (1), 23-hydroxy-3beta-[(O-beta-D-glucopyranosyl-(1-->3)-O-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranosyl)oxy]lup-20(29)-en-28-oic acid (2), 3beta-[(O-alpha-L-rhamnopyranosyl-(1-->2)-O-[O-beta-D-glucopyranosyl-(1-->4)]-alpha-L-arabinopyranosyl)oxy]lup-20(29)-en-28-oic acid (3), 3beta-[(O-beta-D-glucopyranosyl-(1-->3)-O-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranosyl)oxy]lup-20(29)-en-28-oic acid (4), 23-hydroxy-3beta-[(O-beta-D-glucopyranosyl-(1-->4)-alpha-L-arabinopyranosyl)oxy]lup-20(29)-en-28-oic acid (5), and hederagenin 3-O-beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside (6). Their structures were determined on the basis of 1D and 2D NMR ((13)C NMR, (1)H NMR, (1)H-(1)H COSY, HMQC, and HMBC) methods, FABMS, and hydrolysis. All isolated compounds were evaluated for their cytotoxic activity against A-549 human lung carcinoma cells.

Antitumor activity of Pulsatilla koreana saponins and their structure-activity relationship.

Seventeen saponins isolated from the root of Pulsatilla koreana were examined for their in vitro cytotoxic activity against the human solid cancer cell lines, A-549, SK-OV-3, SK-MEL-2, and HCT15, using the SRB assay method, and their in vivo antitumor activity using BDF1 mice bearing Lewis lung carcinoma (LLC). The saponins 5-17, with a free acidic functional group at C-28 of aglycon, exhibited moderate to considerable cytotoxic activity, however, the saponins 1-4, esterified with a trisaccharide at C-28 of aglycon, did not exhibit cytotoxic activity (ED50; >300 microM). Among them, oleanolic acid 3-O-alpha-L-rhamnopyranosyl-(1-->2)-[beta-D-glucopyranosyl-(1-->4)]-alpha-L-arabinopyranoside (10) exhibited the most potent cytotoxic activity (ED50; 2.56, 2.31, 1.57, 8.36 microM, respectively). In vivo test, hederagenin 3-O-alpha-L-rhamnopyranosyl-(1-->2)-[beta-D-glucopyranosyl-(1-->4)]-alpha-L-arabinopyranoside (6, Inhibition Ratio, IR; 66.9%) exhibited more potent antitumor activity than taxol (IR; 35.8%) and doxorubicin (IR; 62.1%). Also, hedragenin 3-O-beta-D-glucopyranosyl-(1-->4)-O-beta-D-glucopyranosyl-(1-->3)-O-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside (17, IR; 50.3%) exhibited potent antitumor activity. These two saponins were identically comprised of a hederagenin aglycon moiety and a sugar sequence O-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside at C-3 of the hederagenin, suggesting that the two elements are essential factors for the antitumor activity.

[The Wormicidal Substance Of Fresh Water Fishes On Clonorchis Sinensis V. Purification And Chemical Characterization Of Clonorchicidal Substance From Epidermal Mucus Of Cyprinus Carpio]

As a series of studies to clarify clonorchicidal substances in body surface mucus of some fresh-water fishes, the substance in the epidermal mucus of Cyprinus carpio was isolated by silica gel column and thin layer chromatography and analyzed for its chemical nature. Wormicidal trial was done in vitro, and the results obtained are summarized as follows: The mucus was extracted by ethyl ether and separated into 4 fractions by column chromatography using benzene as solvent. The second fraction with yellowish red colour among them showed the strongest clonorchicidal effect. The yellowish red fraction obtained by column chromatography was then fractionated into 6 spots by thin layer chromatography with petrol. ether/chloroform(30/70, v/v), and the Rf. 0.714 spot among the 6 spots showed the strongest effect. The Rf. 0.714 spot was further fractionated into 6 spots by thin layer chromatography with benzene/acetone (90/10, v/v), and the Rf. 0.800 spot among the later 6 spots revealed the strongest effect. The Rf. 0.800 spot was chromatographed on column with benzene and 2 fractions were obtained. The second fraction of light brown colour represented the final purified fraction. By these purification procedures, clonorchicidal substance was purified 15-fold with 0.03 percent yield from the mucus of C. carpio, and 10mg of the final fraction killed the cercaria in 26 min, the metacercaria in 115 min, and the adult in 443 min. Infra red and nuclear magnetic resonance spectrometric analysis of the purified substance revealed that the substance belongs to an ethyl ester of unsaturated fatty acid with 2 double bonds, 15 methylene groups and l methyl group.

Inhibitory effects of Vietnamese medicinal plants on tube-like formation of human umbilical venous cells.

Seven of 58 plant materials from Vietnamese medicinal plants showed strong to moderate inhibitory activity on the tube-like formation induced by human umbilical venous endothelial cells in the in vitro angiogenesis assay. These plant materials include the herb of Ephedra sinica, leaves and stem of Ceiba pentandra, seed of Coix lachryma-jobi, rhizome of Drynaria fortunei, fruits and stem of Illicium verum and stem of Bombax ceiba. Of these, the methanol extracts of the herb of Ephedra sinica and stem of -Ceiba pentandra exhibited the strongest activities with inhibition percentages of 89.3% and 87.5% at 30 and 100 microgram/mL, respectively.

Inhibitory effect of Adonis amurensis components on tube-like formation of human umbilical venous cells.

Antiangiogenic activity-guided fractionation and isolation carried out on the methanol extract of Adonis amurensis led to the identification of three compounds, namely cymarin, cymarol, and cymarilic acid. Amongst the three compounds, cymarilic acid was isolated from this plant for the first time. This compound showed no significant cytotoxicity against tumor cell lines but was found to be strongly inhibitory toward tube formation induced by human umbilical venous endothelial (HUVE) cells. Cymarin and cymarol exhibited potent cytotoxicity against a human solid tumor cell line A549 (human lung carcinoma), while being inactive on murine leukemic cells (L1210).

[The Wormicidal Substances Of Fresh Water Fishes On Clonorchis Sinensis: I Preliminary Research On The Wormicidal Substance From Mucous Substance Of Carassius Carassius]

The present work was to observe the wormicidal effects of the external mucous substance of Carassius carassius and Pseudorasbora parva on cercaria, metacercaria (liberating) and adult of Clonorchis sinensis. The mucous substance which was extracted by ether was separated into various spots on the silica gel thin layer chromatography and separated into various fractions in the silica gel column chromatography, using petroleum ether(30%) and chloroform(70%) as solvent. 1. R(f) 0.952 value of the silica gel thin layer chromatography and the first fraction (reddish yellow) frorn the silica gel column chromatography of the ether extracts from C. carassius had the wormicidal effect on all of the developing stages of C. sinensis. Cercaria was killed within 14 minutes, metacercaria within 27 minutes and adult 2.5 hours by the first fraction. 2. But in the normal saline solution (control) and in the other fraction, cercaria and metacercaria were alive more than 24 hours and adult more than 72 hours. 3. The extracts from the mucous substance of P. parva that is well penetrated by cercaria had no wormicidal effect on them.

[The Wormicidal Substances Of Fresh Water Fishes On Clonorchis Sinensis: III. Seasonal variation on the wormicidal substances of mucous substances of Carassius carassius]

The epidermal mucous substances of C. carassius which were extracted by ether were separated into various spots on the silica gel thin layer chromatography and separated into various fractions in the silica gel column chromatography, using petroleum ether (30 %) and chloroform(70 %) as a solvent. The wormicidal effects of the spots and the fractions on the cercaria, the liberating metacercaria and the adult of C. sinensis, were observed by month giving attention to ambient water temperature. 1. Among the four spots of the thin 1ayer chromatography of the ether extracts from C. carassius, the uppermost first spots (Rf. 0.924~0965) had a strong wormicidal effect on the cercaria in May, June, July, August and September, and the cercaria which came in contact with the materials was killed in 29~49 min., but in other months (Rf. 0.612~0.752) it was killed only after 50~115 min. 2. In column chromatography of ether extracts from C. carassius, the first fractions which had a wormicidal effect killed the metacercariae within 38~163 min. in May, June, July, August, and September, but it took about 120~458 min. to kill them in other months, meanwhile the wormicidal time of the fractions on the adults was 445~835 min. in May, June, July, August and September, but was 878~2,130 min. in other months. 3. The Rf. value of the uppermost first spot of the thin layer chromatography of ether extrart from the mucous substance of C. carassius which was reared in the aquarium controlled at 23 degrees C was 0.79, and its first fraction of column chromatography killed the metacercaria and the adult of C. sinensis within 315, and 1,850 min. respectively. As summarized above, the wormicidal materials of the epidermal mucous substances of C. carassius had no relation to ambient water temperature, but the secretion period of this wormicidal materials was the same as the liberating period of the cercariae from Parafossarulus manchouricus.

[The Wormicidal Substances Of Fresh Water Fishes On Clonorchis Sinensis: II. Preliminary Research On The Wormicidal Substances From Mucous Substances Of Various Fresh Water Fishes]

The present work which was investigated in July 1979, was to observe the wormicidal effects of the external mucous substances of 9 species of fresh water fishes (Cyprinus carpio, Parasilurus asotus, Anguilla japonica, Ophicephalus argus, Carassius carassius; golden crusian carp, Misgurnus anguillicaudatus, Zacco platypus, Pseudorasbora parva and Carassius carassius; crusian carp) on cercaria, liberated metacercaria and adult of Clonorchis sinensis. The mucous substances extracted by ether from the above 9 species of fishes were separated into many spots on the silica gel thin layer chromatography, and divided into many fractions in the silica gel column chromatography which used petroleum ether (30%) and chloroform (70%) as a solvent. 1. On the silica gel thin layer chromatography of ether extracts from 9 species of fresh water fishes, each of the Rf. values which had wormicidal effect on the cercaria of C. sinensis were different from others; Rf. value of C. carassius (crusian carp) was 0.937, 0.709 in O. argus, 0.612 in A. japonica, 0.576 in P. asotus, 0.451 in C. carpio, 0.701 in Z. platypus, 0.385 in C. carassius(golden crusian carp) and 0.15 in P. parva. Time for wormicide was different from each other, too. It took 14 min. in a case of C. carassius(crusian carp), 25 min. in Z. platypus, 26 min. in C. carassius(golden crusian carp), 28 min. in C. carpio, 30 min. in P. asotus, 35 min. in O. argus, 40 min. P. parva and 180 min. in A. japonica. But any of the spots of M. anguillicaudatus did not show wormicidal effect on the cercaria within 10 hours. 2. In the silica gel column chromatography of ether extracts from 9 species of fresh water fishes, the fractions which had wormicidal effect on the metacercaria of C. sienesis were different from each other; in a case of C. carassius(crusian carp), the first fraction only had wormicidal effect, the 2nd in P. asotus, the 3rd in O. argus, the 4th in C. carassius(golden crusian carp), the 2nd in C. carpio and the 4th in P. parva. Time for wormicide was 40 min. in C. carassius(crusian carp), 52 min. in P. asotus, 74 min. in C. carassius(golden crusian carp), 92 min. in O. argus, 95 min. in C. carpio and 140 min. in P. parva, but any of the fractions of A. japonicas, M. anguillicaudatus and Z. platypus did not show wormicidal effect within 10 hours. 3. The fractions which had wormicidal effect on the metacercaria of C. sinensis had also the same effect on the adult, but it took longer time to kill them. It took 3 hours in a case of C. carassius(crusian carp), 4 hrs. in C. carassius(golden crusian carp), 5 hrs. in P. asotus, 6 hrs. and 6 min. in O. argus, 6 hrs. and 10 min. in C. carpio and half and 6 hrs. in P. parva. But any of the fractions of A. japonica, M. anguillicaudatus and Z. platypus did not show wormicidal effect within 24 hrs.

[Localization and isolation of anthelmintic activity on Clonorchis sinensis from stem cortex of Machilis thunbergii]

The present work was to localize and isolate the anthelmintic substance from stem cortex of Machilis thunbergii which grows in all parts of the Korea peninsula. The dried M. thunbergii bark was extracted with a solvent mixture (Methanol: Ethyl acetate/1:1) by a Soxhlet extractor at 70 degrees C for 24 hours, and then the extract was separated into solid matter (negative wormicidal) and benzene extract (positive wormicidal) by benzene solvent. The anthelmintic action was observed on the cercaria, the liberated metacercaria and the adult of Clonorchis sinensis. On the silica gel thin layer chromatography with a spot of benzene extract in benzene solvent, nine spots were observed in iodine chamber and the spot of Rf. 0.16 value of them appeared the most strong wormicidal effect on the cercaria and the liberated metacercaria of C. sinensis. Afterward, a 0.3 g colourless solid substance which was equivalent to the Rf. 0.16 value spot was isolated from the benzene extract of 3 kg M. thunbergii stem cortex by means of silica gel column chromatography using benzene carrier, and the adult of C. sinensis was killed within 6 minutes by 0.05 percent solution of the substance.

Antitumor activity of unsaturated fatty acid esters of 4'-demethyldeoxypodophyllotoxin.

Unsaturated fatty acid esters of 4'-demethyldeoxypodophyllotoxin (DDPT) were prepared and tested for antitumor activity. The esters showed increased in vivo antitumor activity despite the lower in vitro activity than DDPT. Especially, the ester (DFE12) of all-cis-11,14-eicosadienoic acid was much better (IR, 83%) than VP-16 (IR, 60%) without loss of body weight. Unsaturated fatty acids could be evaluated to be good carrier vehicles of DDPT.