Once-daily asthma treatment should prevent night-time deterioration, irrespective of the time of dosing. IND/GLY/MF, a fixed-dose combination of inhaled indacaterol acetate (IND, long-acting β 2 -agonist (LABA)), glycopyrronium bromide (GLY, long-acting muscarinic antagonist) and mometasone furoate (MF, inhaled corticosteroid (ICS)) delivered by Breezhaler, is indicated in adult asthma patients inadequately controlled on LABA/ICS. A randomised, double-blind, placebo-controlled, three-period, crossover, phase II study was performed to investigate the bronchodilator effect of IND/GLY/MF (150/50/80 μg) dosed morning and evening versus placebo in patients with mild-moderate asthma. The primary end-point was weighted mean forced expiratory volume in 1 s (FEV 1 ) over 24 h following 14 days of IND/GLY/MF dosed a.m. and p.m. versus placebo. Secondary end-points included the effect of dosing time on peak expiratory flow (PEF) and safety/tolerability. Of 37 randomised patients (age 18-72 years; 21 male, 16 female) 34 completed all three treatment periods. At screening, median (range) pre-bronchodilator FEV 1 was 75.8% (60-96%). Patients were using stable low- (83.8%) or medium-dose (16.2%) ICS. Morning and evening dosing of IND/GLY/MF improved FEV 1 (area under the curve from 0 to 24 h) by 610 mL (90% CI 538-681 mL) and 615 mL (90% CI 544-687 mL), respectively, versus placebo. Mean PEF over 14 days increased by 70.7 L·min -1 (90% CI 60.5-80.9 L·min -1 ) following a.m. dosing, and by 59.7 L·min -1 (90% CI 49.5-69.9 L·min -1 ) following p.m. dosing of IND/GLY/MF versus placebo. IND/GLY/MF demonstrated a safety profile comparable with placebo. Once-daily inhaled IND/GLY/MF was well tolerated and provided sustained lung function improvements over 24 h, irrespective of a.m. or p.m. dosing, in patients with mild-moderate asthma., Competing Interests: Conflict of interest: J. Beier reports personal fees from Novartis during the conduct of the study, and personal fees from AstraZeneca, Berlin Chemie/Menarini and Pohl Boskamp outside the submitted work. Conflict of interest: H. Watz reports grants, personal fees and nonfinancial support from Novartis during the conduct of the study; and grants, personal fees and nonfinancial support from AZ, Berlin Chemie/Menarini, GSK, Chiesi, Bayer and Takeda, outside the submitted work. Conflict of interest: Z. Diamant reports grants from Novartis during the conduct of the study, and personal fees from Acucort, AstraZeneca, ALK, Aquilon, Boehringer Ingelheim, CSL, HAL Allergy, MSD and Sanofi Genzyme outside the submitted work. Conflict of interest: J.M. Hohlfeld reports grants paid to his institution from Novartis during the conduct of the study; and consultancy fees from Boehringer Ingelheim, grants paid to his institution from AstraZeneca AB, Novartis, Janssen Pharmaceutica NV, ALK, Boehringer Ingelheim, LETI, GlaxoSmithKline and grants from Sanofi-Aventis, consultancy fees from Merck & Co, Inc., lecture fees from Novartis, grants paid to his institution from Astellas Pharma and Allergopharma, and lectures fees from HAL, outside the submitted work. Conflict of interest: D. Singh reports personal fees from AstraZeneca, Boehringer Ingelheim, Chiesi, Cipla, Genentech, GlaxoSmithKline, Glenmark, Gossamerbio, Menarini, Mundipharma, Novartis, Peptinnovate, Pfizer, Pulmatrix, Theravance and Verona, outside the submitted work. Conflict of interest: P. Pinot is an employee of Novartis. Conflict of interest: I. Jones is an employee of Novartis. Conflict of interest: H-C. Tillmann is an employee of Novartis., (Copyright ©ERS 2021.)