Fluorescent magnetic nanoprobes, methods of making, and methods of use

• US 8,722,017 B2
• Filed: 07/10/2009
• Issued: 05/13/2014
• Est. Priority Date: 07/10/2008
• Status: Active Grant

• Current Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Original Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Inventors: Fu, Aihua, Wang, Shan X., Gambhir, Sanjiv Sam
• Primary Examiner(s): Hartley, Michael G
• Assistant Examiner(s): Donohue, Sean R
• Application Number:
  US12/460,007
• Publication Number:
  US 20100008862A1
• Time in Patent Office: 1,768 Days
• Field of Search: None
• US Class Current: 424/9.32
• CPC Class Codes: A61K 49/0002 General or multifunctional contrast agents, e.g. chelated agents
• CPC Class Codes: A61K 49/0032 Methine dyes, e.g. cyanine dyes
• CPC Class Codes: A61K 49/0045 the fluorescent agent being a peptide or protein used for imaging or diagnosis in vivo
• CPC Class Codes: A61K 49/005 characterised by the carrier molecule carrying the fluorescent agent
• CPC Class Codes: A61K 49/0093 Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle
• CPC Class Codes: A61K 49/1833 having a (super)(para)magnetic core coated or functionalised with a small organic molecule oligomeric, polymeric, dendrimeric A61K49/1851
• CPC Class Codes: A61K 49/1848 the small organic molecule being a silane
• CPC Class Codes: A61K 49/1866 the nanoparticle having a (super)(para)magnetic core coated or functionalised with a peptide, e.g. protein, polyamino acid
• CPC Class Codes: A61K 51/1244 microparticles or nanoparticles, e.g. polymeric nanoparticles
• CPC Class Codes: A61K 9/5094 Microcapsules containing magnetic carrier material, e.g. ferrite for drug targeting
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: G01N 33/5434 using magnetic particle immunoreagent carriers which constitute new materials per se
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/587 Nanoparticles

Abstract

Embodiments of the present disclosure provide for nanoprobes, methods of imaging, methods of imaging a target, methods of making nanoprobes, and the like.

Claims

1. A nanoprobe, comprising:

a superparamagnetic iron oxide (SPIO) nanoparticle, wherein the SPIO nanoparticle has a silanization coating on the surface of the SPIO nanoparticle, wherein the silanization coating is about 5 nm to 1000 nm thick, does not include silicates or silica, and is made of cross-linked trimethoxyl silanes that are cross-linked with two or more different types of functional groups, such that the functional groups are on the surface of the coated SPIO nanoparticle, wherein the functional groups are selected from the group consisting of;

a mercapto group, an amino group, a PEG group, an alkyl group, a hydroxyl group, and a combination thereof; and

at least ten fluorescent compounds attached to an outside surface of the silanization coating, wherein the silanization coating provides a protective effect for the compounds.

2. The nanoprobe of claim 1, wherein the SPIO nanoparticle has a diameter of about 3 nm to 40 nm.

3. The nanoprobe of claim 1, wherein silanization coating is amorphous and controllable to about 10 nm to 500 nm thick.

4. The nanoprobe of claim 1, wherein the SPIO nanoparticle is a product of an oxidative decomposition of iron pentacarbonyl with a compound selected from the group consisting of:

trimethylamine oxide, oleic acid, hexadecane, and a combination thereof.

5. The nanoprobe of claim 1, wherein the silanization coating includes two or more different types of functional groups on the outside surface of the silanization coating, wherein the functional groups are selected from the group consisting of:

an amino group, a mercapto group, a carboxyl group, a phosphonate group, a hydroxyl group, and a combination thereof, wherein at least one type of fluorescent compound is attached to one type of functional group; and

at least one second type of compound attached to the other type of functional group, wherein the second type of compound is selected from;

a second type of fluorescent compound, a targeting moiety, a drug, a therapeutic agent, a radiological agent, a chemological agent, a small molecule drug, and a biological agent.

6. The nanoprobe of claim 1, wherein the fluorescent compounds are attached to the silanization coating via at least one type of functional group.

7. The nanoprobe of claim 1, wherein the silanization coating includes biotin.

8. The nanoprobe of claim 7, wherein a streptavidin-fluorescent compound is attached to the silanization coating via biotin.

9. The nanoprobe of claim 1, wherein a targeting moiety is attached to the silanization coating, wherein the targeting moiety has an affinity for a target.

10. The nanoprobe of claim 1, wherein the fluorescent compound is a fluorophore.

11. The nanoprobe of claim 1, wherein the iron oxide nanoparticle is composed of a phase selected from the group consisting of:

a maghemite ((γ

-Fe₂O₃) phase, a magnetite (Fe₃O₄) phase, and a combination of both phases.

12. The nanoprobe of claim 1, wherein silanization coating is controlled to about 20 nm to 500 nm thick.

13. The nanoprobe of claim 1, wherein the silanization coating does not change the magnetic properties of the original SPIO nanoparticle.

14. The nanoprobe of claim 1, wherein the nanoprobe includes about 20 to 500 fluorescent compounds attached to the silanization coating.

15. The nanoprobe of claim 1, wherein two or more types of fluorescent compounds are attached to the silanization coating.

16. The nanoprobe of claim 1, wherein the silanization coating includes amino functional groups and mercapto functional groups.

17. The nanoprobe of claim 16, wherein a portion of the amino functional groups are attached to fluorescent compounds, where each amino functional group of the portion is attached to a fluorescent compound;

and a portion of the mercapto functional groups are attached to targeting moieties, wherein each mercapto functional group of the portion is attached to a targeting moiety.

18. The nanoprobe of claim 17, wherein the targeting moiety is a RGD containing peptide.

20. The nanoprobe of claim 1, wherein the SPIO nanoparticles are precipitated with isopropanol prior to silanization.

19. A nanoprobe, comprising:

a superparamagnetic iron oxide (SPIO) nanoparticle, wherein the SPIO nanoparticle has a silanization coating on the surface of the SPIO nanoparticle, wherein the silanization coating is about 5 nm to 1000 nm thick, does not include silicates or silica, and includes two or more different functional groups on the outside surface of the silanization coating, wherein the functional groups are selected from the group consisting of;

an amino group, a mercapto group, a carboxyl group, a phosphonate group, a hydroxyl group, and a combination thereof; and

at least ten fluorescent compounds attached to the functional groups on the outside surface of the silanization coating, wherein each fluorescent compound is attached to the silanization coating via a functional group, and wherein the silanization coating provides a protective effect for the fluorescent compound.

21. The nanoprobe of claim 19, wherein the SPIO nanoparticles are precipitated with isopropanol prior to silanization.