Catalytic delivery nanosubstrates (CDNS) for highly efficient delivery of biomolecules

Patent metadata

Field	Value
Publication number	US-10335491-B2
Application number	US-201214234321-A
Country	US
Kind code	B2
Filing date	Jul 23, 2012
Priority date	Jul 21, 2011
Publication date	Jul 2, 2019
Grant date	Jul 2, 2019

Abstract

This invention relates, e.g., to a molecular delivery system comprising A. a substrate having a nanostructured surface region which comprises a plurality of nanostructures and, covalently attached to the substrate, multiple copies of a first member of a binding pair; and B. at least one vector nanoparticle which comprises, encapsulated therein, a molecule of interest, and on its surface, multiple copies of second member of the binding pair. Methods of using the molecular delivery system to deliver a molecule of interest to a cell are also described.

First claim

We claim:

1. A molecular delivery system, comprising:
a substrate having
a nanostructured surface region which comprises a plurality of nanostructures and, covalently attached to the nanostructured surface region, multiple copies of a first member of a binding pair; and
at least one vector nanoparticle which comprises
encapsulated therein, a molecule of interest, and,
on its surface, multiple copies of a second member of the binding pair,
wherein the plurality of nanostructures are nanofibers or nanowires made of silicon, titanium, aluminum, steel, or an organic oxide, or are organic polymers comprising at least one of polymethacrlate, polysaccharide or polylactide,
wherein the plurality of nanostructures comprise a length and diameter such that the length is greater than the diameter,
wherein the plurality of nanostructures are attached at a first end to the substrate, and
wherein the first member of the binding pair is covalently attached to an outer surface region of the plurality of nanostructures such that the at least one vector nanoparticle is attached to the outer surface region of the plurality of nanostructures when the second member of the binding pair and the first member of the binding pair are engaged, and
wherein each vector nanoparticle further comprises:
a plurality of structural components that are suitable to at least provide some mechanical structure to said vector nanoparticle;
a plurality of binding components, each having a plurality of binding regions adapted to bind to said plurality of structural components; and
a plurality of terminating components, each of which is adapted to bind to a binding region of one of said plurality of binding components,

wherein said plurality of structural components and said plurality of binding components self-assemble when brought into contact to form said vector nanoparticle,
wherein said plurality of terminating components act to occupy binding regions of said plurality of binding components to terminate further binding when said plurality of terminating components are present in a sufficient quantity relative to said plurality of binding regions of said plurality of binding components,
wherein the plurality of binding regions comprise β-cyclodextrin,
wherein each of said plurality of structural components comprises at least one binding element adapted to bind to the binding regions to form a first inclusion complex,
wherein the binding element comprises adamantine,
wherein the first inclusion complex is adamantine-β-cyclodextrin,
wherein each of the plurality of terminating components comprise a single terminating binding element that binds to remaining binding regions of one of said plurality of binding components by forming a second inclusion complex,
wherein said terminating binding element comprises adamantine, and
wherein said second inclusion complex is adamantine-β-cyclodextrin.

2. A method for delivering a molecule of interest into a cell, comprising
contacting the cell with a substrate having a nanostructured surface region which comprises a plurality of nanostructures, wherein multiple copies of a first member of a binding pair are covalently attached to the nanostructured surface region, so that the cell is associated with the nanostructured surface region; then
immobilizing on the nanostructured surface region with which the cell is associated at least one vector nanoparticle, wherein the vector nanoparticle encapsulates the molecule of interest and comprises, on its surface, multiple copies of a second member of the binding pair,
so that the vector nanoparticle is internalized by the cell and the molecule of interest is released from the vector particle and is delivered to the cell,

wherein the plurality of nanostructures are nanofibers or nanowires made of silicon, titanium, aluminum, steel, or an organic oxide, or are organic polymers comprising at least one of polymethacrlate, polysaccharide or polylactide,
wherein the plurality of nanostructures comprise a length and diameter such that the length is greater than the diameter,
wherein the plurality of nanostructures are attached at a first end to the substrate, and
wherein the first member of the binding pair is covalently attached to an outer surface region of the plurality of nanostructures such that the at least one vector nanoparticle is attached to the outer surface region of the plurality of nanostructures when the second member of the binding pair and the first member of the binding pair are engaged, and
wherein the vector nanoparticle further comprises:
a plurality of structural components that are suitable to at least provide some mechanical structure to said vector nanoparticle;
a plurality of binding components, each having a plurality of binding regions adapted to bind to said plurality of structural components; and
a plurality of terminating components, each of which is adapted to bind to a binding region of one of said plurality of binding components,
wherein said plurality of structural components and said plurality of binding components self-assemble when brought into contact to form said vector nanoparticle,
wherein said plurality of terminating components act to occupy binding regions of said plurality of binding components to terminate further binding when said plurality of terminating components are present in a sufficient quantity relative to said plurality of binding regions of said plurality of binding components,
wherein the plurality of binding regions comprise β-cyclodextrin,
wherein each of said plurality of structural components comprises at least one binding element adapted to bind to the binding regions to form a first inclusion complex,
wherein the binding element comprises adamantine,
wherein the first inclusion complex is adamantine-β-cyclodextrin,
wherein each of the plurality of terminating components comprise a single terminating binding element that binds to remaining binding regions of one of said plurality of binding components by forming a second inclusion complex,
wherein said terminating binding element comprises adamantine, and
wherein said second inclusion complex is adamantine-β-cyclodextrin.

3. The method of claim 2 , further comprising delivering more of the molecule of interest or delivering a second molecule of interest into the cell, wherein the method further comprises, after the first molecule of interest is delivered to the cell,
immobilizing on the nanostructured surface region with which the cell is associated at least one vector nanoparticle comprising multiple copies of the second member of the binding pair, wherein the vector nanoparticle encapsulates more of the first molecule or encapsulates a second molecule,
so that the vector nanoparticle is internalized by the cell and the further molecule of interest is released from the vector nanoparticle and is delivered to the cell.

4. A delivery system of claim 1 , wherein the plurality of nanostructures are silicon nanowires.

5. A delivery system of claim 1 , wherein the vector nanoparticle is a liposome, self-assembled nanoparticle based on amphiphilic polymer, an inorganic nanoparticle, polymer-based nanoparticles, or a sol-gel nanoparticle.

6. The molecular delivery system of claim 1 , wherein the plurality of terminating components each have a single binding element that binds to one of the binding regions.

7. A delivery system of claim 1 , wherein the first and second members of the binding pair are antibody-antigen; protein-substrate; protein-inhibitor; protein-protein; a pair of complementary oligonucleotides; or an inclusion complex.

8. A delivery system of claim 1 , wherein the molecule of interest is a nucleic acid, a protein, a polysaccharide, or a small mole

Assignees

• Tseng Hsian Rong
• Wang Hao
• Chen kuan ju
• Univ California
• Nat Institutes Of Health Nih

Inventors

• Tseng Hsian-Rong
• Wang Hao
• Chen Kuan-Ju

Classifications

• A61K48/0008

  characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition · CPC title

• A61K47/6925

  the form being a microcapsule, nanocapsule, microbubble or nanobubble · CPC title

• A61K47/6951

  using cyclodextrin (cyclodextrins used as simple excipients A61K47/40) · CPC title

• B82Y5/00

  Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery · CPC title

• A61K47/6957

  the form being a device or a kit, e.g. stents or microdevices · CPC title