Example compositions of liposomes with hydrophilic polymers on their surface, and containing relatively high concentrations of contrast-enhancing agents for computed tomography are provided. Example pharmaceutical compositions of such liposomes, when administered to a subject, provide for increased contrast of extended duration, as measured by computed tomography, in the bloodstream and other tissues of the subject. Also provided are example methods for making the liposomes containing high concentrations of contrast-enhancing agents, and example methods for using the compositions.

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Patent
   RE45195
Priority
Apr 21 2004
Filed
Feb 15 2013
Issued
Oct 14 2014
Expiry
Apr 21 2024
Inventors
Hoffman, E…
Assg.orig
Marval Pha…
Case Weste…
Cleveland …
Assg.curr
MARVAL PHA…
Entity
Small
Referenced by
0
References
47
Maint.:
EXPIRED
Example 6
In Vivo Images of He…
0. 23. A composition, comprising:
(a) liposomes having an average diameter of less than 150 nanometers, the liposomes comprising:
(i) 1,2-dipalmatoyl-sn-glycero-3-phosphocholine (DPPC);
(ii) N-carbamylmethoxypoly(ethylene glycol)-1,2-distearoyl-sn-glycerol-3-phosphoethanolamine (DSPE-MPEG2000); and
(iii) cholesterol, and
(b) an iodinated nonradioactive contrast enhancing agent, a fraction of the iodinated nonradioactive contrast enhancing agent being encapsulated by the liposomes and a lesser fraction of the iodinated nonradioactive contrast enhancing agent being unencapsulated.
11. A composition for enhancing contrast of one or more areas of a subject for x-ray imaging when administered to the subject, the composition comprising liposomes, the liposomes comprising:
at least one first lipid or phospholipid;
at least one second lipid or phospholipid which is derivatized with one or more polymers; and
at least one sterically bulky excipient capable of stabilizing the liposomes;
wherein the average diameter of the liposomes is less than 150 nanometers, and wherein the liposomes encapsulate at least one iodinated nonradioactive contrast enhancing agent, and wherein the liposomes are contained in a suspension medium, at least some of the iodinated nonradioactive contrast enhancing agent that has not been encapsulated by the liposomes having been removed from the suspension medium.
0. 26. A composition for obtaining x-ray images of a region of interest of a subject when administered to the subject, the composition comprising:
liposomes, a plurality of the liposomes having an internal cavity and a bilayer, the bilayer comprising: cholesterol, a phospholipid, and a phospholipid which is derivatized with a polymer chain; and
an iodinated nonradioactive contrast enhancing agent, at least some of the iodinated nonradioactive contrast enhancing agent being encapsulated in the internal cavity such that the liposomes have an iodine concentration of at least 30 mg I/mL of liposome composition, wherein iodinated nonradioactive contrast enhancing agent that has not been encapsulated has been removed from the composition,
wherein the average diameter of the liposomes is less than 150 nanometers.
1. A composition for enhancing contrast of one or more areas of a subject for x-ray imaging when administered to the subject, the composition comprising:
liposomes, the liposomes encapsulating one or more iodinated nonradioactive contrast-enhancing agents, and the liposomes comprising: cholesterol, at least one phospholipid, and at least one phospholipid which is derivatized with a polymer chain,
wherein the average diameter of the liposomes is less than 150 nanometers, wherein the liposomes encapsulate the one or more iodinated nonradioactive contrast enhancing agents such that the liposomes have an iodine concentration of at least 30 mg I/mL of liposome composition, and wherein the encapsulated one or more iodinated nonradioactive contrast enhancing agents represent at least 90% of the total iodine concentration of the composition.
2. The composition of claim 1, wherein the x-ray imaging is computed tomography.
3. The composition of claim 1, wherein the iodinated nonradioactive contrast-enhancing agents are selected from at least one of: iodinated ionic compounds, iodinated nonionic compounds, and mixtures thereof.
0. 4. The composition of claim 3, wherein a suspension of the liposomes has a concentration of at least 30 milligrams of iodine per milliliter of the suspension.
5. The composition of claim 1, wherein the average diameter of the liposomes is less than 120 nanometers.
6. The composition of claim 1, wherein the composition is capable of being administered to the bloodstream of the subject.
7. The composition of claim 6, wherein the composition provides an enhanced contrast that remains detectable at least 30 minutes after administration.
8. The composition of claim 6, wherein the composition provides an enhanced contrast of at least 50 Hounsfield units in at least part of at least one of a vasculature and an organ of the subject.
9. The composition of claim 1, wherein the liposomes are PEGylated liposomes.
10. The composition of claim 1, wherein the liposomes are targeted liposomes.
12. The composition of claim 11, wherein the at least one first lipid or phospholipid comprises 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC).
13. The composition of claim 11, wherein the at least one second lipid or phospholipid which is derivatized with one or more polymers comprises N-carbamylmethoxypoly(ethylene glycol)-1,2-distearoyl-sn-glycerol-3-phosphoethanolamine (DSPL-MPLG2000).
14. The composition of claim 11, wherein the at least one sterically bulky excipient is selected from at least one of: sterols, fatty alcohols, fatty acids, and mixtures thereof.
15. The composition of claim 11, wherein the at least one sterically bulky excipient is cholesterol.
16. The composition of claim 11, wherein the liposomes are not autoclaved.
0. 17. The composition of claim 11, wherein the liposomes are contained in a suspension medium, at least some of the iodinated nonradioactive contrast enhancing agent that has not been encapsulated by the liposomes having been removed from the suspension medium.
18. The composition of claim 11, wherein the at least one first lipid or phospholipid is present in the amount of from about 55 to about 75 mol %; the at least one second lipid or phospholipid which is derivatized with one or more polymers is present in the amount of from about 1 to about 20 mol %; and the at least one sterically bulky excipient is present in the amount of from about 25 to about 40 mol %.
0. 19. The composition of claim 18, wherein the at least one first lipid or phospholipid is hydrogenated soy phosphatidylcholine; the at least one second lipid or phospholipid which is derivatized with one or more polymers is N-carbamylmethoxypoly(ethylene glycol)-1,2-distearoyl-sn-glycerol-3-phosphoethanolamine (DSPL-MPLG2000); and the at least one sterically bulky excipient is cholesterol.
0. 20. The composition of claim 1, wherein the one or more iodinated nonradioactive contrast-enhancing agents comprise iodixanol and/or iohexol.
0. 21. The composition of claim 1, wherein the at least one phospholipid comprises 1,2-dipalmatoyl-sn-glycero-3-phosphocholine (DPPC).
0. 22. The composition of claim 1, wherein the at least one phospholipid which is derivatized with a polymer chain comprises N-carbamylmethoxypoly(ethylene glycol)-1,2-distearoyl-sn-glycerol-3-phosphoethanolamine (DSPE-MPEG2000).
0. 24. The composition of claim 23, wherein sufficient iodinated nonradioactive contrast enhancing agent is encapsulated by the liposomes such that the liposomes have an iodine concentration of over 30 mg I/mL of liposome composition.
0. 25. The composition of claim 23, with the proviso that the composition is exclusive of Soy PC (HSPC) and pure DSPC (1,2-distearoyl-sn-glycero-3-phosphocholine).
0. 27. The composition of claim 26, wherein the iodinated nonradioactive contrast-enhancing agent comprises at least one of iodixanol and iohexol.
0. 28. The composition of claim 26, wherein the phospholipid comprises 1,2-dipalmatoyl-sn-glycero-3-phosphocholine (DPPC).
0. 29. The composition of claim 26, wherein the phospholipid which is derivatized with a polymer chain comprises N-carbamylmethoxypoly(ethylene glycol)-1,2-distearoyl-sn-glycerol-3-phosphoethanolamine (DSPE-MPEG2000).
, and the pulmonary artery 515 and liver cortex 3.5 hours post contrast injection attenuation was 200 HU (enhancement 130 HU), and in. In the kidney cortex 525 and the back muscle 510, the attenuation was 75 HU (enhancement 25 HU). Attenuation in the blood pool rose rapidly post-injection, and remained virtually constant for the 3.5 hours of the study. A slight increase in attenuation in the liver parenchyma 520 was observed. A transient increase in the kidney core medulla 530 was observed, indicating early clearance with little to no clearance later in the study. The small region of interest placed over the left main coronary artery indicated attenuation of 9 HU at base line and peaked at a value of 118 HU. FIG. 6 shows 0 hour baseline 605 and peak enhanced 610 images obtained 2 hours 18 minutes post liposomal injection at the level of the liver. FIG. 6 also shows 0 hour baseline 615 and peak enhanced 620 images obtained 2 hours 18 minutes post liposomal injection at the level of the mid-heart.

These data indicate the residence time of example PEGylated liposome formulations, which provided contrast enhancement, to be more than 3 hours. Additionally, the data show that contrast enhancement in muscle can be low, indicating the liposomal iohexol can be retained in the blood vessels and does not rapidly extravasate. Additionally, the contrast enhancement in the liver parenchyma indicated that clearance of the composition may substantially be due to the liver, and not the kidneys.

Example 6
In Vivo Images of Heart after Administration of PEGylated Liposomes Containing Iohexol

Additionally, example images of the rabbit heart were analyzed 700 (FIG. 7), 800 (FIG. 8), 900 (FIG. 9) and 1000 (FIG. 10). FIG. 7 shows volume rendered images of the whole rabbit, before 705 and 2 hours 18 minutes after injection of the liposomal iohexol formulation 710. Enhancement to the vasculature 715 due to the liposomes can be seen. The results show that, even more than 2 hours after injection, the blood vessels can be visible 715 while, using the same display and rendering parameters, they may not be visible before liposome administration. This enhancement can persist up until the time that the animal is euthanized at more than 3 hours after injection of the second dose of liposomes.

FIG. 8 shows volume images of the rabbit heart acquired pre-contrast 805 and at 20 minutes 810, 1 hour 15 minutes 815, 1 hour 51 minutes 820, 2 hour 38 minutes 825, and 3 hour 23 minutes 830 after administration of the liposomal iohexol formulation. All display and rendering parameters are identical for all images. The anatomies of all four heart chambers can be distinctly visualized along with the associated great vessels. Note that there may be absence of blood pool in the upper left panel 805 and the persistent enhanced opacity of the blood pool up to the final panel representing 3 hours 23 minutes post injection 830. Visible structures include: right ventricle 835 (RV); left ventricle 840 (LV); Aorta 845 (Ao); pulmonary artery 850 (PA); and the inferior vena cava 855 (IVC). These images demonstrated sustained contrast even 3 hours after administration of the liposomal iohexol.

FIG. 9 shows a thick-slab rendering of the heart obtained at ultrahigh resolution after the rabbit was euthanized and thus cardiac motion was eliminated. Labeled structures include the right ventricle 905 (RV); left ventricle 910 (LV); and aorta 915 (Ao).

FIG. 10 shows images of the left coronary artery of a rabbit under high magnification conditions at 3 hours after the second injection of the liposomal iohexol formulation. The left panel 1005 shows a 1.3 mm thick CT slice of in vivo rabbit heart imaged 3 hours 18 minutes after the second injection of one embodiment of liposomal iohexol. The right panel 1010 shows a volume rendered view of the same data set. The left coronary artery (shown as 1015 in 1010) was enhanced by 109 HU.

The above descriptions have referred to the preferred embodiments and selected alternate embodiments. Modifications and alterations will become apparent to persons skilled in the art upon reading and understanding the preceding detailed description. It is intended that the embodiments described herein be construed as including all such alterations and modifications insofar as they come within the scope of the appended claims or the equivalence thereof.

INVENTORS:

Hoffman, Eric, Bellamkonda, Ravi V., Annapragada, Ananth, Vijayalakshmi, Chandra, Kao, Chen-Yu

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ASSIGNMENT RECORDS    Assignment records on the USPTO
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Feb 15 2013Case Western Reserve University(assignment on the face of the patent)
Feb 15 2013Cleveland State University(assignment on the face of the patent)
Apr 09 2013MARVAL BIOSCIENCES, INC MARVAL PHARMA, INC CHANGE OF NAME SEE DOCUMENT FOR DETAILS 0325890269 pdf
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