Thelvia I. Ramos

There is a critical gap in the preclinical research of recombinant human interferons (rhIFNα-2b and rhIFN-γ), as most studies focus on modified variants, which complicates the understanding of the native molecules’ properties. This study addresses this limitation by comprehensively evaluating the structural stability and intrinsic toxicity of purified IFNs. Our findings confirm that both interferons retain their bioactivity (antiviral, antiproliferative, and immunomodulatory) and exhibit remarkable stability under controlled conditions. Accelerated stability assays showed that neither protein lost biological potency after 18 days at various temperatures, supporting their potential as liquid formulations. Acute and sub-chronic toxicity studies in rodent, non-rodent, and higher-organism animal models showed no signs of toxicity, even at doses 100 to 300 times higher than therapeutic levels. These assays, combined with the absence of pyrogens, support a favorable safety profile for clinical use, with no evidence of systemic or structural damage. This work establishes a reproducible experimental model and lays the groundwork for future preclinical evaluations. We underscore the importance of characterizing the safety profile of active pharmaceutical ingredients from the earliest stages of biopharmaceutical development to ensure a safe and well-founded transition to human clinical trials. Furthermore, these results open the door for the development of advanced formulations and alternative routes of administration, such as the intranasal route, an area with significant potential.

Early embryonic loss is a major cause of reproductive inefficiency in cattle, primarily due to premature luteolysis. Interferon tau (IFN-τ), secreted by the trophoblast, plays a critical role in maternal recognition of pregnancy by maintaining corpus luteum function. However, its practical application has been limited by its rapid degradation and short half-life in vivo. Here, we developed a novel formulation of recombinant bovine IFN-τ, combining chitosan-based microencapsulation with starch–chitosan hydrogel delivery, enabling sustained intrauterine release. This dual-delivery strategy offers a significant improvement over conventional IFN-τ administration methods that rely on repeated intrauterine infusions of soluble protein. The rbIFN-τ was expressed in Pichia pastoris, purified to 90.1% homogeneity, and structurally validated via homology modeling and molecular docking, confirming its interaction with type I interferon receptors. The encapsulated formulation retained antiviral activity, stimulated transcription of interferon-stimulated genes (PKR, OAS1, OAS2), and showed sustained release in vitro for up to 26 days. In vivo evaluation demonstrated safety and biological efficacy, with treated cattle showing inhibited luteolysis, sustained serum progesterone levels, and preserved corpus luteum integrity. This formulation represents a promising biotechnological approach to improve reproductive efficiency through a long-acting, species-specific IFN-τ delivery system.

Early embryonic loss is a significant cause of reproductive inefficiency and economic loss in cattle, largely due to inadequate maternal recognition of pregnancy and premature luteolysis. Interferon tau (IFN-τ), a trophoblast-derived signaling molecule, is essential for maintaining corpus luteum function and embryo survival in ruminants, but its rapid degradation and short half-life have limited its practical use. This study developed a novel delivery system for bovine recombinant IFN-τ (brIFN-τ) produced in Pichia pastoris, utilizing chitosan-based microencapsulation and starch-chitosan hydrogels for sustained intrauterine release. The recombinant protein was successfully expressed, purified, and structurally validated, confirming its interaction with bovine interferon receptors. Encapsulated IFN-τ showed controlled, prolonged in vitro release and retained biological activity, demonstrated by increased viability and antiviral response in bovine cells exposed to Mengo virus. In vivo, the microencapsulated formulation was safe and effectively inhibited luteolysis, as shown by sustained progesterone levels and reduced corpus luteum regression compared to controls. These results indicate that hydrogel-delivered, microencapsulated brIFN-τ is a promising biotechnological strategy to modulate luteolysis, enhance maternal recognition of pregnancy, and reduce early embryonic loss in cattle, potentially improving reproductive efficiency and supporting more sustainable livestock management.

Background: Interferons (IFNs) are cytokines involved in the immune response with a synergistic regulatory effect on the immune response. They are therapeutics for various viral and proliferative conditions, with proven safety and efficacy. Their clinical application is challenging due to the molecules’ size, degradation, and pharmacokinetics. We are working on new drug delivery systems that provide adequate therapeutic concentrations for these cytokines and prolong their half-life in the circulation, such as nanoformulations. Methods: Through nanoencapsulation using electrospray technology and biocompatible and biodegradable polymers, we are developing a controlled release system based on nanoparticles for viral infections of the respiratory tract. Results: We developed a controlled release system for viral respiratory tract infections. A prototype nanoparticle with a core was created, which hydrolyzed the polyvinylpyrrolidone (PVP) shell , releasing the active ingredients interferon-alpha (IFN-α) and interferon-gamma (IFN-γ). The chitosan (QS) core degraded slowly, with a controlled release of IFN-α. The primary and rapid effect of the interferon combination ensured an antiviral and immunoregulatory response from day one, induced by IFN-α and enhanced by IFN-γ. The multilayer design demonstrated an optimal toxicity profile. Conclusions: This formulation is an inhaled dry powder intended for the non-invasive intranasal route. The product does not require a cold chain and has the potential for self-administration in the face of emerging viral infections. This novel drug has applications in multiple infectious, oncological, and autoimmune conditions, and further development is proposed for its therapeutic potential. This prototype would ensure greater bioavailability, controlled release, fewer adverse effects, and robust biological action through the simultaneous action of both molecules.

Interferons (IFNs) are cytokines involved in the immune response with synergistic regulatory action. They are therapeutics for various viral and proliferative conditions, with proven safety and efficacy. Their clinical application presents difficulties due to the molecules' size, degradation, and pharmacokinetics. We developed a controlled release system for viral respiratory tract infections. A core-shell nanoparticle was prototyped, which hydrolyzed the shell (polyvinylpyrrolidone), releasing the active ingredients IFN-α and IFN-γ. The core (chitosan) degraded slowly, with a controlled release of IFN-α. The primary and rapid effect of the combination of interferons ensured an antiviral and immunoregulatory response from day one, induced by IFN-α and enhanced by IFN-γ. The multilayer design demonstrated an optimal toxicity profile. This formulation is an inhaled dry powder targeted for the non-invasive intranasal route. This prototype would ensure greater bioavailability, controlled release, fewer adverse effects, and robust biological action thanks to the simultaneous impact of both molecules.

Nanomedicine plays an essential role in developing new therapies through novel drug delivery systems, diagnostic and imaging systems, vaccine development, antibacterial tools, and high-throughput screening. One of the most promising drug delivery systems are nanoparticles, which can be designed with various compositions, sizes, shapes, and surface modifications. These nanosystems have improved therapeutic profiles, increased bioavailability, and reduced the toxicity of the product they carry. However, the clinical translation of nanomedicines requires a thorough understanding of their properties to avoid problems with the most questioned aspect of nanosystems: safety. The particular physicochemical properties of nano-drugs lead to the need for additional safety, quality, and efficacy testing. Consequently, challenges arise during the physicochemical characterization, the production process, in vitro characterization, in vivo characterization, and the clinical stages of development of these biopharmaceuticals. The lack of a specific regulatory framework for nanoformulations has caused significant gaps in the requirements needed to be successful during their approval, especially with tests that demonstrate their safety and efficacy. Researchers face many difficulties in establishing evidence to extrapolate results from one level of development to another, for example, from an in vitro demonstration phase to an in vivo demonstration phase. Additional guidance is required to cover the particularities of this type of product, as some challenges in the regulatory framework do not allow for an accurate assessment of NPs with sufficient evidence of clinical success. This work aims to identify current regulatory issues during the implementation of nanoparticle assays and describe the major challenges that researchers have faced when exposing a new formulation. We further reflect on the current regulatory standards required for the approval of these biopharmaceuticals and the requirements demanded by the regulatory agencies. Our work will provide helpful information to improve the success of nanomedicines by compiling the challenges described in the literature that support the development of this novel encapsulation system. We propose a step-by-step approach through the different stages of the development of nanoformulations, from their design to the clinical stage, exemplifying the different challenges and the measures taken by the regulatory agencies to respond to these challenges.

Interferons (IFNs) are cytokines involved in the immune response that act on innate and adaptive immunity. These proteins are natural cell-signaling glycoproteins expressed in response to viral infections, tumors, and biological inducers and constitute the first line of defense of vertebrates against infectious agents. They have been marketed for more than 30 years with considerable impact on the global therapeutic protein market thanks to their diversity in terms of biological activities. They have been used as single agents or with combination treatment regimens, demonstrating promising clinical results, resulting in 22 different formulations approved by regulatory agencies. The 163 clinical trials with currently active IFNs reinforce their importance as therapeutics for human health. However, their application has presented difficulties due to the molecules’ size, sensitivity to degradation, and rapid elimination from the bloodstream. For some years now, work has been underway to obtain new drug delivery systems to provide adequate therapeutic concentrations for these cytokines, decrease their toxicity and prolong their half-life in the circulation. Although different research groups have presented various formulations that encapsulate IFNs, to date, there is no formulation approved for use in humans. The current review exhibits an updated summary of all encapsulation forms presented in the scientific literature for IFN-α, IFN-ß, and IFN-γ, from the year 1996 to the year 2021, considering parameters such as: encapsulating matrix, route of administration, target, advantages, and disadvantages of each formulation.

El ligamento cruzado anterior (LCA) es la lesión de la rodilla con mayor prevalencia en los atletas. Los avances en la técnica quirúrgica y la fijación de injertos han permitido a los pacientes participar en una rehabilitación funcional postoperatoria temprana. Este tratamiento está dirigido a lograr un mayor rango de movimiento, progresando hacia la movilización, fortalecimiento y control neuromuscular propioceptivo de la articulación. Existen varios protocolos de rehabilitación con variaciones en ejercicios específicos, progresión a través de fases y componentes claves que permiten la recuperación funcional. El objetivo final de la fisioterapia es devolver al paciente al nivel de rendimiento anterior a la lesión, incluidos el movimiento y la fuerza, sin dañar ni alargar el injerto. Presentamos el reporte de un caso con una lesión de LCA compleja, que recupero la funcionalidad de la articulación por aplicación de plan de tratamiento fisioterapéutico de forma inmediata a su intervención quirúrgica.

El receptor de la lipoproteína de baja densidad oxidado tipo lectina 1 (LOX-1), también conocido como OLR-1, es un receptor scavenger (SR) clase E, que media la absorción del colesterol LDL en su forma oxidada, por las células vasculares. LOX-1 está involucrado en la disfunción endotelial, la adhesión de monocitos, la proliferación, migración y apoptosis de las células del músculo liso, la formación de células espumosas, la activación de plaquetas, así como la inestabilidad a nivel del endotelio vascular; todos eventos críticos en la patogénesis de la aterosclerosis. LOX-1 contribuyen a la inestabilidad de la placa ateroesclerótica y a las últimas secuelas clínicas de ruptura endotelial e isquemia tisular cardíaca potencialmente mortal. No existe en la actualidad ningún fármaco aprobado o en desarrollo clínico a partir de LOX-1, debido a sus complejos mecanismos biológicos no dilucidados completamente. Se han utilizado diversas terapias con el objetivo de inhibir la acción de LOX-1; medicamentos como: antioxidantes, estatinas, agentes antinflamatorios naturales, que actúen sobre su expresión, pero todos con eficacia moderada. También se ha evaluado la administración de anticuerpos anti-LOX-1 inhibe la aterosclerosis al disminuir eventos celulares. El diseño de fármacos enfocados en el conocimiento de las vías de señalización de LOX-1 y la aplicación de herramientas biotecnológicas permite el desarrollo de nuevas dianas terapéuticas basadas en la potencialidad que tienen los anticuerpos monoclonales. Con estos antecedentes el, receptor LOX-1, representa un objetivo terapéutico atractivo para el tratamiento de enfermedades ateroscleróticas humanas. La evidencia reciente indica que la acción sobre este SR es una posible estrategia para el tratamiento de la enfermedad vascular, explorando en esta revisión su papel y posibles futuras aplicaciones en el diagnóstico y la terapéutica.