De novo biosynthetic pathways are designed, assembled and optimized to produce high-value compounds such as drugs and chemical building blocks from renewable resources. Microorganisms are used as synthetic platforms of systems biology where biochemical pathways are engineered into the host metabolic network. Retrosynthetic biology offers a creative pathway design concept that has gained interest because of its potential to identify novel metabolic ways for therapeutic production. Retrosynthetic biology uses the backward search of retrosynthetic analysis to devise and optimize tailor-made pathways. The retrosynthetic process can be seamlessly integrated into a complete circuitry system for therapeutic applications where production, sensing and delivery act as constitutive interconnecting parts. The aim of this review is to highlight recent efforts toward synthetic design for therapeutic development.