Background and Objective: Human monkeypox epidemic represents a public health emergency. In epidemics, medical students act as alert and reliable public information sources. This study aimed to assess knowledge of Monkeypox among medical students recruited from the Eastern Mediterranean Region. Methods: A cross-sectional study was performed between August-October, 2022. Data was collected through an online predesigned questionnaire that was distributed via social media. The questionnaire consists of two sections; the first is on the sociodemographic characteristics (gender, age, educational year), and the second section included 30 items to assess the knowledge on HMPX (five items on the source and nature of HMPX, 7 items on the mode of transmission of HMPX, 6 items on the clinical manifestation, 8 items on the prevention and control, and 4 items on treatment modalities). The level of knowledge was classified into high (80-100%), moderate (60-80%), and low (less than 60%). For analysis, descriptive statistics were performed. Results: A total of 1,740 medical students participated in the study. The mean age of the participants was 21 years, 63% were females, and 47% of them were in the fifth and fourth-year. Only 10% received a training program on human monkeypox. The findings from this study showed that the majority of participants 848 (48.73%) had a moderate level of knowledge regarding HMPX, 274 (15.747%) had poor knowledge, and 618 (35.49%) had high knowledge. Almost one-third (32%) had knowledge of the availability of the licensed monkeypox vaccine. There was no significant association between knowledge and education year, country of residence, or area of residence (urban/rural) (p= 0.23), (p= 0.92), (p= 0.07) respectively. Conclusions: The study emphasized that knowledge towards human monkeypox introduced to currently enrolled undergraduate medical students in the Eastern Mediterranean Region is inadequate and should be increased to enhance the prevention and control of monkeypox.