Early diagnosis of lymph node involvement is essential to determine treatment for most types of cancer. Lymphadenectomy and histological evaluation is currently the gold standard, but is not ideal because it is invasive and can produce false negatives. Magnetic resonance (MR) imaging has become a powerful and non-invasive tool for detecting the spread of cancer to the lymph nodes. Standard MR imaging relies on size and morphology criteria to determine occult lymphoid tissues including lymph node metastasis, which has as low as 60% accuracy. MR contrast enhancing agents are becoming more widely used due to their usefulness in early tumor detection. Contrast enhancing agents diffuse into metastatic lymph nodes and healthy lymph nodes at different rates causing “filling defects.” Predicting lymph node metastasis using filling defects from contrast agents as opposed to size criteria can increase the sensitivity from 29% to 93%.
Generally, there are two types of MR contrast enhancing agents. Superparamagnetic contrast agents have a low r2/r1 ratio and create dark spots in T2- and T2*-weighted images. These are usually based on iron oxide particles and are referred to as negative contrast agents. On the other hand, paramagnetic contrast agents increase the r1 relaxivity and have a high r2/r1 ratio, creating bright spots in T1 weighted MR images. These contrast agents are known as positive contrast agents and are usually complexes of gadolinium (Gd3+).
T1 shortening comes about through dipole-dipole interactions with the protons in water so Gd3+, with its seven unpaired electrons, is the optimal choice for a T1 relaxation agent. Because Gd3+ is a heavy metal toxin, it is commonly delivered as a tightly bound linear or macroscopic chelate. Chelated forms of Gd3+ reduce toxicity by preventing cellular uptake of free Gd3+ and by limiting the clearance almost exclusively to renal filtration that resulted in renal toxicities. Despite reducing toxicity, the rapid clearance and small molecular size of gadolinium chelates mean that low levels of Gd3+ accumulates in the lymph nodes, making these agents a poor choice for MR lymphography. In addition, the FDA posted a warning about the risk of serious nephrogenic systemic fibrosis for all commercially available gadolinium contrast agent to identify well-perfused tissues and organs in subjects with acute or chronic renal insufficiency.
Liposomes and lipid nanoparticles containing Gd3+ have several advantages for MR contrast imaging of lymph nodes. Liposomes as well as lipid nanoparticles can lower the toxicity by encapsulating or binding to their surfaces a large amount of Gd3+. However, the rapid clearance mechanism of intravenously (IV) administered liposomes does not significantly improve liposome-associated Gd3+ accumulation in the lymph nodes. Only a fraction of the lipid nanoparticles in blood are phagocytosed by reticuloendothelial cells, and only a fraction of those cells traverse to lymphatic system. Thus, IV administered liposomes provide indirect targeting of the lymphatic system and lymph nodes. However, this approach results in a majority of Gd3+ carrying liposomes eliminated through reticuloendothelial cells in blood. The pharmacokinetics of liposomes in blood can be optimized for lymph node accumulation through size and surface modification. Reducing the diameter of the liposomes below 200 nm decreases phagocytic dependent clearance and increases the circulation time in the blood. Adding polyethylene glycol (PEG, commonly referred to as PEGylation) to the liposome surface can also increase the circulation time and stability of liposomes.
Currently, gadolinium chelated with diethylenentriaminepentaacetyl (DTPA) provides contrast in magnetic resonance imaging to identify pathogenic tissues. Unfortunately, the soluble Gd-DTPA complexes approved for clinical use, such as OMNISCAN, are cleared within a few minutes and do not provide sufficient concentrations or time in lymphoid tissues. In addition, the residual fraction of gadolinium can lead to fibrosis in patients with renal insufficiency.
Despite the advances in the development of contrast agents, a need exists for improved contrast agents having longer in vivo life, provide sufficient concentration in tissues to be analyzed, and low residual gadolinium concentrations to avoid side effects. The present invention seeks to fulfill this need and provide further related advantages.
The present inventions provides compositions expressing metal ions and methods for using the compositions.
In one aspect, the invention provides a composition, comprising a lipid, a polyalkylene-containing lipid, and a lipid-containing metal chelator.
Representative lipids include phospholipids, sphingolipids, cholesterol and steroid derivatives, bile acids and derivatives, cardilipin, acyl-glycerides and derivatives, glycolipids, acyl-peptides, fatty acids, carbohydrate-based polymers, functionalized silica, polyanhydride polymers, polylactate-glycolate polymers, and bioploymers. In one embodiment, the lipid is a phospholipid. Representative phospholipids include 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC); dipalmitoyl phosphatidylcholine;
dimysristoyl phosphatidyl choline; dioleoyl phosphatidyl choline; trans-esterified phospholipids derived from eggs, soybean, flaxseed, and the like; and phosphatidylcholine substituted with phosphatidyl ethanolamine, phosphatidylglycerol, phosphatidyl serine, and phosphatidic acids.
Representative polyalkylene-containing lipids include polyoxyethylene-containing lipids or polyoxypropylene-containing lipids. In one embodiment, the polyalkylene-containing lipid is a phospholipid functionalized with polyethylene glycol such as N-(carbonyl-methoxypolyethyleneglycol-2000)-1,2-distearoyl-sn-glycero-3-phosphoethanol-amine (mPEG-2000-DSPE).
Representative lipid-containing metal chelators include 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-diethylenetriaminepentaacetyl (DSPE-DTPA), tetraazacyclododecane, tetraacety(gadodiamide)-PE, and lipid-functionalized-[N,N-bis[2-[bis(carboxymethyl)amino]-ethyl]-glycinato-(5″)]. In one embodiment, the lipid-containing metal chelator comprises a PEGylated lipid moiety. In one embodiment, the lipid-containing metal chelator is a PEGylated DTPA. In other embodiments, the lipid-containing metal chelator is a DSPE-BOPTA, a DSPE-DO3A, and a DSPE-DOTA.
In one embodiment, the composition further includes a targeting moiety. Representative targeting moieties include proteins, polypeptides, peptides, antibodies or fragments thereof, small molecules, sugars or polysaccharides or derivatives thereof, and nucleic acids.
In one embodiment, the composition of the invention has the form of a nanoparticle. In one embodiment, the nanoparticle has a diameter of from about 5 nm to about 2 μm. In one embodiment, the nanoparticle has a diameter of from about 50 nm to about 100 μm.
The composition of the invention can further include a metal ion. Suitable metal ions include paramagnetic metal ions and ions of radio-isotopes. Representative paramagnetic metal ions include Gd3+, Cu2+, Fe3+, Fe2+, and Mn2+ ions. Representative ions of radio-isotopes include 68Ga, 55Co, 86Y, 90Y, 177Lu, and 111In ions.
In another aspect, the invention provides administrable compositions including a carrier and a plurality of the nanoparticles of the invention. Suitable carriers include pharmaceutically acceptable carriers such as saline for injection or dextrose for injection.
In further aspect, the invention provides a method for imaging of tissues, comprising administering to a subject to be imaged an effective amount of the composition of the invention. Representative tissues that can be imaged by the method include lymphoid, cardiovascular, liver, kidney, brain, heart, muscle, and gastrointestinal tract tissues, and other tissues accessible by the lymphatic or vascular (blood) systems.
In another aspect, the invention provides a method for delivering a radio-cancer therapeutic agent to a cancer cell, comprising administering to a cancer cell an effective amount of the composition of the invention that includes anion of a radio-isotope.
The foregoing aspects and many of the attendant advantages of this invention will become more readily appreciated as the same become better understood by reference to the following detailed description, when taken in conjunction with the accompanying drawings.
DSPC): pre-dose (5A); 5 min (5B); 14 min (5C); and 24 hr (5D). Contrast enhancement is localized mainly in vasculature and vascularized tissues in lymph nodes at 5 min and 14 min. The arrow in 5C indicates the beginning of gadolinium elimination into the gastrointestinal tract through the biliary route. By 24 hr (5D) most of the contrast agent is eliminated and the residual fraction appears in the intestine.
The present inventions provides compositions expressing metal ions and methods for using the compositions. In one embodiment, the compositions are lipid nanoparticles that include paramagnetic metal ions and are useful for magnetic resonance imaging. In another embodiment, the compositions are lipid nanoparticles that include ions of radio-isotopes and are useful for delivery of radio-cancer therapeutic agents.
In one aspect, the invention provides compositions and methods for magnetic resonance imaging. The compositions and methods enhance gadolinium distribution and accumulation in lymphatics. The invention provides a gadolinium composition (referred to herein as “Gd-DTPA-lipid nanoparticle”) that is suitable for both intravenous and subcutaneous administration. Subcutaneous administration allows direct access to lymphatic system. The composition enhances T1 weighted MR signal in the lymph nodes as well as increases the residence time of the contrast agent in the lymphatics. Upon intravenous administration, the composition exhibits at least 100-fold enhancement over soluble Gd-DTPA as a vascular imaging agent and eliminates predominantly through biliary, rather than renal clearance. The composition was shown to significantly increase signal-to-noise ratio by more than 300-fold for MR visualization of lymph nodes in macaques.
The composition of the invention includes a lipid, a polyalkylene-containing lipid, and a lipid-containing metal chelator. In one embodiment, the composition further includes a chelated metal ion.
In one embodiment, the composition of the invention is a chelator- (or metal chelate-) expressing particle. As used herein, the term “expressing” refers to the particle presenting or having available the chelator or chelated metal for activity. As noted above, in one embodiment, the composition of the invention is a lipid nanoparticle having chelated gadolinium ion (Gd+3) (e.g., Gd-DTPA-lipid nanoparticle). In the lipid nanoparticle, chelated gadolinium ion is expressed.
The lipid nanoparticles of the invention are biocompatible and are readily administered. The nanoparticles have a diameter of from about 5 nm to about 2 μm. In one embodiment, the nanoparticles have a diameter of from about 10 nm to about 100 μm. In one embodiment, the nanoparticles have a diameter of about 70 nm.
As noted above, the composition of the invention (e.g., lipid nanoparticles) includes a lipid, a polyalkylene-containing lipid, and a lipid-containing metal chelator.
Lipids. The lipid component of the nanoparticles of the invention comprise the nanoparticle core.
Representative lipids useful in the compositions include phospholipids, sphingolipids, cholesterol and steroid derivatives, bile acids and derivatives, cardilipin, acyl-glycerides and derivatives, glycolipids, acyl-peptides, fatty acids, carbohydrate-based polymers (e.g., cellulose polymers), suitably functionalized silica, lipophilic polymers (e.g., polyanhydrides, polylactate-glycolate), and lipophilic bioploymers (e.g., proteins, sugar polymers).
In one embodiment, the lipid is disteroylamidomethylamine.
In one embodiment, the lipid is a phospholipid. Representative phospholipids include 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC); dipalmitoyl phosphatidylcholine; dimysristoyl phosphatidyl choline; dioleoyl phosphatidyl choline; trans-esterified phospholipids derived from eggs, soybean, flaxseed, and the like; and phosphatidylcholine substituted with phosphatidyl ethanolamine, phosphatidylglycerol, phosphatidyl serine, and phosphatidic acids. In one embodiment, the phospholipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC).
Polyalkylene-containing lipids. The polyalkylene-containing lipid component of the nanoparticles of the invention serve as surface hydrating agents.
Representative polyalkylene-containing lipids include polyoxyethylene-containing lipids and polyoxypropylene-containing lipids. In one embodiment, the polyalkylene-containing lipid is a phospholipid functionalized with polyethylene glycol (e.g., PEGylated phospholipid). Suitable PEGylated phospholipids include a polyethylene glycol having a number average molecular weight of from about 500 to about 20,000. In one embodiment, the PEGylated phospholipid is N-(carbonyl-methoxypolyethyleneglycol-2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (mPEG-2000-DSPE) (also referred to herein as “mPEG-DSPE” and “mPEG-PE”).
In addition to polyalkylene-containing lipid, in other embodiments, the surface hydrating agent is hydrophilic biomaterials such as a carbohydrate polymer, a polyamine, a polyvinyl pyrrolidone, a poly(aspartate), or a poly(L-amino acid).
Other useful surface hydrating agents include covalent conjugates of polyethoxyl, polymethylene glycol, or propylene glycol and a lipid or other hydrophobic moiety (e.g., long chain hydrocarbon).
The surface hydrating agent is preferably present from about 5 to about 50 mole percent of the composition (i.e., lipid, polyalkylene-containing lipid (surface hydrating agent), and lipid-containing metal chelator).
Lipid-containing metal chelator. The lipid-containing metal chelator component of the nanoparticles of the invention are expressed on the surface of the nanoparticle and serve to chelate metal ions. Suitable lipid-containing metal chelators include two moieties: (1) a lipid moiety and (2) a metal chelator moiety.
Representative lipid-containing metal chelators include 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-diethylenetriaminepentaacetyl (DSPE-DTPA), tetraazacyclododecane, tetraacety(gadodiamide or OMNISCAN)-PE, and lipid-functionalized-[N,N-bis [24bis(carboxymethyl)amino]ethyl]-glycinato-(5″)] (MAGNEVIST).
Representative metal chelators include BOPTA, DO3A, and DOTA chelators.
In one embodiment, the metal chelator includes a PEGylated lipid moiety. Representative PEGylated metal chelators include DSPE-BOPTA, a DSPE-DO3A, and a DSPE-DOTA. In one embodiment, the metal chelator is a PEGylated DTPA (DPTA-PE).
The metal chelator is preferably present from about 5 to about 50 mole percent of the lipid, polyalkylene-containing lipid (surface hydrating agent), and metal chelator.
Chelated metal ion. The compositions of the invention are effective carriers of metal ions. In these embodiments, the composition (e.g., lipid nanoparticles) further includes a chelated metal ion.
For MR applications, useful metal ions include paramagnetic metal ions. Representative paramagnetic metal ions include Gd3+, Cu2+, Fe3+, Fe2+, and Mn2+ ions.
For other applications such as imaging and therapeutic ion delivery, useful metal ions include ions of radio-isotopes. Representative radio-isotopes include ions of 68Ga, 55CO, 86Y, 90Y, 177Lu, and 111In.
For embodiments that include chelated metal ions, the ratio of metal ion: metal chelator is 0.1-1.0:1.0 (less than or equal to 1:1).
Targeting agents. The compositions of the invention can be used to target specific tissues. In these embodiments, the composition (e.g., lipid nanoparticles) further includes a targeting moiety. Representative targeting moieties include proteins, polypeptide, and peptides; antibodies and derivatives (fragments); small molecules; sugars, polysaccharides, and derivatives; and nucleic acids, such as nucleotide polymers (e.g., aptamers), DNAs; and RNAs. Representative targeting moiety targets include cancer cells and virus infected cells.
Lipid nanoparticle formulations. The lipid nanoparticles of the invention can be formulated into compositions for administration. Suitable compositions for administration include a carrier and a plurality of the lipid nanoparticles. Representative carriers include pharmaceutically acceptable carriers, such as saline for injection or dextrose for injection.
The lipid nanoparticle of the invention is not a liposome and does not form liposomes when formulated.
Methods for tissue imaging. In another aspect, the invention provides methods for imaging tissues (e.g., occluded tissues). In one embodiment, the method includes administering to a subject to be imaged a diagnostically effective amount of a composition of the invention. The composition can be administered by a variety of techniques including subcutaneously and intravenously. The method is effective for imaging tissues such as lymphoid, cardiovascular, liver, kidney, brain, heart, muscle, and gastrointestinal tract tissues, and other tissues accessible by the lymphatic or vascular (blood) systems. The method is effective for imaging the tissues above to determine whether the tissues are occluded. For magnetic resonance imaging methods, the composition includes a paramagnetic metal ion (e.g., Gd3+).
In general, the effective amount is from about 0.001 to about 5 mmol metal/kg subject. In one embodiment, the effective amount is from about 0.005 to about 0.050 mmol metal/kg subject. In one embodiment, the effective amount is about 0.010 mmol metal/kg subject.
Methods for radio-cancer therapeutic agent delivery. In another aspect, the invention provides methods for delivering a radio-cancer therapeutic agent to a cancer cell. In one embodiment, the method includes administering to a subject in need thereof a therapeutically effective amount of a composition of the invention in which the chelated metal ion is a radio-isotope (e.g., 68Ga, 55Co, 86Y, 90Y, 177Lu, and 111In). The composition can be administered by a variety of techniques including subcutaneously and intravenously. The method is effective for delivery to tissues such as lymphoid, cardiovascular, liver, kidney, brain, heart, muscle, and gastrointestinal tract tissues, and other tissues accessible by the lymphatic or vascular (blood) systems.
The following is a description of the preparation, characterization, and imaging results for representative lipid nanoparticles of the invention.
Lipid nanoparticles were prepared composed of 10 mole percent of surface-bound DTPA. These lipid nanoparticles contained distearoyl-phosphatidylcholine and PEGylated lipid, mPEG-2000-DSPE. They were allowed to complex with Gd3+ (presented as Gd3+—Cl−) at varying Gd3+-to-DTPA-PE mole ratios. The presence of free Gd3+ in the admixture was determined by the ability of free Gd3+ to quench the fluorescence of calcein. With up to a Gd3+-to-DTPA-PE mole ratio of 4, no free Gd3+ could be detected by the calcein quenching assay. At a 6 or higher Gd3+-to-DTPA-PE mole ratio, free or unbound Gd3+ was detected (see
The contrast properties of the Gd3+-expressed lipid nanoparticles was determined by comparing the effects of the various Gd3+ formulations on the R1 (1/T1) relaxivity of Gd3+. Lipid nanoparticles composed of distearoyl-phosphatidylcholine (DSPC) with or without PEGylated lipid (mPEG-2000-DSPE, referred to herein as “mPEG-DSPE” or “mPEG-PE”) and fixed Gd3+-to-DTPA mole ratio at 1. The T1 and T2 measurements were collected with a 1.5 T MR scanner. A clinically-used Gd-DTPA preparation (OMNISCAN, commercially available from GE Healthcare, Princeton, N.J.) was included as a comparison. As shown in
Both of the nanoparticle formulations containing Gd3+ had higher R1 values than soluble Gd-DTPA. As expected, the DSPC and DSPC plus mPEG-2000-PE control formulations without Gd3+ showed no significant effect on relaxivity. The data indicates that the PEG-containing Gd-DTPA-lipid nanoparticles provide a much greater increase in R1 compared to the other formulations. Up to a 100-fold increase in R1 relaxivity was achieved when compared to the commercially available OMNISCAN. The Gd-DTPA nanoparticle formulation without surface PEG (Gd-DTPA-PE: DSPC) also showed higher R1 values than OMNISCAN, but much less than the PEG-containing Gd-DTPA-lipid nanoparticle formulation (Gd-DTPA-PE: mPEG-PE: DSPC) (see
Because positive contrast generated by Gd3+ in an MR image is dependant on a low R2/R1 ratio, changes of R2 values were also determined.
The use of the Gd-DTPA lipid nanoparticles containing mPEG-2000-PE (Gd-DTPA-PE: mPEG-PE: DSPC or “Gd-DTPA lipid nanoparticles”) for MR imaging studies in primates is described below.
The MR image enhancing property of the Gd-DTPA-lipid nanoparticles can be used to minimize the IV dose need to produce vascular image enhancement and also reduce renal burden. Administration of 0.01 mmole/kg Gd-DTPA nanoparticles (about ⅕ of current dose for human) in rats produced a high quality MR image with clearly discernable central and peripheral vasculature of rat within 5 min (
The PEGylated lipid nanoparticles of the invention having surface-bound gadolinium ion exhibited a great improvement over other preparations in contrast enhanced MR lymphography and vascular imaging. These lipid nanoparticles showed high degree of accumulation in the lymph nodes after subcutaneous injection. The contrast enhancement in lymphoid tissue begins within 20 minutes of injection and is maintained for 24 hours. When given intravenously this agent produced high quality images of vasculature in much higher sensitivity than the current agents. Intravenously administered lipid nanoparticles are cleared almost exclusively through biliary route and appeared to complete within 24 hr. Surface modification by adding mPEG in lipid nanoparticles increased the MR signal of Gd3+ through coordination of water molecules. This leads to a much higher R1 relaxivity and lymph node image enhancement. The lipid nanoparticle formulation may allow using a low dose to achieve a high signal-to-noise MR contrast ratio for increasing the metastatic nodal discrimination and allowing for a much wider time frame for imaging. The potentially lower dose and more favorable elimination route of Gd3+ needed for MR contrast could provide higher safety margin.
The formulations of the invention provide effective contrast at relatively low dose compared to currently available and approved contrast agents.
As noted above, the formulations of the invention offer advantages over currently available and approved contrast agents.
aDTPA is expressed on lipid nanoparticles composed of DSPC, mPEG-PE and DTPA-PE (8:1:0.9 mole ratio, as described in Example 1.
bThe nanoparticles were exposed to GdCl3 at indicated DTPA to Gd3+ mole ratio.
cThe diameter of DTPA expressed nanoparticles were measured by photon correlation spectroscopy and data expressed were mean ± SD of quadruplicate samples at indicated time points.
dThe presence of unbound or free Gd3+ was estimated with calcein fluorescence quenching assay.
The following examples are provided for the purpose of illustrating, not limiting, the invention.
Lipid nanoparticle preparation. 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC, Avanti Polar Lipids, Ala.), N-(carbonyl-methoxypolethyleneglycol-2000)-1,2-distearoyl-sn-glycero-3-phospoethanolamine (mPEG-2000-DSPE, Genzyme, Mass.), and 1,2-distearoyl-sn-glycero-3-phophoethanolamine-N-DTPA (DSPE-DTPA, Avanti Polar Lipids, AL) were combined in chloroform (DSPC: mPEG-DSPE: DSPE-DTPA) in a ratio of 8:2:1 and dried into a thin film under nitrogen and then under high vacuum overnight. The mPEG-DSPE containing PEG polymers of various molecular weights (or chain lengths) are also purchased from Genzyme, Mass. At this point phosphate buffered saline (PBS, pH 7.4) was added to the film and sonicated in a bath-type sonicator (Laboratory Supplies Company, New York). The vesicle diameter, as measured by dynamic light scattering using a Malvern Zetasizer 5000 photon correlation spectroscopy (Malvern Instruments, PA), was 50 nm. The nanoparticles in suspension were mixed with gadolinium (III) chloride hexahydrate (Aldrich, St Louis, Mo.) for 20 minutes at indicated mole ratio to form Gd-DTPA-lipid nanoparticles. To determine the unbound Gd3+, the nanoparticles were incubated with calcein (0.5 μM) (Sigma, St Louis, Mo.) in PBS, pH 7.4, and the fluorescence was measured at 485/535 nm using a Victor3V 1420 multilabel counter (PerkinElmer, Waltham, Mass.). Free ionic Gd3+ quenches calcein fluorescence in [Gd3+] dependent manner. To determine the final Gd concentration, elemental Gd mass was determined using the inductively coupled plasma atomic emission spectrometry. The particles along with control particles without Gd3+ were used in the studies described herein.
Relaxivity studies. Dilutions of Gd-DTPA-lipid nanoparticles were prepared with Gd3+ concentrations between 0-5 μmol/ml. For comparison several samples were prepared from commercial agents such as OMNISCAN (Gd-DTPA-BMA) with Gd3+ concentrations from 0-5 μmol/ml. The relaxation time T1 was measured using the standard spin-echo sequence on a 1.5T MR scanner with a volume head coil as RF receiver. For T1 measurements, TE was fixed to 9 ms and seven TR were 133, 200, 300, 500, 750, 1000 and 2000 ms, respectively. For T2 measurements, TR was fixed to 2000 ms and four TE were 15, 30, 45, and 60 ms, respectively. The imaging intensities were fitted to obtain the corresponding T1 and T2 values, which were plotted versus Gd3+ concentration.
Primate lymphatic MRI Study. In vivo imaging of the lymph nodes using Gd-DTPA-lipid nanoparticles for dynamic contrast enhanced (DCE) MRI was performed in a 1.5T MR scanner. The pigtailed macaque (M. Nemestrina) was anesthetized with inhaled isofluorane (1-2%) and closely monitored during the experiments. A pre-contrast image of the primate was recorded to determine proper lymph node location and fine-tune the imaging parameters. The animal was removed from the MR scanner and injected subcutaneously at four sites. Each injection site received 2, 5, 5, and 8.5 mL, respectively of 6.1 μmol/ml Gd-DTPA-lipid nanoparticles to allow probing of dose effects and contrast diffusion from injection sites. The total dose of Gd is estimated to be 24.4 μmol/kg for the primate studies. The images were recorded on a Signa 1.5T Scanner using a surface coil 12×12 inch. A standard spin-echo imaging sequence was used with TR=500 ms, TE=15 ms, slice thickness of 3 mm, 21 slices, slice gap=0.5 mm, FOV (field of view)=320×320 mm2, matrix size=512×512, which gives an in-plane resolution of 0.63×0.63 mm2 and a temporal resolution is 3.1 min.
Rat vascular MRI study. In vivo imaging of the rat using Gd-DTPA-lipid nanoparticles for dynamic contrast enhanced (DCE) MRI was performed in a 3.0T MR scanner. The rats (SD) was anesthetized with inhaled isofluorane (1-2%) and closely monitored during the experiments. A pre-contrast image of the rat was recorded to determine proper location, orientation and fine-tune the imaging parameters. The animal was removed from the MR scanner and injected with 400 μL of indicated Gd contrast media through femoral vein. The images were recorded on a Signa 1.5T Scanner using a surface coil 12×12 inch. A standard spin-echo imaging sequence was used with TR=500 ms, TE=15 ms, slice thickness of 3 mm, 21 slices, slice gap=0.5 mm, FOV (field of view)=320×320 mm2, matrix size=512×512, which gives an in-plane resolution of 0.63×0.63 mm2 and a temporal resolution is 3.1 min.
While the preferred embodiment of the invention has been illustrated and described, it will be appreciated that various changes can be made therein without departing from the spirit and scope of the invention.
This application is a continuation of International Application No. PCT/US2010/024324, filed Feb. 16, 2010, which application claims the benefit of U.S. Provisional Application No. 61/152,459, filed Feb. 13, 2009, and U.S. Provisional Application No. 61/162,989, filed Mar. 24, 2009. Each application is incorporated herein by reference in its entirety.
This invention was made with Government support under Contract No. AI 52663 awarded by the National Institutes for Health/National Institute of Allergy and Infectious Diseases. The Government has certain rights in the invention.
Number | Date | Country | |
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61152459 | Feb 2009 | US | |
61162989 | Mar 2009 | US |
Number | Date | Country | |
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Parent | PCT/US2010/024324 | Feb 2010 | US |
Child | 13208100 | US |