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[Observations on the mutagenic power of hycanthone in mammals].

Male mice from the C57Bl strain were given 150 mg/kg of hycanthone, an antischistosomial drug. Spermatocytes derived from the treated spermatogonia as well as the spermatocytes from the F1 male offspring sired the first three weeks after treatment were examined for the presence of chromosome rearrangements. Since no reciprocal translocation was recorded, it is concluded that treatment with hycanthone is not susceptible to produce transmissible chromosome anomalies in male mammalian germ cells.

Animals↗

Liver pathology in hycanthone hepatitis.

Histological examination of liver tissue from 3 patients who died 6, 4, and 9 days, respectively, after an intramuscular injection of 3 +/- 0.5 mg of hycanthone per kg of body weight for schistosomiasis, showed features of a toxic hepatitis with massive hepatic necrosis, many acidophil bodies and minimal inflammatory response in 2 patients, and confluent necrosis with early fibrosis in 1. Liver cells showed prominent fatty change. The former 2 patients had evidence of schistosomal granulomatous hepatitis. A follow-up liver biopsy in a 4th patient who had had hycanthone hepatitis with liver failure 1 year before showed good recovery with features of chronic persistent hepatitis.

Chemical and Drug Induced Liver Injury↗

Hycanthone therapy in selected patients with S. mansoni and S. haematobium infections in the Sudan.

The results of a clinical trial of hycanthone in 601 selected Sudanese patients with S. mansoni and S. haematobium infections are reported. Hycanthone was given as a single intramuscular injection at a dose of 3.0 mg per kilogramme body weight. The commonest side effects were nausea and vomiting which occurred in 35 per cent and 32 per cent respectively. No incidence of acute hepatic damage and no deaths were encountered. About one third of patients reported for follow-up. Cure rate for S. mansoni was 90 per cent at 6 weeks of treatment and 97 per cent at three months and six months, while for S. haematobium infection it was 82 per cent at 6 weeks and 91 per cent at 3 months and 6 months. A significant reduction in egg excretion was achieved in those who were not cured.

Adolescent↗

Hepatotoxicity of hycanthone in patients with metastatic breast cancer.

Hycanthone is an antischistosomal drug with promising antitumor activity against experimental animal tumors. In phase I clinical trials, hepatitis weas the dose-limiting toxic effect and a dose of 60-70 mg/m2/day for 5 days was reported as the maximum-tolerated dose. In a phase II study of hycanthone in patients with breast cancer we have recently observed severe hepatotoxicity, even at lower doses, which resulted in two drug-related deaths. From this study and the previously available data, we conclude that this drug is too toxic for human trials at the currently recommended doses. Its radiosensitizing effect may be worth exploring at a lower dose level.

Adult↗

Low mutagenic activity of four hycanthone analogues in Drosophila melanogaster.

Four structural analogues of the antischistosomal drug hycanthone, indicated as IA-3, IA-4, IA-5, and IA-6, were tested for their ability to induce sex-linked recessive lethal mutations in Drosophila melanogaster. The compounds were administered to adult male flies, by abdominal injection or by feeding. Although in some cases a slight enhancement of the mutation frequency was observed, the analogues tested are considerably less mutagenic in this system than hycanthone itself, the concentrations applied being approximately of equal molarity.

Animals↗

Hycanthone resistance: development in Schistosoma mansoni.

Following the administration of relatively high doses of the antischistosomal drug hycanthone to mice and hamsters infected with Schistosoma mansoni, a number of the worms survived. After a period of 6 to 12 months these parasites resumed production of viable eggs that gave rise to schistosomes that proved resistant to hycanthone and to two other related antischistosomal compounds. This drug resistance has remained stable for three subsequent generations of worms.

Animals↗

Hycanthone: a frameshift mutagen.

Rapid spot-test screening of antischistosomal agents reveals that hycanthone is a potent frameshift mutagen while the closely related compound, miracil D, is nonmutagenic in Salmonella. Both hycanthone and miracil D are frameshift mutagens for T4 bacteriophage during growth in Escherichia coli.

Anthelmintics↗

Metabolism of the schistosomicidal agent hycanthone by rats and rhesus monkeys.

The absorption, distribution, and excretion of hycanthone were studied with rats and rhesus monkeys using tritiated materials at the therapeutic dose recommended for man (3.0+/-0.5 mg/kg). Nine pairs of each species received single intramuscular doses of randomly tritiated (specific activity 193.7 mCi/mmol) hycanthone methanesulfonate and were then sacrificed at intervals between 15 min and 72 h after medication. Peak blood and tissue concentrations occurred 30-60 min after administration (plasma half-life-45 min). The highest concentrations were observed in the liver, spleen, kidneys and adrenals, but decreased rapidly (more than 80% of the dose was excreted in 48-72 hours). In monkeys a high concentration of the compound was found in the bile (hours 1-8), probably conjugated to glucuronic acid.Radiochromatography showed only unchanged drug in the blood and tissues, except in the liver where rapid conversion occurred to sulfoxide in the rat and to the deethyl analogue in the monkey.

Adrenal Glands↗

An evaluation of the micronuclei test using triethylenemelamine, trimethylphosphate, hycanthone and niridazole.

To determine the feasibility of the micronuclei procedure for cytogenetic studies, a comparatively weak chromosome breaking agent, trimethylphosphate (TMP) and the potent alkylating agent, triethylenemelamine (TEM) were evaluated. The procedure followed was that of Matter and Schmid with the following modifications: (a) direct flushing of bone marrow with 0.2 ml calf fetal serum. (b) air drying slides for a period of only I h, and (c) the use of pH 6.0 phosphate buffer to dilute both Wright and Giemsa stains. With this technique a dose response curve was generated for both TMP and TEM, using mice as the experimental animal. With TMP, a doubling over background was found when a concentration of 0.5 g/kg per day for five days was administered. To establish a statistically significant doubling dose over the control, a minimum of five animals must be used with 2000 polychromatic cells being analyzed per animal. Of the two antischistosomal agents tested, hycanthone yielded an increase of 20-fold in the number of micronuclei over control at 40 mg/kg administered i.p. for five days, while with niridazole no increase in micronuclei at several concentrations tested both by single and multiple injection was found. The results obtained with these compounds compare favorably with what has been reported for the standard in vivo metaphase analysis.

Alkylating Agents↗

High proportion of multi-locus deletions among hycanthone-induced X-linked recessive lethals in Drosophila melanogaster.

328 X-linked recessive lethal mutations induced in late spermatids by hycanthone methanesulfonate were tested for coverage by duplications that comprised, in total, about 24% of the euchromatic X chromosome; 78 lethals appeared to be covered. Crossover localization tests of a random sample of 38 non-covered lethals revealed 4 chromosomes carrying a lethal within a duplicated segment. Lethals localized to a particular region were crossed to reference deficiencies and single-locus mutations, and inter se, to ascertain their genetic extent. The proportion of multi-locus deletions among these 78 covered and 4 non-covered lethals was 3/48, 1/10 and 13/24 for the distal, medial and proximal regions, respectively. A storage period of 9 days did not noticeably influence these proportions. In the sample of 38 non-covered lethals, and among 17 of the covered single-site lethals, 4 cases of strong crossover suppression were detected. Comparison of these results with data obtained with other mutagens suggests that induction of multi-locus deletions, and possibly of other types of chromosome rearrangement, could in part depend on other mechanisms than those acting in the formation of translocations and chromosome loss. For the purpose of mutagen testing, these findings imply that, in Drosophila, results in the regular genetic tests for chromosome breakage events do not always accurately predict the capacity of a mutagen to induce multi-locus deletions. This is of importance since transmissible multi-locus deletions have been considered a significant source of genetic damage in man.

Animals↗

A genomic change associated with the development of resistance to hycanthone in Schistosoma mansoni.

Ribosomal gene probes were used to investigate the genetic basis of drug resistance in schistosomes in a model where resistance to the anthelmintic hycanthone (HC) is generated by exposing immature worms to the drug. Two strains of Schistosoma mansoni, JHU and NMRI, were used. Drug resistance could be produced in the JHU strain by treatment with HC, but was also found to occur spontaneously. In contrast, it was not possible to detect or produce resistance to HC in the NMRI strain. A genomic alteration accompanied the development of resistance. The change was evidence by the occurrence of restriction fragment length polymorphisms (RFLPs) when Southern blots of genomic DNA from HC-resistant worms were hybridized with the ribosomal probe pSM389, which contains part of the small rRNA gene plus non-transcribed spacer (NTS) sequence. The most reliable marker of HC-resistance was a 3.6-kb BamHI fragment which was present and heritable in 7 drug-resistant lines derived from the JHU strain but absent from the parent JHU population and from NMRI parasites. The universal absence of the 3.6-kb RFLP in HC-sensitive individuals and its presence in the drug-resistant progeny suggest that resistance results from an induced change in the population rather than from selection of HC-resistant parasites. The rRNA gene sequence responsible for detecting the 3.6-kb RFLP appears to be localized either to the NTS or to the 5' end of the small rRNA gene, since hybridization to a probe containing sequence from the rRNA gene contiguous and downstream from the insert of pSM389 failed to reveal the RFLP. These results show that the development of resistance to HC is accompanied by a genomic rearrangement.

Animals↗

Mechanism of hycanthone mutagenesis: the induction of temperature-sensitive mutations in Drosophila melanogaster.

A total of 358 sex-linked recessive lethals induced by hycanthone methane sulphonate (HMS) were checked for temperature sensitivity, along with 239 EMS-induced lethals as a positive control. Only about 1-2% of the HMS-induced lethals were temperature-sensitive, in contrast to about 7-8% for EMS-induced lethals. This can be reasonably explained by assuming that in Drosophila, HMS mainly acts by inducing frame-shift mutations or deletions.

Animals↗

Hycanthone dose-response in Schistosoma mansoni infection in Kenya.

The recommended doses of the drugs now used to cure schistosomiasis mansoni may be associated with toxic side-effects. Since Schistosoma mansoni does not multiply in the human host and the disease seems to be closely associated with the intensity of infection, it may not be necessary to use 100% lethal antischistosomal doses, particularly in endemic areas. A dose-response to the antischistosomal drug, hycanthone was established for three different doses in 169 patients with heavy S. mansoni infections in the Machakos district of Kenya. The highest dose used (1.5 mg/kg or half the recommended package-insert dose) resulted in a 96% decrease in egg output (equivalent ot death of the worms); 0.75 mg/kg in an 85% decrease; and 0.375 mg/kg in an 11% decrease one month after treatment. In contrast to the vomiting common with the package-insert dose (3.0 mg/kg), there were no side-effects with any of the lower doses.

Adolescent↗

Analogues of hycanthone and lucanthone as antitumor agents.

Hycanthone analogues (5 and 6) containing 7-substituted hydroxyl groups were prepared and evaluated as antitumor agents. These compounds were significantly more active than the corresponding unsubstituted derivatives. The 7-hydroxylated 4-(hydroxymethyl)-9H-xanthen-9-ones, 11 and 12, were also active antitumor agents. However, the 7-hydroxy-9H-xanthen-9-one counterparts of the 7-hydroxylucanthones were totally devoid of antitumor activity. Results obtained thus far are consistent with the hypothesis that 4-hydroxymethyl substituents in the 9H-xanthen-9-one and 9H-thioxanthen-9-one series are required for antitumor activity.

Animals↗

Hycanthone and its congeners as bacterial mutagens.

Hycanthone methanesulfonate (HCT) was shown to induce "forward" and "reverse" mutations in Salmonella typhimurium and Escherichia coli. Mutational effects of HCT on S. tyhhimurium TA1532 were concentration and time dependent. A comparison of mutagenic potency for TA1532 was made between HCT and the frame-shift mutagens quinacrine and ICR-191. An investigation of structure-activity relationships revealed the substituent in the 4-position of ring A to be critical for mutagenicity. Activity was found when this group was a hydroxymethyl (i.e., HCT) or an aldehyde (Win 25,315), but the analogues having a carboxyl group (Win 25,850) or methyl group (lucanthone) in this positionwere inactive. Removal of a single ethyl group from the side chain did not affect mutagenic activity inasmuch as the potency of desethyl HCT (Win 27,262) equaled that of HCT on a molar basis. A marginal activity was found with a sample of HCT sulfoxide (win 27,266), but this sample was found to contain traces of HCT. The HCT analogue with a terminal N-oxide in the side chain (Win 29,329) was inactive at the concentration tested.

Bacteria↗

Anti-schistosomal drugs: observations on the mechanism of drug resistance to hycanthone, and on the involvement of host antibodies in the mode of action of praziquantel.

This paper reports recent observations from our laboratory dealing with the anti-schistosome drugs hycanthone (HC) and praziquantel (PZQ). In particular, we discuss a laboratory model of drug resistance to HC in Schistosoma mansoni and show that drug sensitive and resistant lines of the parasite can be differentiated on the basis of restriction fragment length polymorphisms using homologous ribosomal gene probes. In addition, we summarize data demonstrating that effective chemotherapy of S. mansoni infection with PZQ in mice requires the presence of host anti-parasite antibodies. These antibodies bind to PZQ treated worms and may be involved in an antibody-dependent cellular cytotoxicity reactions which result in the clearance of worms from the vasculature.

Animals↗

Genetic complementation analysis of two independently isolated hycanthone-resistant strains of Schistosoma mansoni.

The objective of this study is to determine whether various hycanthone resistant strains of schistosomes which have been independently isolated are all affected in the same gene. A strain obtained from a Brazilian patient was compared with a strain of Puerto Rican origin selected in the laboratory. If the mutation conferring resistance involved two different genes, one would expect that progeny of a cross between the two strains would show complementation, i.e. it would be sensitive to the drug. We have performed such a cross and obtained F1 hybrid worms which were essentially all resistant, thus suggesting that the mutation conferring resistance in the two strains involves the same gene.

Animals↗