Assessing the Antifungal Activity of a New Oral Lipid-Based Amphotericin B Formulation Following Administration to Rats Infected withAspergillus Fumigatus

Abstract

The purpose of this study was to assess the antifungal activity of a new oral amphotericin B (AmpB) lipid-based formulation following administration to rats infected with Aspergillus fumigatus. Aspergillus fumigatus inoculum (2.1-2.5 x 10(7) colony forming units [CFU]) were injected via the jugular vein; 48h later male albino Sprague-Dawley rats (350-400 g) were administered either a single oral dose of AmpB incorporated into Peceol (50 mg AmpB/kg), physiologic saline (nontreated controls) or Peceol alone (vehicle control) once daily for 4 days. To assess antifungal activity Brain, Lung, Heart, Liver, Spleen and Kidney sections were homogenized with normal saline (1 mL/g of tissue) and a 0.1-mL aliquot was spread plated onto a Sabourand dextrose agar plate. The plates were incubated for 48 hr at 37 degrees C, at which time the number of fungal CFU were determined and corrected for tissue weight. In addition, plasma galactomannan antigen concentrations were determined. Data was reported as mean +/- standard error of the mean. The AmpB-Peceol oral formulation significantly decreased total fungal CFU concentrations recovered in all the organs added together, brain CFU concentrations, spleen CFU concentrations and plasma galactomannan antigen concentrations compared to baseline. No significant differences in lung, heart, liver and kidney CFU concentrations between treatment and control groups were observed. Peceol vehicle control did not exhibit any antifungal activity. These findings suggest that a new oral lipid-based formulation of AmpB incorporated into Peceol can significantly decrease brain and spleen CFU concentrations and plasma galactomannan antigen concentrations compared to non-treated controls.

Review and analysis of FDA approved drugs using lipid-based formulations

“…When suspended in Peceol V R (glyceryl monooleate), amphotericin B (AmpB)'s bioavailability is significantly improved [68][69][70]. AmpB in a Peceol/Gelucire 44/14 (lauroyl macrogol-32 glycerides)-based formulation (iCo -010) has demonstrated efficacy in several animal models of systemic fungal infections [71] as well as visceral leishmaniasis [72,73]. Organ distribution of amphotericin B is shifted away from the kidney, the site of dose-limiting toxicity, with a corresponding increased splenic and hepatic uptake when the drug is administered as LBDDS [67].…”

Rodent Models of Invasive Aspergillosis due to Aspergillus fumigatus: Still a Long Path toward Standardization

“…For example, weight ranged from 9 g for very young mice (Li et al, 2014) to around 45 g, when they lived up to 42 months (Khosravi et al, 2012). For rats, it spanned from 26.5 g for 11 day old-pups (Zimmerli et al, 2007) to almost 400 g for oldest animals (Sivak et al, 2004a,b; Risovic et al, 2007; Wasan et al, 2007, 2009). Weight for guinea pigs was more homogeneous: 486.1 g ± 36.9.…”

“…It usually required a smaller A. fumigatus inoculum, regardless the immunocompromised status of the rodents: 2.3 × 10 7 spores in average, extremes ranging from 5.0 × 10 1 (Cutsem et al, 1993) to 2.5 × 10 9 (Bowman et al, 2001), and its usage was reported in 30.0% of the selected articles of this review. Jugular vein (Sivak et al, 2004a,b; Risovic et al, 2007), femoral vein (Wong et al, 1989), and penis vein (Overdijk et al, 1996; Reichard et al, 1997; Odds et al, 1998) were generally chosen for such a purpose in large rodent species, while lateral tail vein was privileged in mice (Odds et al, 1998). Retro-orbital vein was an alternative, but has become less frequent in recent years for ethical considerations (te Dorsthorst et al, 2005; Verweij et al, 2008; Wagener et al, 2008; Mouton et al, 2009; Dirr et al, 2010; Kotz et al, 2010; Heesemann et al, 2011).…”

The Global Access Initiative at the University of British Columbia (UBC): Availability of UBC Discoveries and Technologies to the Developing World

“…One example of a current global access project at UBC is in the laboratory of Dr. Kishor M. Wasan. The development of a lipid‐based amphotericin B formulation for oral administration is currently underway, and has significant implications for treatment of a number of disease states, including systemic fungal infections and kinetoplastid parasitic infections 4–6. The traditional formulation of Amphotericin B (Fungizone®, a colloidal suspension) requires a treatment regime of parenteral administration ranging from 5 to 40 days, and is associated with infusion and drug‐related side‐effects (infection of the indwelling catheter, patient chills and shaking due to RBC haemolysis, dose‐dependent renal toxicity, fever, bone pain, thrombophlebitis) 7.…”

“…Initial data from both cell lines and in vivo research indicate that the oral formulation is highly efficacious and exhibits low toxicity within the dosage range required in order to treat diseases such as disseminated fungal infections and leishmaniasis 4–6. This has significant implications in developed nations, where the rates of opportunistic fungal infections such as candidiasis, histoplasmosis and aspergillosis are climbing, particularly within patient populations affected by cancer, organ transplant recipients, diabetics and HIV/AIDS.…”