Joseph Raymond Winkle

Evaluation and Development of Spinosyns to Control Ectoparasites on Cattle and Sheep

The spinosyns are a novel family of fermentation-derived natural products that exhibit potent insecticidal activities. Spinosad, a naturally-occurring mixture of spinosyn A and spinosyn D, has successfully established its utility for crop protective applications in the agrochemical field. Potential applications of this unique chemical family of macrolides also have been investigated in the field of animal health. Applications for the control of blowfly strike and lice on sheep have now been commercially developed and registered in Australia and potential applications for the control of ectoparasites on cattle are being studied.

Knockdown and Mortality Comparisons Among Spinosad-, Imidacloprid-, and Methomyl-Containing Baits Against Susceptible Musca domestica (Diptera: Muscidae) Under Laboratory Conditions

Abstract The activity of spinosad, imidacloprid, and methomyl baits and technical actives were assessed against susceptible house flies, Musca domestica L. (Diptera: Muscidae). In a feeding assay, imidacloprid affected flies more rapidly than methomyl or spinosad, but spinosad was 2.7 times more potent than methomyl and 8 times more potent than imidacloprid. The profile of technical actives correlated with their respective fly bait formulations in laboratory assays. Although having the most rapid onset of activity in laboratory tests, up to 50% of flies remained alive after exposure to imidacloprid bait. In contrast, <5% of flies survived 24-h exposure to spinosad or methomyl baits. High temperature reduced the knockdown activity of imidacloprid bait and slowed the speed of kill for spinosad and methomyl baits over a 24-h exposure period. Spinosad and methomyl baits were also superior to imidacloprid when applied to the floors of environmentally controlled rooms at label recommended rates, providing good fly control for up to 21 d. The fact that a significant percentage of flies exposed to imidacloprid were rapidly knocked down but subsequently remained alive in all of the assays suggested that flies were recovering from initial exposure to this compound. Given its favorable safety profile, a high degree of initial and residual activity comparable with methomyl and lack of cross-resistance to other chemistries, spinosad bait may be a valuable component of house fly control programs to help control or delay the emergence of resistant populations.

Discovery of an orally bioavailable isoxazoline benzoxaborole (AN8030) as a long acting animal ectoparasiticide

A novel series of isoxazoline benzoxaborole small molecules was designed and synthesized for a structure-activity relationship (SAR) investigation to assess the ectoparasiticide activity against ticks and fleas. The study identified an orally bioavailable molecule, (S)-3,3-dimethyl-5-(5-(3,4,5-trichlorophenyl)-5-(trifluoromethyl)-4,5-dihydroisoxazol-3-yl)benzo[c][1,2]oxaborol-1(3H)-ol (38, AN8030), which was long lasting in dogs (t1/2=22 days). Compound 38 demonstrated 97.6% therapeutic effectiveness within 24 h of treatment, with residual efficacy of 95.3% against American dog ticks (Dermacentor variabilis) on day 30% and 100% against cat fleas (Ctenocephalides felis) on day 32 after a single oral dose at 50 mg/kg in dogs.

Optimization of isoxazoline amide benzoxaboroles for identification of a development candidate as an oral long acting animal ectoparasiticide

Novel isoxazoline amide benzoxaboroles were designed and synthesized to optimize the ectoparasiticide activity of this chemistry series against ticks and fleas. The study identified an orally bioavailable molecule, (S)-N-((1-hydroxy-3,3-dimethyl-1,3-dihydrobenzo[c][1,2]oxaborol-6-yl)methyl)-2-methyl-4-(5-(3,4,5-trichlorophenyl)-5-(trifluoromethyl)-4,5-dihydroisoxazol-3-yl)benzamide (23), with a favorable pharmacodynamics profile in dogs (Cmax=7.42ng/mL; Tmax=26.0h; terminal half-life t1/2=127h). Compound 23, a development candidate, demonstrated 100% therapeutic effectiveness within 24h of treatment, with residual efficacy of 97% against American dog ticks (Dermacentor variabilis) on day 30 and 98% against cat fleas (Ctenocephalides felis) on day 32 after a single oral dose at 25mg/kg in dogs.

Evaluation of hair loss in cats occurring after treatment with a topical flea control product

BACKGROUND A formulation containing 39.6% spinetoram resulted in a higher than anticipated number of reports of alopecia at the site of application in the first months following commercial product launch. HYPOTHESIS/OBJECTIVES To determine the cause of the alopecia using histopathology, including assessment for inflammation, follicular findings of physical trauma (plucking/pulling behaviour) and changes in follicular cycling. ANIMALS Twenty-four flea-free, male and female adult domestic short hair cats within a private research colony. METHODS Cats were treated with a single application of 39.6% spinetoram on day 0; personnel were not blinded. Observations of the skin and hair coat began immediately and were repeated at 30 min and 1, 2, 3, 4, 6, 8 and 12 h post-application and then on subsequent days at the same time as initial dosing and at 2, 4, 6, 8 and 12 h after that time, until day 5. If hair thinning or loss was observed, a skin biopsy sample was collected. Two cats not exhibiting abnormalities were biopsied on day 6. RESULTS Thirty-eight per cent of cats (nine of 24) developed hair thinning and alopecia of sufficient severity within 78 h post-application of the product to warrant skin biopsy. Abnormalities in the skin were limited to the application site and were consistent with physical trauma (pulling or plucking) to the hair. CONCLUSIONS AND CLINICAL IMPORTANCE Microscopic changes in the hair follicles of affected cats were consistent with self-induced trauma or barbering behaviour. All changes were reversible and paralleled findings associated with well-established, topical flea control products.

Evaluation of an 11.2% spinetoram topical spot-on solution for the control of experimental and natural flea (Ctenocephalides felis) infestations on cats in Europe

Spinetoram is the newest member the spinosyn-class of natural products to be commercialized for flea control on cats in the United States under the tradename Cheristin® for cats. This report describes results from two laboratory studies and a multi-center clinical field trial designed to confirm the efficacy of a topical spot-on solution containing spinetoram (11.2% w/w, 130 mg/mL) against European strains of the cat flea, Ctenocephalides felis. In the laboratory studies, cats were allocated to one of two treatment groups of eight animals each: negative control (mineral oil) or spinetoram, both applied as a topical spot-on at the base of the skull on Day 0. Cats were infested with ∼100 newly emerged, unfed adult fleas on Days -2 or -1, 7, 14, 21, 28 and 35. To calculate efficacy, fleas were counted and removed 48 h after treatment, and 48 h after each subsequent infestation through week 5. Spinetoram treatments provided 100% efficacy through at least day 16 and ≥ 97% efficacy (arithmetic mean) for one month. For the field trial, 23 clinics from Northern and Southern Europe participated in the study that compared the effectiveness and safety of spinetoram and fipronil/(S)-methoprene treatments over a period of two months. There were 258 and 248 evaluable efficacy cases for month 1 and month 2, respectively, with 300 total evaluable cases for safety. Treatments were administered on Day 0 and again on Day 30 (±3 days). The effectiveness of treatments was calculated based on reduction in live flea counts on Days 14, 30, 44 and 60 (±3 days) relative to flea counts obtained on Day 0. Efficacy (geometric mean percent flea reduction) on Days 14, 30, 44 and 60 was 97.0%, 95.0%, 99.3% and 99.1% for spinetoram, respectively, and 86.1%, 80.9%, 92.4% and 92.3% for fipronil/(S)-methoprene, respectively. Spinetoram was deemed non-inferior at all intervals and superior to fipronil/(S)-methoprene at Days 30 and 60. Clinical signs of flea allergy dermatitis (FAD) were markedly improved following spinetoram treatment, as demonstrated through statistically significant reductions in severity of FAD scores for most of the clinical signs when compared to fipronil/(S)-methoprene treatment. There was a lower overall adverse event incidence rate for spinetoram (5.1%) versus fipronil/(S)-methoprene treatment (11.5%).