Carlina acaulis L. can provide many valuable phytochemicals. However, the plant has a limited range and is under partial protection in many countries. Therefore, it is necessary to develop a cultivation methodology, including the mode of fertilisation. This study presents answers to the following research hypotheses: 1) C. acaulis plants can be successfully cultivated outside the natural habitat; 2) Nitrogen fertilisation of C. acaulis plants affects the yield of valuable metabolites. A cultivation experiment was carried out in a two-factor scheme with the random block method in five replicates. The research factors included the nitrogen dose (0, 4, and 8 g/m2) and the type of the nitrogen fertiliser, i.e. ammonium sulphate and calcium nitrate. The biomass yield and the metabolite content were determined with the gravimetric method and high performance liquid chromatography, respectively. It was found that C. acaulis plants were able to grow outside their natural habitat and nitrogen fertilisation negatively affected the yield of metabolites per plantation area. One hectare of cultivation without nitrogen fertilisation can provide approx. 18 kg of ursolic acid, 5 kg of oleanolic acid, 70 kg of chlorogenic acid, and 10 kg of carlina oxide. However, more research is needed to determine the effects of plant density, harvest time, and requirements for other nutrients.