JOURNAL OF CHEMICAL INFORMATION AND MODELING pubs. acs. org/jcim Molecular Modeling of the 3D Structure of 5-HT1AR: Discovery of Novel 5-HT1AR Agonists via Dynamic Pharmacophore-Based virtual Screening Lili xut Shanglin Zhon Kunqian Yu. t s Bo Gao, A\ Hualiang jiang *f Xuechu Zhen, and Wei Fu*,t 'Department of Medicinal Chemistry Key Laboratory of Smart Drug Delivery, Ministry of Education, School of Pharmacy,Fudan University, Shanghai 201203, China State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, Department of Pharmacology Soochow University College of Pharmaceuticals Sciences, Suzhou 215123, China S Supporting Information ABSTRACT: The serotonin receptor subtype 1A(5-HTIAR) has een implicated in several neurological conditions, and potent 5- Best Hit: Fwo1 HT1AR agonists have therapeutic potential for the treatment of Ki=51.4 nm EC50=7 nm de Ixiety, schizophrenia, and Parkinsons disease. In the present study, a homology model of 5-HTIaR was built based on the latest released high-resolution crystal structure of the B2AR in its active state(PDB: 3SN6). a dynamic pharmacophore model, which takes the receptor flexibility into account, was constructed, validated, and applied to our dynamic pharmacophore-based virtual y screening approach with the aim to identify potential S-HTIAR agonists. The obtained hits were subjected to S-HTIAR binding and functional assays, and 10 compounds with medium or high K and ECso values were identified. Among them, FWOl(K:=51.9 nM, ECso=7 nM)was evaluated as the strongest agonist for 5 HTLAR The active S-HTIAR model and dynamic pharmacophore model obtained from this study can be used for future discovery and design of novel S-HTIAR agonists. Also, by integrating all computational and available experimental data,a stepwise 5-HTIAR signal transduction model induced by agonist Fwol was proposed. ■ INTRODUCTION antipsychotic), F-13640(highly selective and effective full Serotonin(5-hydroxytrptamine, S-HT), one of the most agonist; undergoing clinical trials for the treatment of pain) important neurotransmitters in the brain, was involved (Table 1). Given the rising interest on 5-HT1AR agonists,new most of the human behavioral and neuropsychological processes, modulating mood, cognition and memory, feeding, Table 1. Pharmacophore Model Validation Using Statistical sexuality, sleep, and pain, etc. Abnormal S-HT transmission isParameters associated with pathogenesis of psychiatric disorders and neurodegenerative disorders. To date, at least 16 serotonin value receptor subtypes have been cloned which are grouped into 1000 seven subfamilies(5-HT1-7)based on their different signaling total number of actives in database(A) echanisms. Among them 5-HTIaR was the first one to be total hits(Ht) fully sequenced and widely studied. Its involvement in anxiety and depression has long been documented -GI evidence has revealed new therapeutic applications of 5- 9 ratio of actives in the hit list HTIAR in the treatment of schizophrenia, Parkinsons disease, enrichment factor or enhancement(E) Accordingly, intensive efforts have been made to explore the fals herapeutic application of 5-HTIAR agonists in the past three H score(goodness of hit list) decades. Representative 5-HTIAR agonists include R-8- E=(Ha/Ht)/(A/D);GH =[((Ha/4HtA)(3A +Ht))(1((HtHa)/ OH-DPAT(full agonist; widely used agonistic tool drug), (DAD))I, GH score of 0.6-0 7 indicates a good model. Buspirone(partial agonist; anxiolytic), Vilazodone(SSRI/S- HTIAR partial agonist; antidepressant),Aripirazole(mixed Received: August 14, 2013 eceptor profile with 5-HTIAR agonistic activity; atypical Published: November 18, ACS Publications o2013 American Chemical Society 3202 dxdoLor/10. 1021/c 00481plJ Chem Inf Model. 2013, 53, 3202-3211Molecular Modeling of the 3D Structure of 5‑HT1AR: Discovery of Novel 5‑HT1AR Agonists via Dynamic Pharmacophore-Based Virtual Screening Lili Xu,†,§ Shanglin Zhou,‡,§ Kunqian Yu,‡,§ Bo Gao,∥ Hualiang Jiang,*,‡ Xuechu Zhen,*,∥ and Wei Fu*,† † Department of Medicinal Chemistry & Key Laboratory of Smart Drug Delivery, Ministry of Education, School of Pharmacy, Fudan University, Shanghai 201203, China ‡ State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China ∥ Department of Pharmacology, Soochow University College of Pharmaceuticals Sciences, Suzhou 215123, China *S Supporting Information ABSTRACT: The serotonin receptor subtype 1A (5-HT1AR) has been implicated in several neurological conditions, and potent 5- HT1AR agonists have therapeutic potential for the treatment of depression, anxiety, schizophrenia, and Parkinson’s disease. In the present study, a homology model of 5-HT1AR was built based on the latest released high-resolution crystal structure of the β2AR in its active state (PDB: 3SN6). A dynamic pharmacophore model, which takes the receptor flexibility into account, was constructed, validated, and applied to our dynamic pharmacophore-based virtual screening approach with the aim to identify potential 5-HT1AR agonists. The obtained hits were subjected to 5-HT1AR binding and functional assays, and 10 compounds with medium or high Ki and EC50 values were identified. Among them, FW01 (Ki = 51.9 nM, EC50 = 7 nM) was evaluated as the strongest agonist for 5- HT1AR. The active 5-HT1AR model and dynamic pharmacophore model obtained from this study can be used for future discovery and design of novel 5-HT1AR agonists. Also, by integrating all computational and available experimental data, a stepwise 5-HT1AR signal transduction model induced by agonist FW01 was proposed. ■ INTRODUCTION Serotonin (5-hydroxytrptamine, 5-HT), one of the most important neurotransmitters in the brain, was involved in most of the human behavioral and neuropsychological processes, modulating mood, cognition and memory, feeding, sexuality, sleep, and pain, etc.1 Abnormal 5-HT transmission is associated with pathogenesis of psychiatric disorders and neurodegenerative disorders. To date, at least 16 serotonin receptor subtypes have been cloned which are grouped into seven subfamilies (5-HT1−7) based on their different signaling mechanisms.2 Among them 5-HT1AR was the first one to be fully sequenced and widely studied. Its involvement in anxiety and depression has long been documented.3−5 Growing evidence has revealed new therapeutic applications of 5- HT1AR in the treatment of schizophrenia,6 Parkinson’s disease,7 and neural damage.8 Accordingly, intensive efforts have been made to explore the therapeutic application of 5-HT1AR agonists in the past three decades.9−11 Representative 5-HT1AR agonists include R-8- OH-DPAT (full agonist; widely used agonistic tool drug),12 Buspirone (partial agonist; anxiolytic),13 Vilazodone (SSRI/5- HT1AR partial agonist; antidepressant),14 Aripirazole (mixed receptor profile with 5-HT1AR agonistic activity; atypical antipsychotic),15 F-13640 (highly selective and effective full agonist; undergoing clinical trials for the treatment of pain)16 (Table 1). Given the rising interest on 5-HT1AR agonists, new Received: August 14, 2013 Published: November 18, 2013 Table 1. Pharmacophore Model Validation Using Statistical Parameters parameter value total compounds in database (D) 1000 total number of actives in database (A) 25 total hits (Ht) 33 active hits (Ha) 21 % yield of actives 63.6 % ratio of actives in the hit list 84 enrichment factor or enhancement (E) a 25.45 false negatives 4 false positives 12 GH score (goodness of hit list)a 0.68 a E = (Ha/Ht)/(A/D); GH = [((Ha/4HtA)(3A +Ht))(1((HtHa)/ (DA)))], GH score of 0.6−0.7 indicates a good model. Article pubs.acs.org/jcim © 2013 American Chemical Society 3202 dx.doi.org/10.1021/ci400481p | J. Chem. Inf. Model. 2013, 53, 3202−3211