Small molecules with novel mechanisms of action are necessary in addressing today’s challenging biomedical problems such as resistance to existing drugs for the treatment of cancer and infectious diseases. One of the approaches to the discovery of new bioactive molecules includes diversity-oriented synthesis (DOS), which primarily implies differences in the molecular scaffold, substituents and stereochemistry. Various drugs containing a morpholine ring in their structure are widely used for the treatment of numerous diseases. It is a privileged pharmacophore with the possibility of forming molecular interactions with diverse biological targets and modifying pharmacokinetic properties. Therefore, morpholine is a structural motif of special interest for the drug design and development. At the same time, interest for the synthesis of less planar bioactive compounds increases, resulting in the introduction of spiro compounds into new studies. Following the stated facts, the aim of this thesis was to synthesize and characterize polysubstituted piperidino-morpholine spiro building block. Furthermore, this scaffold was derivatized in order to obtain novel compounds with different properties, using the diversity-oriented synthesis approach. The scaffold was synthesized via two synthetic routes, in six or four reaction steps, with the last step of intramolecular cyclization under basic conditions. In several subsequent chemical reactions, the corresponding precursors were prepared from the main intermediate 6. Six novel derivatives 9a-c and 13a-c were obtained from compounds 8 and 12 with two free amino groups by N-acetylation, sulfonamidation and Eschweiler-Clarke reaction respectively. Structural diversity was achieved by substituting the scaffold at three different positions. The structures of the novel compounds were confirmed by standard spectroscopic methods (1D and 2D NMR, MS). Noticeable three-dimensionality of the structures of the final compounds 9a-c and 13a-c was proved by means of NOESY NMR spectra. Additionally, the physicochemical properties of final compounds were calculated in silico.