Investigation on Cell Disruption Techniques and Supercritical Carbon Dioxide Extraction of Mortierella alpina Lipid.

Mortierella alpina, an oleaginous fungus, has been shown to be a potential source for arachidonic acid (ARA) production. The recovery of intracellular lipids from M. alpina is an important step for the downstream bioprocessing, and green extraction techniques with a focus on being efficient and eco-friendly have drawn much attention. In this study, different cell disruption techniques (mechanical: high-speed homogenization 10,000 rpm, ultrasonication 20 kHz, high-pressure process (HPP) 200-600 MPa; non- mechanical: acid treatment HCl) were investigated for lipid recovery from M. alpina, and process parameters (A. temperature, B. pressure, C. cosolvent ratio) of supercritical carbon dioxide (SC-CO2) lipid extraction were studied by applying response surface methodology (RSM). Compared with Soxhlet extraction as a control group (100%), high-speed homogenization has the highest lipid recovery (115.40%) among mechanical disruption techniques. Besides, there was no significant difference between high-speed homogenization and 1 M HCl treatment (115.55%) in lipid recovery. However, lipid recovery decreased to 107.36% as the concentration of acid was increased to 3 M, and acid treatment showed a negative effect on the ARA ratio. In HPP treatment, the highest lipid recovery (104.81%) was obtained at 400 MPa, 1 time of treatment and water medium. In the response surface model of SC-CO2 extraction, results showed the major influence of the process parameters to lipid recovery was pressure, and there are interaction effects of AC (temperature and cosolvent ratio) and BC (pressure and cosolvent ratio). Lipid recovery of SC-CO2 extraction reached 92.86% at 201 bar, 58.9 °C and cosolvent ratio 1:15. The microbial lipid recovery process of this study could be used as a reference and an eco-friendly alternative for the future downstream bioprocessing of ARA production by M. alpina.

A robotic pill for oral delivery of biotherapeutics: safety, tolerability, and performance in healthy subjects.

Biotherapeutics are highly efficacious, but the pain and inconvenience of chronic injections lead to poor patient compliance and compromise effective disease management. Despite innumerable attempts, oral delivery of biotherapeutics remains unsuccessful due to their degradation in the gastrointestinal (GI) environment and poor intestinal absorption. We have developed an orally ingestible robotic pill (RP) for drug delivery, which protects the biotherapeutic drug payload from digestion in the GI tract and auto-injects it into the wall of the small intestine as a safe, pain-free injection since the intestines are insensate to sharp stimuli. The payload is delivered upon inflation of a balloon folded within the RP, which deflates immediately after drug delivery. Here we present results from two clinical studies demonstrating the safety, tolerability and performance of the RP in healthy humans. In the first study, three versions of the RP (A, B and C) were evaluated, which were identical in all respects except for the diameter of the balloon. The RP successfully delivered a biotherapeutic (octreotide) in 3 out of 12 subjects in group A, 10 out of 20 subjects in group B and 16 out of 20 subjects in group C, with a mean bioavailability of 65 ± 9% (based on successful drug deliveries in groups A and B). Thus,  reliability of drug delivery with the RP ranged from 25 to 80%, with success rate directly related to balloon size. In a separate study, the deployment of the RP was unaffected by fed or fasting conditions suggesting that the RP may be taken with or without food. These promising clinical data suggest that biotherapeutics currently administered parenterally may be safely and reliably delivered via this versatile, orally ingestible drug delivery platform.

Biphasic transdermal iontophoretic drug delivery platform.

Transdermal iontophoresis is an active drug delivery method that has the potential to transform treatment of conditions such as acute pain that require a succession of on-demand metered-dose drug deliveries. However, current monophasic iontophoresis methods fail to meet these requirements due to their inability to halt the passive diffusion of active agents when therapy is not required. We have developed a biphasic iontophoretic system to overcome these limitations. The viability of this system was assessed in an in vitro porcine skin preparation using FeCl(2) (127 Daltons), a charged molecule which can undergo both active and passive transdermal diffusion. The transport properties of the system were modeled using a Fourier Transform-derived optimum estimate transfer function. Using this model, experimental results showed good correlation to predicted values for both cumulative dose (R(2)=0.912, n=10), and density dose (R(2)=0.802, n=10). Results also showed the ability to effectively deliver the compound during active periods while minimizing delivery during inactive periods. While preliminary, our results suggest biphasic iontophoresis is a viable means of delivering on-demand drug therapy while minimizing unwanted off-demand delivery.