CD 271 possesses a potent anti- inflammatory activity in a large number of models particularly when compared with known reference anti-inflammatory agents. Retinoids (all-trans retinoic acid, 13 cis retinoic acid and Etretinate) show very weak or no activity.

The β-adrenergic receptors, previously shown to be present on the membranes of cultured human epidermal keratinocytes, were found to be functionally coupled to membrane-bound adenylate cyclase. Using membrane preparations, the enzyme could be activated by guanosine triphosphate (GTP), the stable GTP analog GPP(HN)p, and NaF, all of which are known to activate the adenylate cyclase without interacting with membrane receptors. Binding of catecholamine agonists (epinephrine, norepinephrine, and isoproterenol) to the β-adrenergic receptors is followed by an increase in the activity of adenylate cyclase. This activation could be reversed (or prevented) by β-adrenergic antagonists, but was unaffect by the presence of α-adrenergic ligands (either agonists or antagonists). The activation by catecholamines appears to be directly related to receptor occupancy, since the activation constant (Ka) of adenylate cyclase for the three catecholamines was found to be very similar to the equilibrium dissociation constant (Kd) determined from competition binding experiments. The activation of adenylate cyclase under these conditions appears to be restricted to the catecholamine agonists only. The non-catecholamine β-adrenergic agonists (salbutamol, terbutaline) did not show any measurable activation of adenylate cyclase, even though these agonists were shown previously to bind to the β-adrenergic receptors on keratinocyte membranes with the expected affinities.

Keratinocyte/leucocyte interactions have become an area of intense investigations in the last decade. However, few convenient in vitro models are available at present. We have therefore designed a novel in vitro system for autologous human keratinocyte/leucocyte co-culture. Non-invasive epidermal cell sampling was achieved by using outer root sheath cells from hair follicles. After one passage, pure keratinocyte cultures (no Langerhans cells or melanocytes) were obtained. Co-culture experiments were performed on a Transwell system: keratinocytes were grown on the porous cupula, and then laid on to wells containing leucocytes. Alternatively, leucocytes can be added to the cupula when contact interactions between the two cell types are to be investigated. Using this system, we demonstrated that Phaesolus vulgaris phytohaemagglutinin-activated T lymphocytes (with 10% monocytes) in the lower compartment induced intercellular adhesion molecule 1 (ICAM-1) and HLA-DR expression, and inhibited methyl-3H-thymidine incorporation in normal human autologous keratinocytes cultured on the cupula. These changes were mediated by soluble factors (no cell contacts between keratinocytes and leucocytes), and required lymphocyte activation. This is the first direct in vitro evidence for leucocyte-induced ICAM-1 and HLA-DR expression on keratinocytes. This system is a potential tool for the study of keratinocyte/leucocyte interactions.(ABSTRACT TRUNCATED AT 250 WORDS)

We have evaluated a subchronic model of contact hypersensitivity in the guinea pig to mimic human chronic/recurrent eczema. Repeated challenges of the ears of previously sensitized guinea pigs with 0.1% dinitrochlorobenzene (once a week for 4 weeks) induced a typical oedema response, which increased during the first 48 h after each challenge. Crusts were detectable (48 h after challenge) and histological observations (72 h after challenge) revealed hyperplasia, papillomatosis, hyperkeratosis and some mononuclear cell infiltrates in the dermis. In agreement with clinical observations in humans, topical treatment of challenged animals with corticosteroid (1% hydrocortisone) reduced the oedema, hyperplasia, papillomatosis, and leucocyte infiltrates, while application of 5% bufexamac (a non-steroidal drug) was associated with a slight enhancement of the inflammatory response. Thus, this model presents clinical and histological similarities with human eczema. Its pharmacological relevance is also suggested, although further investigations are required to better define its selectivity.

The use of experimental models in cutaneous pharmacology has gained significant importance in recent years. There has been increasing interest in the use of experimental models to investigate the pathophysiology of diseases as well as potential responses to therapeutic agents.

In model systems where anthralin and butanthrone have a high anti-proliferative activity, e.g., inhibition of keratinocyte metabolism in culture or inhibition of ornithine decarboxylase inductionin vivo, CD 003 is inactive. However, in the skin disease psoriasis there is both an inflammatory as well as a differentiation/proliferation component, and consequently we have evaluated anthralin and butanthrone for their potentialin vivo antiinflammatory activities. In addition, we have compared a new analogue, substituted at C10, with these two compound. Our results support the belief that modification of the anthralin structure can result in the formation of less toxic, less irritant and less staining analogues, which maintain or significantly improve upon the topical antiinflammatory properties of anthralin. The possibility that such analogues will be of use in the treatment of skin diseases with a major inflammatory component is at present being investigated.

The fate in vivo of topically applied 1,8-dihydroxy-9-anthrone (dithranol, anthralin) was investigated in the skin of the hairless rat, using a specially designed drug delivery system (film). The film was applied on intact skin as well as on skin with an impaired barrier function (stripped skin). The distribution of the drug was examined either after continuous application or at selected times after short contact periods. The exposed skin was extracted with diisopropylether and free dithranol, its dimer and quinone assayed by quantitative HPLC analysis. The incorporation of trace amounts of 3H-dithranol and 14C-dithranol in the vehicle made it possible to quantify the fraction of penetrated drug which was insoluble in ether. With continuous application to intact skin (up to 24 h), extractable dithranol rapidly reached a plateau level (15 min) and was concentrated in the stratum corneum. Substantial dimer formation occurred in both normal and stripped skin. Ether insoluble material rapidly predominated over soluble material, especially when the stratum corneum was absent. However, on short contact application (0.5-1 h), ether soluble material (dithranol in the stratum corneum) was quantitatively predominant in the intact skin. Removal of the vehicle after a short contact time resulted in the disappearance of dithranol from the skin (normal and stripped). In intact skin, the drug was converted into ether insoluble material. In the stripped skin, this insoluble fraction remained constant over the duration of the experiment (24 h).