Ernest V. Groman

Biologically degradable superparamagnetic materials for use in clinical applications

This invention relates to materials exhibiting certain magnetic and biological properties which make them uniquely suitable for use as magnetic resonance imaging (MRI) agents to enhance MR images of animal organs and tissues. More particularly, the invention relates to the in vivo use of biologically degradable and metabolizable superparamagnetic metal oxides as MR contrast agents. Depending on their preparation, these metal oxides are in the form of superparamagnetic particle dispersoids or superparamagnetic fluids where the suspending medium is a physiologically-acceptable carrier, and may be uncoated or surrounded by a polymeric coating to which biological molecules can be attached. These materials are administered to animals, including humans, by a variety of routes and the metal oxides therein collect in specific target organs to be imaged; in the case of coated particles, the biological molecules can be chosen to target specific organs or tissues. The biodistribution of the metal oxides in target organs or tissues results in a more detailed image of such organs or tissues because the metal oxides, due to their superparamagnetic properties, exert profound effects on the hydrogen nuclei responsible for the MR image. In addition, the dispersoids and fluids are quite stable and, in the case of the fluids, can even be subjected to autoclaving without impairing their utility. Furthermore, the materials are biodegradable and, in the case of iron oxide compounds, can eventually be incorporated into the subjects hemoglobin, making them useful in treating anemia. Thus, the materials are well-suited for in vivo use.

A functionalized superparamagnetic iron oxide colloid as a receptor directed MR contrast agent

We have synthesized a surface functionalized superparamagnetic iron oxide colloid whose clearance from the vascular compartment was inhibited by asialofetuin but not fetuin. Unlike other particulate or colloidal magnetic resonance (MR) contrast agents, the agent of the current communication is not withdrawn from the vascular compartment by cells of the macrophage-monocyte phagocytic system, as indicated by its selective increase in hepatic relaxation rates. Because of this we refer to this colloid as a hepatic selective (HS) MR contrast agent. At 20 mumol Fe/kg the HS MR agent darkened MR images of liver. The HS MR agent exhibited no acute toxicity when injected into rats at 1800 mumol Fe/kg. Based on these observations, surface functionalized superparamagnetic iron oxide colloids may be the basis of MR contrast agents internalized by receptor mediated endocytosis generally, and by the asialoglycoprotein receptor in particular.

Larch arabinogalactan for hepatic drug delivery: isolation and characterization of a 9 kDa arabinogalactan fragment☆

Purified arabinogalactan [AG(37 kDa)] from Larix occidentalis is composed of repeating units of similar molecular weight and composition. A 9 kDa arabinogalactan [AG(9 kDa)] has been obtained in high yield from AG(37 kDa) either by autoclaving at 121 degrees C or by exposure to alkaline solution in the presence of sodium borohydride. The weight average molecular weight of AG(37 (kDa) was determined to be 37 and 38 kDa by intensity light scattering and sedimentation equilibrium, respectively. The weight average molecular weight of AG(9 kDa) was determined to be 9.1 and 9.5 kDa by intensity light scattering and sedimentation equilibrium, respectively. MALDI-TOF mass spectrometry yielded a molecular weight at the peak of the distribution of 8.3 kDa for AG(9 kDa). Both AG(37 kDa) and AG(9 kDa) exhibited narrow molecular-weight distributions (Mw/Mn approximately 1.2). AG(37 kDa) and AG(9 kDa) exhibit nearly identical 13C-NMR spectra, monosaccharide composition, and sugar linkages. It is proposed that AG(37 kDa) is composed of covalently bound subunits of AG(9 kDa). AG(37 kDa) and AG(9 kDa) bind isolated hepatocyte asialoglycoprotein receptor equally well. As a result AG(9 kDa) is a candidate for use in hepatocyte directed drug delivery and may be more desirable for such use than is AG(37 kDa).

Biodegradable superparamagnetic metal oxides as contrast agents for MR imaging

This invention relates to materials exhibiting certain magnetic and biological properties which make them uniquely suitable for use as magnetic resonance imaging (MRI) agents to enhance MR images of animal organs and tissues. More particularly, the invention relates to the in vivo use of biologically degradable and metabolizable superparamagnetic metal oxides as MR contrast agents. Depending on their preparation, these metal oxides are in the form of superparamagnetic particle dispersoids or superparamagnetic fluids where the suspending medium is a physiologically-acceptable carrier, and may be uncoated or surrounded by a polymeric coating to which biological molecules can be attached. These materials are administered to animals, including humans, by a variety of routes and the metal oxides therein collect in specific target organs to be imaged; in the case of coated particles, the biological molecules can be chosen to target specific organs or tissues. The biodistribution of the metal oxides in target organs or tissues results in a more detailed image of such organs or tissues because the metal oxides, due to their superparamagnetic properties, exert profound effects on the hydrogen nuclei responsible for the MR image. In addition, the dispersoids and fluids are quite stable and, in the case of the fluids, can even be subjected to autoclaving without impairing their utility. Furthermore, the materials are biodegradable and, in the case of iron oxide compounds, can eventually be incorporated into the subjects hemoglobin, making them useful in treating anemia. Thus, the materials are well-suited for in vivo use.

Use of USPIO-induced magnetic susceptibility artifacts to identity sentinel lymph nodes and lymphatic drainage patterns. I. dependence of artifact size with subcutaneous Combidex® dose in rats

Subcutaneously administered Combidex contrast agent produced characteristic magnetic susceptibility artifacts in gradient-echo (GE) images of rat brachial and axillary lymph nodes. These artifacts were useful in the rapid location and identification of normal sentinel lymph nodes. A linear dose response was observed with maximum artifact size in transverse images and was used noninvasively to study lymphatic drainage patterns.

Lymphatic mapping and sentinel node location with magnetite nanoparticles

Abstract Subcutaneously administered magnetite nanoparticles were used to locate sentinel lymph nodes in normal rats. Nanoparticles sequestered in brachial and axillary lymph nodes produced magnetic susceptibility artifacts in gradient recall echo magnetic resonance images. The artifact sizes enabled the determination of nanoparticle nodal uptake rates and lymphatic drainage patterns. These studies were confirmed by use of 59Fe labeled magnetite nanoparticles.

NEW MOLECULAR WEIGHT FORMS OF ARABINOGALACTAN FROM LARIX OCCIDENTALIS

Abstract Arabinogalactan fractions with narrow molecular weight distributions were obtained from a crude extract of Larix occidentalis by gel-filtration chromatography. Molecular weight distributions of the fractions were determined by a combination of gel filtration chromatography (GFC) with intensity (Rayleigh) light scattering. Two distinct distributions were observed, with the fractions in the lower distribution as small as 3 kDa. A sensitive immunoassay confirmed that the polysaccharide fractions were arabinogalactans. Compositional analysis revealed that the sub-9 kDa arabinogalactan fractions obtained by gel filtration of the crude extract have significantly lower galactose to arabinose ratios than the previously isolated sub-9 kDa arabinogalactans obtained from arabinogalactan (37 kDa) by chemical methods [J.H. Prescott et al., Carbohydr. Res. , 278 (1995) 113–128]. Arabinogalactan fractions were obtained from a crude extract of Larix Occidentalis . Two distinct molecular weight distributions were observed, with the fractions in the lower molecular weight distribution as small as 3 kDa. This is the first report of such a low molecular weight for Larch arabinogalactan. Fractions were identified as arabinogalactans with a sensitive immunoassay. The sub 9 kDa arabinogalactan fractions obtained from the crude extract have significantly lower galactose to arabinose ratios than the previously isolated sub 9 kDa arabinogalactans obtained from arabinogalactan (37 kDa) by chemical methods.

Cell Tracking Using Nanoparticles

Tracking cells in regenerative medicine is becoming increasingly important for basic cell therapy science, for cell delivery optimization and for accurate biodistribution studies. This report describes nanoparticles that utilize stable-isotope metal labels for multiple detection technologies in preclinical studies. Cells labeled with nanoparticles can be imaged by electron microscopy, fluorescence, and magnetic resonance. The nanoparticle-labeled cells can be quantified by neutron activation, thereby allowing, with the use of standard curves, the determination of the number of labeled cells in tissue samples from in vivo sources. This report describes the characteristics of these nanoparticles and methods for using these nanoparticles to label and track cells.

Development of an immunoassay for larch arabinogalactan and its use in the detection of larch arabinogalactan in rat blood

We describe a sensitive and convenient immunoassay for larch arabinogalactan and demonstrate its specificity for larch arabinogalactan. Anti-larch arabinogalactan antiserum is about 10(4) and 10(6) times more selective for detecting larch arabinogalactan than antiserum binds to branch terminal disaccharides consisting of the terminal beta-D-galactosyl residue and the penultimate branch (1-->6)-beta-D-galactosyl residue. It does not bind L-arabinose. The sensitivity of the assay for larch arabinogalactan is less than 0.1 microgram/mL. The application of the assay for measuring arabinogalactan pharmacokinetics in rat blood is illustrated.

Variability of Complement Response toward Preclinical and Clinical Nanocarriers in the General Population

Opsonization (coating) of nanoparticles with complement C3 component is an important mechanism that triggers immune clearance and downstream anaphylactic and proinflammatory responses. The variability of complement C3 binding to nanoparticles in the general population has not been studied. We examined complement C3 binding to dextran superparamagnetic iron oxide nanoparticles (superparamagnetic iron oxide nanoworms, SPIO NWs, 58 and 110 nm) and clinically approved nanoparticles (carboxymethyl dextran iron oxide ferumoxytol (Feraheme, 28 nm), highly PEGylated liposomal doxorubicin (LipoDox, 88 nm), and minimally PEGylated liposomal irinotecan (Onivyde, 120 nm)) in sera from healthy human individuals. SPIO NWs had the highest variation in C3 binding (n = 47) between subjects, with a 15-30 fold range in levels of C3. LipoDox (n = 12) and Feraheme (n = 18) had the lowest levels of variation between subjects (an approximately 1.5-fold range), whereas Onivyde (n = 18) had intermediate between-subject variation (2-fold range). There was no statistical difference between males and females and no correlation with age. There was a significant correlation in complement response between small and large SPIO NWs, which are similar structurally and chemically, but the correlations between SPIO NWs and other types of nanoparticles, and between LipoDox and Onivyde, were not significant. The calculated average number of C3 molecules bound per nanoparticle correlated with the hydrodynamic diameter but was decreased in LipoDox, likely due to the PEG coating. The conclusions of this study are (1) all nanoparticles show variability of C3 opsonization in the general population; (2) an individuals response toward one nanoparticle cannot be reliably predicted based on another nanoparticle; and (3) the average number of C3 molecules per nanoparticle depends on size and surface coating. These results provide new strategies to improve nanomedicine safety.