A new invention, papain-based wound cleanser is formulated by incorporating papain, a proteolytic enzyme extracted from Carica papaya into the formulation. This cleanser is invented to simplify the methods in wound management by combining wound cleansing and wound debridement using a single formulation. This study describes the preparation and preclinical study of papain-based wound cleanser in accelerating wound healing. In this study, papain-based wound cleanser was used to treat wound incision on Sprague-Dawley rats while distilled water and Betadine were used as negative and positive control. Twenty-seven clinically healthy white rats were randomly divided into three groups and treated accordingly until the 21st day post-incision. Wound reduction rates and histological analysis were obtained to asses the healing pattern. Rats treated with papain-based wound cleanser showed a progressive wound healing based on the wound reduction rates and histological analysis when compared with rats treated with distilled water and Betadine. Better collagen deposition and presence of skin organelles in rats treated with papain-based wound cleanser demonstrated its efficacy in promoting wound healing. In addition to its wound healing effect, papain-based wound cleanser is also integrated with antibacterial properties which make it a complete package for wound management. However, further studies should be carried out to ensure its safety for human usage.