Delivery of compounds with rehydrated blood cells

a technology of rehydrated blood cells and compounds, applied in the field of fixed-dried blood cells, can solve the problems of limited use of cryopreserved platelets and normally liquid-stored for therapeutic delivery, limited practical application of cryopreserved platelets, and limited application of preserved blood cells in general

Inactive Publication Date: 2005-12-08
THE UNIV OF NORTH CAROLINA AT CHAPEL HILL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the practical utility of platelets as therapeutic delivery vehicles has been limited because platelets must be freshly isolated, modified with the therapeutic, and then infused in a short time-span.
The utility of cryopreserved platelets and normally liquid-stored for therapeutic delivery is limited by storage lesion and, in the case of frozen platelets, the need to remove cryopreservatives.
Hence the practical application of cryopreserved platelets, as well as preserved blood cells in general, has been limited.

Method used

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  • Delivery of compounds with rehydrated blood cells
  • Delivery of compounds with rehydrated blood cells
  • Delivery of compounds with rehydrated blood cells

Examples

Experimental program
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examples 1-5

Attachment of Ribavirin to Platelets

[0067] These examples describe methods for the preparation and characterization of reconstituted platelets having ribavirin and rFVIIa coupled thereto.

example 1

Synthesis of Ribavirin-Polylysine Polymers

[0068] Ribavirin is chemically phosphorylated for ribavirin monophosphate (RMP) as detailed by Yoshikawa et al., Tetrahedron Lett. 50, 5065-5068 (1967)). Ribavirin monophosphate (RMP) is coupled to polylysine via a pH-sensitive phosphoramide linkage in accordance with the procedure of Di Stefano and Fiume (Trends in Glycosci. and Glycotech. 50, 461-472 (1997)) or a simplified procedure is based on the formation of an imidazole-ribavirin adduct (see Chu et al., Nuc. Acids. Res. 11, 6513-6529 (1983)). The conjugation chemistry for ribonucleosides polymer synthesis was tested with uracil rather than ribavirin. We do not anticipate that the difference in nitrogenous base structure between uracil and ribavirin will have a large effect on the synthesis.

[0069] Polylysine (=205 kDa for ˜1,400 residues lysine / molecule) was reacted with FITC (fluorescein isothiocyanate) and SANPAH (N-succinimidyl-6-[4′-azido-2′-nitrophenylamino] hexanoate) to respec...

example 2

Attachment of Ribavirin-Polylysine Polymers for Ribavirin-Loaded Lyophilized Platelets

[0073] The procedure for preparing lyophilized platelets involves four steps: Removal of platelets from excess plasma proteins, mild paraformaldehyde cross-linking to stabilize cellular structures, removal of platelets from unreacted cross-linker and lyophilization (Read et al, U.S. Pat. No. 5,651,966). After removal of the excess paraformaldehyde, the platelets are mixed with varying concentrations of the ribavirin polymer and exposed to visible light to activate the SANPAH moieties for the formation of covalent linkage with the platelet surface. The platelets are then lyophilized with standard procedures. Platelets are prepared with different amounts of ribavirin by varying the concentration of ribavirin-copolymer in the coupling step. The ability of the ribonucleoside copolymer to covalently couple to RL platelets was studied by incubating the cells with the delivery polymer in visible-spectrum...

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Abstract

Fixed-dried blood cells carrying an active agent are described, along with methods of making the same, methods of using the same, and compositions containing the same. The blood cells may be red blood cells or blood platelets.

Description

RELATED APPLICATIONS [0001] This application claims the benefit of U.S. Provisional Patent Application Ser. No. 60 / 533,059, filed Dec. 29, 2003, and U.S. Provisional Patent Application Ser. No. 60 / 471,005, filed filed May 16, 2003, the disclosures of which are incorporated by reference herein in their entirety.STATEMENT OF GOVERNMENT SUPPORT [0002] This invention was made with government support from the Department of Defense and the National Institutes of Health under Grant Numbers 1-P20-DE123474, 1-P60-DE 130789, and 1 R21 EB002863. The United States Government has certain rights to this invention.FIELD OF THE INVENTION [0003] The present invention concerns methods and compositions for delivering active agents to a subject in need thereof and more particularly to fixed-dried blood cells that carry an active agent and that are useful to deliver active agents to a site of interest. BACKGROUND OF THE INVENTION [0004] Platelets have been recognized for decades as potential tools for c...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A01N1/02A61K31/00A61K35/14A61K35/18A61K35/19A61K38/18A61K45/06A61K47/48C12N5/07C12N5/0786
CPCA01N1/02A01N1/0215A61K31/00A61K35/18A61K35/19A61K45/06A61K2300/00Y10S977/905A61K31/7088A61K38/02A61K47/6901A61P31/12A61P7/02A61P7/04
Inventor NICHOLS, TIMOTHY C.FISCHERREAD, MARJORIE S.
Owner THE UNIV OF NORTH CAROLINA AT CHAPEL HILL
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