Humulin (LY041001) Page 1 of 1

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(LY041001) These clinical study results are supplied for informational purposes only in the interests of scientific disclosure. They are not intended to substitute for the FDA-approved package insert or other approved labeling. The approved drug label can be found at http://pi.lilly.com/us/humulin-r-u500-pi.pdf Therapeutic Area Alias Indication Trial ID Trial Title Trial Phase Endocrinology B5K-EW-IBHA Diabetes 12774 Comparative Pharmacokinetics and Pharmacodynamics of Common Clinical Doses of Human Regular U-500 Insulin versus Human Regular U-100 Insulin in Healthy Obese Subjects 1 Page 1 of 1

Return to list of studies Page 1 Summary ID# 12774 Clinical Study Summary: Study B5K-EW-IBHA Comparative Pharmacokinetics and Pharmacodynamics of Common Clinical Doses of Human Regular U-500 Insulin versus Human Regular U-100 Insulin in Healthy Obese Subjects Approval Date: 15-Nov-2010 GMT

Return to list of studies Page 2 Title of Study: Comparative Pharmacokinetics and Pharmacodynamics of Common Clinical Doses of Human Regular U-500 Insulin versus Human Regular U-100 Insulin in Healthy Obese Subjects Investigator(s): This single-center study included 1 principal investigator. Study Center(s): This study was conducted at 1 study center in 1 country. Phase of Development: 1 Length of Study: Date of first subject visit: 11 September 2009 Date of last subject visit: 04 December 2009 Objectives: The primary objective of this study was to evaluate the relative exposure (AUC0-t ) for human regular U-500 (U-500R, 500 units [U]/mL) versus human regular U-100 (U-100R, 100 U/mL) insulin after subcutaneous administration of a 50-U dose of each to healthy obese subjects; and to evaluate the relative exposure (AUC0-t ) for both formulations after administration of a 100-U dose. The secondary objectives of the study were: to evaluate the relative exposure AUC0-t' and the overall effect (Gtot) of the subcutaneous administration of a 100-U dose versus a 50-U dose of each formulation to healthy obese subjects; to compare other pharmacokinetic (PK) and pharmacodynamic (PD) parameters between formulations and dose levels after administration of 50- and 100-U subcutaneous doses of U-500R and U-100R insulins to healthy obese subjects; to assess safety and tolerability of the 2 treatments, at the respective doses, in healthy obese subjects. Study Design: Phase 1 (postmarketing), single-center, randomized, 4-period, 4-sequence crossover, investigator- and subject-blinded, euglycemic clamp study. Number of Subjects: Planned: 28 Randomized: 24 Completed: 21 Main Criteria for Inclusion: Healthy subjects, male or female, aged between 21 and 65 years, inclusive, with screening body mass index (BMI) 30 to 40 kg/m2, inclusive (weight 125 kg), participated in this study. Subjects who had a history of diabetes, impaired fasting glucose, or impaired glucose tolerance; significant cardiac, renal, gastrointestinal, or hepatic disease; chronic or recent exposure to glucocorticoid therapy; or known hypersensitivity or allergy to any of the study insulins or recipients of the study insulins were excluded. Test Product, Dose, and Mode of Administration: Human regular U-500 insulin (U-500R, 500 U/mL) at doses of 50 U and 100 U, given by subcutaneous injection on 2 different occasions according to randomized sequence. Reference Therapy, Dose, and Mode of Administration: Human regular U-100 insulin (U-100R, 100 U/mL) at doses of 50 U and 100 U, given by subcutaneous injection on 2 different occasions according to randomized sequence.

Return to list of studies Page 3 Duration of Treatment: Subjects were admitted and randomized to 1 of 4 sequences. Each sequence had a 4-period crossover. In each period, each subject received a single dose of 50 U or 100 U of either formulation of human regular insulin (U-500R or U-100R) and underwent a euglycemic clamp procedure for a period of up to 24 hours. There was a minimum and maximum interval of 7 and 21 days, respectively, between study periods. A follow-up visit was performed between 7 and 28 days after the subject s last dose of study medication. Variables: Bioanalytical: Blood samples were obtained during the euglycemic glucose clamp procedure for analysis of whole blood glucose to adjust glucose infusion rates to maintain euglycemia. Blood samples were collected for the determination of serum immunoreactive insulin (IRI) concentrations for up to 24 hours after dose administration. Pharmacokinetics: The PK parameters included area under the concentration-time curve from time zero to return to baseline (AUC0-t ), from time zero to the last time point with a measurable concentration (AUC0-tlast), maximum serum insulin concentration (Cmax), and the time to maximum insulin concentration (tmax). Pharmacodynamics: The PD parameters included the cumulative amount of glucose infused from dosing until clamp cessation (Gtot) and the maximum glucose infusion rate (Rmax). Other parameters included the time of maximum glucose infusion rate (trmax), time of half maximal glucose infusion rate before or after Rmax (early trmax50 or late trmax50), and times of first measured glucose infusion rate (tonset) and cessation of glucose infusion/clamp (trlast). Safety: Physical examination, vital signs, adverse event (AE) monitoring, and clinical laboratory testing. Evaluation Methods: Bioanalytical: Blood samples collected were analyzed immediately for whole blood glucose concentrations using an automated glucose analyzer. The serum concentrations of IRI were measured by a validated conventional competitive binding radioimmunoassay method. Pharmacokinetic: Pharmacokinetic parameters were evaluated by standard noncompartmental methods of PK analysis using PKS/WinNonlin Enterprise version 5.0.1. Pharmacodynamic: Pharmacodynamic parameters were derived from the glucose clamp data using S-PLUS 7.0 Professional Developer (TIBCO software, Inc.). Statistical: The main PK and PD parameters (AUC0-t, Cmax, Gtot and Rmax) were log-transformed prior to analysis. The model included subject as a random effect, with period, sequence, and treatment as fixed effects. The estimated difference in tmax between the U-100 and U-500 groups for each of the doses together with the associated 90% confidence interval (CI) were analyzed using the nonparametric Wilcoxon signed-rank test. Analyses of trmax, early and late trmax50, tonset and trlast, were performed using similar linear mixed effects model without log-transforming the data. Safety: Summary statistics were tabulated for each safety parameter; AEs were listed and summarized. Summary: The current study was a Phase 1 (postmarketing), single-center, randomized, 4-period, 4-sequence crossover, investigator- and subject-blinded, euglycemic clamp study comparing PK and PD parameters of human regular U-100 (U-100R) insulin versus human regular insulin U-500 (U-500R) in healthy obese subjects. Of the 50 subjects who entered the study, 24 were randomly assigned to treatment and all 24 subjects received at least 1 dose of study drug; 21 subjects completed the study. Table IBHA.1 presents subject demographics.

Return to list of studies Page 4 Table IBHA.1. Patient Characteristics Enrolled N=24 Male n=14 Female n=10 Age range (y) 23 to 59 Primary Analysis: The primary objective of this study was to evaluate the relative exposure (the area under the IRI concentration versus time curve from time 0 to return to baseline [AUC0-t ]) for U-500R versus U-100R insulins after subcutaneous administration of a 50-U dose of each to healthy obese subjects; and to evaluate the relative exposure (AUC0-t ) for both formulations after administration of a 100-U dose. Overall insulin exposure, as assessed by AUC0-t, was not statistically significantly different between formulations at both the 50-U and 100-U doses. Other Pharmacokinetic Evaluations The U-500R peak insulin concentration (Cmax) was statistically significantly lower than that for U-100R at both doses. The time-to-peak insulin concentration (tmax) was statistically significantly longer for U-500R at the 100-U dose only. Pharmacodynamic Evaluations Overall effect (G tot ) for U-500R was not statistically significantly different from U-100R at both doses. Peak effect (R max ) was statistically significantly lower for U-500R versus U-100R at both doses. Time of peak effect (tr max ) was shown to be statistically significantly prolonged for U-500R versus U-100R at the 100-U dose only. Time variables reflective of duration of action (early and late tr max50, tr last ) were statistically significantly prolonged for U-500R versus U-100R at both doses. Table IBHA.2 presents PK and PD parameters.

Return to list of studies Table IBHA.2. Summary of Key Noncompartmental Pharmacokinetic and Pharmacodynamic Parameters [Geometric Mean (CV%)] of Immunoreactive Insulin Following Administration of 50 U and 100 U of U-100R and U-500R Insulin 90% CI U-100R (N=22) 6960 (27%) 50-U Dose Ratio ( ) or Difference U-500R ( ) of LS (N=21) Means 6430 (24%) 0.94 (0.88, 1.00) 12400 (22%) 809 (32%) 548 (22%) 0.69 (0.63, 0.75) 1400a (28%) 3.00 (1.00 8.00) 4.00 (0.50 8.00) 419 (28%) 628 (42%) 0.00 (0.00, 4.00) 3.00a (1.00 8.00) 1.07 0.88 (1.00,1.16) (0.80,0.96) 586 (23%) 966 (22%) PK Parameters U-100R (N=22) AUC0-t (pmol h/l) Cmax (pmol/l) tmax (h) Gtot (g) Rmax (mg/min) trmax (h) Early trmax50 (h) Late trmax50 (h) tonset (h) 387 (33%) 709 (50%) 621 (33%) 826 (37%) PD Parameters 6.17 (20%) 0.78 (0.11,1.45) 5.32 (22%) 6.37 (25%) 2.33 (40%) 0.68 (0.41,0.94) 1.51 (41%) 2.02 (59%) 14.1 (21%) 3.50 (2.64,4.37) 11.7 (16%) 15.1 (16%) 0.230 0.00 (-0.10, 0.10) 0.184 (78%) 0.221 (77%) (96%) trlast (h) 18.3 (22%) 19.7 (18%) 1.38 (0.43, 2.34) 18.3 (15%) 21.5 (11%) Abbreviations: CI = confidence interval; CV = coefficient of variance; PD = pharmacodynamics; PK = pharmacokinetics; U = unit. Parameters are expressed as geometric means (CV%), except for tmax, expressed as median (range). a N=23; bn=24 *p<0.05; Ratio of LS means; Difference of LS means; Median difference 5.30 (25%) 1.69 (33%) 10.5 (25%) 0.270 (64%) 100-U Dose Ratio ( ) or Difference U-500R ( ) of LS (N=23) Means 12300 0.98 (19%) 1020b 0.72 (31%) 8.00b 2.50 (0.50 8.00) 90% CI (0.92, 1.05) (0.66, 0.78) (0.00, 4.00) 1.09 0.87 (1.01, 1.17) (0.80, 0.95) 1.04 0.60 3.39 0.04 (0.38, 1.70) (0.34, 0.87) (2.54, 4.25) (-0.06, 0.14) 3.24 (2.29, 4.19) Page 5

Return to list of studies Page 6 Safety: No death or serious adverse events occurred during this study. No subject discontinuation due to AE occurred during this study. The most common AEs were headache (17 instances reported by 12 subjects) and nausea (5 instances reported by 2 subjects). Most AEs were of mild or moderate severity. No clinically significant alterations in laboratory values were observed during the course of the study.