Introduction: Adolescents are particularly vulnerable to risky behaviour, such as the use of psychoactive substances (alcohol, tobacco and illicit drugs), which can lead to other life-long disorders. As substance consumption has increased over the years, it is important to integrate new perspectives into the culture of prevention and care in the youth and general population. The aim of this study was to assess the relationships between mindfulness Dispositional, spirituality and religion, and their role as factors protecting the use of psychoactive substances in adolescents, as well as analysing differences in mindfulness and spiritual scores between consumers and non-users of substances, and between different religious/spiritual groups. Method: A cross-sectional study was carried out, evaluating 433 adolescents with upper middle level students aged 14-19 (Media = 16.27 ±.91). Statistical correlation tests were performed to assess the relationship between the variables, in addition to Student’s and unidirectional ANOVA’s t-tests to explore differences in mindfulness and spiritual scores between consumer and non-consumer groups of substances, and between religious/spiritual groups, respectively. Finally, by means of a binary logistic regression analysis, the degree of prediction of the use of psychoactive substances was assessed. Results: The sample reported higher use of psychoactive substances compared to the latest national survey. Mindfulness Dispositional was positively correlated with spirituality and religion, and inversely with substance use, with a mild to moderate force. In addition, individuals who used psychoactive substances had significantly lower scores in mindfulness than they had never consumed at all, but no differences were found with regard to spirituality scores. The scores for mindfulness and spirituality were significantly higher for those formally belonging to a religion. Finally, belonging to a religion, followed by mindfulness Dispositional, were the best predictive variables as a factor in protecting alcohol and drug consumption, while spirituality was not a variable that contributed significantly to models and was therefore eliminated. Discussion/Conclusion: Minidfulness Dispositional, spirituality and religion are interrelated variables. Belonging to religion and mindfulness Dispositional could be a better protective factor for the consumption of substances in adolescents than spirituality. Dispositional mindfulness was also higher in those formally practising a religion. Finally, the use of psychoactive substances could alter mindfulness related skills, but more research is needed to complement and clarify these results in the Mexican population.