Multiple approaches to reduce reconstitution time of lyophilized drug products with high protein concentration

Abstract

Background: Lyophilized drug products with high protein concentration often perform long reconstitution time, which is inconvenient for clinical use. The objective of this work is to achieve short reconstitution time with multiple and combined strategies.

Methods: Here, we describe the following approaches that lead to reduction of reconstitution time, including adding annealing step, decreasing headspace pressure, decreasing protein concentration with reducing diluent volume, increasing high surface-area-to-height ratio of the cakes, increasing frequency of swirling and diluent temperature.

Results: Among these strategies, reducing diluent volume to achieve high protein concentration and reducing headspace pressure show markedly reduction of reconstitution time. Moreover, we propose combined strategies to mitigate the reconstitution time, at the same time, to achieve same target dose in clinics.

Conclusions: Therefore, this paper provides insights on the application of multiple strategies to accelerate the reconstitution of lyophilized drug products with high concentration, and facilitates their widespread clinical application.

Keywords: combined strategy; headspace pressure; lyophilized drug products with high concentration; multiple approaches; reconstitution time; reducing diluent volume.

Figure 1

Reconstitution time and reduction ratio of lyophilized formulations (F1, 150 mg/mL protein concentration) without or with annealing process. Formulations in this study were reconstituted following the low-frequency swirling procedure. n = 10 for each analysis. Statistically significant differences between the non-annealing and annealing cakes are denoted by ^**(P < 0.01).

Figure 2

SEM images of freeze-dried cakes obtained from (a) non-annealing, (b) annealing (−3°C), (c) annealing (−10°C) and (d) annealing (−15°C) process. Numbers below images represent specific surface area obtained by Brunauer–Emmett–Teller.

Figure 3

Representative force vs. displacement curves for freeze-dried cakes obtained from (a) non-annealing, (b) annealing (−3°C), (c) annealing (−10°C) and (d) annealing (−15°C).

Figure 4

Reconstitution time (a) and reduction ratio (b) of lyophilized formulations (F1,150 mg/mL protein concentration) with 225 mg protein in 2 mL vial, 450 mg in 8 mL vial and 900 mg in 20 mL vial at headspace vacuum range from 250 Torr to 0.1 Torr. Formulations in this study were reconstituted following the low-frequency swirling procedure. n = 10 for each analysis. Error bars represent one standard deviation. Statistically significant differences between the 250 Torr, 100 Torr, 50 Torr, 10 Torr and 0.1 Torr cakes are denoted by ^*(P < 0.05), ^**(P < 0.01) and ^***(P < 0.001).

Figure 5

Reconstitution time and reduction ratio of lyophilized formulations with protein concentrations range from 150 to 50 mg/mL (protein amount range from 225 to 75 mg). Formulations in this study were reconstituted following the low-frequency swirling procedure. n = 10 for each analysis. Error bars represent one standard deviation. Statistically significant differences between the F1, F2, F3, F4 and F5 cakes are denoted by ^***(P < 0.001).

Figure 6

Reconstitution time and reduction ratio of lyophilized formulations (F1, 150 mg/mL protein concentration) with 600 mg protein in 10 mL vial, 20 mL vial and 50 mL vial. Formulations in this study were reconstituted following the low-frequency swirling procedure. n = 10 for each analysis. Error bars represent one standard deviation. Statistically significant differences between the low SAHR, medium SAHR and high SAHR cakes are denoted by ^**(P < 0.01) and ^***(P < 0.001).

Figure 7

Reconstitution time and reduction ratio of lyophilized formulations (F1, 150 mg/mL protein concentration) with 450 mg protein in 8 mL vial. Reconstitution for frequency of swirling from low to high and diluent temperature from room temperature to 37°C. n = 10 for each analysis. Error bars represent one standard deviation. Statistically significant differences between the RT-low frequency, RT-high frequency, 37°C-low frequency and 37°C-high frequency cakes are denoted by NS (P ˃ 0.05) and ^**(P < 0.01).

Figure 8

Reconstitution time and reduction ratio of lyophilized formulations for combination group. n = 10 for each analysis. Error bars represent one standard deviation. Statistically significant differences between the Control group, combination group1 and combination group2 cakes are denoted by ^*** (P < 0.001).