During beer production, xanthohumol (XN) extracted from hop through wort boiling is largely converted into its isomeric flavanone, isoxanthohumol (IXN), due to the thermal treatment. XN has a great biological interest as a functional compound (being anti-carciogenic, antioxidant, anti-inflammatory, and anti-infective) and its loss leads to beers with very small amounts of XN and reduced functional properties. In this work, we studied the use of high-pressure treatments (HP; 100, 200, 250, 300, 400 and 500 MPa for 5 min and 250 and 400 MPa for 15 min), applied to previously boiled worts with a second hop dosage, as an extractive procedure, to produce XN-enriched beer worts. The results indicated that beer worts processed by HP showed higher levels of XN. The best results were obtained for 250 MPa, 5 min, resulting in an increase of 4-fold in XN, compared with beer worts produced only by boiling, and 2-fold compared with a method mimicking a recently proposed methodology also intended to improve XN in wort (called XAN method). Combinations of lower pressures with short pressurisation periods proved to be more efficient to increase the amount of XN in the wort. Moreover, HP-processed worts showed lower levels of IXN compared with samples produced by the mimicked XAN method. Physical–chemical characteristics of worts (pH, colour, bitterness and original extract) produced by HP, were similar to those produced only by boiling. These results clearly indicate the potential of using HP treatments to increase the XN content in beer wort, together with less isomerisation to IXN, and open promising possibilities to produce beers with higher amounts of XN, with potentially enhanced functional properties and health promoting properties.