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Nucleic acid microparticles for pulmonary delivery

A nucleic acid and particle technology, used in microcapsules, capsule delivery, powder delivery, etc., to solve problems such as inability to interact

Inactive Publication Date: 2010-03-31
BAXTER INT INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although such microparticles can be taken up whole by certain target cells and / or other cells (e.g., macrophages) through endocytosis, these microparticles are insoluble at target sites with an aqueous environment, so nucleic acids in these microparticles cannot interact freely with such target cells

Method used

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  • Nucleic acid microparticles for pulmonary delivery
  • Nucleic acid microparticles for pulmonary delivery
  • Nucleic acid microparticles for pulmonary delivery

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0237] Example 1: Materials Used to Prepare Exemplary Microparticles

[0238] The following materials were used to prepare the exemplary microparticles of the present disclosure. Although specific nucleic acids and siRNAs are provided for illustrative examples, similar microparticles can be prepared using other nucleic acids and oligonucleotides.

[0239] All aqueous solutions were prepared using nuclease-free deionized water that was autoclaved and sterile filtered through a 0.2 micron filter.

[0240] Nucleic acid solutions were prepared in water at a concentration of approximately 15 mg / ml. Exemplary antisense oligonucleotides (anti-CD40, anti-CD80, anti-CD86) used in the methods described herein are commercially available HPLC purified lyophilized preparations. These oligonucleotides are phosphorothioated on the oligonucleotide backbone and are available from Integrated DNA Technologies, Inc. (Coralville, IA).

[0241] Various siRNA compositions were used in the micro...

Embodiment 2

[0247] Example 2: Using Ca 2+ Exemplary antisense oligonucleotide microparticles prepared as cations

[0248] The following examples provide two illustrative process methods for preparing the disclosed Ca-containing 2+ antisense oligonucleotide-based microparticles.

[0249] Preparation process 1: In this process, a series of 6 kinds of reaction mixtures were prepared, wherein each reaction mixture contained non-ionic polymer, salt solution and nucleic acid solution. Briefly, aliquots of nonionic polymer solution A were dispensed into containers such that two-thirds of each final reaction mixture was solution A. Saline solution (5M CaCl 2 stock solution, pH 5.5) and water were added to nonionic polymer aliquots such that the Ca concentrations in the final reaction mixture were 0.1M, 0.17M, 0.33M, 0.67M, 1M and 1.25M, respectively. Aliquots of the antisense nucleic acid solutions were prepared such that when aliquots of these nucleic acid solutions were added to the fin...

Embodiment 3

[0260] Embodiment 3: with Zn 2+ Exemplary antisense oligonucleotide microparticles prepared as cations

[0261] In this process, a series of 7 reaction mixtures were prepared, each of which contained a nonionic polymer solution, a salt solution and a nucleic acid solution. Briefly, aliquots of nonionic polymer solution A were dispensed into containers such that two-thirds of each final reaction mixture contained solution A. Aliquots of the antisense nucleic acid solutions were prepared such that when aliquots of these nucleic acid solutions were added to the final reaction mixtures, the concentration of antisense nucleic acid in each final reaction mixture would be 0.206 mM.

[0262] Use 4M ZnCl 2 Stock solution (pH 4), prepare aliquots of the saline solution by diluting with water such that when the aliquots are added to the reaction mixture, the starting Zn concentrations in the salt and nucleic acid mixtures will be 0.1 M, 0.33 M, respectively. M, 1M, 2M and 3M.

[...

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Abstract

The present disclosure is related to microparticle compositions, in which the microparticles are made of nucleic acids and non-polymeric cations, which are suitable for administration to moist or aqueous target locations (e.g., the lung tissue), where the substantially spherical nucleic acid microparticles release the nucleic acids through dissolution, allowing the released nucleic acids to freely interact with the target cells.

Description

[0001] This application claims priority to U.S. Provisional Patent Application No. 60 / 938,123, filed May 15, 2007, and U.S. Provisional Patent Application No. 60 / 912,320, filed April 17, 2007, the contents of which are at It is hereby incorporated by reference in its entirety. field of invention [0002] In general, the present disclosure relates to the preparation of nucleic acid microparticles. More specifically, the present disclosure relates to pulmonary delivery of nucleic acid-based spherical microparticles prepared using aqueous conditions without the use of polymerizable cations. Background technique [0003] Microparticles are solid or semisolid particles less than 1 mm in diameter, more preferably less than 100 microns, which can be formed from a variety of materials, including synthetic polymers, proteins and polysaccharides. [0004] Exemplary polymers for forming microspheres include U.S. Patent No. 5,213,812 to Ruiz, U.S. Patent No. 5,417,986 to Reid et al., U...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/12A61K9/16
CPCA61K9/5031A61K9/1688A61K38/28A61K9/0075A61K9/1611A61K9/50A61K9/1617
Inventor 拉里·R·布朗金伯利·A·吉利斯迈克尔·加洛
Owner BAXTER INT INC
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