As drug development teams embark on discovering novel solutions to enhance stability and performance of small and large molecules – specifically, biologics, polypeptides, and proteins – lyophilization, or freeze drying, has drawn attention. It can address instability, which constitutes a major barrier in development and manufacturing of commercial drug products. Ascendia Pharmaceutical Solutions has extensive experience in freeze drying techniques and advanced manufacturing capabilities in cGMP lyophilization, as well as lipid nanoparticles (LNPs). to help improve drug performance (figure 1).
Excipients such as saccharides, polymers, solubilizers and amino acids play important roles in stabilizing these drugs in solid lyophilized powder. They overcome their conversion from one form/3D structure to others and minimize generation of new impurities by a phenomenon known as immobilization of components within the frozen state. Freeze drying is also often used to improve solubility of highly crystalline drugs by transformation into an amorphous state.
Over the years, many lyophilized drugs have been approved, including several blockbusters. These include Herceptin® IV (Trastuzumab), Keytruda® (Pembrolizumab), Remicade® (infliximab), Botox® (Daxibotulinumtoxin A), Carimune® NF (Immunoglobin), Xolair® (Omalizumab), Orencia® (Abatacept), Cosentyx® (Secukinumab), and Avonex® (Interferon b-1a), Velcade® (Bortezomib).1
Lyophilization is a process that requires three steps:
The entire lyophilization process takes 24 to 48 hours, as shown in Figure 2.
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(i) FREEZING (ii) PRIMARY DRYING (iii) SECONDARY DRYING (Formation of ice crystals) (Sublimation of ice crystals) (Desoprtion unfrozen water)
ATMOSPHERIC PRESSURE REDUCED PRESSUE REDUCED TEMPERATURE ~1 hour ~ 10-20 hours ~ 3-10 hours |
Freeze drying requires thorough formulation and process characterization. Otherwise, it can lead to more challenges if the appropriate quality by design (QbD) approach and process conditions are not properly set at the onset. A recent article, Ali and Huang (2024), details the lyophilization process and developing the ideal lyophilized cake.2 Small changes in excipient compositions and processing conditions, such as temperature and pressure, or long exposure to elevated temperature can lead to degradation of drugs in the solid form.
Ascendia Pharmaceutical Solutions has the expertise and capabilities for lyophilization of small and large molecules. Either lyophilized as suspensions or as lipid or polymeric nanoparticles, the particle size in the lyophilizate state is as important as maintaining the ideal lyophilized conditions to yield short reconstitution time, low residual moisture content, and drug encapsulation efficiency.
For instance, fish oil encapsulated in polycaprolactone requires freezing temperatures of -30°, -20° and -10o C; whereas, primary drying process occurs at -50o C at 0.05 mbar pressure. In contrast, for encapsulated docetaxel in hyaluronic acid, freezing occurs at liquid nitrogen temperature (ca. -195.8o C) and primary drying occurs at -35o C for 60 hours at 50 m Torr, followed by secondary drying at 0o C for 24 hours.
As evident by figure 2, Ascendia Pharmaceutical Solutions has expertise and capabilities in lyophilization, and microfluidic processing capabilities using the Precision Nanosystems NanoAssemblr systems for generating LNPs with controlled particle size of small molecule therapeutics and vaccines. In a recent study on Covid-19 vaccines, Precision Nanosystems’ NxGen microfluidic system was used for generating LNP suspensions in Tris, PBS at pH 7.4. 3
On dialysis with membrane (MWCO 12-14 kD), ethanol was removed and LNPs were suspended in 25% sucrose or trehalose in Tris or PBS buffer. The LNPs were then lyophilized and encapsulation efficiency of messenger RNA (mRNA)/LNPs was determined by Quanti-iT RiboGreen assay. LNPs encapsulating mRNA after lyophilization at 4o C and 25o C showed good stability.4 Muramatsu et al. used freeze drying temperature of -45o C (VirTis Genesis Pilot Lyophilizer) with primary drying step at -25o C at 20 mTorr and secondary drying step at 30o C at 20 mTorr.5
Precision Nanosystems equipment is part of Ascendia Pharmaceutical Solutions’ expanded 60,000 square-foot manufacturing facility in the New Jersey Bioscience Center. The state-of-the-art facility has Class 10,000 (ISO 7) and Class 100 (ISO 5) cleanrooms for cGMP manufacture of sterile products, and Class 100,000 (ISO 8) manufacturing suites for production of non-sterile dosage forms.
More importantly, the engaged team of scientists at Ascendia Pharmaceutical Solutions is committed to tailoring a custom solution for each drug development project. It is a unique approach not shared by many CDMOs and is part of the Ascendia Difference.
To learn more and to discuss how we can provide solutions to your drug development project, contact us.