Anti-aggregative activity of doxycycline hyclate loaded-nanostructured lipid carriers against neurodegeneration-related pathogenic proteins

• Grant number: 24/13159-2
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate (Direct)
• Start date: January 15, 2025
• End date: December 14, 2025
• Field of knowledge: Biological Sciences - Pharmacology - Biochemical and Molecular Pharmacology
• Principal Investigator: Maria Palmira Daflon Gremião
• Grantee: Mariana Conceição
• Supervisor: Tiago Fleming de Oliveira Outeiro
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Institution abroad: University Medical Center Göttingen, Germany
• Associated to the scholarship: 23/04282-2 - Evaluation of the potential of nanostructured lipid carriers containing doxycycline functionalized with anti-beta-amyloid protein antibody for intranasal administration in the treatment of Alzheimer's Disease, BP.DD

Abstract

Neurodegenerative diseases such as Alzheimer's, Parkinson's, Huntington's, and Amyotrophic lateral sclerosis share the common feature of pathological protein aggregation that become toxic to brain tissue leading to cell death. While in Alzheimer's beta-amyloid (²A) and Tau proteins aggregate in insoluble amyloid plaques and neurofibrillary tangles, in Parkinson's disease (PD) alfa-synuclein (aSyn), a presynaptic neuronal protein, is the one to provoke degeneration through a prion mechanism. Doxycycline, a second-generation tetracycline antibiotic, showed anti-aggregative activity against ²A, Tau, and aSyn both in vitro and in vivo. It represents a promising molecule to be applied to Alzheimer's and Parkinson's diseases as it prevents the formation of protein aggregates and disrupts the already formed ones. Aiming to a repurposed use of doxycycline against neurodegeneration in a nanotechnological approach, this research intends to evaluate doxycycline's anti-aggregation capacity when encapsulated in nanostructured lipid carriers. These are lipid nanoparticles whose size and constitution allows controlled drug delivery with improved ability to reach brain areas. In the research, aSyn will be challenged with doxycycline and its encapsulated version to access whether the nanoparticles can improve aggregate disruption and prevent its formation. The analysis includes cell-free assays such as Thioflavin T, seeding, immunoblotting, dynamic light scattering, and fluorescence analysis to assess aSyn aggregation, and cell-based ones as immunocytochemistry, cytotoxicity, and microscopy.