title: CHF6001 I: a novel highly potent and selective phosphodiesterase 4 inhibitor with robust anti-inflammatory activity and suitable for topical pulmonary administration.
abstract: This study examined the pharmacologic characterization of CHF6001 [(S)-3,5-dichloro-4-(2-(3-(cyclopropylmethoxy)-4-(difluoromethoxy)phenyl)-2-(3-(cyclopropylmethoxy)-4-(methylsulfonamido)benzoyloxy)ethyl)pyridine 1-oxide], a novel phosphodiesterase (PDE)4 inhibitor designed for treating pulmonary inflammatory diseases via inhaled administration. CHF6001 was 7- and 923-fold more potent than roflumilast and cilomilast, respectively, in inhibiting PDE4 enzymatic activity (IC50 = 0.026 ± 0.006 nM). CHF6001 inhibited PDE4 isoforms A-D with equal potency, showed an elevated ratio of high-affinity rolipram binding site versus low-affinity rolipram binding site (i.e., >40) and displayed >20,000-fold selectivity versus PDE4 compared with a panel of PDEs. CHF6001 effectively inhibited (subnanomolar IC50 values) the release of tumor necrosis factor-α from human peripheral blood mononuclear cells, human acute monocytic leukemia cell line macrophages (THP-1), and rodent macrophages (RAW264.7 and NR8383). Moreover, CHF6001 potently inhibited the activation of oxidative burst in neutrophils and eosinophils, neutrophil chemotaxis, and the release of interferon-γ from CD4(+) T cells. In all these functional assays, CHF6001 was more potent than previously described PDE4 inhibitors, including roflumilast, UK-500,001 [2-(3,4-difluorophenoxy)-5-fluoro-N-((1S,4S)-4-(2-hydroxy-5-methylbenzamido)cyclohexyl)nicotinamide], and cilomilast, and it was comparable to GSK256066 [6-((3-(dimethylcarbamoyl)phenyl)sulfonyl)-4-((3-methoxyphenyl)amino)-8-methylquinoline-3-carboxamide]. When administered intratracheally to rats as a micronized dry powder, CHF6001 inhibited liposaccharide-induced pulmonary neutrophilia (ED50 = 0.205 μmol/kg) and leukocyte infiltration (ED50 = 0.188 μmol/kg) with an efficacy comparable to a high dose of budesonide (1 μmol/kg i.p.). In sum, CHF6001 has the potential to be an effective topical treatment of conditions associated with pulmonary inflammation, including asthma and chronic obstructive pulmonary disease.
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Nadia
authors.initials: N
authors.lastname: Moretto
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Paola
authors.initials: P
authors.lastname: Caruso
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Raffaella
authors.initials: R
authors.lastname: Bosco
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Gessica
authors.initials: G
authors.lastname: Marchini
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Fiorella
authors.initials: F
authors.lastname: Pastore
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Armani
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Gabriele
authors.initials: G
authors.lastname: Amari
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Andrea
authors.initials: A
authors.lastname: Rizzi
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Eleonora
authors.initials: E
authors.lastname: Ghidini
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Renato
authors.initials: R
authors.lastname: De Fanti
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Carmelida
authors.initials: C
authors.lastname: Capaldi
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Laura
authors.initials: L
authors.lastname: Carzaniga
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Emilio
authors.initials: E
authors.lastname: Hirsch
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Carola
authors.initials: C
authors.lastname: Buccellati
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Angelo
authors.initials: A
authors.lastname: Sala
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Chiara
authors.initials: C
authors.lastname: Carnini
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Riccardo
authors.initials: R
authors.lastname: Patacchini
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Maurizio
authors.initials: M
authors.lastname: Delcanale
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Maurizio
authors.initials: M
authors.lastname: Civelli
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.).
authors.forename: Gino
authors.initials: G
authors.lastname: Villetti
authors.affiliation: Corporate Pre-Clinical R & D, Chiesi Farmaceutici S.p.A., Parma, Italy (N.M., P.C., R.B., G.M., F.P., E.A., G.A., A.R., E.G., R.D.F., Ca.C., L.C., Ch.C., R.P. M.D., M.C., G.V., F.F.); Molecular Biotechnology Center, University of Turin, Turin, Italy (E.H.); and Dipartimento di Scienze Farmacologiche e Biomolecolari, Milan, Italy (C.B., A.S.) f.facchinetti@chiesi.com.
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Facchinetti
mesh_terms: D000280:Administration, Inhalation; D000287:Administration, Topical; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000008:administration & dosage* / Q000378:metabolism*; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4 / Q000378:metabolism*; D005289:Ferrets; D008297:Male; D008810:Mice, Inbred C57BL; D058988:Phosphodiesterase 4 Inhibitors / Q000008:administration & dosage* / Q000378:metabolism*; D051381:Rats; D011914:Rats, Inbred BN; D017207:Rats, Sprague-Dawley
chemical_list: D000893:Anti-Inflammatory Agents; D058988:Phosphodiesterase 4 Inhibitors; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4