METHODS TO REGULATE POLARIZATION AND ENHANCE FUNCTION OF CELLS

• US 20140056815A1
• Filed: 11/01/2013
• Published: 02/27/2014
• Est. Priority Date: 08/05/2005
• Status: Active Grant

• Current Assignee: Gholam A. Peyman
• Original Assignee: Gholam A. Peyman
• Inventors: Peyman, Gholam A.
• Granted Patent: US 9,962,558 B2
• Time in Patent Office: Days
• US Class Current: 424/9.2
• CPC Class Codes: A61F 9/0017 implantable in, or in contact with, the eye, e.g. ocular inserts
• CPC Class Codes: A61F 9/00727 Apparatus for retinal reattachment
• CPC Class Codes: A61K 31/7105 Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 41/0047 Sonopheresis, i.e. ultrasonically-enhanced transdermal delivery, electroporation of a pharmacologically active agent
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 48/0033 the non-active part being non-polymeric
• CPC Class Codes: A61K 49/00 Preparations for testing in vivo
• CPC Class Codes: A61K 49/0067 quantum dots, fluorescent nanocrystals
• CPC Class Codes: A61K 49/085 conjugated systems
• CPC Class Codes: A61K 9/0009 involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis microelectromechanical systems A61K9/0097
• CPC Class Codes: A61K 9/0048 Eye, e.g. artificial tears
• CPC Class Codes: A61K 9/1611 Inorganic compounds
• CPC Class Codes: A61K 9/5115 Inorganic compounds
• CPC Class Codes: A61N 1/36046 of the eye
• CPC Class Codes: A61N 1/37205 Microstimulators, e.g. implantable through a cannula
• CPC Class Codes: A61N 5/062 Photodynamic therapy, i.e. excitation of an agent
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery

Abstract

Minimally invasive delivery with intercellular and/or intracellular localization of nano- and micro-particle solar cells within and among excitable biological cells to controllably regulate membrane polarization and enhance function of such cells. The cells include retinal and other excitable cells.

Claims

1. A method for providing and/or monitoring therapy for a condition in a patient, the method comprising

administering to the patient, the patient having a pathology, genetic disease, symptom of a genetic disease, degenerative disease, or symptom of a degenerative disease, a complex comprising a quantum dot, an agent to target the complex to particular cells or cell types in the patient, and a biomolecule,

after accumulation at the target, providing the biomolecule to provide a therapeutic effect at the target, and thereafter performing at least one of (a) and (b):

(a) applying light to the quantum dots under conditions sufficient to controllably activate the quantum dot to controllably regulate plasma membrane polarization of target cells in the patient to result in repolarizing, hyperpolarizing, or hypopolarizing the target cells to regulate the target cell'"'"'s plasma membrane polarization,

(b) imaging, tracking, and/or monitoring the complex in the patient using a sensor, wherein the complex provides therapy for the underlying pathology and/or symptom of the genetic and/or degenerative disease in the patient.

2. The method of claim 1 where light is applied to controllably activate the quantum dots by controlling at least one of exposure time, exposure intensity, and exposure site.

3. The method of claim 1 where the biomolecule is directly or indirectly associated with, or covalently conjugated to, the quantum dots.

4. The method of claim 1 where the biomolecule comprises a nucleic acid selected from DNA, RNA, and synthetic congeners thereof.

5. The method of claim 4 where the nucleic acid is selected from plasmid DNA encoding a protein or an inhibitory RNA that controls at least one of protein, peptide, and nucleic acid production.

6. The method of claim 4 where the nucleic acid is selected from the group consisting of genomic DNA, linear DNA, plasmid DNA, cDNA, RNAi, siRNA, shRNA, mRNA, tRNA, rRNA, microRNA, and DNA/RNA, the nucleic acid being either naturally occurring or synthetic.

7. The method of claim 4 wherein the nucleic acid encodes for a therapeutic protein selected from the group consisting of an anti-cancer agent, a growth factor, a hypoglycemic agent, an anti-angiogenic agent, a bacterial antigen, a viral antigen, a tumor antigen, and a metabolic enzyme.

8. The method of claim 7 where the anti-cancer agent is selected from the group consisting of interleukin-2, interleukin-4, interleukin-7, interleukin-12, interleukin-15, interferon-α

, interferon-β

, interferon-γ

, colony stimulating factor, granulocyte-macrophage stimulating factor, anti-angiogenic agents, tumor suppressor genes, thymidine kinase, eNOS, iNOS, p53, p16, TNF-α

, Fas-ligand, mutated oncogenes, tumor antigens, viral antigens, and bacterial antigens.

9. The method of claim 6 where the plasmid DNA encodes for an RNAi molecule designed to inhibit protein(s) involved in tumor or hyperproliferative cell growth or maintenance.

10. The method of claim 6 where the plasmid DNA simultaneously encodes a therapeutic protein and at least one RNAi molecules.

11. The method of claim 4 where the nucleic acid comprises plasmid DNA and synthetic RNA.

12. The method of claim 1 where the complex is administered to an individual with a tumor, to reduce the tumor burden, ameliorate tumor effects, and/or treat the tumor.

13. The method of claim 1 where the therapy is curative, palliative, or remediative, and is either therapeutic or preventive.

14. The method of claim 1 where the complex is used in gene targeting or knockout of specific gene(s), and provides gene therapy.

15. The method of claim 1 where the complex contains a wild-type or non-mutated form of a gene or part of a gene, and is introduced into a cell or cells, with the wild-type or non-mutated form of the nucleic acid replacing a defective and/or mutated form of the nucleic acid.

16. The method of claim 14 where the gene therapy is somatic gene therapy or germ line gene therapy.

17. The method of claim 14 wherein the gene therapy comprises delivery of a single gene or multiple genes, where the multiple genes are provided with a single quantum dot complex, or provided in multiple quantum dot complexes.

18. The method of claim 1 further comprising an additional therapy selected from the group consisting of stem cell therapy, immunomodulators, anti-VEGF agents, anti-integrins, anti-inflammatory agents, antibiotics, anti-viral agents, anti-fungal agents, anti-proliferative agents, and/or anti-cancer agents.

19. The method of claim 1 where the condition is selected from the group of a retinal, CNS, and cardiovascular disease.

20. The method of claim 1 where the complex is targeted and/or directed to a specific region of the body selected from a specific organ, tissue type, and/or cell type, where the targeted location is the site of a disease or a location affected by a disease.

21. The method of claim 1 where the quantum dot contains or is provided with a coating to enhance or impart biocompatibility and/or cell selectivity using an antibody or a receptor that directs the complex to the target.

22. The method of claim 1 where when the condition is an ocular disease, the complex is injected intravitreally or introduced into the cornea, choroid, or retina, and the ocular disease is selected from the group consisting of retinitis pigmentosa, color blindness, wet age related macular degeneration, dry age related macular degeneration, diabetic retinopathy, corneal dystrophies, Meesman syndrome, Fuchs syndrome, granular and macular corneal dystrophies, keratoconous, Sejorgen'"'"'s syndrome, inherited glaucoma, retinohyaloidopathies, congenital cataract, Marfan syndrome, choridermia x-linked retinoschisis, and/or achromatopsia.

23. The method of claim 1 performed on a patient having a condition selected from the group consisting of

spinal muscular dystrophy,

muscular dystrophy,

diseases affecting myeloid cells,

chronic lymphocytic leukemia,

multiple myeloma,

malignant tumors,

melanomas,

carcinoma of various organs including breast, intestine, prostate, CNS

glioblastoma,

sarcoma,

cystic fibrosis and the biomolecule is a nucleic acid encoding Cystic Fibrosis Transmembrane

Conductance Regulator (CFTR),

hemophilia B and the biomolecule is a nucleic acid encoding Factor IX;

sickle cell disease

anemia and the biomolecule is a nucleic acid encoding erythropoietin;

vasospasm and the biomolecule is a nucleic acid encoding inducible nitric oxide synthase (iNOS);

diminished cell survival and the biomolecule is a nucleic acid encoding a heat shock protein;

restenosis of a blood vessel and the biomolecule is a nucleic acid encoding a heat shock protein;

diminished growth from a hair follicle and the biomolecule is a nucleic acid encoding a telomerase or an immunosuppressive protein;

an X-linked agammaglobulinemia and the biomolecule is a nucleic acid encoding a Bruton'"'"'s tyrosine kinase;

adenosine deaminase severe combined immunodeficiency (ADA SCID) and the biomolecule is a nucleic acid encoding an ADA;

spinal muscular atrophy and the biomolecule is a nucleic acid encoding SMN-1;

exudative age related macular degeneration (AMD) and the biomolecule is a nucleic acid encoding an anti-VEGF protein sFlt-1;

choriodemia and the biomolecule is a nucleic acid encoding Rab-Escort Protein (REP-1);

Leber'"'"'s congenital amaurosis (LCA) and the biomolecule is a nucleic acid encoding RPE-specific 65-kDA protein (RPE65);

retinitis pigmentosa and the biomolecule is a nucleic acid encoding MERTK;

Stargardt'"'"'s syndrome and the biomolecule is a nucleic acid encoding ABC4;

Usher'"'"'s syndrome (1B) and the biomolecule is a nucleic acid encoding MY07A;

advanced and/or metastatic pancreatic cancer and the biomolecule is a nucleic acid encoding two genes, somatostatin receptor subtype 2 (sst2) and deoxycitidine kinase uridylmonophosphate kinase (dck;

;

umk);

cirrhotic liver and the biomolecule is a nucleic acid encoding IGF-I;

type II diabetes and the biomolecule is a nucleic acid encoding GLP-1 or a GLP-1 analog, where the GLP-1 analog is selected from the group consisting of GLP-1(7-34), GLP-1(7-35), GLP-1(7-36), Val⁸-GLP-1(7-37), Gln⁹-GLP-1(7-37), D-Gln⁹-GLP-1(7-37), Thr¹⁶-Lys¹⁸-GLP-1(7-37), and Lys¹⁸-GLP-1(7-37);

or

characterized by dysregulation of the immune system and the biomolecule is a nucleic acid encoding human interferon α

2b (hINFα

).

24. The method of claim 1 where the biomolecule is a tumor suppressor gene or genes selected from the group consisting of p53 gene, APC gene, DPC-4/Smad4 gene, BRCA-1 gene, BRCA-2 gene, WT-1 gene, retinoblastoma gene, MMAC-1 gene, adenomatouspolyposis coil protein gene, deleted in colorectal cancer (DCC) gene, MMSC-2 gene, NF-1 gene, nasopharyngeal carcinoma tumor suppressor gene that maps at chromosome 3p21.3, MTS1 gene, CDK4 gene, NF-1 gene, NF-2 gene and VHL gene.

25. The method of claim 1 where the complex comprises a therapeutic gene selected from genes which encode enzymes, blood derivatives, hormones, lymphokines, interleukins, interferons, tumor necrosis factor, growth factors, neurotransmitters or their precursors or synthetic enzymes, trophic factors selected from BDNF, CNTF, NGF, IGF, GMF, α

FGF, β

FGF, NT3, NT5, and HARP/pleiotrophin, apolipoproteins selected from ApoAI, ApoAIV, ApoE, dystrophin or a minidystrophin, the CFTR protein associated with cystic fibrosis, intrabodies, tumor-suppressing genes selected from p53, Rb, Rap1A, DCC, k-rev, genes encoding coagulation factors selected from factors VII, VIII, IX, genes participating in DNA repair, suicide genes defined as genes whose products cause cell death, cytosine deaminase, pro-apoptic genes, prodrug converting genes defined as genes encoding enzymes that convert prodrugs to drugs, and anti-angiogenic genes or genes that promote angiogenesis.

26. The method of claim 1 where the biomolecule provides for gene silencing.

27. The method of claim 26 where the gene silencing biomolecule is an inhibitory RNA.

28. The method of claim 27 where the condition is myotonic dystrophy and the biomolecule is an RNAi molecule which knocks down DMPK with multiple CUG repeats;

the condition is Huntington'"'"'s disease and the biomolecule is a plasmid expressing shRNA that reduce the mutant Huntington gene (htt);

the condition is Alzheimer'"'"'s disease and the biomolecule is an RNAi which targets BACE-1;

the condition is cancer and the biomolecule siRNA or shRNAs which targets ras, c-myc and/or VEGFR-2;

the condition Parkinson'"'"'s disease and the biomolecule is an RNAi which targets α

-synuclein;

or

the condition is ischemic damage and the biomolecule is an RNAi which targets caspase-3 used in therapy of stroke to reduce ischemic damage.

29. The method of claim 1 where in (b) the complex is imaged by a method selected from the group consisting of photography, optical coherence tomography (OCT), magnetic resonance imaging (MRI), spectroscopy, microscopy of a body fluid or tissue, and combinations thereof.

30. The method of claim 29 where the complex comprises a targeting moiety and diagnosis of a patient condition is based on at least one of complex concentration or complex localization at a target site in the patient.

31. The method of claim 1 where the agent to target the complex is at least one binding moiety that interacts with at least one specific protein marker of a tumor and an antibody directed to a tumor marker and conjugated with a reporter molecule.

32. The method of claim 31 where the complex is administered systemically to the patient and the method provides diagnosis of the tumor.

33. The method of claim 32 where, following complex administration, the reporter is assayed to qualitatively and/or quantitatively assess the tumor marker.

34. The method of claim 31 where the quantum dot and the reporter molecule have different excitation and emission maxima, where activation is performed at the different maxima, and where determination of binding by the quantum dot and the anti-tumor marker antibody assess tumor presence in the patient.

35. The method of claim 31 where specificity of the complex for a tumor is increased by associating a plurality of tumor marker binding proteins to the complex.

36. The method of claim 1 where the complex further comprises a magnetic or paramagnetic nanoparticle facilitating introduction of the complex into a cell.

37. The method of claim 1 further comprising a magnetic and/or paramagnetic nanoparticle conjugated with the targeting agent.

38. The method of claim 1 where the targeting agent is selected from the group consisting of an antibody, a ligand for a receptor, and combinations thereof.

39. The method of claim 37 where the quantum dot ranges from about 3 nm to about 50 nm and the magnetic or paramagnetic nanoparticle ranges from about 70 nm to about 200 nm.

40. The method of claim 37 further comprising applying at least one of an alternating magnetic field or electromagnetic radiation resulting in a temperature increase in the magnetic or paramagnetic nanoparticles that sufficiently perturbs a membrane of a cell at a target site to result in facilitated entry of the complex via the perturbed membrane.

41. The method of claim 37 further comprising the quantum dot associated with a thermosensitive polymer that, upon application of an external energy source, results in a temperature increase in the quantum dot to about 39°

C. to about 43°

C. facilitating release of the biomolecule from the thermosensitive quantum dot.

42. The method of claim 1 where the complex comprises a quantum dot of a first size and a quantum dot of a second size different from that of the first size, at least one size of the quantum dot having enhanced susceptibility to an external energy source capable of increasing the temperature of the enhanced susceptibility quantum dot, the increase in temperature providing enhanced access to a cell.

43. The method of claim 1 further comprising implanting an apparatus providing an energy source in a patient to provide localized internal activation of the complex.

44. The method of claim 43 where the apparatus comprises an LED light source and a rechargeable battery, and the light source provides a light pulse activating the complex.

45. The method of claim 44 where the light source further comprises a fiber optic tube or a flexible silicone tube to direct the light to a desired area(s).

46. The method of claim 43 used in a patient with a condition affecting the central nervous system.

47. The method of claim 1 where the complex comprises at least a first and a second particle where the first and the second particles absorb energy at different wavelengths, and the first and second particles are activated by different wavelengths.

48. The method of claim 47 where selective activity of the complex is by selective wavelength exposure.

49. The method of claim 1 where the quantum dot is made from organic molecules selected from organic nanocrystal and/or crystalline forms of carbon.

50. The method of claim 49 where the crystalline form of carbon is selected from the group consisting of fullerene, graphene, carbon nanotube, and combinations thereof.

51. The method of claim 49 where the quantum dot is coated with at least one of organic molecule, biocompatible protein, peptide, phospholipid, polymer, and/or polyethylene glycol polymers, the coating increasing biocompatibility of the quantum dot.