Transdermal drug delivery (TDD) is a painless method of delivering drug systemically by applying a drug formulation onto intact and healthy skin. Serratiopeptidase is a proteolytic enzyme which shows effective treatment for inflammation, pain and has been used for various treatment namely arthritis, atherosclerosis, post - operative swelling and bronchitis, sinusitis, fibrocystic breast disease. Cubosomes are increasingly being recognized and utilized as a drug delivery system, particularly in the treatment of diseases like cancer. Topical gel formulation of serratiopeptidase was prepared using polymer such as pluronic F- 127 and lipid such as a Glyceryl monooleate. The formulation was optimized using 32 full factorial design and was evaluated for various parameters such as particle size, polydispersity index, zeta potential, invitro drug release and surface morphology was analysed by transmission electron microscopy. The optimized cubosomal formulation was incorporated into a gel and was evaluated for various parameters such as pH, spreadability, washability, viscosity, drug content, and in-vitro drug diffusion with franz diffusion cell. The optimized formulation F1 cubosomal formulation showed an uniform and homogenous spherical shaped structure as confirmed by TEM imaging, the particle size of the compound was between 200-500nm, it showed Entrapment efficiency 87.3%, polydispersity index was 0.123cps, and zeta potential was 87.73%. The optimized gel showed better results due to its small particles size and high entrapment efficiency leading to its better topical delivery of serratiopeptidase cubosomal gel in arthritis.