Pharmacokinetic and Pharmacodynamic Characteristics of BioChaperone® Combo, the First Fixed Combination of Glargine and Lispro, in Type 1 Diabetes

Ulrike Hövelmann1, Bertrand Alluis2, Grégory Meiffren2, Aymeric Ranson2, Olivier Soula*2, Gérard Soula2, Cyril Seroussi2, Birgit Kronshage1, Leszek Nosek1, Susanne Famulla1, Tim Heise1

1 Profil, Neuss, Germany; 2 Adocia, Lyon, France; * corresponding author (o.soula@adocia.com)

Abstract Methods Efficacy Results Figure 2: PD GIR profiles of BioChaperone Combo and MIX

In this double-blind, crossover study we investigated the PK/PD characteristics of BC Combo 25, a novel formulation combining insulins lispro (25%) and glargine (75%) in a limpid pH-neutral solution. Twenty people with type 1 diabetes participated in automated euglycemic clamps (ClampArt®, target blood glucose (BG) 100 mg/dL, clamp duration 30h) and received 0.8 U/kg of BC Combo or Humalog® Mix 25 (MIX). Mean glucose infusion rate (GIR) curves (Figure) illustrate an earlier onset of action (25±11 vs. 40±13 min) consistent with earlier Tmax (2.8±0.8 vs. 3.4±0.8 h) of BC Combo vs. MIX. Likewise, the early metabolic effect (AUCGIR 0-2h 504±210 vs. 325±183 mg/kg) and the early PK exposure (AUCPK 0-1h 86±39 vs. 34±19 h*mU/L) were higher, and BC Combo showed a more pronounced late metabolic effect (AUCGIR 12-30h 1480±900 vs. 961±553 mg/kg) consistent with a higher basal exposure (AUCPK 12-30h 563±409 vs. 286±233 h*mU/L). Duration of action (time to BG >118 mg/dL, 29.8±0.7 vs. 25.5±4.3 h) and half-life (17.6±8.7 vs.

7.7±3.0 h, p<0.05 for all comparisons) were longer with BC Combo indicating the potential for once daily dosing. Both formulations were well tolerated, no local reactions occurred. Both PK and PD demonstrate faster prandial and longer basal action for BC Combo supporting the potential for improved BG control vs. MIX with only one daily injection.

o This was a randomized, single-center, double-blind, two-way crossover glucose clamp study.
o Eligible study participants: Male subjects with type 1 diabetes mellitus (T1DM), age 18 - 64 years, body mass index (BMI) 18.0 - 28.0 kg/m2, HbA1c ≤ 9.0 %, total daily insulin dose < 1.2 U/kg/day.
o Patients were randomly assigned to two single-dose administrations of 0.8 U/kg BioChaperone® Combo or
Humalog® Mix 25 under euglycemic clamp conditions at two separate visits.
o After an overnight fast, patients were connected in the morning to ClampArt® (a modern clamp device developed by Profil, Neuss, Germany). A variable intravenous infusion of insulin aspart was started to reach a target blood glucose level (BG) of 100 mg/dl (run-in).
o Glucose infusion rate (GIR) was automatically adjusted by ClampArt® to maintain a BG level close to target for
30 hours post-dosing. The clamp experiment was stopped earlier if BG increased to >200 mg/dl without any
glucose infusion in the past 30 min.
o Blood samples for determination of pharmacokinetics (PK) were drawn at pre-dose and in regular intervals until 30 h post-dose.
o Serum insulin concentrations were determined with an immunoradiometric sandwich assay (BI-INSULIN- IRMA, Cisbio Bioassays). Pre-experiment validation showed a cross-reactivity of approximately 80% to insulin lispro and approximately 100% to insulin glargine.

o Compared with MIX, BC Combo showed a significantly faster onset of action, reached maximum action (GIR tmax)
earlier and had a higher glucose-lowering effect in the first two hours post-dosing (AUCGIR0-2h) [Table 2, Figure 2].
o Only 2 subjects on BC Combo (in contrast to 14 subjects on MIX) reached end of action (BG >118 mg/dL without any GIR in the last 15 min) during the 30 hour clamp duration indicating a significantly longer duration of action of BC Combo.
o The longer duration of action of BC Combo is also supported by a better blood glucose control in the last hour of the clamp [Table 2, Figure 5] and a significantly higher late metabolic effect (higher AUCGIR12-30h).
o Total insulin concentrations were nearly twice as high with BC Combo compared with MIX [AUCINS0-30h, Table 2].
The difference of cross-reactivity of the assay to lispro and glargine (80 and 100%, respectively) does not fully
explain the difference in total AUC. Therefore a trend to higher bioavailability for BC Combo compared to MIX is
conceivable. That is supported by the strong trend toward a higher bio-efficacy seen in GIR (total AUC in GIR in favour to BC Combo, p=0.05).
o Even when AUC-parameters were corrected for total insulin AUCs, parameters for duration of absorption
(AUCINS12-30h, t½) were higher with BC Combo and were in line with the pharmacodynamic results.

Table 2: PK/PD Parameters Aim


o To investigate the pharmacodynamic and pharmacokinetic properties and safety of BioChaperone Glargine
Lispro and HumalogMix 25 in subjects in patients with type 1 diabetes

Figure 1: Study Design

Visit 1 (Screening visit)

Visit 2 (Dosing visit)

BC Combo (0.8 U/kg)

Visit 3 (Dosing visit)

Visit 4 (Follow-up visit)

b #

Figure 3: PD BG profiles of BioChaperone Combo and MIX Introduction

o Pre-mixed insulins are widely used as they provide both prandial and basal insulin coverage in one formulation. While short-acting insulin analogues with improved pharmacodynamic characteristics for prandial insulin coverage are available, it has not yet been possible to combine short-acting analogues with

2−21 days

M IX (0.8 U/kg)

5−14 days wash-out

insulin glargine, the most widely used basal insulin analogue.
o To date all currently available pre-mixed insulins contain protamine-retarded insulin as basal component which often show a peaked activity profile with rather high effects 6-12 h post-dosing and a high variability. (Heise T, et al. Diabetes 2004; 53: 1614-20).
o Furthermore, all these premixed insulins require at least a twice daily injection to fully cover basal insulin requirements over 24 hours.
o Therefore a premixed insulin combining the long-acting insulin glargine with a fast-acting insulin may provide the option of once daily use together with improved glycaemic control (in terms of better HbA1c values) and a reduction in hypoglycaemic episodes.

BioChaperone Combo

BioChaperone Combo 25 is an injectable combination at pH 7 of insulin glargine (U300) and insulin lispro (U100).
This combination is made possible by the use of BioChaperone, Adocia's proprietary platform technology.
The BioChaperone technology does not modify the physical, chemical and biological integrity of either insulin
lispro (U100) or insulin glargine (U300).
o The study insulins were given s.c. into a lifted skinfold around the umbilicus by means of an insulin syringe.
All injections were performed by a competent unblinded person not otherwise involved in the study conduct
to keep the double-blind character of the study.

Statistical Analysis

o The pharmacodynamic (PD) endpoints were derived from the individual GIR profiles which were smoothed using a locally weighted regression technique (LOESS, smoothing factor 0.25). Time-related parameters were derived from the smoothed curves, AUCs from the unsmoothed data.
o Key endpoints including early (0-1h and 0-2h) and late (12-30h) AUCs under the PK/PD curves were analyzed using a mixed effect linear model with treatment, period and sequence as fixed effects and subjects within sequence as random effect. Endpoints that were not normally or log normally-distributed were analyzed non- parametrically using Wilcoxon Signed Rank Test.

Table 1: Baseline Characteristics (N=21*)

*21 patients were exposed, 2 patients withdrew consent, and 19 patients completed the trial.
For the analysis of PD and PK parameters, data from the Full Analysis Set which included 20 subjects was used.
For safety analysis of safety data from the Safety Analysis Set which included 21 subjects was used.

ᴥGeometric means and (CV%) for log-normally distributed parameters

§ Arithmetic means and (CV%) for normally distributed parameters

#Medians and (Min/Max) for parameters that were neither normally nor log-normally distributed

aTime from dosing until blood glucose concentrations decreased by 5 mg/dL from baseline

bTime from dosing until End Of Action defined as an increase in blood glucose concentrations to >118 mg/dL without any

glucose infusion in the last 15 min (Bolli GB, et al. Diabetes Care 2012; 35:2626-30). Duration Of Action was only reached in 2 clamps with BC Combo and 14 clamps with MIX. The p-value is based on a comparison of number of clamps reaching end of action (Chi-Square Test)

cMean blood glucose (BG) values [mg/dL] in the last hour of the glucose clamp (29-30 hours after dosing). The p-value is based on

a non-parametric analysis

Safety

o No safety issues were identified.
o Both insulin formulations were well tolerated and no injection site reactions occurred.

Conclusions

o This is the first study providing a "proof-of-concept" that glargine can be combined with lispro in one insulin product at neutral pH using the BioChaperone technology.

o BioChaperone Combo 25 shows a faster onset, a more pronounced early metabolic effect as well as a longer duration of action and a higher basal metabolic effect than Humalog® Mix25.

o BioChaperone Combo 25 has the potential to improve glycemic control compared to HumalogMix.

o The current data suggest that BioChaperone Combo 25 has the potential to be used as either once-a-day or twice-a-day treatment.

This study was funded by Adocia and performed by Profil.
NCT trial number: NCT01981031
Presented at the American Diabetes Association, 13-17 June 2014, San Francisco, USA.

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