Analysis of Usnea fasciata crude extracts with antineoplastic activity.

Different fractions, isolated from the lichen Usnea fasciata, were analyzed by PC, TLC, and RP-HPLC. Analysis of the organic phases, mainly containing phenolics, revealed that usnic acid is the main product from secondary metabolites, whereas the polysaccharides isolichenin and raffinose are the most abundant water-soluble carbohydrates. Fractions containing usnic acid, as well as those containing isolichenin, showed moderate activity against sarcoma 180 and Ehrlich tumor cells. High antitumoral activity, near 90% inhibition, was found associated with the fraction containing raffinose.

Determination of 8-methoxypsoralen kinetics: a relevant factor in the treatment of psoriasis by psoralen plus ultraviolet A therapy.

We compared the efficacy of psoralen plus ultraviolet A (UVA) therapy in 2 groups of psoriatic patients: a group of patients treated by a protocol adapted to the results of 8-methoxypsoralen (8-MOP) plasma kinetics (kinetics group) versus a control group of patients treated according to Pathak's standard protocol (UVA exposure 2 h after oral administration of a dose of 8-MOP equal to 0.6 mg/kg) (control group). The 8-MOP plasma kinetics were determined before the beginning of treatment. The parameter for comparison is the rapidity of clearing, taking into account the number of UVA exposures and UVA joules received. The analysis of the results observed show a 26.6% decrease in the number of UVA exposures and a 38.4% decrease in the dose of UVA received. These results are confirmed by the individual analysis of the rapidity with which the clearance of psoriatic lesions was obtained in patients who were treated with the standard PUVA protocol during the first attack and with the adapted protocol during the second attack.

Antitumor activity of patulin and structural analogs.

A comparison between the cytotoxicity and the antitumor activity of patulin and five structural analogs (isopatulin, dehydroisopatulin, dimethylisopatulin, trimethylisopatulin and isopropylisopatulin) has been established. In vitro assays using L 1210 and P 388 cells showed that the structure of the pyranic ring as well as the nature of the substituents influenced the observed activities. Among the five structural analogs of patulin assayed in vivo against Ehrlich carcinoma, L 1210 and P 388 leukemias, dehydroisopatulin was the only one to be active on all 3 types of tumors at a dose of 100 mg.kg-1.d-1. The ratio between the LD50 in mice and the active dose was 5 while with patulin it was 10. It can be assumed that the lactone function is not solely responsible for the activity of patulin and its structural analogs.

[Annomonysvin: a new cytotoxic gamma-lactone-monotetrahydrofuranyl acetogenin from Annona montana]. / Annomontacine: une nouvelle acétogénine gamma-lactone-monotetrahydrofurannique cytotoxique de l'Annona montana.

The structure of annomontacin [I], a novel monotetrayhydrofuran fatty acid gamma-lactone (acetogenin) isolated from the seeds of Amnona montana, was determined by spectral analysis. The cytotoxicities in vitro of annomontacin [I], annonacinone [2], and annonacin were measured against murine leukemia L1210, human breast adenocarcinoma MDA-MB231, and human breast carcinoma MCF7 cell lines and compared with adriamycin.

[Influence of retinoids on the bioavailability of methoxy-8-psoralen]. / Influence des rétinoïdes sur la biodisponibilité du méthoxy-8-psoralène.

The retinoid-PUVA combination has been recognized as a satisfactory treatment of psoriasis. The absorption of 8-methoxypsoralen (8-MOP) is subject to wide interindividual variations under the influence of factors that are not yet known with certainty but are independent of age, sex and food taken at the same time as the psoralen. Whether concomitant retinoid administration influences the bioavailability of 8-MOP was considered an interesting question. The pharmacokinetics of 8-MOP were studied and compared in two populations of psoriatic patients: 119 patients treated with PUVA alone (psoralen-ultraviolet A) and 40 patients treated with the etretinate-PUVA combination (RePUVA). 8-MOP was assayed by the modified Ljunggren method 1 h, 1 h 30, 2 h, 2 h 30, 3 h and 4 h after ingestion of 8-MOP. The pharmacokinetic values recorded were: time and peak value of maximum plasma 8-MOP concentration (Tmax, Cmax) and area under the curve of time-related 8-MOP concentrations (AUC). The results obtained were as follows: Tmax PUVA 2 h 02 +/- 53 min RePUVA 1 h 56 +/- 50 min Cmax PUVA 159.12 +/- 88.85 ng/ml RePUVA 163.63 +/- 92.85 ng/ml AUC PUVA 343.33 +/- 211.06 ng*h/ml RePUVA 388.12 +/- 251.03 ng*h/ml Statistical analysis showed no significant difference in pharmacokinetic values between patients on PUVA alone and patients on RePUVA. Taking etretinate therefore does not alter the pharmacokinetics of 8-MOP and should not require any change in PUVA treatment.

In vitro and in vivo antitumoral activity of free, and encapsulated taxol.

The anti-tumoral activity of taxol encapsulated either in liposomes or in nanocapsules was compared with that of free taxol, using the P388 and L1210 leukaemia test systems. The in vitro inhibition of cell growth was measured after 48 h and 96 h exposure to various concentrations of taxol. With P388 cells, the inhibitory activities of the three forms of the drug were similar. With the L1210 cells, however, the concentrations required for a 50 per cent inhibition of cell growth (IC50) after 48 h exposure to the drug were greater for nanocapsules than for liposomes or free taxol, the values being 0.060, 0.043 and 0.035 micrograms ml-1, respectively. However, a greater efficiency of nanocapsules was observed after 96 h exposure. Using cytomorphometric analysis, no difference was found between L1210 cells treated either with free or encapsulated taxol. In vivo, mice bearing P388 leukaemia, and treated either with taxol solubilized with 5 per cent DMSO + 5 per cent cremophor in saline solution, or with taxol encapsulated in liposomes (IP daily dose of 12.5 mg Kg-1 body weight x 4 days) showed ILS values of 65.8% and 67.9% respectively. Nanocapsules proved to be toxic, apparently due to their composition: this problem is currently under investigation.

Diels-Alder reactions between dienamides and quinones: stereochemistry of the cycloadditions and cytotoxic activity of the adducts.

Tetrahydronaphthoquinones and tetrahydroanthraquinones bearing an amido group have been prepared by Diels-Alder reactions between (E)-1-(N-carbobenzyloxyamino)-1,3-butadiene (2) or (E)-1-(N-benzoyl-N-benzylamino)-1,3-butadiene (5) and benzoquinone or 5-substituted naphthoquinones. The stereochemistry of the cycloadditions was investigated. A high regioselectivity was observed in the reaction of the diene carbamate 2 with 5-methoxy and 5-acetoxy naphthoquinones. This latter gave the unexpected 1,8-regioisomer 3d. The cycloadditions of the dienamide 5 with naphthoquinones 1 (R = OH, OMe, OAc) are regiospecific. Assignment of the structure of the tetrahydroanthraquinone 6b is in good agreement with the known directing effect of the 5-hydroxy group of juglone 1b in analogous Diels-Alder reactions. With 5-methoxy and 5-acetoxy naphthoquinones, the opposite regiochemistry observed is consistent with the electron-donating influence of the methoxy or acetoxy group, making the C-3 carbon atom more electron deficient. Aromatization of the adducts 6b and 7c was accompanied by an unusual elimination of the amido moiety. Thus, 1-hydroxy and 1-methoxy anthraquinones were obtained. Reactions of the dienes 2 and 5 with benzoquinone gave the tetrahydronaphthoquinones 9 and 10 with an endo stereospecificity. Oxidation of 9 by activated manganese dioxide gave the naphthoquinone 11. These compounds were submitted to in vitro cytotoxic assays towards murine L 1210 leukemia cells and clonogenic human tumor cell line MDA-MB 231. The naphthoquinone derivatives 9, 10 and 11 had significant activities with IC50 less than or equal to 0.4 microgram/ml towards these two tumor cell systems.

Antibacterial, antifungal and antitumoral activities of Micromycetes. I. Preliminary study.

The ability of 211 strains of Micromycetes to produce antibiotic, antifungal and antitumoral compounds has been investigated in vitro using test strains and P 388 leukemia cells. Cytotoxicity was determined on Vero cells. Convenient activities were obtained depending on the taxonomic group. Finally, 17 strains of Micromycetes were selected for their antibacterial or antifungal activities and 12 for their antitumoral properties. Investigations are in progress concerning these activities.

Antibacterial, antifungal and antitumoral activities of 5-carbamoyloxy and 5-acyloxynaphthalene-1,8-carbolactones.

Naphthalene-1,8 carbolactone derivatives have been investigated in order to compare their activities using antitumoral, antibacterial and antifungal tests in vitro. The effect of 5-substitution is the reduction of the toxicity and the suppression of the antibiotic activity. The best results were obtained with the ester series (5-acetoxy and 5-propionyloxy) on both antitumoral and antifungal tests.

Clinical value of an amplified electrophoretic determination of enolase isoenzymes in small cell lung carcinoma patients.

Neurone specific enolase (NSE); alpha gamma and gamma gamma isoenzymes of the glycolytic enzyme enolase, is found in considerable quantity in serum of patients with small cell lung cancer (SCLC). The spectrophotometric measurement of serum enolase activity, plus the electrophoretic separation of isoenzymes (alpha alpha, alpha gamma and gamma gamma) using an amplification reaction constitute a simple method, the application of which has not yet been demonstrated. In patients, serum values of higher than 25 U/l of total enolase activity, with more than 10% NSE, were considered to be positive. Seventy patients diagnosed as SCLC were classified before treatment as having either limited (LD) or extensive (ED) disease, and after chemotherapy as being in complete (CR) or partial remission (PR), stable state (SS) or in relapse (R). The levels of enolase activity and NSE (M +/- SE) in these patients (enolase: 67 +/- 7 U/l, NSE 27 +/- 2%) were different from those in a control group of 19 patients with non-small cell lung cancer (enolase: 29 +/- 2 U/l, NSE: 7 +/- 1%) (p less than 0.001) at the time of diagnosis. Mean enolase and NSE levels in patients with SCLC were seen to differ significantly according to the clinical stage. The results of those patients with ED differed from those of patients with LD (p less than 0.001). The results of the group of patients that achieved remission differed from that of patients during relapse (p less than 0.0001). Serial measurements demonstrated a good correlation between enolase and NSE serum levels and the progression of the disease. The usefulness of this method in the early assessment of treatment was also demonstrated. The clinical usefulness of the dosage of NSE with that of two other tumour markers CKBB and mitochondrial CK was compared.

Antineoplastic activity of two taxol derivatives on an ovarian tumor xenografted into nude mice.

In order to compare the antineoplastic activities of taxol A, taxol B, a mixture of the two (taxol A 72%) and vinblastine, a human ovarian tumor serially transplanted into 104 female athymic mice was used. In the first experiment (11th passage), the antineoplastic activities of taxol A, taxol B and the mixture taxol AB were tested. The same dose was used in each case (12.5 mg/kg i.e. 1/20 of the evaluated LD50 value). It was administered subcutaneously for 5 consecutive days. Three courses of treatment were performed, with 2 rest periods of 1 week in between. All the taxol derivatives produced a statistically significant delay in the tumor growth. However, taxol B had the lowest chemotherapeutic response. In the second experiment (18th passage), different dose levels were administered (mixture 12.5 mg/kg/day x 4 - taxol A 8.8. mg/kg/day x 4 - taxol B 3.5 mg/kg/day x 4 - vinblastine 0.5 mg/kg/day x 2). For all the taxol derivatives 4 treatment courses with 3 rest periods of 4 days were used, and for vinblastine 4 treatment courses with 3 rest periods of 1 week. At the end of the second experiment, vinblastine, taxol A and a mixture of the two showed similar significant activity, whereas no objective antitumor response was observed following the taxol B treatment at the dose level chosen. The experimental results obtained clearly demonstrate that, in the taxane system, the greatest degree of antineoplastic activity can be attributed to taxol A.

Isoenzyme pattern of enolase in human lung tumor xenografts in nude mice.

A simple method is described which allows easy determination of neuroendocrine (NE) differentiation in human broncho-pulmonary tumor models grown in heterotransplanted nude mice. Enolase (EC 4.2.1.11) isoenzyme composition is studied using the electrophoretic method in xenograft tumor homogenates. The relatively large amount of alpha gamma and gamma gamma isoenzymes (neuron-specific enolase (NSE] is indicative of the neuroendocrine differentiation level of these tumors. The gamma gamma isoenzyme is present at a high level (M +/- SE: 10 +/- 2%) in all NE tumor models and absent in non NE tumor models. The alpha gamma isoenzyme is found in a significantly higher proportion in NE tumor models (30 +/- 2%) than in non NE tumor models (9 +/- 2%) (p less than 0.001). Moreover it is possible to discriminate between human and mice isoenzymes to estimate the proportion of mouse tissue hat is present in the xenograft.

Isoenzyme pattern of enolase in the diagnosis of neuroendocrine bronchopulmonary tumors.

Electrophoretic separation of enolase isoenzymes and the measurement of enolase activity were performed in 25 lung tumor extracts. In 13 neuroendocrine (NE) tumors (nine small cell lung carcinoma [SCLC], three atypical NE tumors, and one carcinoid tumor), the NE differentiation was assessed by ultrastructural determination of neurosecretory granule (NSG) density. Twelve non-NE lung tumors also were studied (three adenocarcinomas, four epidermoid, two composite, two large cell undifferentiated carcinomas, and one lymphoma). Four normal lung tissues and 1 human brain were used as controls. The gamma gamma isoenzyme was present at a high level (mean +/- SE, 12 +/- 3%) in all NE carcinomas and consistently absent in all non-NE tumors as well as in normal lung. The alpha gamma isoenzyme was found in significantly higher proportion in NE carcinomas (mean +/- SE, 29 +/- 2%) than in non-NE tumors (mean +/- SE, 8 +/- 1%) (P less than 0.0001), despite an equally high level of total enolase activity in both groups of tumor. The separation of alpha gamma and gamma gamma isoenzymes of enolase allows for the accurate diagnosis of NE tumors and NE components of atypical NE carcinomas, and the gamma gamma isoenzyme, in contrast to gamma chain detection by immunoassay, can be considered to be a specific marker in itself of NE differentiation in lung neoplasms.

Therapeutic response to taxol of six human tumors xenografted into nude mice.

To test the antineoplastic activity of taxol, a natural product isolated from yew (Taxus baccata L.), six human tumors transplanted into athymic mice were used (primary tumors of breast, endometrium, ovary, brain, lung and a recurrence of tongue tumor). While the growth rates varied with the histopathological characteristics of different tumor types, all mice were treated at a mean tumor volume of 200 +/- 8 mm3. Taxol was given SC at a dose level of 12.5 mg/kg per injection per day for 5 consecutive days out of 7 over a period of 3 weeks. With this schedule antitumor responses were obtained in all of the six neoplasms xenografted into nude mice. In the case of the ductal carcinoma of the breast total tumor regressions were observed in four of the five treated animals. In the five other experimental models taxol produced significant growth delays. We believe that the results of these initial tests on the nude mouse--human tumor xenograft system are convincing and justify clinical assessment of this drug.

Influence of the diet on the plasma kinetics of 8-methoxypsoralen.

The plasma kinetics of 8-methoxypsoralen (8-MOP) have been determined in 103 patients treated by PUVA in the routine conditions in which PUVA therapy is carried out in hospital. 8-MOP is taken with breakfast; four types of breakfast were available, which differed by their lipid content; this study showed that the diet has no influence on the bioavailability of 8-MOP.