Investigation of some amino acid analogues and metabolites as inhibitors of wool and hair growth.

Sheep were given intravenous infusions of ethionine together with cycloleucine or reduced glutathione, in attempts to prevent the inhibition of wool growth by ethionine. Other sheep were given cycloleucine alone to measure effects on wool growth. Twenty-two compounds related to cystine, methionine, ethionine, lysine, phenylalanine and tyrosine were given as intravenous infusions to sheep to investigate their potential as depilatory agents. Nineteen of these compounds were also tested in mice during their first cycle of hair growth. The concurrent administration of cycloleucine with ethionine prevented the weakening of wool fibres caused by ethionine, but reduced glutathione was ineffective. Cycloleucine weakened wool fibres, as judged subjectively, and caused a small reduction in fibre diameter. Selenocystine and selenomethionine caused some hair loss in mice but selenocystine was also toxic. Both seleno-amino acids were toxic for sheep; selenocystine was lethal at 0.025 mmol kg-0.75 and selenomethionine at 0.09 mmol kg-0.75. Doses that permitted survival of sheep did not have depilatory effects. However, the presence of autophagic vacuoles in the cytoplasm of follicle bulb cells of sheep indicated that a toxic dose of selenocystine had potential depilatory activity. Other compounds investigated did not induce loss of wool or hair. Some compounds, notably 3-methylthiopropionic acid and S-(2-aminoethyl)-L-cysteine, were toxic to mice but not sheep. The methionine analogue, methoxinine (O-methyl-DL-homoserine), caused a substantial reduction in the strength of wool fibres and a prolonged alteration of the crimp pattern. It is suggested tentatively that cycloleucine inhibits methionine adenosyltransferase and thereby reduces or prevents the formation of S-adenosylethionine. The failure of various compounds related to methionine and ethionine to have any depilatory activity in sheep supports the view that ethionine influences wool growth via the formation of S-adenosylethionine.

Inhibitory effects of ethionine, an analogue of methionine, on wool growth.

Varying amounts of DL-, L- or D-ethionine were administered intravenously to sheep, either as a continuous infusion, usually over 2 days, or as a single injection. Groups of sucking mice and rats, in their first cycle of hair growth, were given subcutaneous injections of DL-ethionine at several dose rates. Ethionine was a potent inhibitor of wool growth in sheep; the L- and D-isomers appeared equally effective. An infusion of 20 mg/kg DL-ethionine (c. 50 mg/kg0.75) given at a daily rate of 10 mg/kg for 2 days, or an injection of 40 mg/kg DL-ethionine (c. 100 mg/kg0.75), were sufficient to cause the growth of very weak wool and allow the fleece to be readily removed by hand within 3 weeks after dosing. The inhibition of wool growth was usually associated with a concentration of ethionine in blood plasma, during intravenous infusion, of 10 mumol/l or higher. An infusion of DL-ethionine at a daily rate of 1 mg/kg for 12 days caused the growth of weak fibres and substantially reduced both length growth rate and diameter of fibres. The toxicity of ethionine to sheep was dependent on the total dose and the duration of administration. An infusion of 40 mg/kg (20 mg/kg daily for 2 days) produced severe effects, but the sheep recovered; a dose of 14 mg/kg (2 mg/kg daily for 7 days) was lethal. The effects of ethionine on wool growth were reduced or prevented by the concurrent infusion of methionine (10-15 mol/mol ethionine). Doses of DL-ethionine as high as 460 mg/kg0.75 failed to cause hair loss in sucking mice. While body growth was severely retarded at this dose, no deaths occurred. Likewise, DL-ethionine failed to cause hair loss in sucking rats, but was lethal to some rats at a dose of 360 mg/kg0.75.

Depilatory effects of certain chemicals during the first hair growth cycle in sucking mice.

The chemicals were administered, subcutaneously, orally or topically. Generally, the depilation produced in the mice by mimosine or cyclophosphamide differed from that produced by the steroid analogues tested. In the first 2 cases almost completely naked mice were commonly seen, while in the steroid-treated groups the complete inhibition of all hair fibres was rare. This is discussed in relation to the effects of the same compounds on wool growth in sheep. When related to bodyweight, the doses of cyclophosphamide (62 mg/kg0.75) and dexamethasone (5--10 mg/kg0.75), that depilated mice in our experiments were in good agreement with those reported to inhibit the growth of wool fibres in some sheep. An example of synergism in depilatory effect between dexamethasone and cyclophosphamide is also presented. The time of onset and the initial spread over the body of the 2nd hair cycle in depilated mice was observed.

Fate of mimosine administered orally to sheep and its effectiveness as a defleecing agent.

Mimosine was administered orally to Merino sheep once daily for periods of 1-3 days, either as the isolated compound or in the foliage of Leucaena leucocephala. A single daily dose of mimosine of 450 or 600 mg/kg body weight was effective for defleecing sheep. A daily dose rate of 300 mg/kg was effective for defleecing sheep if given on two successive days. The effectiveness of a treatment for defleecing sheep was related to the concentration of mimosine in plasma following dosing; defleecing ensued when the concentration of mimosine in plasma was maintained above 0-1 mmol/l for at least 30 h. The main products excreted in urine were mimosine and 3,4-dihydroxypyridine (DHP); small amounts of mimosinamine were also excreted. During the first day following dosing, the major excretory product was mimosine; DHP was an important component during the second and third days. In the three days following the start of dosing, between 32 and 53% of the mimosine given was accounted for as mimosine in the urine. Following an intravenous infusion of mimosine, no DHP was detected in urine; most of the mimosine was excreted intact but a small amount (c. 9%) was excreted as mimosinamine.

Effects of mimosine, a potential chemical defleecing agent, on wool growth and the skin of sheep.

Twenty-two Merino sheep were dosed with various amounts of L-mimosine, given either as an intravenous or an intraperitoneal injection, or as a continuous intravenous infusion for periods of 1-4 days. Single injections of mimosine (1-16 g) had no effect on the strength of wool, and wool growth rates were not appreciably altered by injections of small amounts (4 g or less). Injections of larger amounts slightly reduced both length growth rate and diameter of tibres during the 4 days after dosing. The effects of intravenous infusions of mimosine depended on the rate and the duration of administration. Small amounts (0.5 or 1 g/day given for 4 days) has no effects on the strength of wool or on wool growth rates. Infusions of a total of 8 g, either at the rate of 2 or 8 g/day, weakened the wool but not sufficiently to allow the sheep to be defleeced. Both these treatments caused a temporary reduction in length growth rate and in diameter of fibres, and transient degenerative changes were observed in wool follicles. A region of the fibres representing 1-2 days' growth was constricted to about half the pre-infusion diameter when 8 g was given for 1 day. Infusions of at least 8 g mimosine over a period of 1 1/2-2 days were effective for defleecing all sheep dosed. This corresponded to a daily rate of infusion of about 80 mg/kg. No toxic effects were observed with infusions given for periods of 2 days. Defleecing was judged to be possible by 6-7 days after the start of infusion, and was readily carried out by about 14 days. Defleecing was associated with follicle retrogression and an abrupt cessation of wool growth within 2 days of the start of the infusions. It was estimated that fibre growth stopped for about 10 dyas; regrowth was first observed 17-18 days from the beginning of dosing. Low rates of infusion of mimosine (up to 2 g/day) resulted in plasma levels below 0.1 mmol/l. Infusion at the rate of 4 g/day or above, which produced defleecing, quickly resulted in levels of mimosine in plasma above 0.1 mmol/l; after 2 days the concentration was steady at aboug 0.2 mmol/l. Injections of 8 or 16 g mimosine resulted in very large, but transient, rises of the level in plasma.