As part of the study of pyrrole containing biheterocyclic derivatives we have synthesized different pyrrole-azole biheterocyclic derivatives: pyrrole-pyrazolones, pyrrole-pyrazoles and pyrrole 1,3,4 oxadiazoles have been synthesized using cyclization of pyrrolyl hydrazide-hydrazone intermediates in the presence of selected reagents. This work deals with structural clarification, NLO analysis and antimicrobial / anticancer activity evaluation of synthesized compounds. The structure of the pyrrole-azole derivatives were confirmed by modern spectroscopic methods. All quantum chemical calculations have been performed by density functional theory (DFT), using B3LYP functional and 6-311++G(d, p) as basis set. The calculated data corroborate well with the experimental data. The FT-IR analysis shows red shifts in vN–H and vCO stretching as result of dimer formation. The binding energies of dimer are found in the range of 12.75 - 11.21 kcal/mol after basis set superposition error. The UV–vis result shows major transparent over visible range enabling their suitability for application in the visible range. The optical energy gaps of these derivatives were found to be less than 3.2 eV. The β0 values for compounds (5a-b, 6a-b, 7a-b) are calculated as 24.61 × 10−30, 24.61 × 10−30, 42.510 × 10−30, 10.065 × 10−30, 19.146 × 10−30, 6.432 × 10−30 esu, respectively, indicating suitability for better non–linear optical response. All compounds (5a-b, 6a-6b, 7a-7b) have been screened against microbes and exhibited good antimicrobial activity. All compounds evaluated for their antiproliferative activity against two human cancer cell lines, i.e., HCT-116 (colon), and HL-60 (leukemia) cells. The compounds showed significant cytotoxicity against types of human cancer cell lines. In addition, the molecular docking studies have been achieved using 5b0e, 1i6w, 1q8i, 6p8u, bacterial, 6t1u, 1KS5 fungal and cancer cell protein 3QX3.