PMID- 35956947 OWN - NLM STAT- MEDLINE DCOM- 20220815 LR - 20220831 IS - 1420-3049 (Electronic) IS - 1420-3049 (Linking) VI - 27 IP - 15 DP - 2022 Aug 5 TI - Anti-Inflammatory Effects of Auranamide and Patriscabratine-Mechanisms and In Silico Studies. LID - 10.3390/molecules27154992 [doi] LID - 4992 AB - Auranamide and patriscabratine are amides from Melastoma malabathricum (L.) Smith. Their anti-inflammatory activity and nuclear factor erythroid 2-related factor 2 (NRF2) activation ability were evaluated using Escherichia coli lipopolysaccharide (LPSEc)-stimulated murine macrophages (RAW264.7) and murine hepatoma (Hepa-1c1c7) cells, respectively. The cytotoxicity of the compounds was assessed using a 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. The anti-inflammatory activity was determined by measuring the nitric oxide (NO) production and pro-inflammatory cytokines (Interleukin (IL)-1beta, Interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha, and IL-6) and mediators (NF-kappaB and COX-2). NRF2 activation was determined by measuring the nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) quinone oxidoreductase 1 (NQO1), nuclear NRF2 and hemeoxygenase (HO)-1. In vitro metabolic stability was assessed using the mouse, rat, and human liver microsomes. The compounds were non-toxic to the cells at 10 muM. Both compounds showed dose-dependent effects in downregulating NO production and pro-inflammatory cytokines and mediators. The compounds also showed upregulation of NQO1 activity and nuclear NRF2 and HO-1 levels. The compounds were metabolically stable in mouse, rat and human liver microsomes. The possible molecular targets of NRF2 activation by these two compounds were predicted using molecular docking studies and it was found that the compounds might inhibit the Kelch domain of KEAP1 and GSK-3beta activity. The physicochemical and drug-like properties of the test compounds were predicted using Schrodinger small molecule drug discovery suite (v.2022-2). FAU - Mak, Kit-Kay AU - Mak KK AUID- ORCID: 0000-0001-6939-5280 AD - Department of Pharmaceutical Chemistry, School of Pharmacy, International Medical University, Kuala Lumpur 57000, Malaysia. AD - Centre of Excellence for Bioactive Molecules and Drug Delivery, Institute for Research, Development & Innovation, International Medical University, Kuala Lumpur 57000, Malaysia. AD - School of Postgraduate Studies, International Medical University, Kuala Lumpur 57000, Malaysia. FAU - Shiming, Zhang AU - Shiming Z AD - School of Postgraduate Studies, International Medical University, Kuala Lumpur 57000, Malaysia. FAU - Low, Jun Sheng AU - Low JS AUID- ORCID: 0000-0003-1142-9593 AD - School of Postgraduate Studies, International Medical University, Kuala Lumpur 57000, Malaysia. FAU - Balijepalli, Madhu Katyayani AU - Balijepalli MK AD - Department of Pharmacology, Faculty of Medicine, MAHSA University, Jenjarom 42610, Malaysia. FAU - Sakirolla, Raghavendra AU - Sakirolla R AD - Department of Chemistry, Central University of Karnataka, Gulbarga 585367, India. FAU - Dinkova-Kostova, Albena T AU - Dinkova-Kostova AT AUID- ORCID: 0000-0003-0316-9859 AD - Jacqui Wood Cancer Centre, Division of Cellular Medicine, School of Medicine, University of Dundee, Dundee DD1 4HN, UK. AD - Departments of Medicine and Pharmacology and Molecular Sciences, School of Medicine, Johns Hopkins University, Baltimore, MD 21205, USA. FAU - Epemolu, Ola AU - Epemolu O AD - Charles River Laboratories Edinburgh Ltd., Tranent EH33 2NE, UK. FAU - Mohd, Zulkefeli AU - Mohd Z AD - Department of Pharmaceutical Chemistry, School of Pharmacy, International Medical University, Kuala Lumpur 57000, Malaysia. AD - Centre of Excellence for Bioactive Molecules and Drug Delivery, Institute for Research, Development & Innovation, International Medical University, Kuala Lumpur 57000, Malaysia. FAU - Pichika, Mallikarjuna Rao AU - Pichika MR AD - Department of Pharmaceutical Chemistry, School of Pharmacy, International Medical University, Kuala Lumpur 57000, Malaysia. AD - Centre of Excellence for Bioactive Molecules and Drug Delivery, Institute for Research, Development & Innovation, International Medical University, Kuala Lumpur 57000, Malaysia. LA - eng GR - FRGS/1/2018/WAB13/MAHSA/02/1/Ministry of Higher Education/ GR - PMHS I-2018 (02)/International Medical University/ PT - Journal Article DEP - 20220805 PL - Switzerland TA - Molecules JT - Molecules (Basel, Switzerland) JID - 100964009 RN - 0 (Amides) RN - 0 (Anti-Inflammatory Agents) RN - 0 (Cytokines) RN - 0 (Flavonoids) RN - 0 (Kelch-Like ECH-Associated Protein 1) RN - 0 (Lipopolysaccharides) RN - 0 (NF-E2-Related Factor 2) RN - 0 (NF-kappa B) RN - 0 (patriscabratine) RN - 31C4KY9ESH (Nitric Oxide) RN - EC 1.14.14.18 (Heme Oxygenase-1) RN - EC 2.7.11.1 (Glycogen Synthase Kinase 3 beta) SB - IM MH - *Amides/pharmacology MH - Animals MH - *Anti-Inflammatory Agents/pharmacology MH - Cytokines/metabolism MH - *Flavonoids/pharmacology MH - Glycogen Synthase Kinase 3 beta/metabolism MH - *Heme Oxygenase-1/metabolism MH - Humans MH - Kelch-Like ECH-Associated Protein 1/metabolism MH - Lipopolysaccharides/pharmacology MH - Mice MH - Molecular Docking Simulation MH - *NF-E2-Related Factor 2/metabolism MH - NF-kappa B/metabolism MH - Nitric Oxide/metabolism MH - Rats PMC - PMC9370761 OTO - NOTNLM OT - KEAP1 OT - Melastoma malabathricum OT - NRF2 OT - anti-inflammatory OT - auranamide OT - patriscabratine COIS- The authors have no conflict of interest to declare. EDAT- 2022/08/13 06:00 MHDA- 2022/08/16 06:00 PMCR- 2022/08/05 CRDT- 2022/08/12 01:20 PHST- 2022/07/11 00:00 [received] PHST- 2022/08/02 00:00 [revised] PHST- 2022/08/02 00:00 [accepted] PHST- 2022/08/12 01:20 [entrez] PHST- 2022/08/13 06:00 [pubmed] PHST- 2022/08/16 06:00 [medline] PHST- 2022/08/05 00:00 [pmc-release] AID - molecules27154992 [pii] AID - molecules-27-04992 [pii] AID - 10.3390/molecules27154992 [doi] PST - epublish SO - Molecules. 2022 Aug 5;27(15):4992. doi: 10.3390/molecules27154992.