Reversibly Masked Polymers

Inactive Publication Date: 2008-11-13
ARROWHEAD MADISON
View PDF44 Cites 170 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0013]In a preferred embodiment, a polynucleotide is attached to the polymer in the presence of an excess of polymer. The excess polymer may aid in formulation of the polynucleotide-polymer conjugate. The excess polymer may reduce aggregation of the conjugate during formulation of the conjugate. The polynucleotide-polymer conjugate may be separated from the excess polymer prior to administration of the conjugate to the cell or organism. Alternatively, the polynucleotide-polymer conjugate may be co-administered with the excess polymer to the cell or organism. The excess polymer may be the same as the polymer or it may be different, a helper or boost polymer.
[0014]In a preferred embodiment, the described membrane active amph

Problems solved by technology

Drugs used in antisense and gene therapies are relatively large hydrophilic polymers and are frequently highly negatively charged as well.
However, in vivo delivery of polynucleotides is complicated by toxicity, serum interactions, and poor targeting of transfection reagents that are effective in vitro.
Transfection reagents that work well in vitro, cationic polymers and lipids, typically destabilize cell membranes and form large particles.
These properties render transfection reagents ineffective or toxic in vivo.
Cationic charge results in interaction with serum components, which causes destabiliza

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Reversibly Masked Polymers
  • Reversibly Masked Polymers
  • Reversibly Masked Polymers

Examples

Experimental program
Comparison scheme
Effect test

Example

Example 1

Polynucleotide Synthesis and Assembly

[0168]The following synthetic RNA oligonucleotides (Dharmacon, Lafayette Colo.) were used. Sense strand RNAs contained a primary amine with a six carbon spacer at the 5′ to allow conjugation to the delivery vehicle. The 2′ACE protected RNA oligonucleotides were deprotected prior to the annealing step according to the manufacturer's instructions. The siRNAs had the following sequences:

apoB (Ensembl# ENSMUST00000037520);apoB-1 siRNAsenseSEQ ID 15′-NH4-GAAmUGmUGGGmUGGmCAAmCmUmUmUmA*G,,antisenseSEQ ID 25′-P-AmAAGUUGCCACCCACAUUCmA*G,;apoB-2 siRNAsenseSEQ ID 35′-NH4-GGAmCAmUGGGmUmUCCAAAmUmUAmC*G,,antisenseSEQ ID 45′-P-UmAAUUUGGAACCCAUGUCCmC*G,;ppara (GenBank# NM_011144);ppara-1 siRNA,senseSEQ ID 55′-NH4-mUmCAmCGGAGmCmUmCAmCAGAAmUmUmC*U-3′,,antisenseSEQ ID 65′-P-AmAUUCUGUGAGCUCCGUGAmC*U-3′,;ppara-2 siRNAsenseSEQ ID 75′-NH4-mUmCCCAAAGCmUCCmUmUmCAAAAmU*U-3′,,antisenseSEQ ID 85′-P-mUmUUUGAAGGAGCUUUGGGAmA*G-3′,;Control siRNA (GL-3 luciferase report...

Example

Example 2

Poly(Vinyl Ether) Random Copolymers

[0170]A. Synthesis of a vinyl ether monomer. 2-Vinyloxy Ethyl Phthalimide was prepared via reacting 2-chloroethyl vinyl ether (25 g, 0.24 mol) and potassium phthalimide (25 g, 0.135 mol) in 100° C. DMF(75 ml) using tetra n-butyl ammonium bromide (0.5 g) as the phase transfer catalyst. This solution was heated for 6 h and then crashed out in water and filtered. This solid was then recrystallized twice from methanol to give white crystals.

B. Amine+lower alkyl poly(vinyl ether) polymers. A series of copolymers was synthesized from vinylether monomers with varying alkyl to amine group ratios and with alkyl groups having from one to four carbons (FIG. 1, R and R′ may be the same or different). Membrane activity was dependent on the size (alkyl chain length) and ratio of hydrophobic monomers (Wakefield 2005). Propyl and butyl-derived polymers were found to be membrane lytic using model liposomes while methyl and ethyl containing polymers were no...

Example

Example 3

Masking Agents

[0175]A. Synthesis of 2-propionic-3-methylmaleic anhydride (carboxydimethylmaleic anhydride or CDM). To a suspension of sodium hydride (0.58 g, 25 mmol) in 50 ml anhydrous tetrahydrofuran was added triethyl-2-phosphonopropionate (7.1 g, 30 mmol). After evolution of hydrogen gas had stopped, dimethyl-2-oxoglutarate (3.5 g, 20 mmol) in 10 ml anhydrous tetrahydrofuran was added and stirred for 30 min. Water, 10 ml, was then added and the tetrahydrofuran was removed by rotary evaporation. The resulting solid and water mixture was extracted with 3×50 ml ethyl ether. The ether extractions were combined, dried with magnesium sulfate, and concentrated to a light yellow oil. The oil was purified by silica gel chromatography elution with 2:1 ether:hexane to yield 4 g (82% yield) of pure triester. The 2-propionic-3-methylmaleic anhydride was then formed by dissolving of this triester into 50 ml of a 50 / 50 mixture of water and ethanol containing 4.5 g (5 equivalents) of p...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Solubility (mass)aaaaaaaaaa
Structureaaaaaaaaaa
Electric chargeaaaaaaaaaa
Login to view more

Abstract

The present invention is directed to reversibly inactivation of membrane active polymers useful for cellular delivery of compounds. Described are polyconjugates systems that incorporate targeting, anti-opsonization, anti-aggregation, and transfection activities into small biocompatible in vivo delivery conjugates. The use of multiple reversible linkages connecting component parts provides for physiologically responsive activity modulation.

Description

BACKGROUND OF THE INVENTION[0001]The delivery of polynucleotide and other membrane impermeable compounds into living cells is highly restricted by the complex membrane systems of the cell. Drugs used in antisense and gene therapies are relatively large hydrophilic polymers and are frequently highly negatively charged as well. Both of these physical characteristics preclude their direct diffusion across the cell membrane. For this reason, the major barrier to polynucleotide delivery is the delivery of the polynucleotide to the cellular interior. Numerous transfection reagents have been developed to deliver polynucleotides to cells in vitro. However, in vivo delivery of polynucleotides is complicated by toxicity, serum interactions, and poor targeting of transfection reagents that are effective in vitro. Transfection reagents that work well in vitro, cationic polymers and lipids, typically destabilize cell membranes and form large particles. The cationic charge of transfection reagent...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C08G83/00
CPCA01K2217/058A01K2267/0362A61K47/48176A61K48/0041C12N15/87A61K47/58A61K47/60
Inventor ROZEMA, DAVID B.WAKEFIELD, DARREN H.WOLFF, JON A.HAGSTROM, JAMES E.
Owner ARROWHEAD MADISON
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap