OBJECTIVES: Digitalization, flexibilization, boundaryless work, participation, and meaningfulness are characteristics of new ways of work ("Work 4.0") and have an effect on the health and well-being of employees. For workplace health promotion (WHP) measures to be effective, companies must know what challenges they face or what opportunities are available. The present study examines challenges and opportunities of digitalization for health and well-being at work, interrelations with key aspects of Work 4.0, and differences between companies. *** METHODS: We chose a mixed-methods approach and followed 20 companies along a WHP project management cycle. We conducted semistructured group interviews, document analyses of the developed WHP catalogs of measures, and an online survey to quantify and validate the results. *** RESULTS: We identified challenges and opportunities for health and wellbeing at work resulting from Work 4.0. The main challenges were the sensitization of managers and the constant availability of employees as a health risk; the main opportunities were the optimization of knowledge transfer, communication processes, documentation, and work information materials. The companies surveyed were similar in most outcomes, only two opportunities varied in relevance across companies. Regarding the key aspects of Work 4.0, only "participation" was associated with some of the identified opportunities. *** CONCLUSIONS: Companies can benefit from addressing Work 4.0 and health and well-being at work together. To realize meaningful and target-oriented solutions, companies should identify the challenges and opportunities perceived as most important before implementing measures. Considering these basic principles, WHP can support the digital transformation change that companies face today.