Relating to the successful use of griseofulvin as a systemic antifungal agent, it has been thought to be important to develop investigation on the drug in the skin. But lack of the method to extract griseofulvin completely from skin may have caused our poor knowledge on this field, even on the skin levels after oral administration.
In the present work, skin griseofulvin was extracted with a new method. Each skin sample measured was finely divided, heated with ether-ethanol and filterd. Then the filterd solution was evaporated. In order to isolate griseofulvin in residue, a chromatographic method was convenient. In this work, residue was redissolved in ether, applied on a piece of chromatostrip (made by silica-gel) and developed with methanol. Visualised by means of its ultraviolet fluorescence, the separated griseofulvin was eluded with 10ml of ethanol and measured for fluorescence with Farrand's spectrophotofluorometer.
The results obtained were as follows:
1) Skin griseofulvin levels in rabbits: orally administered 125mg fine particle griseofulvin showed the peak level of 2.8mcg/g (in epidermis and dermis) and 8.2mcg/g (in subcutaneous fat tissue) at 4 hours after administration. As the levels reduced slowly, the drug was measured in the concentration of 0.9mcg/g (in epidermis and dermis) and 2.9mcg/g (in subcutaneous fat tissue). On the other hand, serum level showed 1.8mcg/ml at 4 hours after administration and a trace at 24 hours.
2) Skin griseofulvin levels in human subjects: As it was difficult to obtain skin samples repeatedly and to know nonspecific fluorescence in each subject, only the average skin level at 4 hours after single oral dose of fine particle griseofulvin 500mg was estimated. The estimated level was 12.1 mcg/g (in epidermis and dermis) and 17.5mcg/g (in subcutaneous fat tissue).
3) In vitro uptake of griseofulvin measured in the skin sample was 20.3mcg/g (epidermis and dermis) and 36.0mcg/g (subcutaneous fat tissue) in 1% ethanol solution with griseofulvin 0.5mcg/ml.
4) Griseofulvin demonstrated a strong affinity to subcutaneous fat tissue in vitro and in vivo. These results, together with many other reports (2, 4, 8, 10, 15, 16), may suggest a possible hypothesis for coexistence of griseofulvin and fat throughout its course from duodenum until stratum coenrum.

抄録全体を表示

There exists a considerable literature on the tricarboxylic acid cycle (TCA cycle) in bacteria and Candida, but little work has been done on that in Trichophyton. On the other hand, sufficient knowledge concerning the acting mechanism of antifungal agents on fungi is not yet obtained.
This report deals with the biochemical assay of cell free extract of Trichophytons on some dehydrogenase and fumarase activities which concern with TCA cycle, and the effects of several antifungal agents on these enzyme activities.
The strains of Trichophyton used in this study were Tr. mentagrophytes strain 840 obtained from the Department of Dermatology, Juntendo University, and Tr. rubrum isolated from patient in Tonan Hospital. The material which was scraped from two-months' Sabouraud culture was removed to 200ml Sabouraud glucose broth and then shake culture was made at 25°C for one week. The pellets of organism were washed and dried, then ground in mortar with sea sand. The supernatant of centrifuged material was dialysed, then used as cell free extract. The enzyme activities such as iso-citric dehydrogenase (ICDH), alpha-ketoglutaric dehydrogenase (α-KG), succinic dehydrogenase (SDH) and malic dehydrogenase (MDH) were measured colorimetrically with TTC reduction system. The fumarase activity alone was measured qualitatively with paper chromatography. Several antifungal agents such as bisethylmercuri phosphate (BEMP), salicylic acid, trichomycin, griseofulvin, naphthiomate N and vitamin K₃ were added respectively to cell free extract in order to observe the effect of these agents on enzyme activities.
The endogenous respiration of each sample was makedly inhibited though dialysis procedure, while enzyme activities were not influenced. Therefore, every cell free extract were dialysed in this study. Activities of all four dehydrogenases tested were found in both Tr. rubum and mentagrophytes. The values of each enzyme were as follows: 1) Tr. rubrum: The average±standand error of 7 samples. Endogenous respiration (ER) 0.4±0.05 milimicromoles/mg protein/hour, SDH 0.9±0.07, MDH 1.6±0.21, α-KG 1.2±0.20, ICDH 2.9±0.42. 2) Tr. metagrophytes: The average±standard error or 6 samples. ER 0.4±0.08, SDH 1.0±0.18, MDH 2.2±0.41, α-KG 1.0±0.16, ICDH 4.2±0.59.
As to Tr. mentagrophytes, the spot of malic acid (Rf 41) was identified on paper chromatography as the proof of presence of fumarase activity. The changes of enzyme activities of two species induced by six different antifungal agents were as follows: 1) 30, 000×solution of BEMP: No effect was observed on ER in both species, while the activities of SDH, MDH, α-KG and ICDH were markedly inhibited in both species. 2) 600×solution of salicylic acid: The MDH activity in both species was increased while activities of SDH, α-KG, ICDH and ER were inhibited. 3) 9, 000×solution of trichomycin: No effect of the solution was observed in both species, while ER of Tr. mentagrophytes was slightly inhibited and MDH activity of that species was slightly increased. 4) 9, 000×solution of griseofulvin: No effect of the solution was observed in both species, while ER of Tr. rubrum was slightly increased. 5) 9, 000×solution of napthiomate N: No effect was observed in both species, while ER of Tr. mentagrophytes was inhibited and MDH of that species was slightly increased. 6) 15, 000×solution of vitamin K₃: α-KG activity of Tr. rubrum was inhibited, while SDH, MDH and ICDH activities of two species were markedly increased.

抄録全体を表示