Reconciling clinical data with risk: the Kathon biocide story.

This chapter discusses the approach employed by Rohm and Haas Company to shepherd a microbicide through the stages of risk analysis and management, with special emphasis on the role of clinical evaluation. The risk analysis process for a microbicide with broad applications as well as varied human exposure patterns involves assiduous planning along with development and implementation of appropriate actions to monitor and reduce risk levels. The cooperation of industry and the dermatology community was instrumental in achieving the goals of the program.

The irritancy and allergenicity of 2-n-octyl-4-isothiazolin-3-one (Skane M-8), with recommendations for patch test concentration.

Preparations of 2-n-octyl-4-isothiazolin-3-one (Skane M-8) at 1000 p.p.m. a.i. and 500 p.p.m. a.i. in pet. induced allergy in a cumulative insult human patch test. Challenge patch testing using 250 p.p.m. a.i. Skane M-8 elicited allergy in 6 out of 6 sensitized individuals. Sensitized individuals did not cross react with Kathon biocide, (Kathon CG), a mixture of 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-4-isothiazolin-3-one. All pretrolatum preparations to which Tween 85 was added at 2.5% produced mild irritation in cumulative insult patch tests, but no irritation was observed with Tween 85 at 0.625%. We conclude that Skane M-8, 250 p.p.m. a.i. with Tween 85 0.625% in petrolatum is appropriate for patch testing for Skane M-8 allergy.

Human safety study of body lotion containing Kathon CG.

The safety of Kathon CG biocide as a preservative in leave-on body lotions was assessed by 2 double-blind studies, using similar protocols. A total of 209 healthy male and female subjects aged 18 to 65 years, 100 in California (72 test subjects, 28 controls) and 109 in Florida (88 test subjects, 21 controls) completed the studies which included pre- and post-use phase diagnostic patch testing with Kathon CG 100 ppm active ingredient, and 13 weeks daily applications of either a test lotion containing Kathon CG 15 ppm active ingredient or a control lotion without Kathon CG. No evidence of irritation or sensitization attributable to use of the biocide was found during regular dermatological examinations during the use phase. Post-use phase patch testing produced negative results in all subjects with the exception of 1 control subject in Florida who had positive readings at the 2- and 4-week post-use phase patch testing. Overall, these studies show there is minimal, if any, risk of adverse effects associated with the use of Kathon CG 15 ppm active ingredient in a leave-on application.

Absorption and disposition of 14C-labelled Kathon biocide, a mixture of 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-4-isothiazolin-3-one, following intravenous or dermal administration to male Sprague-Dawley rats.

Kathon biocide (KB), a 75:25 mixture of 5-chloro-2-methyl-4-isothiazolin-3-one (KI) and 2-methyl-4-isothiazolin-3-one (KII), is a broad-spectrum antimicrobial agent. The absorption and disposition of 14C label were studied in male Sprague-Dawley rats following iv or dermal administration of KB 14C labelled in the carbonyl carbon of either KI or KII. KI-labelled Kathon was distributed rapidly following an iv dose (0.8 mg/kg). Total 14C label in the plasma was rapidly eliminated, with the data best described by a three-compartment model. Total 14C label concentration in the blood, however, remained constant at 3 ppm from 6 to 96 hr after administration and represented 29% of the dose, indicating that KI-labelled Kathon and/or metabolites was sequestered by the cellular fraction of blood. Elimination of 14C label from the tissues examined was biphasic, with a terminal half-life of more than 4 days; by 96 hr, faeces, urine and CO2 accounted for 35, 31 and 4% of the dose, respectively. Following a single 24-hr dermal application of 0.2 ml of an aqueous solution containing 2000 ppm [14C]KB (400 micrograms), rats absorbed 94% of KI-labelled KB and 82% of KII-labelled KB. However, the systemic bioavailability of KB was substantially less than this, since approximately half of the absorbed KB was associated with the skin at the application site 24 hr after the application. Percutaneous absorption was not affected by concentration over the range 500-4000 ppm. As the concentration of KI-labelled KB in the applied solution increased from 500 to 4000 ppm, the relative amount of 14C label associated with the skin decreased, while that in the excreta increased, indicating greater systemic penetration at higher concentrations. Low amounts of 14C label were found in the testes (less than 2 ppb) and blood (24 ppb) 28 days after a single dermal application of KI-labelled KB. Four consecutive daily applications of KI-labelled KB did not influence the proportion of the dose absorbed from the skin. However, the proportion of the dose excreted was higher than after a single application of an equivalent amount of KB.

Kathon biocide: manifestation of delayed contact dermatitis in guinea pigs is dependent on the concentration for induction and challenge.

The potential of Kathon biocide, an aqueous solution containing, as active ingredients (a.i.), a mixture of 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-4-isothiazolin-3-one (14.4% a.i.), to produce delayed contact dermatitis, a sensitization response, was evaluated in outbred Hartley guinea pigs by a modified Buehler's occluded epicutaneous patch technique. The relationship of the response as a function of induction/elicitation concentrations was investigated. Groups of guinea pigs received 9 induction doses of the biocide, 3 times a week, at concentrations ranging from 25-2000 ppm a.i. These guinea pigs were challenged with the biocide at concentrations ranging from 20-2000 ppm a.i., and the application sites were scored for erythema 24 and 48 h after the challenge. The incidence of delayed contact dermatitis in induced guinea pigs was dependent on both the induction and challenge concentrations. The EC50 (concentration at which delayed contact dermatitis was seen in 50% of the population) for induction at a challenge concentration of 2000 ppm a.i., a nonirritating concentration, was estimated to be 88 ppm a.i. with a slope of 3.47 probits/unit log concentration. The EC50 for elicitation at an induction concentration of 1000 ppm a.i. was estimated to be 429 ppm a.i. with a slope of 2.74 probits/unit log concentration. These data demonstrate that for Kathon biocide, there is an induction/elicitation concentration dependency for delayed contact dermatitis response, and there is a "no response concentration" zone where the biocide can be used without concern for clinically significant delayed contact dermatitis. In comparison with a previous study, these data also suggest that the number of induction doses may be an important factor in demonstrating the sensitization potential of a chemical.

The genetic toxicology of Kathon biocide, a mixture of 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-4-isothiazolin-3-one.

Kathon biocide, an aqueous solution containing a mixture of 5-chloro-2-methyl-4-isothiazolin-3-one and 2-methyl-4-isothiazolin-3-one in an approximate ratio of 3:1, was tested for mutagenic activity in Salmonella typhimurium, L5178Y mouse lymphoma cells in culture and Drosophila melanogaster. Tests also were conducted for chromosome aberrations in vivo on mouse bone marrow cells, for DNA damage/repair in primary rat hepatocytes in culture, and for morphological transformation in C3H 10T1/2 cells in culture. Kathon biocide produced point mutations in the absence of a rat-liver metabolizing system in bacteria (strain TA 100) and mammalian cells in culture. In the presence of rat-liver metabolizing system a 10-fold higher concentration was required to induce point mutations in mammalian cells in culture. No mutagenic activity was observed with the metabolizing system and S. typhimurium. Negative results were obtained in the sex-linked recessive lethal assay in Drosophila, the in vivo cytogenetic assay in mice, the unscheduled DNA synthesis assay in cultured rat hepatocytes, and the in vitro cell transformation assay.

GI pharmacology of polyethyleneimine I: effects on gastric emptying in rats.

Experiments were conducted to determine the effect of ingested polyethyleneimine upon gastric emptying of the fasted rat. Emptying was evaluated by the phenolsulfonphthalein and resin bead methods. The two techniques gave comparable results; both showed that this agent inhibited gastric emptying. A delay in gastric emptying could be detected within 15 min of intubation. The effect was dose related, quite long lasting (approximately 4 hr), and reversible. Commercially available, branched polyethyleneimines were highly active, but the linear polyethyleneimine was without observable effect. A branched polyethyleneimine derivative with all primary amine sites selectively acetylated also was inactive.

GI pharmacology of polyethyleneimine II: motor activity in anesthetized dogs.

The effects of orally and intravenously administered doses of polyethyleneimine were observed in 18 chloralose-urethan-anesthetized dogs. Polyethyleneimine produced an initial augmentation of rhythmic segmenting gastric antral contractions, a copious flow of gastric mucus, increased segmenting and propulsive activities of the small and large intestines, and occasional micturition and defecation. The gastric corpus and fundic regions became relaxed and enlarged. These events were associated with the prompt appearance of retching. The retching response to oral administration could only be abolished by bilateral vagotomy or bilateral sympathectomy. The skeletal muscle component of retching was blocked by tubocurarine. Intravenous administration of chlorpromazine blocked the retching response and gastric corporal atonia to either intravenous or oral doses of polyethyleneimine. Either oral or intravenous administration of polyethyleneimine produced no detectable changes in the lead II ECG but was associated with marked transient reductions in both mean and pulsatile arterial blood pressures. These depressor effects showed clear tachyphylaxis. In all cases where GI effects were noted, respiration was augmented and erratic in a manner associated with the retching responses.