The Antidepressant 5-HT2A Receptor Antagonists Pizotifen and Cyproheptadine Inhibit Serotonin-Enhanced Platelet Function
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{"title"=>"The antidepressant 5-HT2Areceptor antagonists pizotifen and cyproheptadine inhibit serotonin-enhanced platelet function", "type"=>"journal", "authors"=>[{"first_name"=>"Olivia A.", "last_name"=>"Lin", "scopus_author_id"=>"55860019400"}, {"first_name"=>"Zubair A.", "last_name"=>"Karim", "scopus_author_id"=>"12805828600"}, {"first_name"=>"Hari Priya", "last_name"=>"Vemana", "scopus_author_id"=>"56149927100"}, {"first_name"=>"Enma V.P.", "last_name"=>"Espinosa", "scopus_author_id"=>"54681725900"}, {"first_name"=>"Fadi T.", "last_name"=>"Khasawneh", "scopus_author_id"=>"14621469500"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-84899875224", "sgr"=>"84899875224", "pui"=>"373020818", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"24466319", "doi"=>"10.1371/journal.pone.0087026"}, "id"=>"1c91b264-28c2-34ec-9707-8acc90a4764d", "abstract"=>"There is considerable interest in defining new agents or targets for antithrombotic purposes. The 5-HT2A receptor is a G-protein coupled receptor (GPCR) expressed on many cell types, and a known therapeutic target for many disease states. This serotonin receptor is also known to regulate platelet function. Thus, in our FDA-approved drug repurposing efforts, we investigated the antiplatelet activity of cyproheptadine and pizotifen, two antidepressant 5-HT2A Receptor antagonists. Our results revealed that cyproheptadine and pizotifen reversed serotonin-enhanced ADP-induced platelet aggregation in vitro and ex vivo. And the inhibitory effects of these two agents were found to be similar to that of EMD 281014, a 5-HT2A Receptor antagonist under development. In separate experiments, our studies revealed that these 5-HT2A receptor antagonists have the capacity to reduce serotonin-enhanced ADP-induced elevation in intracellular calcium levels and tyrosine phosphorylation. Using flow cytometry, we also observed that cyproheptadine, pizotifen, and EMD 281014 inhibited serotonin-enhanced ADP-induced phosphatidylserine (PS) exposure, P-selectin expression, and glycoprotein IIb-IIIa activation. Furthermore, using a carotid artery thrombosis model, these agents prolonged the time for thrombotic occlusion in mice in vivo. Finally, the tail-bleeding time was investigated to assess the effect of cyproheptadine and pizotifen on hemostasis. Our findings indicated prolonged bleeding time in both cyproheptadine- and pizotifen-treated mice. Notably, the increases in occlusion and bleeding times associated with these two agents were comparable to that of EMD 281014, and to clopidogrel, a commonly used antiplatelet drug, again, in a fashion comparable to clopidogrel and EMD 281014. Collectively, our data indicate that the antidepressant 5-HT2A antagonists, cyproheptadine and pizotifen do exert antiplatelet and thromboprotective effects, but similar to clopidogrel and EMD 281014, their use may interfere with normal hemostasis.", "link"=>"http://www.mendeley.com/research/antidepressant-5ht2areceptor-antagonists-pizotifen-cyproheptadine-inhibit-serotoninenhanced-platelet", "reader_count"=>18, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>3, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>5, "Student > Master"=>2, "Other"=>3, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>3, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>5, "Student > Master"=>2, "Other"=>3, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>1, "Medicine and Dentistry"=>7, "Agricultural and Biological Sciences"=>4, "Neuroscience"=>1, "Veterinary Science and Veterinary Medicine"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>3}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>7}, "Neuroscience"=>{"Neuroscience"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>4}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}, "Unspecified"=>{"Unspecified"=>1}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>3}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1357465"], "description"=>"<p>(A) Human PRP was stimulated with submaximal concentration of ADP (1 µM) in the presence or absence of serotonin (15 µM). (B) Human PRP was pre-incubated with increasing doses of cyproheptadine (0.1–10 nM) for 1 min and activated with ADP (1 µM) and serotonin (15 µM). (C) Human PRP was pre-incubated with increasing doses of pizotifen (0.01–1 nM) for 1 min and activated with ADP and serotonin. (D) Human PRP was pre-incubated with increasing doses of EMD 281014 (10–40 nM) for 1 min and activated with ADP and serotonin. (E) Human PRP was treated with the highest concentrations of cyproheptadine (10 nM), pizotifen (1 nM), and EMD 281014 (40 nM) used in previous experiments, in the absence of agonists. (F) Human PRP was pre-incubated for 1 min with cyproheptadine (10 nM), pizotifen (1 nM), and EMD 281014 (40 nM) for 1 min and activated with serotonin (15 µM). (G) Human PRP was pre-incubated with cyproheptadine (10 nM), pizotifen (1 nM), and EMD 281014 (40 nM) for 1 min and activated with ADP (1 µM). Inset shows quantification of the data. Each experiment was repeated at least 3 times, with blood obtained from three separate donors.</p>", "links"=>[], "tags"=>["Molecular cell biology", "Signal transduction", "Membrane receptor signaling", "systems biology", "toxicology", "Anatomy and physiology", "cardiovascular", "Cardiovascular pharmacology", "Drugs and devices", "Drug research and development", "Pharmacodynamics", "hematology", "pizotifen", "inhibit", "serotonin-enhanced", "adp-induced", "platelet", "aggregation"], "article_id"=>909795, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Olivia A. Lin", "Zubair A. Karim", "Hari Priya Vemana", "Enma V. P. Espinosa", "Fadi T. Khasawneh"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0087026.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cyproheptadine_and_pizotifen_inhibit_serotonin_enhanced_ADP_induced_human_platelet_aggregation_in_vitro_/909795", "title"=>"Cyproheptadine and pizotifen inhibit serotonin-enhanced ADP-induced human platelet aggregation <i>in vitro.</i>", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-23 03:03:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/1357468"], "description"=>"<p>(A) Human PRP was stimulated with submaximal concentration of U46619 (0.125 µM) in the presence or absence of serotonin (15 µM). (B) Human PRP was pre-incubated with increasing doses of cyproheptadine (0.1–250 nM) for 1 min and activated with U46619 (0.125 µM) and serotonin (15 µM). (C) Human PRP was pre-incubated with increasing doses of pizotifen (0.1–30 nM) for 1 min and activated with 0.125 µM U46619 and 15 µM serotonin. (D) Human PRP was pre-incubated with increasing doses of EMD 281014 (5–20 nM) for 1 min and activated with 0.125 µM U46619 and 15 µM serotonin. (E) Human PRP was pre-incubated for 1 min with the highest concentrations of cyproheptadine (250 nM), pizotifen (30 nM), and EMD 281014 (20 nM) used in this set of experiments, and activated with U46619 (0.125 µM). Inset shows quantification of the data. Each experiment was repeated at least 3 times, with blood obtained from three separate donors.</p>", "links"=>[], "tags"=>["Molecular cell biology", "Signal transduction", "Membrane receptor signaling", "systems biology", "toxicology", "Anatomy and physiology", "cardiovascular", "Cardiovascular pharmacology", "Drugs and devices", "Drug research and development", "Pharmacodynamics", "hematology", "pizotifen", "inhibit", "serotonin-enhanced", "u46619-induced", "platelet", "aggregation"], "article_id"=>909797, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Olivia A. Lin", "Zubair A. Karim", "Hari Priya Vemana", "Enma V. P. Espinosa", "Fadi T. Khasawneh"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0087026.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cyproheptadine_and_pizotifen_inhibit_serotonin_enhanced_U46619_induced_human_platelet_aggregation_in_vitro_/909797", "title"=>"Cyproheptadine and pizotifen inhibit serotonin-enhanced U46619-induced human platelet aggregation <i>in vitro</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-23 03:03:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/1357469"], "description"=>"<p>(A) Human PRP was treated with different combinations of cyproheptadine (10 nM), pizotifen (1 nM), and EMD 281014 (40 nM) in the absence of agonists. (B) Human PRP was pre-incubated with different combinations of cyproheptadine (10 nM), pizotifen (1 nM), and EMD 281014 (40 nM) for 1 min and activated with serotonin (15 µM). (C) Human PRP was pre-incubated with different combinations of cyproheptadine (10 nM), pizotifen (1 nM), and EMD 281014 (40 nM) for 1 min and activated with ADP (1 µM). (D) Human PRP was pre-incubated with different combinations of cyproheptadine (10 nM), pizotifen (1 nM), and EMD 281014 (40 nM) for 1 min and activated with ADP (1 µM) and serotonin (15 µM). Each experiment was repeated at least 3 times, with blood obtained from three separate donors.</p>", "links"=>[], "tags"=>["Molecular cell biology", "Signal transduction", "Membrane receptor signaling", "systems biology", "toxicology", "Anatomy and physiology", "cardiovascular", "Cardiovascular pharmacology", "Drugs and devices", "Drug research and development", "Pharmacodynamics", "hematology", "cyproheptadine", "pizotifen", "inhibits", "serotonin-enhanced", "adp-induced", "platelet", "aggregation"], "article_id"=>909798, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Olivia A. Lin", "Zubair A. Karim", "Hari Priya Vemana", "Enma V. P. Espinosa", "Fadi T. Khasawneh"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0087026.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Combination_of_Cyproheptadine_and_pizotifen_inhibits_serotonin_enhanced_ADP_induced_human_platelet_aggregation_in_vitro_/909798", "title"=>"Combination of Cyproheptadine and pizotifen inhibits serotonin-enhanced ADP-induced human platelet aggregation <i>in vitro</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-23 03:03:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/1357471"], "description"=>"<p>(A) Mouse PRP was stimulated with submaximal concentration of ADP (1 µM) in the presence or absence of serotonin (15 µM). (B) Mouse PRP was pre-incubated with increasing doses of cyproheptadine (0.01–100 nM) for 1 min and activated with ADP (1 µM) and serotonin (15 µM). (C) Mouse PRP was pre-incubated with increasing doses of pizotifen (5–100 nM) for 1 min and activated with 1 µM ADP and 15 µM serotonin. (D) Mouse PRP was pre-incubated with increasing doses of EMD 281014 (1–40 nM) for 1 min and activated with 1 µM ADP and 15 µM serotonin. Inset shows quantification of the data. Each experiment was repeated at least 3 times, with blood pooled from at least eight mice each time.</p>", "links"=>[], "tags"=>["Molecular cell biology", "Signal transduction", "Membrane receptor signaling", "systems biology", "toxicology", "Anatomy and physiology", "cardiovascular", "Cardiovascular pharmacology", "Drugs and devices", "Drug research and development", "Pharmacodynamics", "hematology", "pizotifen", "inhibit", "serotonin-enhanced", "adp-induced", "platelet", "aggregation"], "article_id"=>909800, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Olivia A. Lin", "Zubair A. Karim", "Hari Priya Vemana", "Enma V. P. Espinosa", "Fadi T. Khasawneh"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0087026.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cyproheptadine_and_pizotifen_inhibit_serotonin_enhanced_ADP_induced_mouse_platelet_aggregation_in_vitro_/909800", "title"=>"Cyproheptadine and pizotifen inhibit serotonin-enhanced ADP-induced mouse platelet aggregation <i>in vitro</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-23 03:03:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/1357472"], "description"=>"<p>(A) Human platelets were loaded with Fura-2/AM to measure intracellular [Ca2+]<sub>i</sub>, in the presence or absence of Cyproheptadine (10 nM), pizotifen (1 nM) or EMD 281014 (40 nM) and activated with ADP (1 µM), serotonin (15 µM) or ADP and serotonin together. (B) Human platelets were incubated in the presence or absence of Cyproheptadine (10 nM), pizotifen (1 nM) or EMD 281014 (40 nM) for 5 minutes and then stimulated with ADP (1 µM), serotonin (15 µM) or ADP and serotonin together for 3 minutes, and subjected to immunoprecipitation followed by immunoblotting with anti-Src and anti-phosphotyrosine antibodies; upper panel shows quantification of the data using densitometry analysis. Each experiment was repeated at least 3 times, with blood obtained from three separate donors.</p>", "links"=>[], "tags"=>["Molecular cell biology", "Signal transduction", "Membrane receptor signaling", "systems biology", "toxicology", "Anatomy and physiology", "cardiovascular", "Cardiovascular pharmacology", "Drugs and devices", "Drug research and development", "Pharmacodynamics", "hematology", "pizotifen", "inhibit", "intracellular", "calcium", "elevation", "src", "activation", "platelets"], "article_id"=>909801, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Olivia A. Lin", "Zubair A. Karim", "Hari Priya Vemana", "Enma V. P. Espinosa", "Fadi T. Khasawneh"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0087026.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cyproheptadine_and_pizotifen_inhibit_intracellular_calcium_elevation_and_Src_activation_in_human_platelets_in_vitro_/909801", "title"=>"Cyproheptadine, and pizotifen inhibit intracellular calcium elevation and Src activation in human platelets <i>in vitro</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-23 03:03:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/1357473"], "description"=>"<p>(A) Mouse PRP obtained from vehicle-injected mice (once daily for 5 days) was stimulated with submaximal concentration of ADP (1 µM) in the presence or absence of serotonin (15 µM). (B) Mouse PRP obtained from cyproheptadine-injected mice (1 mg/kg IP once daily for 5 days) was stimulated with submaximal concentration of ADP in the presence or absence of serotonin. (C) Mouse PRP obtained from pizotifen-injected mice (3 mg/kg IP once daily for 5 days) was stimulated with submaximal concentration of ADP in the presence or absence of serotonin. (D) Mouse PRP obtained from EMD 281014 injected mice (5 mg/kg IP once daily for 5 days) was stimulated with submaximal concentration of ADP in the presence or absence of serotonin. Inset shows quantification of the data. Each experiment was repeated at least 3 times, with blood pooled from at least eight mice each time.</p>", "links"=>[], "tags"=>["Molecular cell biology", "Signal transduction", "Membrane receptor signaling", "systems biology", "toxicology", "Anatomy and physiology", "cardiovascular", "Cardiovascular pharmacology", "Drugs and devices", "Drug research and development", "Pharmacodynamics", "hematology", "pizotifen", "inhibit", "serotonin-enhanced", "adp-induced", "platelet", "aggregation"], "article_id"=>909802, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Olivia A. Lin", "Zubair A. Karim", "Hari Priya Vemana", "Enma V. P. Espinosa", "Fadi T. Khasawneh"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0087026.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cyproheptadine_and_pizotifen_inhibit_serotonin_enhanced_ADP_induced_mouse_platelet_aggregation_ex_vivo_/909802", "title"=>"Cyproheptadine and pizotifen inhibit serotonin-enhanced ADP-induced mouse platelet aggregation <i>ex vivo</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-23 03:03:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/1357475"], "description"=>"<p>Washed platelets were incubated in the presence or absence of Cyproheptadine (10 nM), pizotifen (1 nM) or EMD 281014 (40 nM) for 5 minutes and then stimulated with ADP (1 µM), serotonin (15 µM) or ADP and serotonin together for 3 minutes. The reactions were stopped by fixing the platelets with 2% formaldehyde for 30 min at room temperature. (A–C): Platelets were incubated with FITC-conjugated Annexin V antibody, the fluorescent intensities were measured by flow cytometry, and the data were plotted as histogram. (D, E): Platelets were incubated with FITC-conjugated anti–P-selectin antibody, the fluorescent intensities were measured by flow cytometry, and the data were plotted as histogram. (F, G): Platelets were incubated with FITC-conjugated PAC-1 antibody, the fluorescent intensities were measured by flow cytometry, and the data were plotted as histogram. Each experiment was repeated at least 3 times, with blood obtained from three separate donors.</p>", "links"=>[], "tags"=>["Molecular cell biology", "Signal transduction", "Membrane receptor signaling", "systems biology", "toxicology", "Anatomy and physiology", "cardiovascular", "Cardiovascular pharmacology", "Drugs and devices", "Drug research and development", "Pharmacodynamics", "hematology", "pizotifen", "inhibit", "platelet", "ps", "gpiib-iiia", "activation"], "article_id"=>909804, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Olivia A. Lin", "Zubair A. Karim", "Hari Priya Vemana", "Enma V. P. Espinosa", "Fadi T. Khasawneh"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0087026.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cyproheptadine_and_pizotifen_inhibit_human_platelet_PS_exposure_Annexin_V_P_selectin_and_GPIIb_IIIa_PAC_1_binding_activation_in_vitro_/909804", "title"=>"Cyproheptadine and pizotifen inhibit human platelet PS exposure (Annexin V), P-selectin, and GPIIb-IIIa (PAC-1 binding) activation <i>in vitro</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-23 03:03:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/1357477"], "description"=>"<p>Each point represents the occlusion time or tail bleeding time of a single animal. Mice were treated IP with vehicle, 6/kg clopidogrel, 1 mg/kg cyproheptadine, 3 mg/kg pizotifen, or 5 mg/kg EMD 281014 once daily for 5 days before experiments. (A) Mean occlusion times for mice treated with: vehicle = 375.3±31.89 sec (n = 11), clopidogrel = 987.6±196.5 sec (n = 8), cyproheptadine = 787.4±84.08 sec (n = 8), pizotifen = 1199±253.1 sec (n = 8), and EMD 281014 = 879.9±270.0 sec (n = 8). **p<0.0022; ****p<0.0001; **p<0.0014; *p<0.0431. (B) Mean bleeding times for mice treated with: vehicle = 213.1±81.03 sec (n = 9), clopidogrel = 793.6±58.17 sec (n = 9), cyproheptadine = 643.1±97.78 sec (n = 9), pizotifen = 714.4±96.78 sec (n = 9), and EMD 281014 = 558.9±67.12 sec (n = 9). ****p<0.0001; **p<0.0038; **p<0.0011; **p<0.0047.</p>", "links"=>[], "tags"=>["Molecular cell biology", "Signal transduction", "Membrane receptor signaling", "systems biology", "toxicology", "Anatomy and physiology", "cardiovascular", "Cardiovascular pharmacology", "Drugs and devices", "Drug research and development", "Pharmacodynamics", "hematology", "pizotifen", "prolong", "occlusion", "times", "bleeding"], "article_id"=>909806, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Olivia A. Lin", "Zubair A. Karim", "Hari Priya Vemana", "Enma V. P. Espinosa", "Fadi T. Khasawneh"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0087026.g008"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cyproheptadine_and_pizotifen_prolong_occlusion_times_and_tail_bleeding_times_in_mice_/909806", "title"=>"Cyproheptadine and pizotifen prolong occlusion times and tail bleeding times in mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-23 03:03:57"}

PMC Usage Stats | Further Information

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