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Biosci. (Landmark Ed)"],"abstract":"<jats:p>Background: Immunomodulators play a central role in the treatment of cancer and immune-mediated disorders. Small-molecule immunomodulators are particularly attractive due to their structural diversity, pharmacological versatility, and potential for oral administration. This study aimed to investigate the immunomodulatory potential of newly synthesized aza-spirocyclic derivatives. Methods: A series of novel aza-spirocyclic compounds was evaluated for cytotoxicity in RAW264.7 murine macrophages. Immunomodulatory activity was assessed by quantifying nitric oxide (NO) production and profiling pro- and anti-inflammatory cytokines in macrophages and murine splenocytes. In vitro anti-inflammatory effects were further examined using a lipopolysaccharide (LPS)-induced inflammation model. Mechanistic insights were explored using molecular docking and immuno-informatics analyses. Results: All compounds were non-toxic at the tested concentrations. Most derivatives failed to induce NO production, indicating an absence of danger-associated molecular pattern (DAMP)-like activity. Several compounds significantly reduced the pro-inflammatory cytokine interleukin-6 (IL-6) while increasing the anti-inflammatory cytokine IL-10 in vitro. In the LPS-induced inflammation model, four compounds notably suppressed IL-6 and enhanced IL-10 expression. The binding affinities obtained from docking, along with in vitro validation of these compounds on Janus kinases (JAKs) signaling, suggested that compounds AS8 and AS10 modulated inflammatory signaling pathways. Conclusion: The synthesized aza-spirocyclic derivatives exhibit promising anti-inflammatory profiles, characterized by IL-6 suppression and IL-10 enhancement. These findings position aza-spirocyclic scaffolds as candidates for the development of next-generation small-molecule immunomodulators.<\/jats:p>","DOI":"10.31083\/fbl48867","type":"journal-article","created":{"date-parts":[[2026,5,30]],"date-time":"2026-05-30T07:25:36Z","timestamp":1780125936000},"source":"Crossref","is-referenced-by-count":0,"title":["Immunomodulatory Effect of Aza-Spirocyclic Frameworks"],"prefix":"10.31083","volume":"31","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-3855-1178","authenticated-orcid":true,"given":"Anjali","family":"Prajapati","sequence":"first","affiliation":[{"name":"Vaccine Immunology Laboratory, Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, 500007 Hyderabad, Telangana, India"},{"name":"Academy of Scientific and Innovative Research (AcSIR), 201002 Ghaziabad, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1007-5888","authenticated-orcid":true,"given":"Rajitha Rajeshwar","family":"Tatikonda","sequence":"additional","affiliation":[{"name":"Vaccine Immunology Laboratory, Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, 500007 Hyderabad, Telangana, India"},{"name":"Academy of Scientific and Innovative Research (AcSIR), 201002 Ghaziabad, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1491-7381","authenticated-orcid":true,"given":"Raji Reddy","family":"Chada","sequence":"additional","affiliation":[{"name":"Academy of Scientific and Innovative Research (AcSIR), 201002 Ghaziabad, India"},{"name":"Department of Organic Synthesis & Process Chemistry, CSIR-Indian Institute of Chemical Technology, 500007 Hyderabad, Telangana, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2888-2418","authenticated-orcid":true,"given":"Srinivasa Reddy","family":"Bonam","sequence":"additional","affiliation":[{"name":"Vaccine Immunology Laboratory, Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, 500007 Hyderabad, Telangana, India"},{"name":"Academy of Scientific and Innovative Research (AcSIR), 201002 Ghaziabad, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"16242","published-online":{"date-parts":[[2026,5,18]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","unstructured":"Kim MK, Shin H, Park KS, Kim H, Park J, Kim K, et al. 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