Background: Implementation and uptake of novel and cost-effective medicines can improve patient health outcomes and healthcare efficiency. However, the uptake of new medicines into practice faces a wide range of obstacles Earlier reviews provided insight into determinants for new medicine uptake (such as medicine, prescriber, patient, organization, and external environment factors), but the methodological approaches used had limitations (e.g., single author, narrative review, narrow search, no quality assessment of reviewed evidence).This systematic review aims to identify barriers and facilitators affecting the uptake of new medicines into clinical practice and identify areas for future research. Method: A systematic search was undertaken within seven databases. Eligible qualitative, quantitative, and mixed-methods studies focused on adult participants (18 years and older) requiring or taking new medicine(s) for any condition, in the context of healthcare organizations and identified factors affecting the uptake of new medicines. The methodological quality was assessed using QASTDD tool. A narrative synthesis of reported factors was conducted using framework analysis and conceptual framework was utilised to group them. Results: A total of 66 studies were included. Most studies (n=62) were quantitative and used secondary data (n=46) from various databases, e.g., insurance databases. The identified factors had a varied impact on the uptake of the different studied new medicines. Differently from earlier reviews, patient factors (patient education, engagement with treatment, therapy preferences), cost of new medicine, reimbursement and formulary conditions, and guidelines were suggested to influence the uptake. Also, the review highlighted that health economics, wider organizational factors, and underlying behaviours of adopters were not or under explored. Conclusion: This systematic review identifies additional factors affecting new medicine use not reported in earlier reviews, which included patient influence and education level, cost of new medicines, formulary and reimbursement restrictions, and guidelines. Further research employing determinant frameworks or implementation theories is needed to gain a better understanding of factors, especially patient, prescriber, and organizational, affecting the uptake of new medicines into clinical practice.Registration: PROSPERO database (CRD42018108536)