FLUORESCENT SILICA-BASED NANOPARTICLES

• US 20130039848A1
• Filed: 07/02/2010
• Published: 02/14/2013
• Est. Priority Date: 07/02/2009
• Status: Active Grant

• Current Assignee: Cornell University, Sloan-Kettering Institute For Cancer Research (Memorial Sloan-Kettering Cancer Center)
• Original Assignee: Cornell University, Sloan-Kettering Institute For Cancer Research (Memorial Sloan-Kettering Cancer Center)
• Inventors: Wiesner, Ulrich, Burns, Andrew, Larson, Steven, Bradbury, Michelle, Penate Medina, Oula, Ow, Hoosweng, Lewis, Jason
• Granted Patent: US 9,625,456 B2
• Time in Patent Office: Days
• US Class Current: 424/1.37
• CPC Class Codes: A61K 49/0019 characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
• CPC Class Codes: A61K 49/0032 Methine dyes, e.g. cyanine dyes
• CPC Class Codes: A61K 49/0054 Macromolecular compounds, i.e. oligomers, polymers, dendrimers
• CPC Class Codes: A61K 49/0056 Peptides, proteins, polyamino acids
• 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 51/08 Peptides, e.g. proteins , carriers being peptides, polyamino acids, proteins
• CPC Class Codes: A61K 51/082 the peptide being a RGD-containing peptide
• CPC Class Codes: A61K 51/086 the peptide being alphaMSH, alpha melanocyte stimulating hormone
• CPC Class Codes: A61K 51/1244 microparticles or nanoparticles, e.g. polymeric nanoparticles
• CPC Class Codes: A61K 51/1255 Granulates, agglomerates, microspheres
• CPC Class Codes: A61K 9/5115 Inorganic compounds
• CPC Class Codes: A61K 9/5146 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, polyamines, polyanhydrides
• CPC Class Codes: A61K 9/5169 Proteins, e.g. albumin, gelatin
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 39/06 Free radical scavengers or antioxidants
• CPC Class Codes: A61P 7/02 Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 9/08 : Vasodilators for multiple indications
• 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 33/54346 : Nanoparticles
• CPC Class Codes: G01N 33/552 : Glass or silica
• CPC Class Codes: G01N 33/574 : for cancer
• CPC Class Codes: G01N 33/582 : with fluorescent label
• CPC Class Codes: G01N 33/587 : Nanoparticles
• CPC Class Codes: G01N 33/60 : involving radioactive labelled substances
• CPC Class Codes: Y10S 977/773 : Nanoparticle, i.e. structure having three dimensions of 100 nm or less
• CPC Class Codes: Y10S 977/927 : Diagnostic contrast agent

Abstract

The present invention provides a fluorescent silica-based nanoparticle that allows for precise detection, characterization, monitoring and treatment of a disease such as cancer The nanoparticle has a fluorescent compound positioned within the nanoparticle, and has greater brightness and fluorescent quantum yield than the free fluorescent compound To facilitate efficient urinary excretion of the nanoparticle, it may be coated with an organic polymer, such as polyethylene glycol) (PEG) The small size of the nanoparticle, the silica base and the organic polymer coating minimizes the toxicity of the nanoparticle when administered in vivo The nanoparticle may further be conjugated to a ligand capable of binding to a cellular component associated with the specific cell type, such as a tumor marker A therapeutic agent may be attached to the nanoparticle Radionuclides/radiometals or paramagnetic ions may be conjugated to the nanoparticle to permit the nanoparticle to be detectable by various imaging techniques.

Claims

1. A fluorescent silica-based nanoparticle comprising:

a silica-based core comprising a fluorescent compound positioned within the silica-based core;

a silica shell surrounding at least a portion of the core;

an organic polymer attached to the nanoparticle;

from about 1 to about 20 ligands attached to the nanoparticle; and

a contrast agent or a chelate attached to the nanoparticle.

2. The nanoparticle of claim 1, wherein the nanoparticle has a diameter between about 1 nm and about 25 nm.

3. The nanoparticle of claim 2, wherein the nanoparticle has a diameter between about 1 nm and about 8 nm.

4. The nanoparticle of claim 1, wherein the organic polymer is selected from the group consisting of poly(ethylene glycol) (PEG), polylactate, polylactic acids, sugars, lipids, polyglutamic acid (PGA), polyglycolic acid, poly(lactic-co-glycolic acid) (PLGA), polyvinyl acetate (PVA), and the combinations thereof.

5. The nanoparticle of claim 1, wherein the ligand is capable of binding to at least one cellular component.

6. The nanoparticle of claim 5, wherein the cellular component is a tumor marker.

7. The nanoparticle of claim 1, wherein the number of ligands attached to the nanoparticle ranges from about 1 to about 10.

8. The nanoparticle of claim 1, wherein the ligand is selected from the group consisting of peptide, protein, biopolymer, synthetic polymer, antigen, antibody, microorganism, virus, receptor, hapten, enzyme, hormone, chemical compound, pathogen, toxin, surface modifier, and combinations thereof.

9. The nanoparticle of claim 8, wherein the peptide is selected from the group consisting of tripeptide RGD, cyclic peptide cRGD, octreotate, EPPT1 and peptide analogs of alpha-MSH.

10. The nanoparticle of claim 1, wherein the contrast agent is a radionuclide.

11. The nanoparticle of claim 10, wherein the radionuclide is selected from the group consisting of ⁸⁹Zr, ⁶⁴Cu, ⁶⁸Ga, ⁸⁶Y, ¹²⁴I and ¹⁷⁷Lu.

12. The nanoparticle of claim 1, wherein the chelate is adapted to bind a radionuclide.

13. The nanoparticle of claim 1, wherein the chelate is selected from the group consisting of DFO, DOTA, TETA and DTPA.

14. The nanoparticle of claim 1, wherein the nanoparticle is detectable by PET, SPECT, CT, MRI, optical imaging, bioluminescence imaging, or combinations thereof.

15. The nanoparticle of claim 14, wherein the optical imaging is fluorescence imaging.

16. The nanoparticle of claim 1, wherein a therapeutic agent is attached to the nanoparticle.

17. The nanoparticle of claim 16, wherein the therapeutic agent is selected from the group consisting of antibiotics, antimicrobials, antiproliferatives, antineoplastics, antioxidants, endothelial cell growth factors, thrombin inhibitors, immunosuppressants, anti-platelet aggregation agents, collagen synthesis inhibitors, therapeutic antibodies, nitric oxide donors, antisense oligonucleotides, wound healing agents, therapeutic gene transfer constructs, extracellular matrix components, vasodialators, thrombolytics, anti-metabolites, growth factor agonists, antimitotics, statin, steroids, steroidal and non-steroidal anti-inflammatory agents, angiotensin converting enzyme (ACE) inhibitors, free radical scavengers, PPAR-gamma agonists, small interfering RNA (siRNA), microRNA, and anti-cancer chemotherapeutic agents.

18. The nanoparticle of claim 16, wherein the therapeutic agent is radio labeled.

19. The nanoparticle of claim 18, wherein the therapeutic agent is bound to radiofluorine ¹⁸F.

20. The nanoparticle of claim 16, wherein the therapeutic agent is a radionuclide.

21. The nanoparticle of claim 20, wherein the radionuclide is selected from the group consisting of ⁹⁰Y, ¹³¹I and ¹⁷⁷Lu.

22. The nanoparticle of claim 1, wherein blood residence half-time of the nanoparticle after administration of the nanoparticle to a subject ranges from about 2 hours to about 25 hours.

23. The nanoparticle of claim 22, wherein the blood residence half-time of the nanoparticle ranges from about 3 hours to about 15 hours.

24. The nanoparticle of claim 23, wherein the blood residence half-time of the nanoparticle ranges from about 4 hours to about 10 hours.

25. The nanoparticle of claim 1, wherein tumor residence half-time of the nanoparticle after administration of the nanoparticle to a subject ranges from about 5 hours to about 5 days.

26. The nanoparticle of claim 25, wherein the tumor residence half-time of the nanoparticle ranges from about 10 hours to about 4 days.

27. The nanoparticle of claim 26, wherein the tumor residence half-time of the nanoparticle ranges from about 15 hours to about 3.5 days.

28. The nanoparticle of claim 1, wherein the ratio of tumor residence half-time to blood residence half-time of the nanoparticle after administration of the nanoparticle to a subject ranges from about 2 to about 30.

29. The nanoparticle of claim 28, wherein the ratio of the tumor residence half-time to the blood residence half-time of the nanoparticle ranges from about 3 to about 20.

30. The nanoparticle of claim 29, wherein the ratio of the tumor residence half-time to the blood residence half-time of the nanoparticle ranges from about 4 to about 15.

31. The nanoparticle of claim 1, wherein renal clearance of the nanoparticle after administration of the nanoparticle to a subject ranges from about 10% ID (initial dose) to about 100% ID in about 24 hours.

32. The nanoparticle of claim 31, wherein the renal clearance of the nanoparticle ranges from about 30% ID to about 80% ID in about 24 hours.

33. The nanoparticle of claim 32, wherein the renal clearance of the nanoparticle ranges from about 40% ID to about 70% ID in about 24 hours.

34. The nanoparticle of claim 1, wherein blood residence half-time of the nanoparticle ranges from about 2 hours to about 25 hours, tumor residence half-time of the nanoparticle ranges from about 5 hours to about 5 days, and renal clearance of the nanoparticle ranges from about 30% ID to about 80% ID in about 24 hours after the nanoparticle is administered to a subject.

35. The nanoparticle of claim 1, wherein substantially no anemia, weight loss, agitation, increased respiration, GI disturbance, abnormal behavior, neurological dysfunction, abnormalities in hematology, abnormalities in clinical chemistries, drug-related lesions in organ pathology, mortality, or combinations thereof is observed in a subject in about 10 to about 14 days after the nanoparticles in the amount of about 100 times of the human dose equivalent are administered to the subject.

36. The nanoparticle of claim 1, wherein multivalency enhancement of the nanoparticle ranges from about 2 fold to about 4 fold.

37. A fluorescent silica-based nanoparticle comprising:

a silica-based core comprising a fluorescent compound positioned within the silica-based core;

a silica shell surrounding at least a portion of the core;

an organic polymer attached to the nanoparticle; and

a ligand attached to the nanoparticle,

wherein the nanoparticle has a diameter between about 1 nm and about 15 nm,

and after administration of the nanoparticle to a subject, blood residence half-time of the nanoparticle ranges from about 2 hours to about 25 hours,

tumor residence half-time of the nanoparticle ranges from about 5 hours to about 5 days, and

renal clearance of the nanoparticle ranges from about 30% ID to about 80% ID in about 24 hours.

38. The nanoparticle of claim 37, wherein the number of ligands attached to the nanoparticle ranges from about 1 to about 20.

39. The nanoparticle of claim 38, wherein the number of ligands attached to the nanoparticle ranges from about 1 to about 10.

40. The nanoparticle of claim 37, wherein the nanoparticle has a diameter between about 1 nm and about 8 nm.

41. The nanoparticle of claim 37, wherein the organic polymer is selected from the group consisting of poly(ethylene glycol) (PEG), polylactate, polylactic acids, sugars, lipids, polyglutamic acid (PGA), polyglycolic acid, poly(lactic-co-glycolic acid) (PLGA), Polyvinyl acetate (PVA), and the combinations thereof.

42. The nanoparticle of claim 37, wherein the ligand is capable of binding to at least one cellular component.

43. The nanoparticle of claim 42, wherein the cellular component is a tumor marker.

44. The nanoparticle of claim 37, wherein the ligand is selected from the group consisting of peptide, protein, biopolymer, synthetic polymer, antigen, antibody, microorganism, virus, receptor, hapten, enzyme, hormone, chemical compound, pathogen, toxin, surface modifier, and combinations thereof.

45. The nanoparticle of claim 44, wherein the peptide is selected from the group consisting of tripeptide RGD, cyclic peptide cRGD, octreotate, EPPT1 and peptide analogs of alpha-MSH.

46. The nanoparticle of claim 37, wherein a contrast agent is attached to the nanoparticle.

47. The nanoparticle of claim 46, wherein the contrast agent is a radionuclide.

48. The nanoparticle of claim 47, wherein the radionuclide is selected from the group consisting of ⁸⁹Zr, ⁶⁴Cu, ⁶⁸Ga, ⁸⁶Y, ¹²⁴I and ¹⁷⁷Lu.

49. The nanoparticle of claim 37, wherein a chelate is attached to the nanoparticle.

50. The nanoparticle of claim 49, wherein the chelate is adapted to bind a radionuclide.

51. The nanoparticle of claim 49, wherein the chelate is selected from the group consisting of DFO, DOTA, TETA and DTPA.

52. The nanoparticle of claim 37 or 46, wherein the nanoparticle is detectable by PET, SPECT, CT, MRI, optical imaging, bioluminescence imaging, or combinations thereof.

53. The nanoparticle of claim 52, wherein the optical imaging is fluorescence imaging.

54. The nanoparticle of claim 37, wherein a therapeutic agent is attached to the nanoparticle.

55. The nanoparticle of claim 54, wherein the therapeutic agent is selected from the group consisting of antibiotics, antimicrobials, antiproliferatives, antineoplastics, antioxidants, endothelial cell growth factors, thrombin inhibitors, immunosuppressants, anti-platelet aggregation agents, collagen synthesis inhibitors, therapeutic antibodies, nitric oxide donors, antisense oligonucleotides, wound healing agents, therapeutic gene transfer constructs, extracellular matrix components, vasodialators, thrombolytics, anti-metabolites, growth factor agonists, antimitotics, statin, steroids, steroidal and non-steroidal anti-inflammatory agents, angiotensin converting enzyme (ACE) inhibitors, free radical scavengers, PPAR-gamma agonists, small interfering RNA (siRNA), microRNA, and anti-cancer chemotherapeutic agents.

56. The nanoparticle of claim 54, wherein the therapeutic agent is radio labeled.

57. The nanoparticle of claim 56, wherein the therapeutic agent is bound to radiofluorine ¹⁸F.

58. The nanoparticle of claim 54, wherein the therapeutic agent is a radionuclide.

59. The nanoparticle of claim 58, wherein the radionuclide is selected from the group consisting of ⁹⁰Y, ¹³¹I and ¹⁷⁷Lu.

60. The nanoparticle of claim 37, wherein blood residence half-time of the nanoparticle after administration of the nanoparticle to a subject ranges from about 3 hours to about 15 hours.

61. The nanoparticle of claim 60, wherein the blood residence half-time of the nanoparticle ranges from about 4 hours to about 10 hours.

62. The nanoparticle of claim 37, wherein tumor residence half-time of the nanoparticle after administration of the nanoparticle to a subject ranges from about 10 hours to about 4 days.

63. The nanoparticle of claim 62, wherein the tumor residence half-time of the nanoparticle ranges from about 15 hours to about 3.5 days.

64. The nanoparticle of claim 37, wherein the ratio of tumor residence half-time to blood residence half-time of the nanoparticle after administration of the nanoparticle to a subject ranges from about 2 to about 30.

65. The nanoparticle of claim 64, wherein the ratio of the tumor residence half-time to the blood residence half-time of the nanoparticle ranges from about 3 to about 20.

66. The nanoparticle of claim 65, wherein the ratio of the tumor residence half-time to the blood residence half-time of the nanoparticle ranges from about 4 to about 15.

67. The nanoparticle of claim 37, wherein renal clearance of the nanoparticle ranges from about 40% ID to about 70% ID in about 24 hours after administration of the nanoparticle to a subject.

68. The nanoparticle of claim 37, wherein substantially no anemia, weight loss, agitation, increased respiration, GI disturbance, abnormal behavior, neurological dysfunction, abnormalities in hematology, abnormalities in clinical chemistries, drug-related lesions in organ pathology, mortality, or combinations thereof is observed in a subject in about 10 to about 14 days after the nanoparticles in the amount of about 100 times of the human dose equivalent are administered to the subject.

69. The nanoparticle of claim 37, wherein multivalency enhancement of the nanoparticle ranges from about 2 fold to about 4 fold.

70. A fluorescent silica-based nanoparticle comprising:

a silica-based core comprising a fluorescent compound positioned within the silica-based core;

a silica shell surrounding at least a portion of the core;

an organic polymer attached to the nanoparticle; and

a ligand attached to the nanoparticle,

wherein the nanoparticle has a diameter between about 1 nm and about 8 nm,

and after administration of the nanoparticle to a subject, the ratio of tumor residence half-time to blood residence half-time of the nanoparticle ranges from about 2 to about 30,

and renal clearance of the nanoparticle ranges from about 30% ID to about 80% ID in about 24 hours.

71. A method for detecting a component of a cell comprising the steps of:

contacting the cell with a fluorescent silica-based nanoparticle comprising;

a silica-based core comprising a fluorescent compound positioned within the silica-based core;

a silica shell surrounding at least a portion of the core;

an organic polymer attached to the nanoparticle;

from about 1 to about 20 ligands attached to the nanoparticle; and

a contrast agent or a chelate attached to the nanoparticle; and

monitoring the binding of the nanoparticle to the cell or a cellular component by at least one imaging technique.

72. The method of claim 71, wherein the imaging technique is selected from the group consisting of PET, SPECT, CT, MRI, optical imaging, bioluminescence imaging, and combinations thereof.

73. A method for targeting a tumor cell comprising administering to a cancer patient an effective amount of a fluorescent silica-based nanoparticle comprising:

a silica-based core comprising a fluorescent compound positioned within the silica-based core;

a silica shell surrounding at least a portion of the core;

an organic polymer attached to the nanoparticle;

a ligand attached to the nanoparticle and capable of binding a tumor marker; and

at least one therapeutic agent.

74. The method of claim 73, wherein the nanoparticle is administered orally, intravenously, nasally, subcutaneously, intramuscularly or transdermally.

75. The method of claim 73, wherein the nanoparticle is radiolabeled.