Summary Background  Retinoids and active vitamin D3 analogues regulate the proliferation and differentiation of keratinocytes and are effective in the treatment of psoriasis. Retinoids are known to be effective against acne vulgaris through comedolysis. However, the comedolytic effect of active vitamin D3 analogues has not been reported. Objectives  To investigate whether maxacalcitol, one of the active vitamin D3 analogues, has a comedolytic effect by using spontaneously comedogenic rhino mice. Methods  Rhino mice were treated topically with tretinoin and maxacalcitol once daily for 2 and 4 weeks, respectively. The dermal side of the epidermal sheet was observed to determine the size of the utricle. Haematoxylin and eosin‐stained vertical sections were used to measure utricle diameter and density and to evaluate histological changes. Results  Maxacalcitol (25 μg g−1) and tretinoin (0·1%) significantly decreased the size and the diameter of the utricle after 1 week of treatment. However, maxacalcitol did not affect the density while tretinoin did have an effect. Histopathologically, maxacalcitol and tretinoin markedly induced epidermal hyperplasia accompanied by a minor accumulation of inflammatory cells in the dermis, with and without hypercornification, respectively. Conclusions  These results indicate that maxacalcitol has a prominent effect on comedolysis and that its mechanism of action may be different from that of retinoids.