Preclinical testing and validation of therapeutic strategies developed for patients with type 1 diabetes (T1D) require a cohort of virtual patients (VPs). However, current simulators provide a limited number of VPs, lack real-life scenarios, and inadequately represent intra- and inter-day variability in insulin sensitivity and blood glucose (BG) profile. The generation of a realistic scenario was achieved by using the meal patterns, insulin profiles (basal and bolus), and exercise sessions estimated as disturbances using clinical data from a cohort of 14 T1D patients using the Medtronic 640G insulin pump provided by the Hospital Clínic de Barcelona. The UVa/Padova’s cohort of adult patients was used for the generation of a new cohort of VPs. Insulin model parameters were optimized and adjusted in a day-by-day fashion to replicate the clinical data to create a cohort of 75 VPs. All primary and secondary outcomes reflecting the BG profile of a T1D patient were analyzed and compared to the clinical data. The mean BG 166.3 versus 162.2 mg/dL ( = 0.19), coefficient of variation 32% versus 33% ( = 0.54), and percent of time in range (70 to 180 mg/dL) 59.6% versus 66.8% ( = 0.35) were achieved. The proposed methodology for generating a cohort of VPs is capable of mimicking the BG metrics of a real cohort of T1D patients from the Hospital Clínic de Barcelona. It can adopt the inter-day variations in the BG profile, similar to the observed clinical data, and thus provide a benchmark for preclinical testing of control techniques and therapy strategies for T1D patients.