ABSTRACT
Optimization starting with our lead compound 1 (IC(50)=4.9 nM) led to the identification of pyrrolidinyl phenylurea derivatives. Further modification toward improvement of the bioavailability provided (R)-1-(1-((6-fluoronaphthalen-2-yl)methyl)pyrrolidin-3-yl)-3-(2-(2-hydroxyethoxy)phenyl)urea 32 (IC(50)=1.7 nM), a potent and orally active CCR3 antagonist.
Subject(s)
Phenylurea Compounds/chemistry , Pyrrolidines/chemistry , Receptors, CCR3/antagonists & inhibitors , Administration, Oral , Animals , Biological Availability , Half-Life , Macaca fascicularis , Phenylurea Compounds/chemical synthesis , Phenylurea Compounds/pharmacokinetics , Pyrrolidines/chemical synthesis , Pyrrolidines/pharmacokinetics , Receptors, CCR3/metabolismABSTRACT
The synthesis and structure-activity relationships of ureas as CCR3 antagonists are described. Optimization starting with lead compound 2 (IC(50)=190 nM) derived from initial screening hit compound 1 (IC(50)=600 nM) led to the identification of (S)-N-((1R,3S,5S)-8-((6-fluoronaphthalen-2-yl)methyl)-8-azabicyclo[3.2.1]octan-3-yl)-N-(2-nitrophenyl)pyrrolidine-1,2-dicarboxamide 27 (IC(50)=4.9 nM) as a potent CCR3 antagonist.
Subject(s)
Receptors, CCR3/antagonists & inhibitors , Urea/analogs & derivatives , Drug Evaluation, Preclinical , Humans , Naphthalenes/chemical synthesis , Naphthalenes/chemistry , Naphthalenes/metabolism , Proline/chemistry , Protein Binding , Pyrrolidines/chemical synthesis , Pyrrolidines/chemistry , Pyrrolidines/metabolism , Receptors, CCR3/metabolism , Structure-Activity Relationship , Urea/chemical synthesis , Urea/metabolismABSTRACT
Our laboratory has identified several acrylamide derivatives with potent CCR3 inhibitory activity. In the present study, we evaluated the in vitro metabolic stability (CL(int); mL/min/kg) of these compounds in human liver microsomes (HLMs), and assessed the relationship between their structures and CL(int) values. Among the compounds identified, N-{(3R)-1-[(6-fluoro-2-naphthyl)methyl]pyrrolidin-3-yl}-2-[1-(2-hydroxybenzoyl)piperidin-4-ylidene]acetamide (30j) was found to be a potent inhibitor (IC(50)=8.4nM) with a high metabolic stability against HLMs.
Subject(s)
Acetamides/chemical synthesis , Acrylamides/chemistry , Anti-Allergic Agents/chemical synthesis , Naphthalenes/chemical synthesis , Receptors, CCR3/antagonists & inhibitors , Acetamides/chemistry , Acetamides/pharmacology , Acrylamides/chemical synthesis , Acrylamides/pharmacokinetics , Animals , Anti-Allergic Agents/chemistry , Anti-Allergic Agents/pharmacokinetics , Haplorhini , Humans , Mice , Microsomes, Liver/metabolism , Naphthalenes/chemistry , Naphthalenes/pharmacology , Piperidines/chemical synthesis , Piperidines/chemistry , Piperidines/pharmacology , Protein Isoforms/antagonists & inhibitors , Protein Isoforms/metabolism , Receptors, CCR3/metabolism , ThermodynamicsABSTRACT
In our previous study on discovering novel types of CCR3 antagonists, we found a fluoronaphthalene derivative (1) that exhibited potent CCR3 inhibitory activity with an IC(50) value of 20 nM. However, compound 1 also inhibited human cytochrome P450 2D6 (CYP2D6) with an IC(50) value of 400 nM. In order to reduce its CYP2D6 inhibitory activity, we performed further systematic structural modifications on 1. In particular, we focused on reducing the number of lipophilic moieties in the biphenyl part of 1, using ClogD(7.4) values as the reference index of lipophilicity. This research led to the identification of N-{(3-exo)-8-[(6-fluoro-2-naphthyl)methyl]-8-azabicyclo[3.2.1]oct-3-yl}-3-(piperidin-1-ylcarbonyl)isonicotinamide 1-oxide (30) which showed comparable CCR3 inhibitory activity (IC(50)=23 nM) with much reduced CYP2D6 inhibitory activity (IC(50)=29,000 nM) compared with 1.
Subject(s)
Cytochrome P-450 CYP2D6 Inhibitors , Drug Design , Hydrocarbons, Fluorinated/pharmacology , Naphthalenes/pharmacology , Receptors, CCR3/antagonists & inhibitors , Calcium/chemistry , Calcium/metabolism , Cytoplasm/chemistry , Cytoplasm/metabolism , Humans , Hydrocarbons, Fluorinated/chemical synthesis , Inhibitory Concentration 50 , Naphthalenes/chemical synthesis , Receptors, CCR3/metabolism , Structure-Activity RelationshipABSTRACT
A novel class of potent CCR3 receptor antagonists were designed and synthesized starting from N-{1-[(6-fluoro-2-naphthyl)methyl]piperidin-4-yl}benzamide (1),which was found by subjecting our chemical library to high throughput screening (HTS). The CCR3 inhibitory activity of the synthesized compounds against eotaxin-induced Ca(2+) influx was evaluated using CCR3-expressing preB cells. Systematic chemical modifications of 1 revealed that the 6-fluoro-2-naphthylmethyl moiety was essential for CCR3 inhibitory activity in this new series of CCR3 antagonists. Further structural modifications of the benzamide and piperidine moieties of 1 led to the identification of exo-N-{8-[(6-fluoro-2-naphthyl)methyl]-8-azabicyclo[3.2.1]oct-3- yl}biphenyl-2-carboxamide [corrected] (31) as a potent CCR3 antagonist with an IC(50) value of 0.020 microM.
Subject(s)
Benzamides/pharmacology , Receptors, CCR3/antagonists & inhibitors , Benzamides/chemical synthesis , Calcium/metabolism , Chemokine CCL11 , Humans , Inhibitory Concentration 50 , Precursor Cells, B-Lymphoid , Small Molecule Libraries , Structure-Activity RelationshipABSTRACT
Eosinophilic chemokines and histamine play distinct but important roles in allergic diseases. Inhibition of both eosinophilic chemokines and histamine, therefore, is an ideal strategy for the treatment of allergic inflammation, such as asthma, allergic rhinitis, and atopic dermatitis. YM-344484 was found to potently inhibit both the CCL11-induced Ca2+ influx in human CCR3-expressing cells (Kb=1.8 nM) and histamine-induced Ca2+ influx in histamine H1 receptor-expressing PC3 cells (Kb=47 nM). YM-344484 also inhibited the CCL11-induced chemotaxis of human CCR3-expressing cells (IC50=6.2 nM) and CCL11-induced eosinophil-derived neurotoxin release from human eosinophils (IC50=19 nM). Orally administered YM-344484 inhibited the increase in histamine-induced vascular permeability in mice (82% inhibition at a dose of 10 mg/kg) and the accumulation of eosinophils in a mouse asthma model (74% at a dose of 300 mg/kg). These results indicate that YM-344484, a novel and functional dual antagonist for chemokine CCR3 receptor and histamine H1 receptor, is an attractive candidate for development as a novel anti-allergic inflammation drug.
Subject(s)
Anti-Allergic Agents/pharmacology , Anti-Inflammatory Agents/pharmacology , Histamine Antagonists/pharmacology , Piperidines/pharmacology , Pyridazines/pharmacology , Receptors, Chemokine/antagonists & inhibitors , Receptors, Histamine H1/drug effects , Animals , Anti-Allergic Agents/therapeutic use , Anti-Inflammatory Agents/therapeutic use , Asthma/chemically induced , Asthma/complications , Calcium Signaling/drug effects , Capillary Permeability/drug effects , Cell Line, Tumor , Chemotaxis/drug effects , Disease Models, Animal , Dose-Response Relationship, Drug , Eosinophil-Derived Neurotoxin/metabolism , Eosinophils/drug effects , Eosinophils/metabolism , Female , Histamine/pharmacology , Histamine Antagonists/therapeutic use , Humans , Mice , Mice, Inbred BALB C , Ovalbumin , Pneumonia/etiology , Pneumonia/prevention & control , Pulmonary Eosinophilia/etiology , Pulmonary Eosinophilia/prevention & control , Rats , Receptors, CCR3 , Receptors, Chemokine/genetics , Receptors, Chemokine/metabolism , Receptors, Histamine H1/metabolism , Skin/blood supply , TransfectionABSTRACT
Eosinophils play a prominent proinflammatory role in a broad range of diseases, including atopic dermatitis and asthma. Eotaxin-1 and its receptor CCR3 are implicated in the recruitment of eosinophils from blood into inflammatory tissues, therefore inhibition of Eotaxin-1/CCR3 interaction may have therapeutic potential for allergic inflammation with eosinophil infiltration. YM-344031, a novel and selective small molecule CCR3 antagonist, potently inhibited ligand binding (IC(50)=3.0nM), ligand-induced Ca(2+) flux (IC(50)=5.4nM), and the chemotaxis of human CCR3-expressing cells (IC(50)=19.9nM). YM-344031 (1-10mg/kg) orally administered to cynomolgus monkeys significantly inhibited Eotaxin-1-induced eosinophil shape change in whole blood. Additionally, orally administered YM-344031 (100mg/kg) prevented both immediate- and late-phase allergic skin reactions in a mouse allergy model. YM-344031 therefore has potential as a novel and orally available compound for the treatment of allergic inflammation, such as atopic dermatitis and asthma.
Subject(s)
Amides/administration & dosage , Dermatitis/immunology , Dermatitis/prevention & control , Pyridinium Compounds/administration & dosage , Receptors, Chemokine/agonists , Receptors, Chemokine/immunology , Spleen/immunology , Administration, Oral , Animals , Cells, Cultured , Dermatitis/pathology , Dose-Response Relationship, Drug , Humans , Macaca fascicularis , Male , Mice , Mice, Inbred BALB C , Receptors, CCR3 , Spleen/drug effects , Treatment OutcomeABSTRACT
CC chemokine ligand 11 (CCL11/eotaxin) and other CC chemokine receptor 3 (CCR3) ligands (CCL24/eotaxin-2, CCL26/eotaxin-3, CCL13/monocyte chemotactic protein-4, etc.) play important roles in the chemotaxis and activation of eosinophils and other CCR3-expressing cells (basophils, mast cells, and CD4(+) T helper 2 cells) in allergic inflammation incidents, including asthma and rhinitis. A newly synthesized compound, N-{(3R)-1-[(6-fluoro-2-naphthyl)methyl]pyrrolidin-3-yl}-2-{1-[(5-hydroxy-3-methylpyridin-2-yl)carbonyl]piperidin-4-ylidene}-acetamide hemifumarate (YM-355179), inhibited the binding of CCL11 and CCL5/regulated on activation normal T cell expressed and secreted to CCR3-expressing B300-19 cells with IC(50) values of 7.6 and 24 nM, respectively. In contrast, YM-355179 did not affect the binding of CCL5 to CCR1 or CCR5. In functional assays, YM-355179 inhibited CCL11-induced, intracellular Ca(2+) influx, chemotaxis, and eosinophil degranulation with IC(50) values of 8.0, 24, and 29 nM, respectively. YM-355179 did not, however, affect any CC chemokine receptor (CCR1, CCR2, CCR4, or CCR5)-mediated Ca(2+) influx signals. Furthermore, oral administration of YM-355179 (1 mg/kg) inhibited CCL11-induced shape change of whole blood eosinophils in cynomolgus monkeys. Intravenous injection of YM-355179 (1 mg/kg) also inhibited eosinophil infiltration into airways of cynomolgus monkeys after segmental bronchoprovocation with CCL11. These results indicate that YM-355179 is a novel, selective, and orally available CCR3 antagonist with therapeutic potential for treating eosinophil-related allergic inflammatory diseases.