Research on new biological active substances of natural origin is also a significant field of research, due to the growing number of adverse effects of synthetic drugs. For this reason, any new research on the from nature isolated compounds can contribute to the pharmaceutical industry, or the development of new drugs. Inhibition of important enzymes is one of the fundamental therapies for treating diseases such as Alzheimer's dementia (AD) and Diabetes mellitus (DM). Genus Centaurea L. is one of the most important families of the Asteraceae family, with a large number of species. In Croatia there are around 80 species of this genus, of which 27 are endemic. In this study, the chemical and biological profile of the hydrolysates of five plants of the genus Centaurea was determined. The chemical profile of the hydrolysates was determined by qualitatively proving some secondary metabolites and quantitatively determining the content of the total phenol and flavonoid components of the hydrolates. Biological potential was investigated in terms of the inhibition of acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and α-glucosidase enzymes. Glycosides and saponins have been shown to be beneficial in plant hydrolytes, while alkaloids, tandines, steroids and terpenoids have not been proven. The results show that all samples of the Centaurea plant hydrolysates contain a relatively low content of phenolic and flavonoid compounds. C. scabiosa and C. rhenana hydrolytes inhibit AChE, C. triumfetti and C. ragusina show low inhibiting of AChE, whereas C. alba does not inhibit AChE. C. alba, C. rhenana and C. scabiosa samples are poorly inhibiting BChE, while C. triumfettii and C. ragusina do not inhibit this enzyme. C. scabiosa inhibits the enzyme α-glucosidase noticeably, C. rhenana sample weakly inhibits the α-glucosidase enzyme while the other hydrolysates do not inhibit this enzyme.