Advanced biotherapeutic systems such as gene therapy, mRNA lipid nanoparticles, antibody–drug conjugates, fusion proteins, and cell therapy have proven to be promising platforms for delivering targeted biologic therapeutics. Preserving the intrinsic stability of these advanced therapeutics is essential to maintain their innate structure, functionality, and shelf life. Nevertheless, various challenges and obstacles arise during formulation development and throughout the storage period due to their complex nature and sensitivity to various stress factors. Key stability concerns include physical degradation and chemical instability due to various factors such as fluctuations in pH and temperature, which results in conformational and colloidal instabilities of the biologics, adversely affecting their quality and therapeutic efficacy. This review emphasizes key stability issues associated with these advanced biotherapeutic systems and approaches to identify and overcome them. In gene therapy, the brittleness of viral vectors and gene encapsulation limits their stability, requiring the use of stabilizers, excipients, and lyophilization. Keeping cells viable throughout the whole cell therapy process, from culture to final formulation, is still a major difficulty. In mRNA therapeutics, stabilization strategies such as the optimization of mRNA nucleotides and lipid compositions are used to address the instability of both the mRNA and lipid nanoparticles. Monoclonal antibodies are colloidally and conformationally unstable. Hence, buffers and stabilizers are useful to maintain stability. Although fusion proteins and monoclonal antibodies share structural similarities, they show a similar pattern of instability. Antibody–drug conjugates possess issues with conjugation and linker stability. This review outlines the stability issues associated with advanced biotherapeutics and provides insights into the approaches to address these challenges.

Background/Objectives: Nilotinib (NTB) a tyrosine kinase inhibitor, helps CML patients with the Philadelphia chromosome. An increasingly rigorous dose regimen and a boxed warning limit the drug's use. NTB should be taken orally on an empty stomach two hours before and one hour after the dosage. A new reduced-dose nanoparticle formulation of NTB reduces the effect of food on PK and achieves equivalent bioavailability (BA) while fasting. The study investigated if nanostructured lipid carriers could make BCS class II medication NTB more soluble and easier to swallow. Methods: The formulation was optimized using the "Box-Behnken design (3-factor, 3-level)". Polydispersity index, Zeta potential, EE, and particle size were dependent variables, while total lipid concentration, surfactant concentration, and sonication time were independent. Investigation of the optimized formulation's in vivo pharmacokinetics, tissue absorption, and in vitro release. Results: In the optimized formulation, PDI, particle size, EE%, and ZP were 135.26±3.86 nm, 0.126±0.09, 89.62±1.26%, and -42.16±1.75mV, respectively. Transmission electron microscopy revealed circular particles. In vitro, NTB released 96.15% with pure NTB dispersion and 56.34% with NTB-NLCs after 24 hours. The data was acquired using a zero order. NTB was ad-ministered to acute myeloid, acute lymphoblastic, and acute T cell leukaemia at IC50 values of 251, 302, 398, 218, 248, and 225 µg/mL, respectively. NTB had IC50 values of 124, 118, 146, 125, 139, and 135 µg/mL against acute myeloid, acute lymphoblastic, and acute T cell leukaemia cell lines. Conclusions: The malignant leukaemia cell line's vitality decreased dose-dependently with NLCs and NTB. Our cytotoxicity experiments in various cell lines and Wistar rats enhanced oral bioavailability of NTB suggest that the optimized NTB-loaded NLCs.

Hypophosphatasia (HPP), a rare metabolic disorder caused by mutations in the ALPL gene, leads to deficient alkaline phosphatase activity and presents unique clinical challenges for adult patients, including fractures, chronic pain, and dental issues. This disorder is often underdiagnosed due to its variable clinical presentation and overlap with other conditions, further complicating timely intervention. Early diagnosis is critical for effective management; however, current diagnostic criteria have limitations, often resulting in delays. Insights from the Global HPP Registry reveal significant disease burden and treatment gaps in treated and untreated adults. These insights underscore the importance of multidisciplinary approaches in addressing HPP-related complications, including musculoskeletal and systemic manifestations. Enzyme replacement therapy (ERT) with asfotase alfa has proven effective in reducing disease burden and improving quality of life. Recent clinical evidence suggests that ERT not only alleviates symptoms but may also prevent disease progression when initiated early. Emerging therapies and alternative regimens like teriparatide dosing adjustments are being explored for their potential benefits. In addition, advanced imaging modalities and biomarker studies are improving diagnostic accuracy and monitoring of therapeutic outcomes. This comprehensive review highlights the significant challenges and advances in diagnosing and treating HPP in adults. Ongoing research aims to enhance diagnosis and treatment through genetic testing and personalized medicine, focusing on identifying and addressing knowledge gaps to improve care for adult HPP patients. Collaborative efforts between researchers, clinicians, and patient advocacy groups are crucial for driving innovation and improving access to care. Continued research and innovation are essential, and healthcare professionals must stay informed about the latest advancements in HPP diagnosis and treatment to ensure optimal patient care. By addressing these challenges, the field can move closer to improving the lives of adults living with HPP.