{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T11:03:36Z","timestamp":1789383816862,"version":"build-2803163510"},"reference-count":27,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T00:00:00Z","timestamp":1789344000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Beijing Life Science Academy Research Project","award":["2025100CE0030"],"award-info":[{"award-number":["2025100CE0030"]}]},{"name":"Beijing Postdoctoral Research Foundation Project","award":["2025100AS0020"],"award-info":[{"award-number":["2025100AS0020"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Persistent inflammation driven by excessive AKR1B1 activation contributes to the progression of multiple chronic inflammatory diseases. Natural chlorogenic acid (CGA) exhibits anti-inflammatory activity but suffers from poor membrane permeability, while its laurate ester derivative (CGL) shows clear dose-dependent cytotoxicity in macrophages. Here, we synthesized a novel chlorogenic acid amide derivative (CGAA) by selective amidation of the aliphatic carboxyl group on the quinic acid moiety of CGA, fully preserving the chiral polyphenol scaffold. Structural identity was confirmed by HSQC, HMBC, FTIR and HR-ESI-MS. CCK-8 assays demonstrated that CGAA maintained &gt; 95% viability of RAW264.7 cells at 100 \u03bcM, displaying markedly superior cellular safety relative to CGL. In LPS-stimulated RAW264.7 macrophages, CGAA dose-dependently suppressed NO, TNF-\u03b1 and IL-6 production and exhibited stronger antioxidant activity than both CGA and CGL. Temperature-gradient CETSA, DARTS and enzymatic assays established direct binding and potent inhibition of AKR1B1 by CGAA (IC50 = 7.85 \u03bcM). Molecular docking revealed that the newly introduced amide forms an additional hydrogen bond with ILE-260, enhancing ligand\u2013target affinity. This carboxyl-amidation strategy simultaneously improves the cellular safety and AKR1B1 inhibitory potency of CGA, providing a rational approach for developing low-toxicity polyphenol-based anti-inflammatory leads.<\/jats:p>","DOI":"10.3390\/molecules31183247","type":"journal-article","created":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T10:19:41Z","timestamp":1789381181000},"page":"3247","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Carboxyl Amidation of Chlorogenic Acid Improves Anti-Inflammatory Activity and Biosafety via Potent AKR1B1 Inhibition in LPS-Induced Macrophages"],"prefix":"10.3390","volume":"31","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-1544-7731","authenticated-orcid":false,"given":"Faliu","family":"Yang","sequence":"first","affiliation":[{"name":"Beijing Life Science Academy, Beijing 102200, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"}]},{"given":"Chunmei","family":"Guang","sequence":"additional","affiliation":[{"name":"Beijing Life Science Academy, Beijing 102200, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"}]},{"given":"Huan","family":"Chen","sequence":"additional","affiliation":[{"name":"Beijing Life Science Academy, Beijing 102200, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"conceptualization","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]},{"given":"Hongwei","family":"Hou","sequence":"additional","affiliation":[{"name":"Beijing Life Science Academy, Beijing 102200, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"supervision","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"8599","DOI":"10.18632\/aging.205827","article-title":"Senolytic effects of exercise in human muscles require acute inflammation","volume":"16","author":"Jean","year":"2024","journal-title":"Aging"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"428","DOI":"10.1038\/nature07201","article-title":"Origin and physiological roles of inflammation","volume":"454","author":"Medzhitov","year":"2008","journal-title":"Nature"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Oxombre, B., Madouri, F., Journ\u00e9, A.S., Ravez, S., Woitrain, E., Odou, P., Duhal, N., Ninni, S., Montaigne, D., and Delhem, N. (2022). Safe and Efficient Sigma1 Ligand: A Potential Drug Candidate for Multiple Sclerosis. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms231911893"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"7204","DOI":"10.18632\/oncotarget.23208","article-title":"Inflammatory responses and inflammation-associated diseases in organs","volume":"9","author":"Chen","year":"2018","journal-title":"Oncotarget"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"889","DOI":"10.1038\/ni.1937","article-title":"Macrophage plasticity and interaction with lymphocyte subsets: Cancer as a paradigm","volume":"11","author":"Biswas","year":"2010","journal-title":"Nat. Immunol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"958","DOI":"10.1038\/nri2448","article-title":"Exploring the full spectrum of macrophage activation","volume":"8","author":"Mosser","year":"2008","journal-title":"Nat. Rev. Immunol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Ferreira-Santos, P., Badim, H., Salvador, \u00c2.C., Silvestre, A.J.D., Santos, S.A.O., Rocha, S.M., Sousa, A.M., Pereira, M.O., Wilson, C.P., and Rocha, C.M.R. (2021). Chemical Characterization of Sambucus nigra L. Flowers Aqueous Extract and Its Biological Implications. Biomolecules, 11.","DOI":"10.3390\/biom11081222"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Huang, J., Xie, M., He, L., Song, X., and Cao, T. (2023). Chlorogenic acid: A review on its mechanisms of anti-inflammation, disease treatment, and related delivery systems. Front. Pharmacol., 14.","DOI":"10.3389\/fphar.2023.1218015"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Santana-G\u00e1lvez, J., Cisneros-Zevallos, L., and Jacobo-Vel\u00e1zquez, D.A. (2017). Chlorogenic Acid: Recent Advances on Its Dual Role as a Food Additive and a Nutraceutical against Metabolic Syndrome. Molecules, 22.","DOI":"10.3390\/molecules22030358"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Zalewska, K., Kulawik, M., Gierszewska, J., Gramala, Z., Kalus, O., Karpi\u0144ski, M., Ma\u0107kowiak, J., Staniewski, A., Szyma\u0144ska, Z., and Zalewska, B. (2025). Chlorogenic Acid\u2019s Role in Metabolic Health: Mechanisms and Therapeutic Potential. Nutrients, 17.","DOI":"10.3390\/nu17203303"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Liang, N., and Kitts, D.D. (2015). Role of Chlorogenic Acids in Controlling Oxidative and Inflammatory Stress Conditions. Nutrients, 8.","DOI":"10.3390\/nu8010016"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"292","DOI":"10.1016\/j.cbpa.2008.03.009","article-title":"Peptidomimetics, a synthetic tool of drug discovery","volume":"12","author":"Vagner","year":"2008","journal-title":"Curr. Opin. Chem. Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"129904","DOI":"10.1016\/j.foodchem.2021.129904","article-title":"Antioxidant activities of chlorogenic acid derivatives with different acyl donor chain lengths and their stabilities during in vitro simulated gastrointestinal digestion","volume":"357","author":"Wang","year":"2021","journal-title":"Food Chem."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Wang, L.N., Wang, W., Hattori, M., Daneshtalab, M., and Ma, C.M. (2016). Synthesis, Anti-HCV, Antioxidant and Reduction of Intracellular Reactive Oxygen Species Generation of a Chlorogenic Acid Analogue with an Amide Bond Replacing the Ester Bond. Molecules, 21.","DOI":"10.3390\/molecules21060737"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"606","DOI":"10.1039\/B701677H","article-title":"Amide bond formation: Beyond the myth of coupling reagents","volume":"38","author":"Valeur","year":"2009","journal-title":"Chem. Soc. Rev."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"250","DOI":"10.1016\/j.cbi.2008.11.007","article-title":"Posttranslational glutathiolation of aldose reductase (AKR1B1): A possible mechanism of protein recovery from S-nitrosylation","volume":"178","author":"Baba","year":"2009","journal-title":"Chem. Biol. Interact."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.biocel.2009.09.009","article-title":"Aldose reductase: A novel therapeutic target for inflammatory pathologies","volume":"42","author":"Ramana","year":"2010","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"110856","DOI":"10.1016\/j.cbi.2024.110856","article-title":"Aldose reductase is a potential therapeutic target for neurodegeneration","volume":"389","author":"Rao","year":"2024","journal-title":"Chem. Biol. Interact."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"117942","DOI":"10.1016\/j.bmc.2024.117942","article-title":"Inhibition of LPS-induced inflammatory response in RAW264.7 cells by natural Chlorogenic acid isomers involved with AKR1B1 inhibition","volume":"114","author":"Han","year":"2024","journal-title":"Bioorg. Med. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1002\/9780470559277.ch110180","article-title":"Target identification using drug affinity responsive target stability (DARTS)","volume":"3","author":"Lomenick","year":"2011","journal-title":"Curr. Protoc. Chem. Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1255784","DOI":"10.1126\/science.1255784","article-title":"Tracking cancer drugs in living cells by thermal profiling of the proteome","volume":"346","author":"Savitski","year":"2014","journal-title":"Science"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Saeed, A., Ejaz, S.A., Sarfraz, M., Tamam, N., Siddique, F., Riaz, N., Qais, F.A., Chtita, S., and Iqbal, J. (2022). Discovery of Phenylcarbamoylazinane-1,2,4-Triazole Amides Derivatives as the Potential Inhibitors of Aldo-Keto Reductases (AKR1B1 & AKRB10): Potential Lead Molecules for Treatment of Colon Cancer. Molecules, 27.","DOI":"10.3390\/molecules27133981"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Ali, M.Y., Zaib, S., Jannat, S., Khan, I., Rahman, M.M., Park, S.K., and Chang, M.S. (2022). Inhibition of Aldose Reductase by Ginsenoside Derivatives via a Specific Structure Activity Relationship with Kinetics Mechanism and Molecular Docking Study. Molecules, 27.","DOI":"10.3390\/molecules27072134"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Gu, T., Zhang, Z., Liu, J., Chen, L., Tian, Y., Xu, W., Zeng, T., Wu, W., and Lu, L. (2023). Chlorogenic Acid Alleviates LPS-Induced Inflammation and Oxidative Stress by Modulating CD36\/AMPK\/PGC-1\u03b1 in RAW264.7 Macrophages. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms241713516"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1122","DOI":"10.4049\/jimmunol.1402101","article-title":"IRAK4 as a molecular target in the amelioration of innate immunity-related endotoxic shock and acute liver injury by chlorogenic acid","volume":"194","author":"Park","year":"2015","journal-title":"J. Immunol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1016\/j.cbpa.2008.03.016","article-title":"The impact of natural products upon modern drug discovery","volume":"12","author":"Ganesan","year":"2008","journal-title":"Curr. Opin. Chem. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1189","DOI":"10.1016\/j.lwt.2007.08.006","article-title":"Scavenging and antioxidant properties of compound derived from chlorogenic acid in South-China honeysuckle","volume":"41","author":"Xiang","year":"2008","journal-title":"LWT-Food Sci. Technol."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/18\/3247\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T10:20:10Z","timestamp":1789381210000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/18\/3247"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,14]]},"references-count":27,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["molecules31183247"],"URL":"https:\/\/doi.org\/10.3390\/molecules31183247","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,14]]}}}