Raman spectroscopy is a very promising technique increasingly used in the pharmaceutical industry. Due to its development and improved instrumental versatility achieved over recent decades and through the application of chemometric methods, this technique has become highly precise and sensitive for the quantification of drug substances. Thus, it has become fundamental in identifying critical variables and their clinical relevance in the development of new drugs. In process monitoring, it has been used to highlight in-line real-time analysis, and it has been used more commonly since 2004 when the Food and Drug Administration (FDA) launched Process Analytical Technology (PAT), integrated with the concepts of Pharmaceutical Current Good Manufacturing Practices (CGMPs) for the 21st Century. The present review presents advances in the application of this tool in the development of pharmaceutical products and processes in the last six years.

Bacterial conjunctivitis significantly impacts public health, including more than one-third of eye diseases reported worldwide. It is an infection caused by various aerobic and anaerobic bacteria and is highly contagious. Therefore, it has a high incidence of bacterial resistance to the antibiotics commonly used for treatment. Among the most recent antibiotics, besifloxacin is a fourth-generation fluoroquinolone antibiotic indicated exclusively for topical ophthalmic use. Due to its importance in treating bacterial conjunctivitis and its low solubility in water, limiting its efficacy, a nanotechnology-based drug delivery preparation was developed to overcome this hurdle. Besifloxacin nanocrystals were prepared by small-scale wet milling and response surface methodology, using Povacoat® as a stabilizer. The particle’s average hydrodynamic diameter (Z-ave) was approximately 550 nm (17 times smaller than raw material), with a polydispersity index (PdI) of less than 0.2. The saturation solubility increased about two times compared to the raw material, making it possible to increase the dissolution rate of this drug substance, potentially improving its bioavailability and safety. The optimized preparation was stable under an accelerated stability study (90 days). The Z-ave, PZ, PdI, and content did not alter significantly during this period. Furthermore, the 0.6% m/m besifloxacin nanocrystals at the maximum dose and the Povacoat® stabilizer did not show toxicity in Galleria mellonella larvae. The innovative ophthalmic preparation minimum inhibitory concentration (MIC) was 0.0960 µg/mL and 1.60 µg/mL against Staphylococcus aureus and Pseudomonas aeruginosa, respectively, confirming in vitro efficacy. Therefore, besifloxacin nanocrystals revealed the potential for reduced dosing of the drug substance, with a minor occurrence of adverse effects and greater patient adherence to treatment.

À minha esposa, com amor e gratidão por sua compreensão, aos meus filhos, à minha família, amigos e colegas por todo apoio e torcida. AGRADECIMENTOSEm primeiro lugar, agradeço a Deus por iluminar e me prover de coragem para realizar esse sonho, Ele que abençoa a minha vida em todas as circunstâncias.Agradeço à Profa. Dra. Nádia Araci Bou Chacra por me aceitar como aluno de pósgraduação, por toda dedicação, aconselhamentos, paciência e confiança.Aos meus colegas de laboratório, por todos ensinamentos, parcerias e discussões técnicas.À toda equipe do Departamento de Farmácia do Programa de Pós-graduação em Fármaco e Medicamentos. À Profa. Dra. Iolanda Midea Cuccovia do Departamento de Bioquímica (IQ/USP) que abriu as portas do seu laboratório, possibilitando a conclusão de etapas essenciais dessse estudo, em tempo adequado. Ao Prof. Dr. Humberto Gomes Ferraz e à toda equipe e colegas do DEINFAR, pelo compartilhamento do laboratório e equipamentos. À Profa. Dra. Kelly Ishida e a doutoranda Thayná L. Barreto, pelos ensinamentos e parceria referente às análises de toxicidade. Ao Prof. Dr. Felipe Rabelo Lourenço pelo apoio na realização das análises microbiológicas. À Profa. Dra. Priscyla Daniely M. Gaspari e ao Prof. Dr. Newton Andréo Filho pelos comentários valiosos durante meu exame de qualificação. Ao Grupo Eurofarma, principalmente, à unidade Momenta e aos meus gestores que me deram flexibilidade para concluir esse projeto. À minha família, colegas e amigos que torceram por mim. Especialmente, a minha esposa pela compreensão e paciência e aos meus filhos Victor e Heloísa que me proporcionam motivação todos os dias.