Retinoid chemoprevention studies in upper aerodigestive tract and lung carcinogenesis.

Chemoprevention is a clinical strategy to block or reverse carcinogenesis before the development of invasive cancer. Studies of chemoprevention in the lungs and upper aerodigestive tract have relied on the field carcinogenesis hypothesis, which predicts that diffuse epithelial injury will result from exposure of that epithelium to carcinogens. This hypothesis is supported by the frequent occurrence of multiple primary tumors within the exposed field. In addition, the understanding of carcinogenesis as a multistep process supports the use of interventions in damaged epithelium before the development of clinically invasive cancer. Current efforts are focused on applying to chemoprevention studies the increasing knowledge of the molecular events in carcinogenesis. In our program, clinical trials in lung and head and neck chemoprevention have focused on individuals with evidence of field carcinogenesis, i.e., a history of previous epithelial cancer or the presence of premalignant lesions. These trials include studies to develop clinically applicable intermediate markers of carcinogenesis and large Phase III trials to evaluate the efficacy of the retinoid isotretinoin in preventing second primary tumors following head and neck or lung cancers.

Percutaneous retinoid absorption and embryotoxicity.

A single application of 17 micrograms/kg or 8.7 mg/kg all-trans-[10,11-3H2]-retinoic acid dissolved in acetone to shaved dorsal hamster skin resulted in rapid absorption and dose-dependent rates of elimination. An equation describing a two-compartment open model with a very brief lag time and first-order uptake and elimination was used to describe the central plasma compartment kinetics. Unchanged all-trans-retinoic acid represented less than or equal to 4% of the total circulating radio-activity. Peak circulating concentrations of parent all-trans-retinoic acid were less than those observed after an equivalent oral dose, but prolonged absorption from the skin appears to contribute to high total bioavailability of topical retinoid. Topical administration to intact skin of up to three consecutive doses of 10.5 mg/kg/d all-trans-retinoic acid or a single 5 mg/kg dose of etretinate (Ro 10-9359) during a critical stage of embryogenesis in hamsters caused erythema and/or dose-dependent epidermal hyperplasia at the site of application, but failed to induce a significant teratogenic response. Topical application of 0.01-1.0 mg/kg arotinoid Ro 13-6298 resulted in dose-dependent mucocutaneous toxicity and an increase in the numbers of dead embryos and malformed offspring. The marked skin toxicity and attenuated concentrations in maternal blood, compared to the oral route, limit the amounts of retinoid that can reach the hamster embryo. It is thus more important to compare the retinoid systemic values (absorbed dose) than it is to compare the oral or topical (applied) dose, when interpreting the results of conventional teratogenicity bioassays. The data suggest that in the human it is skin toxicity that limits the amounts of retinoid that can be applied and subsequently reach the embryo. In the rodent, overt skin toxicity under continued dosing could increase the amounts of retinoid penetrating the skin and reaching the embryo.

Morphological and biochemical alterations in growing rats induced by etretinate.

Etretinate is an aromatic retinoid extensively used on Dermatology. Its toxic effects, however, reduce its application from a clinical point of view. In the present paper, we study etretinate intoxication of 48 growing Wistar rats. The intoxication was for 12 weeks using etretinate doses of 0.5 and 6 (mg/kg)/day. The concentrations of etretinate in plasma and liver were determined. Total seric cholesterol and triglyceride concentrations were analyzed. Structural and ultrastructural histological studies of liver samples were carried out. Continuous etretinate ingestions seem to produce an alteration in the detoxication of enzymatic complexes in the growing rats with both the concentrations, due to the increase in etretinate blood plasma observed during the study. There is a relationship between the etretinate dose and its blood plasma concentration and toxic effect, but there is not with etretinate concentration in the liver. The blood plasma concentration of cholesterol and triglycerides is not related to histological liver lesions. The histological study confirms hepatotoxicity with both doses. Nevertheless, the anatomopathological lesions observed do not seem to be related to the blood plasma and liver etretinate concentrations.

Retinoid-induced ear malformations.

Treatment of pregnant Sprague-Dawley rats with retinoic acid or etretinate pregnancy day 8.5 to 9.0 resulted in craniofacial defects in 100% of the embryos. A morphologic investigation of the malformations occurring in the ear was performed. Outer ears were missing, microtic, low-placed, and dorsally situated. External acoustic meatus was short or absent. Middle ear structures were delayed in differentiation, middle ear ossicle primordia were hypoplastic and malformed, the stapedial artery and facial nerve were hypoplastic, and their relation to the stapes was variable. In the inner ear, the otic capsule was thick, the cochlea had fewer turns and the semicircular ducts showed poor differentiation.

[A toxicological study of etretinate and its effect on blood and tissue (author's transl)].

10 mg/kg of etretinate was administrated orally for 4 weeks to a Sprague-Dawley rat (male, approx. 140 g in weight) and its toxicity was checked at 2 and 4 weeks after the start of administration. Recovery was checked 2 and 4 weeks after end of administration. Three weeks after administration of etretinate, a decline in the increase of body weight and a decline in movement due to abnormality in the lower half of the body was seen. Two weeks after administration, the thigh bone was observed to thin and to have fragility; 4 weeks after administration the changes in the thigh bone became more eminent and the forearm was seen to thin 4 weeks after administration of etretinate, the hematological test showed an increase of leukocytes and the biochemical test showed an increase of triglycerides and phospholipids. The calcium in the thigh bone showed a notable decrease 2 and 4 weeks after administration. The decline in the increase of body weight; the increase of leukocytes, triglycerides and phospholipids; and the decrease of calcium in the thigh bone recovered to normal 2 weeks after the end of administration. Blood retinol decreased and a decrease in retinyl palmitate and retinol in the liver was induced by the administration of etretinate. The toxicity of etretinate was seen to be comparatively lower than that of retinoid. A more detailed study is thought to be needed concerning the changes in bone.

Effect of etretinate (aromatic retinoid) treatment during gestation and lactation periods on viability and somatic growth in F1 rats.

When female SD rats were continuously treated with Etretinate throughout pre-mating, gestation, and lactation periods, the resulting F1 pups exhibited low viability and inhibition of somatic growth after birth (Hummler et al., 1981). Nevertheless, these pups showed no notable change in body weight and external appearance at birth. We used the cross-fostering (between control and treated groups) method and investigate the neonatal viability and the growth hormonal changes in order to assess which treatment period of gestation or lactation was mainly involved in these effects and what changes were actually induced in the F1 pups. The results showed that low viability and inhibition of somatic growth after birth were mainly related to treatment during the gestation period, and these effects were augmented by treatment during the lactation period. Serum GH and IGF-I levels were increased on day 21 in F1 pups groups in which inhibition of somatic growth was observed. These results indicated that treatment with Etretinate during the gestation period might induce a decrease in the number of receptors of GH and IGF-I or other changes, such as a poor-response in target tissues due to a down-regulation, with an increase of serum GH and IGF-I levels.

Male fertility in rats treated with etretinate for 4 weeks.

The toxicity of Etretinate, a retinoid compound, on the male reproductive system was studied in male rats. The drug was administered for four weeks at the dose levels of 0 (control: Vehicle, Peanut oil), 5 and 25 mg/kg/day. The animals were then allowed to mate, and their male reproductive functions and organs were examined in detail. No significant changes due to toxicity were observed in male reproductive functions and organs in the 5 mg/kg/day group after the 4-week treatment. In contrast, males in the 25 mg/kg/day group showed drug-related changes in their reproductive performance (decrease of mating ability and fertility rate), testosterone blood level, sperm head counts, sperm viability and number in the caudal epididymis, organ weight and in the histopathology of their reproductive organs (atrophy of seminiferous tubules, necrosis of spermatocytes and spermatids, vacuolation of nuclei of spermatocytes and spermatids). Even though Etretinate belong to the retinoid group of compounds, the changes seen in the 25 mg/kg/day group were almost the same as those observed in Vitamin A-deficient animals. In conclusion, there is a correlation between changes due to toxicity observed for parameters of male fertility and for histopathological evaluation of the testis of rats that receiving high dose, treatment with Etretinate for 4 weeks.

Prenatal diagnosis by isoenzymic differentiation of Treacher Collins' syndrome induced by retinoids in rats.

A series of branchial arch malformations was induced in 618 embryos from 72 pregnant rats by a single intraperitoneal injection of 10 mg/kg etretinate at 8.5 days of gestation. The litters developed several malformations, including microtia, low set and dorsally placed outer ears, defective middle ear ossicles, short cochleas, defectively differentiated Meckel's cartilages, micrognathia, rudimentary malar bones, lateral facial clefts, fistulas and skin tags, all of which were similar to Treacher Collins' syndrome in man. The defects were accompanied by a pathological differentiation pattern of various isoenzymes in maxillary and mandibular processes. These isoenzymes could be detected in amniotic fluid from the 9th to the 20th days of pregnancy and showed a pathological differentiation pattern here as well. We conclude that a teratogenically induced syndrome affecting the first and second branchial arches is accompanied by a pathological differentiation pattern that can be traced by determinations of isoenzymes in the branchial arches as well as in amniotic fluid.

The effects of alcohol on the metabolism and toxicology of anti-psoriasis drugs.

INTRODUCTION: Alcohol has long been suspected to be a triggering and precipitating factor of psoriasis. Alcohol misuse is common in patients with moderate-to-severe psoriasis and appears to impair treatment outcome. AREAS COVERED: In this article, the authors review the available data regarding the metabolic and toxicological interactions between anti-psoriasis systemic drugs and ethanol and/or alcoholic beverages. Special attention is given to the influence of alcohol consumption on the hepatotoxic risk of some anti-psoriasis drugs. The article was prepared using a MEDLINE literature search. EXPERT OPINION: The available knowledge highlights the existence of a few significant pharmacological interactions, such as the reduced exposure to cyclosporine by red wine, the possible increase of cyclosporine levels following a heavy acute alcohol intake, and, especially, the conversion of acitretin to etretinate, in the presence of ethanol, with important implications in females of child-bearing potential. There are limited data on the contributing role of alcohol in the hepatotoxicity induced by some anti-psoriasis drugs and the existing information on this topic is still controversial. However, further investigation is needed to assess the relevance of interactions between alcohol consumption and drug therapy for psoriasis, under both pharmacological and toxicological perspectives. Long-term prospective studies on large cohorts of patients are warranted to disclose the actual significance of such potential interactions in clinical practice.

Teratogenic dose-response relationships of etretinate in the golden hamster.

Etretinate (Ro 10-9359; Tigason; 4-methoxy-2,3,6-trimethylphenyl analog of retinoic acid, ethyl ester) was evaluated for teratogenic activity in the Syrian golden hamster. Groups of pregnant hamsters were given a single oral dose of 2.8-88 mg/kg etretinate during the early primitive streak stage of gestation. No signs of maternal intoxication were observed in any of the hamsters given the retinoid and maternal body weight changes throughout gestation were not significantly different from those of the vehicle-treated group. Etretinate administration was associated with a dose-dependent increase in the incidence and severity of malformations. The average fetal body weight was significantly less in litters recovered from dams given 44 or 88 mg/kg of etretinate when compared to the average body weight of fetuses recovered from dams given an equivalent volume of the vehicle. The average crown-rump lengths also were significantly shorter in fetuses taken from the dams given 44 or 88 mg/kg etretinate as compared to the control group. The malformations induced by etretinate administration were similar to those noted following an oral dose of all-trans-retinoic acid (Willhite and Shealy, 1984). A comparison of the dose-response curves for induction of terata following treatment with etretinate or all-trans-retinoic acid revealed that etretinate was twice as potent as a teratogen in the hamster as all-trans-retinoic acid. Teratogenic activity of etretinate in the hamster was achieved at doses (mg/kg body wt) used in patients at current clinical therapeutic levels.

[Liposomes--drug carriers for retinally active drugs?]. / Liposomen--"Drug Carrier" für retinal wirksame Medikamente?

Small lipid droplets, the so-called liposomes, can serve as drug carriers and can in theory be targeted directly at diseased tissue. Thus, comparatively smaller quantities of drugs than usual are necessary. However, liposomes are recognized by the immune system and are absorbed by the reticuloendothelial system and therefore also by the Kupffer cells in the liver. These cells are storage compartments for esterified vitamin A. As about 80% of the liposomes are metabolized in the liver, retinoids form a pool for substances transported with retinoid-binding proteins. Whether retinoids reach the retina via direct liposome contact or through the endocytotic process is still not clear. In rabbits, however, retinoids applied in small unilamellar liposomes (SUV) produce photoreceptor malfunctioning (isotretinoin and etretinate) and photoreceptor outer segment damage with an increase in phagocytotic activity of the retinal pigment epithelium (etretinate).

Toxic effects of systemic retinoids on meibomian glands.

Systemic use of retinoids is common in the treatment of various dermatological disorders. Blepharitis and conjunctivitis have been reported in 20-45% of the patients following systemic treatment with 13-cis-retinoic acid. Our purpose was to study the histopathological changes in the eyelids caused by long-term systemic treatment of female New Zealand rabbits with isotretinoin (2 mg/kg) and etretinate (2 mg/kg). The histopathological evaluation showed degenerative changes in the meibomian gland acini, leading to cell necrosis and a decrease in the basaloid cells lining the acini walls. No evidence of acute or chronic inflammatory reaction was noted.

Preventative effect of etretinate therapy on multiple actinic keratoses.

Multiple actinic keratoses (MAK) are premalignant lesions that may produce a significant treatment problem not always controllable by destructive therapy. Etretinate therapy has been shown to produce both prophylactic and therapeutic effects on premalignant epithelial tumors of mice. Results are presented on an 8-month double blind crossover trial (4 months placebo/4 months etretinate, randomly assigned) in 15 patients with severe MAK. Fourteen patients improved significantly on etretinate, whereas five of six patients who had placebo in the first 4-month period became worse as measured by the number of lesions and diameter of larger lesions. However, eight of nine patients who had placebo in the second 4-month period (ie, following etretinate therapy) showed no significant increase in the number of actinic keratoses, suggesting that etretinate may prevent the appearance of new lesions during, and for some time after, its administration. Moderate dose-dependent mucocutaneous side effects were frequent. It is proposed that an annual 3-month course of etretinate may help control severe MAK patients, minimizing the need for destructive therapies.

Biotransformation of etretinate and developmental toxicity of etretin and other aromatic retinoids in teratogenesis bioassays.

Developmental toxicity of the anti-psoriatic drug etretinate (Tegison) and some features of its metabolic conversion to etretin and isoetretin were investigated in in vivo and in vitro teratogenesis bioassays. We found that a single dose of etretinate administered orally to pregnant mice on day 11 of gestation was a potent teratogen (ED50 = 26 mg/kg). Etretin (acitretin, Neotigason), given as a single dose, was about 8-fold less active as a teratogen than etretinate. A ring substituted congener of etretinate, Ro 11-4768, was essentially inactive under similar conditions. Although the mechanisms which operate to make Ro 11-4768 inactive in teratogenesis are unknown and intriguing, it is suggested that the differences between etretinate and etretin may be dependent on individual pharmacokinetic characteristics. The in vitro chondrogenesis bioassay confirmed previous reports that the presence of an acidic endgroup was necessary for suppression of chondrogenesis, and that on that basis etretin was an active inhibitor of chondrogenesis, whereas etretinate was not. Introduction of esterase into the culture medium resulted in complete hydrolysis of etretinate and a quantitative conversion to acid congeners sufficient to account for an appropriate suppression in chondrogenesis. Although limb bud cells were virtually incapable of converting etretinate to etretin in the absence of exogenous esterase, they did influence the metabolism so that in the presence of esterase, isoetretin rather than etretin was the major endproduct of etretinate hydrolysis. Since etretinate therapy endangers the conceptus for a prolonged period of time even after cessation of therapy, further studies are necessary to determine the nature and the extent of hazard posed by the storage and/or metabolism of etretinate.

Ultrastructural survey and tissue analysis of human livers after a 6-month course of etretinate.

A prospective study of the histology and ultrastructure of liver biopsies and analysis of liver tissue for retinoid was performed in twenty psoriasis patients treated with etretinate for 6 months. Nonspecific ultrastructural changes were noted in several liver specimens. Etretinate was detected in all samples. We find no significant hepatotoxicity after a 6 month course of etretinate. Body fat is probably a more important site than the liver for storage of etretinate.

The first stage and complete promoting activity of retinoic acid but not the analog RO-10-9359.

Retinoic acid has the ability to act as either a weak first stage promoter or a weak complete promoter in the initiation-promotion protocol for skin carcinogenesis in the SENCAR mouse. The retinoid analog RO-10-9359 lacks this tumor promoting activity. Both retinoids however inhibit 12-O-tetradecanoylphorbol-13-acetate (TPA) promotion. Additional comparisons revealed that retinoic acid alone can induce dark keratinocytes, a characteristic of tumor promoters, while RO-10-9359 cannot. Retinoic acid but not RO-10-9359 can induce an immediate chemiluminescence response in human polymorphonuclear cells. Both retinoids, however, inhibit a TPA-induced response. Since the chemiluminescence response is believed to be due to oxygen free radical generation, the data suggest that the ability of retinoic acid but not RO-10-9359 to promote tumors and induce dark cells may be due to initial oxidative reactions at the cell membrane.

Evaluation of hypervitaminosis A in the rat by measurement of tibial bone breaking strain.

Using etretinate as a model compound, the bone changes characteristic of hypervitaminosis A were evaluated in the rat in terms of tibial bone-breaking strain. Dose-related effects were observed in the dose range of 5-30 mg/kg p.o. for 15 days. The model proved a simple and precise means of assessing hypervitaminosis A in this species. Isotretinoin also showed a small but significant reduction in tibial breaking strain, but with a shallow dose-response curve in the range of 50-150 mg/kg.