title: Tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide and N-benzyl-6',7'-dihydrospiro[piperidine-4,4'-thieno[3,2-c]pyran] analogues with bactericidal efficacy against Mycobacterium tuberculosis targeting MmpL3.
abstract: Mycobacterium tuberculosis is a major human pathogen and the causative agent for the pulmonary disease, tuberculosis (TB). Current treatment programs to combat TB are under threat due to the emergence of multi-drug and extensively-drug resistant TB. As part of our efforts towards the discovery of new anti-tubercular leads, a number of potent tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide (THPP) and N-benzyl-6',7'-dihydrospiro[piperidine-4,4'-thieno[3,2-c]pyran] (Spiro) analogues were recently identified against Mycobacterium tuberculosis and Mycobacterium bovis BCG through a high-throughput whole-cell screening campaign. Herein, we describe the attractive in vitro and in vivo anti-tubercular profiles of both lead series. The generation of M. tuberculosis spontaneous mutants and subsequent whole genome sequencing of several resistant mutants identified single mutations in the essential mmpL3 gene. This 'genetic phenotype' was further confirmed by a 'chemical phenotype', whereby M. bovis BCG treated with both the THPP and Spiro series resulted in the accumulation of trehalose monomycolate. In vivo efficacy evaluation of two optimized THPP and Spiro leads showed how the compounds were able to reduce >2 logs bacterial cfu counts in the lungs of infected mice.
authors.affiliation: Diseases of the Developing World, GlaxoSmithKline, Tres Cantos, Madrid, Spain.
authors.forename: Modesto J
authors.initials: MJ
authors.lastname: Remuiñán
authors.forename: Esther
authors.initials: E
authors.lastname: Pérez-Herrán
authors.forename: Joaquín
authors.initials: J
authors.lastname: Rullás
authors.forename: Carlos
authors.initials: C
authors.lastname: Alemparte
authors.forename: María
authors.initials: M
authors.lastname: Martínez-Hoyos
authors.forename: David J
authors.initials: DJ
authors.lastname: Dow
authors.forename: Johnson
authors.initials: J
authors.lastname: Afari
authors.forename: Nalini
authors.initials: N
authors.lastname: Mehta
authors.forename: Jorge
authors.initials: J
authors.lastname: Esquivias
authors.forename: Elena
authors.initials: E
authors.lastname: Jiménez
authors.forename: Fátima
authors.initials: F
authors.lastname: Ortega-Muro
authors.forename: María Teresa
authors.initials: MT
authors.lastname: Fraile-Gabaldón
authors.forename: Vickey L
authors.initials: VL
authors.lastname: Spivey
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Loman
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Pallen
authors.forename: Chrystala
authors.initials: C
authors.lastname: Constantinidou
authors.forename: Douglas J
authors.initials: DJ
authors.lastname: Minick
authors.forename: Mónica
authors.initials: M
authors.lastname: Cacho
authors.forename: María José
authors.initials: MJ
authors.lastname: Rebollo-López
authors.forename: Carolina
authors.initials: C
authors.lastname: González
authors.forename: Verónica
authors.initials: V
authors.lastname: Sousa
authors.forename: Iñigo
authors.initials: I
authors.lastname: Angulo-Barturen
authors.forename: Alfonso
authors.initials: A
authors.lastname: Mendoza-Losana
authors.forename: David
authors.initials: D
authors.lastname: Barros
authors.forename: Gurdyal S
authors.initials: GS
authors.lastname: Besra
authors.forename: Lluís
authors.initials: L
authors.lastname: Ballell
authors.forename: Nicholas
authors.initials: N
authors.lastname: Cammack
mesh_terms: D000818:Animals; D000995:Antitubercular Agents / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology* / Q000627:therapeutic use; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology* / Q000627:therapeutic use; D002855:Chromatography, Thin Layer; D003311:Cord Factors; D004195:Disease Models, Animal; D004285:Dogs; D024881:Drug Resistance, Bacterial; D005838:Genotype; D056945:Hep G2 Cells; D006801:Humans; D007700:Kinetics; D051379:Mice; D008826:Microbial Sensitivity Tests; D050296:Microbial Viability / Q000187:drug effects; D009154:Mutation / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D011720:Pyrazoles / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology* / Q000627:therapeutic use; D051381:Rats; D013141:Spiro Compounds / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology* / Q000627:therapeutic use; D016896:Treatment Outcome; D014376:Tuberculosis / Q000188:drug therapy / Q000382:microbiology
chemical_list: D000995:Antitubercular Agents; D001426:Bacterial Proteins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D003311:Cord Factors; D011720:Pyrazoles; D013141:Spiro Compounds; C021099:trehalose monomycolate