title: DF2755A, a novel non-competitive allosteric inhibitor of CXCR1/2, reduces inflammatory and post-operative pain.
abstract: The activation of CXCR1/2 has been implicated in the genesis of inflammatory and postoperative pain. Here, we investigated a novel orally acting allosteric inhibitor of CXCR1/2 (DF2755A) and evaluated its antinociceptive effect in several models of inflammatory and post-operatory pain. DF2755A was tested in vitro for efficacy in the chemotaxis assay, selectivity and toxicity. In vivo, C57Bl/6 mice were treated orally with DF2755A and the following experiments were performed: pharmacokinetic profile; inflammatory hyperalgesia models using electronic pressure meter test; neutrophil migration assay assessed by myeloperoxidase assay. DF2755A selectively inhibited neutrophil chemotaxis induced by CXCR1/2 ligands without effect on CXCL8 binding to neutrophils. A single mutation of the allosteric site at CXCR1 abrogated the inhibitory effect of DF2755A on CXCL8-induced chemotaxis. DF2755A given orally was well absorbed (88.2%), and it was able to reduce, in a dose (3-30mg/kg)-dependent manner, inflammatory hyperalgesia induced by carrageenan, LPS and CXCL1/KC as well as neutrophil recruitment and IL-1β production. In addition, DF2755A was able to reduce post-incisional nociception. Therapeutic treatment with DF2755A reduced CFA-induced inflammatory hyperalgesia even when injected intrathecally. The present results indicate that DF2755A is a novel selective allosteric inhibitor of CXCR1/2 with a favorable oral pharmacokinetic profile. Furthermore, the results might suggest that DF2755A might be a candidate of a novel therapeutic option to control inflammatory and post-operative pain.
authors.affiliation: Department of Pharmacology, Ribeirao Preto Medical School, University of Sao Paulo, Av. Bandeirantes, 3900, 14049-900, Ribeirão Preto, SP, Brazil; Departamento de Bioquímica e Immunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, 31270901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Alexandre H
authors.initials: AH
authors.lastname: Lopes
authors.affiliation: R&D Department, Dompé Farmaceutici s.p.a., via Campo di Pile, 67100 L'Aquila, Italy; Departamento de Bioquímica e Immunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, 31270901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Laura
authors.initials: L
authors.lastname: Brandolini
authors.affiliation: R&D Department, Dompé Farmaceutici s.p.a., via Campo di Pile, 67100 L'Aquila, Italy; Departamento de Bioquímica e Immunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, 31270901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Andrea
authors.initials: A
authors.lastname: Aramini
authors.affiliation: R&D Department, Dompé Farmaceutici s.p.a., via Campo di Pile, 67100 L'Aquila, Italy; Departamento de Bioquímica e Immunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, 31270901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Gianluca
authors.initials: G
authors.lastname: Bianchini
authors.affiliation: Department of Pharmacology, Ribeirao Preto Medical School, University of Sao Paulo, Av. Bandeirantes, 3900, 14049-900, Ribeirão Preto, SP, Brazil; Departamento de Bioquímica e Immunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, 31270901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Rangel L
authors.initials: RL
authors.lastname: Silva
authors.affiliation: Departamento de Patologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Rod. Celso Garcia Cid PR445 KM380, 86051-990 Londrina, Paraná, Brazil; Departamento de Bioquímica e Immunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, 31270901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Ana C
authors.initials: AC
authors.lastname: Zaperlon
authors.affiliation: Departamento de Patologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Rod. Celso Garcia Cid PR445 KM380, 86051-990 Londrina, Paraná, Brazil; Departamento de Bioquímica e Immunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, 31270901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Waldiceu A
authors.initials: WA
authors.lastname: Verri
authors.affiliation: Department of Pharmacology, Ribeirao Preto Medical School, University of Sao Paulo, Av. Bandeirantes, 3900, 14049-900, Ribeirão Preto, SP, Brazil; Departamento de Bioquímica e Immunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, 31270901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: José C
authors.initials: JC
authors.lastname: Alves-Filho
authors.affiliation: Department of Pharmacology, Ribeirao Preto Medical School, University of Sao Paulo, Av. Bandeirantes, 3900, 14049-900, Ribeirão Preto, SP, Brazil; Departamento de Bioquímica e Immunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, 31270901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Fernando Q
authors.initials: FQ
authors.lastname: Cunha
authors.affiliation: Departamento de Patologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Rod. Celso Garcia Cid PR445 KM380, 86051-990 Londrina, Paraná, Brazil; Departamento de Bioquímica e Immunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, 31270901 Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Mauro M
authors.initials: MM
authors.lastname: Teixeira
authors.affiliation: R&D Department, Dompé Farmaceutici s.p.a., via Campo di Pile, 67100 L'Aquila, Italy; Departamento de Bioquímica e Immunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, 31270901 Belo Horizonte, Minas Gerais, Brazil. Electronic address: marcello.allegretti@dompe.it.
authors.forename: Marcello
authors.initials: M
authors.lastname: Allegretti
authors.affiliation: Department of Pharmacology, Ribeirao Preto Medical School, University of Sao Paulo, Av. Bandeirantes, 3900, 14049-900, Ribeirão Preto, SP, Brazil; Departamento de Bioquímica e Immunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antonio Carlos, 6627, 31270901 Belo Horizonte, Minas Gerais, Brazil. Electronic address: thicunha@fmrp.usp.br.
authors.forename: Thiago M
authors.initials: TM
authors.lastname: Cunha
mesh_terms: D000328:Adult; D000700:Analgesics / Q000493:pharmacokinetics / Q000494:pharmacology / Q000627:therapeutic use*; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000493:pharmacokinetics / Q000494:pharmacology / Q000627:therapeutic use*; D002351:Carrageenan; D045744:Cell Line, Tumor; D002465:Cell Movement; D002478:Cells, Cultured; D054360:Chemokine CXCL1 / Q000378:metabolism; D015232:Dinoprostone / Q000378:metabolism; D006168:Guinea Pigs; D006801:Humans; D006930:Hyperalgesia / Q000139:chemically induced / Q000188:drug therapy*; D016209:Interleukin-8 / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000502:physiology; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D010149:Postoperative Pain / Q000188:drug therapy*; D010648:Phenylacetates / Q000493:pharmacokinetics / Q000494:pharmacology / Q000627:therapeutic use*; D010812:Physical Stimulation; D011817:Rabbits; D051381:Rats; D023062:Receptors, Interleukin-8A / Q000037:antagonists & inhibitors; D023063:Receptors, Interleukin-8B / Q000037:antagonists & inhibitors; D013844:Thiazoles / Q000493:pharmacokinetics / Q000494:pharmacology / Q000627:therapeutic use*
chemical_list: D000700:Analgesics; D000893:Anti-Inflammatory Agents; D054360:Chemokine CXCL1; C000612078:DF2755A; D016209:Interleukin-8; D010648:Phenylacetates; D023062:Receptors, Interleukin-8A; D023063:Receptors, Interleukin-8B; D013844:Thiazoles; D002351:Carrageenan; D015232:Dinoprostone
keywords: Allosteric; CXCR1/2; Chemokines; DF2755A; Inflammatory pain; Neutrophil; Neutrophil chemotaxis