Gerhard Pütz

Molecular MRI in the Earth’s Magnetic Field Using Continuous Hyperpolarization of a Biomolecule in Water

In this work, we illustrate a method to continuously hyperpolarize a biomolecule, nicotinamide, in water using parahydrogen and signal amplification by reversible exchange (SABRE). Building on the preparation procedure described recently by Truong et al. [ J. Phys. Chem. B , 2014 , 118 , 13882 - 13889 ], aqueous solutions of nicotinamide and an Ir-IMes catalyst were prepared for low-field NMR and MRI. The (1)H-polarization was continuously renewed and monitored by NMR experiments at 5.9 mT for more than 1000 s. The polarization achieved corresponds to that induced by a 46 T magnet (P = 1.6 × 10(-4)) or an enhancement of 10(4). The polarization persisted, although reduced, if cell culture medium (DPBS with Ca(2+) and Mg(2+)) or human cells (HL-60) were added, but was no longer observable after the addition of human blood. Using a portable MRI unit, fast (1)H-MRI was enabled by cycling the magnetic field between 5 mT and the Earths field for hyperpolarization and imaging, respectively. A model describing the underlying spin physics was developed that revealed a polarization pattern depending on both contact time and magnetic field. Furthermore, the model predicts an opposite phase of the dihydrogen and substrate signal after one exchange, which is likely to result in the cancelation of some signal at low field.

Elimination of Liposomes by Different Separation Principles Used in Low-density Lipoprotein Apheresis

Abstract:  Clinical success of many therapies is impaired by dose limiting toxicities. Nanoscale particle‐based drug delivery systems such as liposomes show unique pharmacokinetic properties and improved toxicity profiles. Liposomes accumulate in tumor tissue, but only a small fraction of a total dose reaches the target site. The overwhelming amount of a given dose is needed only to build up a diffusion gradient for effective accumulation at the target site. In order to find a way to detoxify this predominant fraction after accumulation is completed, the different separation principles used for the apheresis of lipoproteins were evaluated for the extracorporeal elimination of liposomes. Appropriate radiolabeled model liposomes were prepared by extrusion. Separation efficacy, leakage of liposomal content and influence of plasma contact were measured. Membranes with pore sizes between 25 and 400 nm were used to investigate filtration properties of liposomes. Liposomes were precipitated by adding heparin and Ca2+. Adsorption chromatography was investigated using dextran sulfate, heparin sepharose and functionalized polyacrylamide beads. Membrane filtration allowed the elimination of various liposomes, while precipitation and adsorption were only useful for positively charged liposomes. Leakage of liposomal content was not induced by adsorption, but precipitation induced leakage. Leakage during filtration was dependent on liposomal membrane lipids. Plasma contact reduced precipitation and adsorption efficacy of positively charged liposomes, while filtration properties of liposomes remained unchanged. For extracorporeal elimination of liposomal drug delivery systems, filtration‐based techniques are presumably more convenient and versatile than precipitation‐ or adsorption‐based apheresis technologies.

Controlled application and scheduled removal of nanoparticle based chemotherapeutics (CARL) will reduce dose limiting adverse events in anticancer chemotherapy

Clinical success of cancer chemotherapy is impaired by dose limiting toxicities. Nanoscale particle based drug delivery systems (DDS) like long circulating liposomes show improved toxicity profiles. Nevertheless, their unique pharmacokinetic properties lead to new dose limiting adverse events such as elevated skin toxicity. Though DDS accumulate in tumor tissue, only a very small fraction of the total dose reaches the target site. The overwhelming amount of a given dose is needed only to build up a diffusion gradient for effective accumulation at the target site. Due to the altered endothelial barrier, accumulation of DDS in tumor tissue is much faster than accumulation in other tissues, where dose limiting side effects occur. On the basis of these pharmakinetic data we hypothesize, that once accumulation in the tumor tissue is completed, rapid elimination of the DDS fraction still circulating in the plasma may diminish otherwise dose limiting toxicities. Rapid elimination of circulating DDS might be performed by extracorporeal apheresis treatment. Within this paper the principle of kinetic targeting by scheduled extracorporeal elimination of long circulating DDS is presented in detail. Benefits for patients are as well discussed as possible criticisms and future developments. In conclusion, the combination of DDS and scheduled apheresis may allow the development of new chemotherapy regiments with higher impact and/or less toxicity.

Letter by Winkler et al Regarding Article, “Pilot Study of Extracorporeal Removal of Soluble Fms-Like Tyrosine Kinase 1 in Preeclampsia”

To the Editor: We read with interest the paper by Thadhani et al.1 The authors describe a pilot study using dextran sulfate cellulose (DSC) apheresis to address soluble fms-like tyrosine kinase 1 (sFlt-1), which is associated with the clinical symptoms of preeclampsia. The authors were able to reduce sFLT-1 plasma levels by 20% to 30%. Pregnancy in 3 very preterm preeclamptic women treated at least twice with DSC apheresis was continued between 15 and 23 days.1 However, this is not the first successful approach to prolong pregnancy in preeclampsia with extracorporeal treatment: In 2006, Wang and colleagues2 published a study with 9 very preterm preeclamptic women …

Optimizing Antitumor Efficacy and Adverse Effects of Pegylated Liposomal Doxorubicin by Scheduled Plasmapheresis: Impact of Timing and Dosing

Background: Nanoscale drug delivery systems accumulate in solid tumors preferentially by the enhanced permeation and retention effect (EPR-effect). Nevertheless, only a miniscule fraction of a given dosage reaches the tumor, while >90% of the given drug ends up in otherwise healthy tissues, lead-ing to the severe toxic reactions observed during chemotherapy. Once accumulation in the tumor has reached its maximum, extracorporeal elimination of circulating nanoparticles by plasmapheresis can dimin-ish toxicities. Objective: In this study, we investigated the effect of dosing and plasmapheresis timing on adverse events and antitumor efficacy in a syngeneic rat tumor model. Methods: MAT-B-III cells transfected with a luciferase reporter plasmid were inoculated into female Fisher rats, and pegylated liposomal doxorubicin (PLD) was used for treatment. Plasmapheresis was performed in a discontinuous manner via centrifugation and subsequent filtration of isolated plasma. Results: Bioluminescence measurements of tumor growth could not substitute caliper measurements of tumor size. In the control group, raising the dosage above 9 mg PLD/kg body weight did not increase therapeutic efficacy in our fully immunocompetent animal model. Plasmapheresis was best done 36 h after injecting PLD, leading to similar antitumor efficacy with significantly less toxicity. Plasmapheresis 24 h after injection interfered with therapeutic efficacy, while plasmapheresis after 48 h led to fewer side effects but also to increased weight loss. Conclusion: Long-circulating nanoparticles offer the unique possibility to eliminate the excess of circulat-ing particles after successful accumulation in tumors by EPR, thereby reducing toxicities and likely toxici-ty-related therapeutic limitations

Interference of phosphatidylcholines with in-vitro cell proliferation — no flock without black sheep

According to early experiments with natural extracts, phosphatidylcholines (PCs) are widely considered essentially non-toxic. In addition to these physiological mixed-chain PCs, many different synthetic diacyl-PCs are currently available, but they have never been systematically evaluated for any interference with cell proliferation. We thus investigated the cell proliferation of several cell lines in the presence of various liposomes consisting of a single PC component and cholesterol. Most of the PCs investigated did not interfere with cell proliferation, supporting the notion that most PCs are safe excipients. Significant IC50 values below 0.5mM were detected for PC(12:0/12:0), PC(14:1/14:1)trans and all diacyl-PCs containing two polyunsaturated fatty acids (PUFAs). The ω-3 PC(22:6/22:6) was the most toxic PC assessed, revealing IC50 values below 100 μM, but no rule concerning ω-3/6 configuration or acyl chain length could be observed. Physiological mixed-chain PCs containing PUFAs were much less toxic than respective non-physiological diacyl-PCs. All trans fatty acids in diacyl-PCs interfered more with proliferation than their respective cis-configured counterparts. Depending on the concentration, those diacyl-PCs not only inhibited proliferation but also induced cell death. Unlike the non-toxic PCs usually used for liposomal drug delivery, the elucidated diacyl-PCs may be worthy of further examination to eventually construct a toxic shell for toxic drugs, thereby enhancing anticancer drug delivery via lipid particles.