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Formulation of the microbicide INP0341 for in vivo protection against a vaginal challenge by Chlamydia trachomatis.

Pedersen C, Slepenkin A, Andersson SB, Fagerberg JH, Bergström CA, Peterson EM - PLoS ONE (2014)

Bottom Line: Gelling agents (polymers) with inherent antimicrobial properties were chosen to maximize the total antimicrobial effect of the gel.Formulations with and without INP0341 afforded protection, but the inclusion of INP0341 increased the protection.The lack of toxicity was confirmed by in vitro assays using EpiVaginal tissues, which showed that a 24 h exposure to the gel formulation did not decrease the cell viability or the barrier function of the tissue.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, Uppsala University, Biomedical Center, Uppsala, Sweden.

ABSTRACT
The salicylidene acylhydrazide (SA) compounds have exhibited promising microbicidal properties. Previous reports have shown the SA compounds, using cell cultures, to exhibit activity against Chlamydia trachomatis, herpes simplex virus and HIV-1. In addition, using an animal model of a vaginal infection the SA compound INP0341, when dissolved in a liquid, was able to significantly protect mice from a vaginal infection with C. trachomatis. To expand upon this finding, in this report INP0341 was formulated as a vaginal gel, suitable for use in humans. Gelling agents (polymers) with inherent antimicrobial properties were chosen to maximize the total antimicrobial effect of the gel. In vitro formulation work generated a gel with suitable rheology and sustained drug release. A formulation containing 1 mM INP0341, 1.6 wt% Cremophor ELP (solubility enhancer) and 1.5 wt% poly(acrylic acid) (gelling and antimicrobial agent), was chosen for studies of efficacy and toxicity using a mouse model of a vaginal infection. The gel formulation was able to attenuate a vaginal challenge with C. trachomatis, serovar D. Formulations with and without INP0341 afforded protection, but the inclusion of INP0341 increased the protection. Mouse vaginal tissue treated with the formulation showed no indication of gel toxicity. The lack of toxicity was confirmed by in vitro assays using EpiVaginal tissues, which showed that a 24 h exposure to the gel formulation did not decrease the cell viability or the barrier function of the tissue. Therefore, the gel formulation described here appears to be a promising vaginal microbicide to prevent a C. trachomatis infection with the potential to be expanded to other sexually transmitted diseases.

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Gel viscosity as a function of shear rate.Gels with PAA concentrations of 1.0–2.0 wt% were measured by continuous rotational viscometry at 37°C.
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pone-0110918-g001: Gel viscosity as a function of shear rate.Gels with PAA concentrations of 1.0–2.0 wt% were measured by continuous rotational viscometry at 37°C.

Mentions: Figure 1 shows the viscosity as a function of shear rate for gels with 1.0–2.0 wt% PAA. The reported viscosities are the steady shear viscosities, calculated from the torque measured after 1 minute of continuous shear. All gels exhibit shear thinning rheology, i.e. decreasing viscosity with increasing shear rate, which is a warranted feature of vaginal gels [18]. The viscosity values of the PAA gels are within the viscosity range of commercial vaginal gels [18]. The elastic modulus as function of strain (Figure S1 in File S1) and the shear stress as function of shear rate (Figure S2 in File S1) are reported in the Supporting Information. All PAA gels exhibited yield stress, i.e. positive shear stress values at zero shear rate, as shown in Table S1 in the File S1. The yield stress of the INP0341 gel studied in vivo (1.5 wt% PAA) was determined to be 11.0 Pa, which was slightly lower than observed for the commercial vaginal gels Crinone (16.2 Pa) and Replens (14.3 Pa). The rheological data was used in combination with a squeezing flow model, after which 1.5 wt% PAA was chosen as a suitable polymer concentration for in vivo use (File S1).


Formulation of the microbicide INP0341 for in vivo protection against a vaginal challenge by Chlamydia trachomatis.

Pedersen C, Slepenkin A, Andersson SB, Fagerberg JH, Bergström CA, Peterson EM - PLoS ONE (2014)

Gel viscosity as a function of shear rate.Gels with PAA concentrations of 1.0–2.0 wt% were measured by continuous rotational viscometry at 37°C.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4214720&req=5

pone-0110918-g001: Gel viscosity as a function of shear rate.Gels with PAA concentrations of 1.0–2.0 wt% were measured by continuous rotational viscometry at 37°C.
Mentions: Figure 1 shows the viscosity as a function of shear rate for gels with 1.0–2.0 wt% PAA. The reported viscosities are the steady shear viscosities, calculated from the torque measured after 1 minute of continuous shear. All gels exhibit shear thinning rheology, i.e. decreasing viscosity with increasing shear rate, which is a warranted feature of vaginal gels [18]. The viscosity values of the PAA gels are within the viscosity range of commercial vaginal gels [18]. The elastic modulus as function of strain (Figure S1 in File S1) and the shear stress as function of shear rate (Figure S2 in File S1) are reported in the Supporting Information. All PAA gels exhibited yield stress, i.e. positive shear stress values at zero shear rate, as shown in Table S1 in the File S1. The yield stress of the INP0341 gel studied in vivo (1.5 wt% PAA) was determined to be 11.0 Pa, which was slightly lower than observed for the commercial vaginal gels Crinone (16.2 Pa) and Replens (14.3 Pa). The rheological data was used in combination with a squeezing flow model, after which 1.5 wt% PAA was chosen as a suitable polymer concentration for in vivo use (File S1).

Bottom Line: Gelling agents (polymers) with inherent antimicrobial properties were chosen to maximize the total antimicrobial effect of the gel.Formulations with and without INP0341 afforded protection, but the inclusion of INP0341 increased the protection.The lack of toxicity was confirmed by in vitro assays using EpiVaginal tissues, which showed that a 24 h exposure to the gel formulation did not decrease the cell viability or the barrier function of the tissue.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, Uppsala University, Biomedical Center, Uppsala, Sweden.

ABSTRACT
The salicylidene acylhydrazide (SA) compounds have exhibited promising microbicidal properties. Previous reports have shown the SA compounds, using cell cultures, to exhibit activity against Chlamydia trachomatis, herpes simplex virus and HIV-1. In addition, using an animal model of a vaginal infection the SA compound INP0341, when dissolved in a liquid, was able to significantly protect mice from a vaginal infection with C. trachomatis. To expand upon this finding, in this report INP0341 was formulated as a vaginal gel, suitable for use in humans. Gelling agents (polymers) with inherent antimicrobial properties were chosen to maximize the total antimicrobial effect of the gel. In vitro formulation work generated a gel with suitable rheology and sustained drug release. A formulation containing 1 mM INP0341, 1.6 wt% Cremophor ELP (solubility enhancer) and 1.5 wt% poly(acrylic acid) (gelling and antimicrobial agent), was chosen for studies of efficacy and toxicity using a mouse model of a vaginal infection. The gel formulation was able to attenuate a vaginal challenge with C. trachomatis, serovar D. Formulations with and without INP0341 afforded protection, but the inclusion of INP0341 increased the protection. Mouse vaginal tissue treated with the formulation showed no indication of gel toxicity. The lack of toxicity was confirmed by in vitro assays using EpiVaginal tissues, which showed that a 24 h exposure to the gel formulation did not decrease the cell viability or the barrier function of the tissue. Therefore, the gel formulation described here appears to be a promising vaginal microbicide to prevent a C. trachomatis infection with the potential to be expanded to other sexually transmitted diseases.

Show MeSH
Related in: MedlinePlus