TY - JOUR
T1 - Preformulation and stability in biological fluids of the retrocyclin RC-101, a potential anti-HIV topical microbicide
AU - Sassi, Alexandra B.
AU - Bunge, Katherine E.
AU - Hood, Brian L.
AU - Conrads, Thomas P.
AU - Cole, Alexander M.
AU - Gupta, Phalguni
AU - Rohan, Lisa C.
N1 - Funding Information:
The project described was supported by Grant Number NIH 1U19 AI065430-01 and AI082623 from the National Institute of Allergy and Infectious Diseases (NIAID). Its contents are solely the responsibility of the author and do not necessarily represent the official views of the NIAID. Funding was also provided by the James B. Pendleton Charitable Trust. The authors would like to thank Dr. Michael Cascio at the Molecular Genetics and Biochemistry Department at the University of Pittsburgh for the use of the Circular Dichroism spectrophotometer and the assistance provided with the experimental design. Dr. Billy W. Day and Dr. Manimalha Balasubramani at the Genomics and Proteomics Core Laboratories at the University of Pittsburgh for the assistance provided for the MALDI-TOF MS analysis. Lorna Rabe and her team for the microbiological assessment of the biological fluids. Phillip Graebing at the Magee-Womens Research Institute for the analytical help and support provided. Ingrid Macio, Patricia Barcic, Mary McQueen, Kathy Laychak, and Cindy Schatzman from the Magee-Womens Clinical & Translational Research Center (CTRC) for all the assistance provided. Lindsay Ferguson, Yardlee Kauffman, Gargi Bajpayee, and Lin Wang for the help provided during enrolment of the volunteers.
PY - 2011/7/29
Y1 - 2011/7/29
N2 - Background: RC-101, a cationic peptide retrocyclin analog, has in vitro activity against HIV-1. Peptide drugs are commonly prone to conformational changes, oxidation and hydrolysis when exposed to excipients in a formulation or biological fluids in the body, this can affect product efficacy. We aimed to investigate RC-101 stability under several conditions including the presence of human vaginal fluids (HVF), enabling the efficient design of a safe and effective microbicide product. Stability studies (temperature, pH, and oxidation) were performed by HPLC, Circular Dichroism, and Mass Spectrometry (LC-MS/MS). Additionally, the effect of HVF on formulated RC-101 was evaluated with fluids collected from healthy volunteers, or from subjects with bacterial vaginosis (BV). RC-101 was monitored by LC-MS/MS for up to 72 h.Results: RC-101 was stable at pH 3, 4, and 7, at 25 and 37°C. High concentrations of hydrogen peroxide resulted in less than 10% RC-101 reduction over 24 h. RC-101 was detected 48 h after incubation with normal HVF; however, not following incubation with HVF from BV subjects.Conclusions: Our results emphasize the importance of preformulation evaluations and highlight the impact of HVF on microbicide product stability and efficacy. RC-101 was stable in normal HVF for at least 48 h, indicating that it is a promising candidate for microbicide product development. However, RC-101 stability appears compromised in individuals with BV, requiring more advanced formulation strategies for stabilization in this environment.
AB - Background: RC-101, a cationic peptide retrocyclin analog, has in vitro activity against HIV-1. Peptide drugs are commonly prone to conformational changes, oxidation and hydrolysis when exposed to excipients in a formulation or biological fluids in the body, this can affect product efficacy. We aimed to investigate RC-101 stability under several conditions including the presence of human vaginal fluids (HVF), enabling the efficient design of a safe and effective microbicide product. Stability studies (temperature, pH, and oxidation) were performed by HPLC, Circular Dichroism, and Mass Spectrometry (LC-MS/MS). Additionally, the effect of HVF on formulated RC-101 was evaluated with fluids collected from healthy volunteers, or from subjects with bacterial vaginosis (BV). RC-101 was monitored by LC-MS/MS for up to 72 h.Results: RC-101 was stable at pH 3, 4, and 7, at 25 and 37°C. High concentrations of hydrogen peroxide resulted in less than 10% RC-101 reduction over 24 h. RC-101 was detected 48 h after incubation with normal HVF; however, not following incubation with HVF from BV subjects.Conclusions: Our results emphasize the importance of preformulation evaluations and highlight the impact of HVF on microbicide product stability and efficacy. RC-101 was stable in normal HVF for at least 48 h, indicating that it is a promising candidate for microbicide product development. However, RC-101 stability appears compromised in individuals with BV, requiring more advanced formulation strategies for stabilization in this environment.
UR - http://www.scopus.com/inward/record.url?scp=79960857058&partnerID=8YFLogxK
U2 - 10.1186/1742-6405-8-27
DO - 10.1186/1742-6405-8-27
M3 - Article
AN - SCOPUS:79960857058
SN - 1742-6405
VL - 8
JO - AIDS Research and Therapy
JF - AIDS Research and Therapy
M1 - 27
ER -