Background: Semaglutide is a long-acting glucagon-like peptide-1 receptor agonist used in the management of type 2 diabetes mellitus. Broader access to semaglutide therapy may be supported by clinically suitable alternative formulations if pharmacokinetic comparability with the reference product is established. Aim: To evaluate the pharmacokinetic bioequivalence of semaglutide 0.5 mg solution for injection developed by Dr. Reddy’s Laboratories Limited, India, compared with Ozempic® (semaglutide) 0.5 mg solution for injection in a pre-filled pen under fasting conditions in healthy adult subjects. Methods: This was an open-label, balanced, randomized, two-treatment, single-dose, parallel-group bioequivalence study. Eighty healthy volunteers were enrolled, 78 were dosed, and 74 were included in the pharmacokinetic and statistical analyses. Plasma semaglutide concentrations were measured using a validated liquid chromatography–tandem mass spectrometry method across 31 sampling time points up to 816 hours after dosing. Primary pharmacokinetic endpoints were Cmax, area under the curve (AUC)0–t, and AUC0–∞. Bioequivalence was concluded if the 90% confidence intervals for the test/reference ratio were within 80% to 125%. Results: The 90% confidence intervals were 93.11%–107.22% for Cmax, 93.76%–112.65% for AUC0–t, and 91.48%–104.47% for AUC0–∞, meeting bioequivalence criteria for all primary parameters. Forty-six non-serious adverse events were reported in 28 subjects. No serious adverse events or deaths occurred. Conclusion: The test semaglutide formulation was bioequivalent to the reference product and was generally well tolerated in healthy adult subjects under fasting conditions. Relevance for patients: A pharmacokinetically comparable semaglutide injection may help expand access to long-term semaglutide treatment where innovator product cost or availability may limit patient use.