Segmented intravaginal ring for the combination delivery of hydroxychloroquine and anti-CCR5 siRNA nanoparticles as a potential strategy for preventing HIV infection

Vaginal drug delivery has been shown to be a promising strategy for the prevention of sexually transmitted infections. Therapy delivered at the site of infection has many advantages including improved therapeutic efficacy, reduction in systemic toxicity, and reduced potential for development of drug resistance.

We developed a “smart” combination intravaginal ring (IVR) that will (1) provide continuous release of hydroxychloroquine (HCQ) to induce T cell immune quiescence as the first-line of defense and (2) release nanoparticles containing anti-CCR5 siRNA only during sexual intercourse when triggered by the presence of seminal fluid as the second-line of defense. The IVR was capable of releasing HCQ over 25 days with a mean daily release of 31.17 ± 3.06 µg/mL. In the presence of vaginal fluid simulant plus seminal fluid simulant, over 12 × more nanoparticles (5.12 ± 0.9 mg) were released over a 4-h period in comparison to IVR segments that were incubated in the presence of vaginal fluid simulant alone (0.42 ± 0.19 mg).

Anti-CCR5 siRNA nanoparticles were able to knockdown 83 ± 5.1% of CCR5 gene expression in vitro in the CD4⁺ T cell line Sup-T1. The IVR system also demonstrated to be non-cytotoxic to VK2/E6E7 vaginal epithelial cells.

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Materials: Hydrophilic thermoplastic polyurethane (PU) (Tecophilic™ HP-60D-35) was purchased from Lubrizol Advanced Materials (Cleveland, OH, USA). Eudragit L100 (methacrylic acid-methyl acrylate copolymer; anionic pH-sensitive polymer) was kindly donated by Evonik Industries (Essen, Germany). Hydroxypropyl methylcellulose (HPMC) K100M was donated by Dow Chemical Company (New Milford, CT, USA). Polyethylene glycol 400, NF (PEG-8) was acquired from Medisca (Saint-Laurent, QC, Canada). HCQ was purchased from Thermo Fisher Scientific (Burlington, ON, Canada). CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS) was purchased from Promega (Madison, USA). Glyceryl monostearate (molecular weight 358.56) was purchased from Sigma-Aldrich (Ontario, Canada). L-α-phosphatidylcholine (Soy-95%) (molecular weight 770.123 g/mol) was purchased from Avanti Polar Lipids (AL, USA). Polyvinyl alcohol (PVA; 31 − 50 kDa) and polyethyleneimine (PEI; branched, MW 25 K) were obtained from Sigma-Aldrich (Ontario, Canada). Tris–EDTA was purchased from ThermoFisher Scientific (Ontario, Canada). 2-(N-morpholino) ethanesulfonic acid (MES) was purchased from Sigma-Aldrich (Ontario, Canada). 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC; 200 mg/mL) N-hydroxysuccinimide (NHS; 275 mg/mL) were purchased from G-Biosciences (Missouri, USA). Anti-Human CCR5 siRNA (sense: 5′-GUUCAGAAACUACCUCUUAdTdT-3′, antisense: 3′-dTdTCAAGUVUUUGAUGGAGAAU-5′) was purchased from Dharmacon (ON, Canada). Human CCR5 primers (Forward: 5′-TTCATCATCCTCCTGACAATCG-3′; Reverse: 5′-GCCACCACCCAAGTGATCAC-3′) and human GAPDH primer (Forward: 5′-AAGAAGGTGGTGAAGCAGGCG-3′; Reverse: 5′-AGACAACCTGGTCCTCAGTGTAGC-3′) were purchased from Thermo Fisher (ON, Canada). E.Z.N.A.® RNA isolation kit was purchased from Sigma Aldrich (ON, Canada). PerfeCTa SYBR® Green SuperMix and qScript™ cDNA were purchased from Quanta (ON, Canada). Anti-CD4 antibody was purchased from Abcam (ON, Canada).

Article information: Traore, Y.L., Chen, Y., Padilla, F. et al. Segmented intravaginal ring for the combination delivery of hydroxychloroquine and anti-CCR5 siRNA nanoparticles as a potential strategy for preventing HIV infection. Drug Deliv. and Transl. Res. (2021). https://doi.org/10.1007/s13346-021-00983-w