Liposomal Drug Delivery against Helicobacter pylori Using Furazolidone and N-Acetyl Cysteine in Augmented Therapy

Abstract

Helicobacter pylori (H. pylori) infection is a significant global health concern, affecting approximately 50% of the world's population and leading to gastric ulcers, gastritis, and gastric cancer. The increase in antibiotic resistance has compromised the efficacy of existing therapeutic regimens, necessitating novel approaches for effective eradication. This study aimed to develop a targeted liposomal drug delivery system incorporating furazolidone and N-acetylcysteine (NAC) to enhance mucopenetration and improve Helicobacter pylori eradication. Liposomes were formulated with furazolidone, NAC, and Pluronic F-127 using a modified reverse-phase evaporation technique. The formulations were categorized based on charge as neutral, negative, and positive and tested for mucopenetration using a modified silicon tube method with coumarin-6 as a fluorescent marker. The encapsulation efficiency and particle size were analyzed using HPLC and an Izon q-nano particle size analyzer. The results indicated that charged liposomes showed a higher encapsulation efficiency than neutral liposomes with Pluronic F-127. Notably, combining furazolidone with 1% NAC achieved complete eradication of H. pylori in 2.5 h, compared to six hours without NAC. The findings of this study suggest that incorporating NAC and Pluronic F-127 into liposomal formulations significantly enhances mucopenetration and antimicrobial efficacy.

Keywords: H. pylori; N-acetyl cysteine; augmented therapy; furazolidone; liposomes; mucus penetration.

Scheme 1

The coding for mucopenetrative liposomal formulation compositions (MP1-MP6).

Figure 1

Spot plating technique for different dilutions from (A). 10⁻¹ through 10⁻³ and (B). from 10⁻¹ to 10⁻⁶.

Figure 2

Transmission electron microscopic, TEM, images of mucopenetrative liposomes (A). MP3, cationic liposomes with Pluronic F-127 (B). MP1, neutral liposomes with Pluronic F-127. For formulations’ composition, refer to Table 1.

Figure 3

Diffusion of mucopenetrative liposomal particles from MP1 to MP6 through 1 mm thick sigma mucin type I in a silicon tube maintained at pH 6.0 at 37 °C at 1, 2, and 3 hours’ time (n = 3, mean ± SD shown). Abbreviations: MP1 (neutral with Pluronic F-127), MP4 (neutral without Pluronic F-127), MP2 (negative with Pluronic F-127), MP5 (negative without Pluronic F-127), MP3 (positive with Pluronic F-127), MP6 (positive without Pluronic F-127). For formulations’ composition, refer to Table 1.

Figure 4

(A). In vitro drug release of furazolidone from the mucopenetrative formulation MP1 and MP4 up to a 4 h time at pH 6.0 (n = 3, mean ± SD shown). Abbreviations: MP1 (neutral with Pluronic F-127), MP4 (neutral without Pluronic F-127). (B). In vitro drug release of furazolidone from the mucopenetrative formulation MP3 and MP6 up to a 4 h time at pH 6.0 (n = 3, mean ± SD shown). Abbreviations: MP3 (positive with Pluronic F-127), MP6 (positive without Pluronic F-127). (C). In vitro drug release of furazolidone from the mucopenetrative formulation MP2 and MP5 for up to a 4 h time at pH 6.0 (n = 3, mean ± SD shown). Abbreviations: MP2 (negative with Pluronic F-127), MP5 (negative without Pluronic F-127).

Figure 4

(A). In vitro drug release of furazolidone from the mucopenetrative formulation MP1 and MP4 up to a 4 h time at pH 6.0 (n = 3, mean ± SD shown). Abbreviations: MP1 (neutral with Pluronic F-127), MP4 (neutral without Pluronic F-127). (B). In vitro drug release of furazolidone from the mucopenetrative formulation MP3 and MP6 up to a 4 h time at pH 6.0 (n = 3, mean ± SD shown). Abbreviations: MP3 (positive with Pluronic F-127), MP6 (positive without Pluronic F-127). (C). In vitro drug release of furazolidone from the mucopenetrative formulation MP2 and MP5 for up to a 4 h time at pH 6.0 (n = 3, mean ± SD shown). Abbreviations: MP2 (negative with Pluronic F-127), MP5 (negative without Pluronic F-127).

Figure 5

(A). In vitro drug release of NAC from the mucopenetrative formulation from MP1 and MP4 for up to 4 h time at pH 6.0 (n = 3, mean ± SD shown). Abbreviations: MP1 (neutral with Pluronic F-127), MP4 (neutral without Pluronic F-127). (B). In vitro drug release of NAC from the mucopenetrative formulation from MP2 and MP5 for up to 4 h time at pH 6.0 (n = 3, mean ± SD shown). Abbreviations: MP2 (negative with Pluronic F-127), MP5 (negative without Pluronic F-127). (C). In vitro drug release of NAC from the mucopenetrative formulation from MP3 and MP6 for up to 4 h time at pH 6.0 (n = 3, mean ± SD shown). Abbreviations: MP3 (positive with Pluronic F-127), MP6 (positive without Pluronic F-127).

Figure 5

(A). In vitro drug release of NAC from the mucopenetrative formulation from MP1 and MP4 for up to 4 h time at pH 6.0 (n = 3, mean ± SD shown). Abbreviations: MP1 (neutral with Pluronic F-127), MP4 (neutral without Pluronic F-127). (B). In vitro drug release of NAC from the mucopenetrative formulation from MP2 and MP5 for up to 4 h time at pH 6.0 (n = 3, mean ± SD shown). Abbreviations: MP2 (negative with Pluronic F-127), MP5 (negative without Pluronic F-127). (C). In vitro drug release of NAC from the mucopenetrative formulation from MP3 and MP6 for up to 4 h time at pH 6.0 (n = 3, mean ± SD shown). Abbreviations: MP3 (positive with Pluronic F-127), MP6 (positive without Pluronic F-127).

Figure 6

A time–kill curve experiment of furazolidone augmented with NAC at a concentration of 1% of its minimum inhibitory concentration (MIC) against H. pylori inoculated on a blood agar plate and incubated at 37 °C for 24 h in an anaerobic jar with CampyGen gas packs before counting the colonies. The mean value of the log number of cfu per milliliter was plotted against time (n = 6, mean ± SD).