NANOPARTICLES FOR SUSTAINED OPHTHALMIC DRUG DELIVERY AND METHODS OF USE

Abstract

Disclosed is a compound having the Formula (I): X-[NHCHR.sup.1C(O)NHCHR.sup.2C(O)].sub.xY (I) or a pharmaceutically acceptable salt or tautomer thereof, wherein R.sup.1 is H or the side chain of a neutral amino acid; R.sup.2 is the side chain of a basic amino acid R.sup.3; x is inclusive; X is H or a residue of a therapeutic agent; Y is OH, or a residue of a therapeutic agent; R.sup.3 is: [Formula should be inserted here]; R.sup.5 is a residue of a therapeutic agent; and provided that when R.sup.2 is R.sup.3, X is H and Y is OH. Also disclosed is a method of treating an ocular disorder, comprising: (a) intravitreal administration to an eye of a subject in need thereof with an effective amount of a therapeutic nanoparticle composition, the therapeutic nanoparticle composition comprising (i) at least one population of nanostructures and (ii) at least one peptide attached to the at least one population of nanostructures. The nanostructures may be exposed to light in the eye thereby electrostimulating the eye and treating the ocular disorder. Also disclosed is a method of treating an ocular disorder, comprising contacting the eye of a subject in need thereof with an effective amount of a therapeutic nanoparticle composition, the therapeutic nanoparticle composition comprising (i) at least one population of nanostructures, (ii) a peptide attached to the at least at least one population of nanostructures, (iii) a therapeutic agent useful for the treatment of the ocular disorder attached to the at least one population of nanostructures or to the peptide; and (iv) optionally, a linkage between the at least one population of nanostructures or the peptide and the therapeutic agent.

##STR00001##

Claims

1-32. (canceled)

33. A method of treating an ocular disorder, comprising contacting the eye of a subject in need thereof with an effective amount of a therapeutic nanoparticle composition, the therapeutic nanoparticle composition comprising (i) at least one population of nanostructures, (ii) a peptide attached to the at least one population of nanostructures, and (iii) a therapeutic agent useful for the treatment of the ocular disorder attached to the at least one population of nanostructures or to the peptide.

34. The method of claim 33, wherein the therapeutic agent is selected from the group consisting of an antibody, a protein, a nucleic acid and a small organic molecule.

35. The method of claim 33, wherein the therapeutic agent is selected from the group consisting of an anti-inflammatory, an anti-infective, an anti-viral, a calcium channel blocker, a neuroprotective agent, a growth factor, a growth factor antagonist, an intraocular pressure lowering drug, and an antineoplastic drug.

36. The method of claim 33, wherein the ocular disorder is selected from the group consisting of glaucoma including Open Angle Glaucoma, Angle Closure Glaucoma, Aniridic Glaucoma, Congenital Glaucoma, Juvenile Glaucoma, Lens-Induced Glaucoma, Neovascular Glaucoma, Post-Traumatic Glaucoma, Steroid-Induced Glaucoma, Sturge-Weber Syndrome Glaucoma, and Uveitis-Induced Glaucoma, diabetic retinopathy, macular degeneration, choroidal neovascularization, vascular occlusion, vascular leak, retinal edema, bacterial conjunctivitis, fungal conjunctivitis, viral conjunctivitis, allergic conjunctivitis, uveitis, keratic precipitates, macular edema, inflammation response after intra-ocular lens implantation, uveitis syndromes, retinal vasculitis, sarcoidosis, Eales disease, acute retinal necrosis, Vogt Koyanaki Harada syndrome, ocular toxoplasmosis, radiation retinopathy, proliferative vitreoretinopathy, endophthalmitis, ocular glaucomas, optic neuritis, ischemic optic neuropathy, thyroid associated orbitopathy, orbital pseudotumor, pigment dispersion syndrome, scleritis, episcleritis choroidopathies, retinopathies, retinal vascular disease, retinal artery occlusions, retinal vein occlusions, retinopathy of prematurity, retinitis pigmentosa, familial exudative vitreoretinopathy (FEVR), idiopathic polypoidal choroidal vasculopathy, epiretinal macular membranes and cataracts, and keratoconjunctivitis sicca (KCS).

37. The method of claim 36, wherein the ocular disorder is macular edema, Neovascular Glaucoma, diabetic retinopathy, or choroidal neovascularization.

38. The method of claim 37, wherein the therapeutic agent is (i) Vascular Endothelial Growth Factor (VEGF) decoy, Pigment Derived Growth Factor (PDGF), Endostatin, Angiostatin, or Angiopoietin-1 or (ii) a nucleotide molecule coding for VEGF decoy, PDGF, Endostatin, Angiostatin, or Angiopoietin-1.

39. The method of claim 36, wherein the ocular disorder is macular degeneration.

40. The method of claim 39, wherein the therapeutic agent is (i) VEGF decoy, PDGF, Endostatin, Angiostatin, Angiopoietin-1, or ATP Binding Cassette Subfamily A Member 4 or (ii) a nucleotide molecule coding for VEGF decoy, PDGF, Endostatin, Angiostatin, Angiopoietin-1, ATP Binding Cassette Subfamily A Member 4, glutamate agonist, or glutamate antagonist.

41. The method of claim 36, wherein the ocular disorder is ischemic optic neuropathy.

42. The method of claim 41, wherein the therapeutic agent is (i) Allotopic NADH dehydrogenase Unit 4 or (ii) a nucleotide molecule coding for Allotopic NADH dehydrogenase Unit 4.

43. The method of claim 36, wherein the ocular disorder is a retinopathy.

44. The method of claim 43, wherein the therapeutic agent is (i) Glial Cell Derived Neurotropic Factor or Peripherin-2 or (ii) a nucleotide molecule coding for Glial Cell Derived Neurotropic Factor or Peripherin-2.

45. The method of claim 36, wherein the ocular disorder is retinitis pigmentosa.

46. The method of claim 45, wherein the therapeutic agent is (i) Retinal Pigment Specific 65 kDa protein or (ii) a nucleotide molecule coding for Retinal Pigment Specific 65 kDa protein or (iii) a source of electrical stimulation such as a quantum dot.

47. The method of claim 36, wherein the ocular disorder is a viral infection of the eye.

48. The method of claim 47, wherein the therapeutic agent is an antisense oligonucleotide that inhibits viral replication.

49. The method of claim 48, wherein the antisense oligonucleotide inhibits cytomegalovirus (CMV) replication.

50. The method of claim 33, wherein the peptide has Formula (I):
X[NHCHR.sup.1C(O)NHCHR.sup.2C(O)].sub.xY (I) or a pharmaceutically acceptable salt or tautomer thereof, wherein R.sup.1 is H or the side chain of a neutral amino acid; R.sup.2 is the side chain of a basic amino acid or R.sup.3; x is 1-5 inclusive; X is H or a residue of the therapeutic agent; Y is OH, or a residue of the therapeutic agent; R.sup.3 is: ##STR00074## R.sup.5 is a residue of the therapeutic agent; and provided that when R.sup.2 is R.sup.3, X is H and Y is OH.

51-91. (canceled)

Description

DETAILED DESCRIPTION OF THE INVENTION

[0071] As used herein the specification, a or an may mean one or more. As used herein in the claim(s), when used in conjunction with the word comprising, the words a or an may mean one or more than one.

[0072] The term targeted as used herein encompasses the use of antigen-antibody binding, ligand-receptor binding, and other chemical and/or biochemical binding interactions to direct the binding of a chemical species to a specific site.

[0073] As used herein, light means electromagnetic radiation, which includes but is not limited to infrared, visible, and ultraviolet radiation. The wavelength of the light may be in the range of 600-2000 nm. In one embodiment, the light has a wavelength of 700-1200 nm. In another embodiment, the light has a wavelength of 750-1100 nm.

[0074] As used herein, a core/shell nanoparticle is a nanoparticle having a discrete core section surrounded by one or more shell layers.

[0075] As used herein, nanoparticle means one or more nanoparticles. As used herein, core/shell nanoparticle means one or more core/shell nanoparticles. As used herein, shell means one or more shells.

[0076] As used herein, localized means substantially limited to a desired area with only minimal, if any, dissemination outside of such area.

[0077] The nanoparticles may be administered to an animal using standard methods. Animals that may be treated include, but are not limited to, humans, non-human primates, cows, horses, pigs, dogs, cats, sheep, goats, rabbits, rats, mice, birds, chickens or fish.

[0078] Nanometer is 10.sup.9 meter and is used interchangeably with the abbreviation nm.

[0079] A nanostructure has at least one region or characteristic dimension with a dimension of less than about 500 nm, and down to on the order of less than about 1 nm. The nanostructure may have any shape or morphology.

[0080] When referring to any numerical value, about means a value of 10% of the stated value (e.g. about 100 nm encompasses a range of sizes from 90 nm to 110 nm, inclusive).

[0081] As used herein, the term nanocrystal refers to a nanostructure that is substantially monocrystalline. The terms nanocrystal, nanodot, dot and quantum dot are understood by the ordinarily skilled artisan to represent like structures and are used herein interchangeably. The present invention also encompasses the use of polycrystalline or amorphous nanocrystals. As used herein, the term nanocrystal also encompasses luminescent nanocrystals. As used herein, the term luminescent nanocrystals means nanocrystals that emit light when excited by an external energy source (suitably light).

[0082] Typically, the region of characteristic dimension will be along the smallest axis of the structure. Nanocrystals can be substantially homogenous in material properties, or in certain embodiments, can be heterogeneous. The optical properties of nanocrystals can be determined by their particle size, chemical or surface composition. In one embodiment, the luminescent nanocrystal size ranges between about 1 nm and about 15 nm.

[0083] Nanostructures for use herein can be produced using any method known to those skilled in the art. Suitable methods and exemplary nanocrystals are disclosed in Published U.S. patent application No. 2008/0237540; U.S. Pat. No.7,374,807; U.S. patent application Ser. No. 10/796,832, filed Mar. 10, 2004; U.S. Pat. No. 6,949,206; and U.S. Provisional Patent Application No. 60/578,236, filed Jun. 8, 2004. The nanocrystals for use in the present invention can be produced from any suitable material, including an inorganic material, and more suitably an inorganic conductive or semiconductive material. Suitable materials include those disclosed in U.S. patent application Ser. No. 10/796,832, and include any type of semiconductor, including group II-VI, group III-V, group IV-VI and group IV semiconductors. Suitable semiconductor materials include, but are not limited to, Si, Ge, Sn, Se, Te, B, C (including diamond), P, BN, BP, BAs, AIN, AlP, AlAs, AlSb, GaN, GaP, GaAs, GaSb, InN, InP, InAs, InSb, ZnO, ZnS, ZnSe, ZnTe, CdS, CdSe, CdTe, HgS, HgSe, HgTe, BeS, BeSe, BeTe, MgS, MgSe, GeS, GeSe, GeTe, SnS, SnSe, SnTe, PbO, PbS, PbSe, PbTe, CuF, CuCl, CuBr, Cul, Si.sub.3N.sub.4, Ge.sub.3N.sub.4, Al.sub.2O.sub.3, (Al, Ga, In).sub.2(S, Se, Te).sub.3, Al.sub.2CO, and an appropriate combination of two or more such semiconductors.

[0084] In certain aspects, semiconductor nanocrystals may comprise a dopant from the group consisting of: a p-type dopant or an n-type dopant. The nanocrystals useful in the present invention can also comprise II-VI or III-V semiconductors. Examples of II-VI or III-V semiconductor nanocrystals include any combination of an element from Group II, such as Zn, Cd and Hg, with any element from Group VI, such as S, Se, Te, Po, of the Periodic Table; and any combination of an element from Group III, such as B, Al, Ga, In, and Tl, with any element from Group V, such as N, P, As, Sb and Bi, of the Periodic Table.

[0085] The nanocrystals, including luminescent nanocrystals, useful in the present invention can also further comprise ligands conjugated, cooperated, associated or attached to their surface as described throughout. Suitable ligands include any group known to those skilled in the art, including those disclosed in U.S. Pat. No. 7,374,807, U.S. Pat. No. 6,949,206 and U.S. Provisional Patent Application No. 60/578,236.

[0086] In one embodiment, the peptide of Formula II can be synthesized from a peptide containing the basic amino acid sidechain (imidazol-4-yl)methyl (his), the method comprising: [0087] i) reacting the peptide with methylacrylate in the presence of base; [0088] ii) removing a methyl group from the methyl acrylate substituent by treatment with a base to expose a carboxylic acid group; [0089] iii) coupling a therapeutic agent to the exposed carboxylic acid group with a coupling reagent, optionally in the presence of an additive.

[0090] Examples of base include, but are not limited to, 2,6-Di-tert-butylpyridine, N,N-diisopropylethylamine, 1,8-Diazabicyclo[5.4.0]undec-7-ene, sodium hydroxide, potassium hydroxide, and lithium hydroxide.

[0091] Examples of coupling reagents include, but are not limited to, dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), and (N-(3-Dimethylaminopropyl)-N-ethylcarbodiimide. HCl (EDAC).

[0092] Examples of additives include, but are not limited to, 1-Hydroxybenzotriazole (HOBt), hydroxy-3,4-dihydro-4-oxo-1,2,3-benzotriazine (HOOBt), N-hydroxysuccinimide (HOSu), 1-hydroxy-7-aza-1H-benzotriazole (HOAt), (4-(N,N-Dimethylamino)pyridine (DMAP)

[0093] In one embodiment, the peptide has Formula (I):

X[NHCHR.sup.1C(O)NHCHR.sup.2C(O)].sub.xY (I) [0094] or a pharmaceutically acceptable salt or tautomer thereof, wherein [0095] R.sup.1 is H or the side chain of a neutral amino acid; [0096] R.sup.2 is the side chain of a basic amino acid; [0097] x is 1-5 inclusive; [0098] X is H or a residue of the therapeutic agent; and

[0099] Y is OH, or a residue of the therapeutic agent.

[0100] Examples of side chains of neutral amino acids include methyl (ala), isopropyl (val), 2-methylpropyl (leu), and 1-methylpropyl (ile).

[0101] Examples of side chains of basic amino acids include 4-aminobutyl (lys), 4-guanidinobutyl (arg) and (imidazol-4-yl)methyl (his).

[0102] Particular examples of peptides that may be linked to a therapeutic agent to give a compound of Formula (I) include, but are not limited to, ala-his, ala-his-ala-his (SEQ ID NO: 1), ala-his-ala-his-ala-his (SEQ ID NO: 2), ala-his-ala-his-ala-his-ala-his (SEQ ID NO: 3), gly-his, gly-his-gly-his (SEQ ID NO: 4), gly-his-gly-his-gly-his (SEQ ID NO: 5), gly-his-gly-his-gly-his-gly-his (SEQ ID NO: 6), gly-his-gly-his-gly-his-gly-his-gly-his (SEQ ID NO: 7), val-his, val-his-val-his (SEQ ID NO: 8), val-his-val-his-val-his (SEQ ID NO: 9), val-his-val-his-val-his-val-his (SEQ ID NO: 10), ile-his, ile-his-ile-his (SEQ ID NO: 11), ile-his-ile-his-ile-his (SEQ ID NO: 12), and ile-his-ile-his-ile-his-ile-his (SEQ ID NO: 13).

[0103] Therapeutic agents that may be derivatized with a peptide include, without limitation, anti-inflammatories, anti-infectives, anti-virals, calcium channel blockers, neuroprotective agents, growth factors, growth factor antagonists, intraocular pressure lowering drugs, and antineoplastic drugs. Particular examples of therapeutic agents that are useful for the treatment of ocular disorders that may be derivatized with the peptide include acyclovir, betamethasone, bimatoprost, brinzolamide, carteolol, ciprofloxacin, dexamethasone, dorzolamide, epinastine, fluorometholone, fusidic acid, gentamicin, levobunolol, lodoxamide, moxifloxicin, nepaphenac, olopatadine, acetylcysteine, atropine, azithromycin, betaxolol, bromfenac, chloramphenicol, diclofenac, flurbiprofen, ganciclovir, homatropine, ketorolac, latanoprost, levofloxacin, loteprednol, nedocromil, ofloxacin, rimexolone, timolol, travoprost, tafluprost, tobramycin, tropicamide, cyclosporine, fexofenadine, terfenadine, cetirizine, levocetirizine, desloratadine, and hydroxyzine.

[0104] The derivitized therapeutic agents are exemplified by the following:

##STR00003## [0105] 1.

2-(2-aminopropanamido)-N-(9-((2-hydroxyethoxy)methyl)-6-oxo-6,9-dihydro-1H-purin-2-yl)-3-(1H-imidazol-4-yl)propanamide

[0106] ##STR00004## [0107] 2.

2-((8S,9R,10S,13S,14S,16S,17R)-9-fluoro-17-hydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-17-yl)-2-oxoethyl alanylhistidinate

[0108] ##STR00005## [0109] 3.

(1R,2R,3R,4S)-3-((Z)-7-(ethylamino)-7-oxohept-2-en-1-yl)-4-hydroxy-2-((S,E)-3-hydroxy-5-phenylpent-1-en-1-yl)cyclopentyl alanylhistidinate

[0110] ##STR00006## [0111] 4.

2-(2-aminopropanamido)-N-ethyl-3-(1H-imidazol-4-yl)-N-((R)-2-(3-methoxypropyl)-1,1-dioxido-6-sulfamoyl-3,4-dihydro-2H-thieno[3,2-e][1,2]thiazin-4-yl) propanamide

[0112] ##STR00007## [0113] 5.

1-(tert-butylamino)-3-((2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)oxy)propan-2-yl alanylhistidinate

[0114] ##STR00008## [0115] 6.

(1-cyclopropyl-6-fluoro-4-oxo-7-(piperazin-1-yl)-1,4-dihydroquinoline-3-carbonyl)alanylhistidine

[0116] ##STR00009## [0117] 7.

2-((8S,9R,10S,11S,13S,14S,16R,17R)-9-fluoro-11,17-dihydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-17-yl)-2-oxoethyl alanylhistidinate

[0118] ##STR00010## [0119] 8.

2-(2-aminopropanamido)-N-ethyl-3-(1H-imidazol-4-yl)-N-((4S,6S)-6-methyl-7,7-dioxido-2-sulfamoyl-5,6-dihydro-4H-thieno[2,3-b]thiopyran-4-yl)propanamide

[0120] ##STR00011## [0121] 9.

2-(2-aminopropanamido)-N-(9,13b-dihydro-1H-dibenzo[c,f]imidazo[1,5-a]azepin-3-yl)-3-(1H-imidazol-4-yl)propanamide

[0122] ##STR00012## [0123] 10.

(6S,8S,9R,10S,11S,13S,14S,17R)-17-acetyl-9-fluoro-17-hydroxy-6,10,13-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-11-yl alanylhistidinate

[0124] ##STR00013## [0125] 11.

((Z)-2-((3R,4S,5S,8S,9S,10S,11R,13R,14S,16S)-16-acetoxy-3,11-dihydroxy-4,8,10,14-tetramethylhexadecahydro-17H-cyclopenta[a]phenanthren-17-ylidene)-6-methylhept-5-enoyl)alanylhistidine

[0126] ##STR00014## [0127] 12.

N-((R)-1-((2S,5R,6R)-5-amino-6-(((1R,2S,3S,4R,6S)-4,6-diamino-3-(((2R,3R,4R,5R)-3,5-dihydroxy-5-methyl-4-(methylamino)tetrahydro-2H-pyran-2-yl)oxy)-2-hydroxycyclohexyl)oxy)tetrahydro-2H-pyran-2-yl)ethyl)-2-(2-aminopropanamido)-3-(1H-imidazol-4-yl)propanamide

[0128] ##STR00015## [0129] 13.

(R)-1-(tert-butylamino)-3-((5-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)oxy)propan-2-yl alanylhistidinate

[0130] ##STR00016## [0131] 14.

(2-((3-(carboxyformamido)-2-chloro-5-cyanophenyl)amino)-2-oxoacetyl)alanylhistidine

[0132] ##STR00017## [0133] 15.

(1-cyclopropyl-6-fluoro-8-methoxy-7-((4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl)-4-oxo-1,4-dihydroquinoline-3-carbonyl)alanylhistidine

[0134] ##STR00018## [0135] 16.

N-(2-(2-amino-2-oxoethyl)-6-benzoylphenyl)-2-(2-aminopropanamido)-3-(1H-imidazol-4-yl)propanamide

[0136] ##STR00019## [0137] 17.

(Z)-(2-(11-(3-(dimethylamino)propylidene)-6,11-dihydrodibenzo[b,e]oxepin-2-yl)acetyl)alanylhistidine

[0138] ##STR00020## [0139] 18.

acetyl-L-cysteinylalanylhistidine

[0140] ##STR00021## [0141] 19.

3-((1R,3s,5S)-8-methyl-8-azabicyclo[3.2.1]octan-3-yl)oxy)-3-oxo-2-phenylpropyl alanylhistidinate

[0142] ##STR00022## [0143] 20.

(2S,3S,4R)-6-(((2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-(((2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2-ethyl-3,4,10-trihydroxy-3,5,6,8,10,12,14-heptamethyl-15-oxo-1-oxa-6-azacyclopentadecan-13-yl)oxy)-4-methoxy-2,4-dimethyltetrahydro-2H-pyran-3-yl alanylhistidinate

[0144] ##STR00023## [0145] 21.

2-(2-aminopropanamido)-N-(3-(4-(2-(cyclopropylmethoxy)ethyl)phenoxy)-2-hydroxypropyl)-3-(1H-imidazol-4-yl)-N-isopropylpropanamide

[0146] ##STR00024## [0147] 22.

(2-(2-amino-3-(4-bromobenzoyl)phenyl)acetyl)alanylhistidine

[0148] ##STR00025## [0149] 23.

(2R,3R)-2-(2,2-dichloroacetamido)-3-hydroxy-3-(4-nitrophenyl)propyl alanylhistidinate

[0150] ##STR00026## [0151] 24.

(2-(3-((2,6-dichlorophenyl)amino)phenyl)acetyl)alanylhistidine

[0152] ##STR00027## [0153] 25.

(2-(2-fluoro-[1,1-biphenyl]-4-yl)propanoyl)alanylhistidine

[0154] ##STR00028## [0155] 26.

2-(2-aminopropanamido)-N-(9-(((1,3-dihydroxypropan-2-yl)oxy)methyl)-6-oxo-6,9-dihydro-1H-purin-2-yl)-3-(1H-imidazol-4-yl)propanamide

[0156] ##STR00029## [0157] 27.

2-(((1R,3s,5S)-8-methyl-8-azabicyclo[3.2.1]octan-3-yl)oxy)-2-oxo-1-phenylethyl alanylhistidinate

[0158] ##STR00030## [0159] 28.

(5-benzoyl-2,3-dihydro-1H-pyrrolizine-1-carbonyl)alanylhistidine

[0160] ##STR00031## [0161] 29.

isopropyl (Z)-7-((1R,2R,3R,5S)-3-((alanylhistidyl)oxy)-5-hydroxy-2-((R)-3-hydroxy-5-phenylpentyl)cyclopentyl)hept-5-enoate

[0162] ##STR00032## [0163] 30

((S)-9-fluoro-3-methyl-10-(4-methylpiperazin-1-yl)-7-oxo-2,3-dihydro-7H-[1,4]oxazino[2,3,4-ij]quinoline-6-carbonyl)alanylhistidine

[0164] ##STR00033## [0165] 31.

chloromethyl (8S,9S,10R,11S,13S,14S,17R)-11-((alanylhistidyl)oxy)-17-((ethoxycarbonyl)oxy)-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthrene-17-carboxylate

[0166] ##STR00034## [0167] 32.

2-((1-((1-carboxy-2-(1H-imidazol-4-yl)ethyl)amino)-1-oxopropan-2-yl)carbamoyl)-9-ethyl-4,6-dioxo-10-propyl-6,9-dihydro-4H-pyrano[3,2-g]quinoline-8-carboxylic acid

[0168] ##STR00035## [0169] 33.

(9-fluoro-3-methyl-10-(4-methylpiperazin-1-yl)-7-oxo-2,3-dihydro-7H-[1,4]oxazino[2,3,4-ij] quinoline-6-carbonyl)alanylhistidine

[0170] ##STR00036## [0171] 34.

(8S,9S,10R,11S,13S,14S,16R,17S)-10,13,16,17-tetramethyl-3-oxo-17-propionyl-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-11-yl alanylhistidinate

[0172] ##STR00037## [0173] 35.

(S)-1-(tert-butylamino)-3-((4-morpholino-1,2,5-thiadiazol-3-yl)oxy)propan-2-yl alanylhistidinate

[0174] ##STR00038## [0175] 36.

isopropyl (Z)-7-((1R,2R,3R,5 S)-5-((alanylhistidyl)oxy)-3-hydroxy-2-(( R,E)-3-hydroxy -4-(3-(trifluoromethyl)phenoxy)but-1-en-1-yl)cyclopentyl)hept-5-enoate

[0176] ##STR00039## [0177] 37.

isopropyl (Z)-7-((1R,2R,3R,5S)-3-((alanylhistidyl)oxy)-2-((E)-3,3-difluoro-4-phenoxybut-1-en-1-yl)-5-hydroxycyclopentyl)hept-5-enoate

[0178] ##STR00040## [0179] 38.

N-(((2R,3S,5R,6R)-5-amino-6-((1R,2S,3S,4R,6S)-4,6-diamino-3-(((2S,3R,4S,5S,6R)-4-amino-3,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-2-hydroxycyclohexyl)oxy)-3-hydroxytetrahydro-2H-pyran-2-yl)methyl)-2-(2-aminopropanamido)-3-(1H-imidazol-5-yl)propanamide

[0180] ##STR00041## [0181] 39.

3-(ethyl(pyridin-4-ylmethyl)amino)-3-oxo-2-phenylpropyl alanylhistidinate

[0182] ##STR00042## [0183] 40.

(1R,2R,E)-1-((2S,5S,11S,14S,17S,20S,23R,26S,29S,32S)-5-ethyl-11,17,26,29-tetraisobutyl-14,32-diisopropyl-1,7,10,16,20,23,25,28,31-nonamethyl-3,6,9,12,15,18,21,24,27,30,33-undecaoxo-1,4,7,10,13,16,19,22,25,28,31-undecaazacyclotritriacontan-2-yl)-2-methylhex-4-en-1-yl alanylhistidinate

[0184] ##STR00043## [0185] 41.

(2-(4-(1-hydroxy-4-(4-(hydroxydiphenylmethyl)piperidin-1-yl)butyl)phenyl)-2-methylpropanoyl)alanylhistidine

[0186] ##STR00044## [0187] 42.

1-(4-(tert-butyl)phenyl)-4-(4-(hydroxydiphenylmethyl)piperidin-1-yl)butyl alanylhistidinate

[0188] ##STR00045## [0189] 43.

2-(2-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-1-yl)ethoxy)ethyl alanylhistidinate

[0190] ##STR00046## [0191] 44.

2-amino-N-(1-(4-(8-chloro-5,6-dihydro-11H-benzo[5,6]cyclohepta[1,2-b]pyridin-11-ylidene)piperidin-1-yl)-3-(1H-imidazol-5-yl)-1-oxopropan-2-yl)propanamide

[0192] ##STR00047## [0193] 45.

N-alanyl-N-(3-(2-((8S,9R,10S,11S,13S,14S,16S,17R)-9-fluoro-11,17-dihydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-17-yl)-2-oxoethoxy)-3-oxopropyl)histidine

[0194] ##STR00048## [0195] 46.

Na-alanyl-Nt-(3-(((1R,2R,3R,4S)-3-((Z)-7-(ethylamino)-7-oxohept-2-en-1-yl)-4-hydroxy-2-((S,E)-3-hydroxy-5-phenylpent-1-en-1-yl)cyclopentyl)oxy)-3-oxopropyl)histidine

[0196] ##STR00049## [0197] 47.

Na-alanyl-Nt-(3-(2-((8S,9R,10S,11S,13S,14S,16R,17R)-9-fluoro-11,17-dihydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-17-yl)-2-oxoethoxy)-3-oxopropyl)histidine

[0198] ##STR00050## [0199] 48.

Na-alanyl-Nt-(3-oxo-3-(2-((6-oxo-1,6-dihydro-9H-purin-9-yl)methoxy)ethoxy)propyl)histidine

[0200] ##STR00051## [0201] 49.

Na-alanyl-Nt-(3-((1-(tert-butylamino)-3-((2-oxo-1,2,3,4-tetrahydroquinolin-5-yl)oxy)propan-2-yl)oxy)-3-oxopropyl)histidine

[0202] ##STR00052## [0203] 50.

Nt-(3-(((6S,8S,9R,10S,11S,13S,14S,17R)-17-acetyl-9-fluoro-17-hydroxy-6,10,13-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]lphenanthren-11-yl)oxy)-3-oxopropyl)-Na-alanylhistidine

[0204] ##STR00053## [0205] 51.

(Z)-2-((3R,4S,5S,8S,9S,10S,11R,13R,14S,16S)-16-acetoxy-3-((3-(alanyl-Nt-histidino)propanoyl)oxy)-11-hydroxy-4,8,10,14-tetramethylhexadecahydro-17H-cyclopenta[a]phenanthren-17-ylidene)-6-methylhept-5-enoic acid

[0206] ##STR00054## [0207] 52.

Na-alanyl-Nt-(3-(((2R,3R,4R,5R)-2-((((1S,2S,3R,4S,6R)-4,6-diamino-3-(((2R,3R,6S)-3-amino-6-((R)-1-aminoethyl)tetrahydro-2H-pyran-2-yl)oxy)-2-hydroxycyclohexyl)oxy)-5-hydroxy-5-methyl-4-(methylamino)tetrahydro-2H-pyran-3-yl)oxy)-3-oxopropyl)histidine

[0208] ##STR00055## [0209] 53.

Na-alanyl-Nt-(3-(((S)-1-(tert-butylamino)-3-((5-oxo-5,6,7,8-tetrahydronaphthalen-1-yl)oxy)propan-2-yl)oxy)-3-oxopropyl)histidine

[0210] ##STR00056## [0211] 54.

Na-alanyl-Np-(3-(3-(((1R,3s,5S)-8-methyl-8-azabicyclo[3.2.1]octan-3-yl)oxy)-3-oxo-2-phenylpropoxy)-3-oxopropyl)histidine

[0212] ##STR00057## [0213] 55.

Na-alanyl-Nt-(3-(((2S,3S,4R)-6-(((2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-(((2S,3R,4S,6R)-4-(dimethylamino)-3-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2-ethyl-3,4,10-trihydroxy-3,5,6,8,10,12,14-heptamethyl-15-oxo-1-oxa-6-azacyclopentadecan-13-yl)oxy)-4-methoxy-2,4-dimethyltetrahydro-2H-pyran-3-yl)oxy)-3-oxopropyl)histidine

[0214] ##STR00058## [0215] 56.

Na-alanyl-Nt-(3-((1-(4-(2-(cyclopropylmethoxy)ethyl)phenoxy)-3-(isopropylamino)propan-2-yl)oxy)-3-oxopropyl)histidine

[0216] ##STR00059## [0217] 57.

Na-alanyl-Nt-(3-((2R,3R)-2-(2,2-dichloroacetamido)-3-hydroxy-3-(4-nitrophenyl)propoxy)-3-oxopropyl)histidine

[0218] ##STR00060## [0219] 58.

Na-alanyl-Nt-(3-((R)-3-hydroxy-2-((6-oxo-1,6-dihydro-9H-purin-9-yl)methoxy)propoxy)-3-oxopropyl)histidine

[0220] ##STR00061## [0221] 59.

Na-alanyl-Nt-(3-(2-(((1R,3s,5S)-8-methyl-8-azabicyclo[3.2.1]octan-3-yl)oxy)-2-oxo-1-phenylethoxy)-3-oxopropyl)histidine

[0222] ##STR00062## [0223] 60.

Na-alanyl-Nt-(3-(((1R,2R,3R,4S)-4-hydroxy-2-((R)-3-hydroxy-5-phenylpentyl)-3-((Z)-7-isopropoxy-7-oxohept-2-en-1-yl)cyclopentyl)oxy)-3-oxopropyl)histidine

[0224] ##STR00063## [0225] 61.

Na-alanyl-Nt-(3-(((8S,9S,10R,11S,13S,14S,17R)-17-((chloromethoxy)carbonyl)-17-((ethoxycarbonyl)oxy)-10,13-dimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-11-yl)oxy)-3-oxopropyl)histidine

[0226] ##STR00064## [0227] 62.

Na-alanyl-Nt-(3-oxo-3-(((8S,9S,10R,11S,13S,14S,16R,17S)-10,13,16,17-tetramethyl-3-oxo-17-propionyl-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-11-yl)oxy)propyl)histidine

[0228] ##STR00065## [0229] 63.

Na-alanyl-Nt-(3-(((S)-1-(tert-butylamino)-3-((4-morpholino-1,2,5-thiadiazol-3-yl)oxy)propan-2-yl)oxy)-3-oxopropyl)histidine

[0230] ##STR00066## [0231] 64.

Na-alanyl-Nt-(3-(((1S,2R,3R,4R)-4-hydroxy-3-((R,E)-3-hydroxy-4-(3-(trifluoromethyl)phenoxy)but-1-en-1-yl)-2-((Z)-7-isopropoxy-7-oxohept-2-en-1-yl)cyclopentyl)oxy)-3-oxopropyl)histidine

[0232] ##STR00067## [0233] 65.

Na-alanyl-Nt-(3-(((1R,2R,3R,4S)-2-((E)-3,3-difluoro-4-phenoxybut-1-en-1-yl)-4-hydroxy-3-((Z)-7-isopropoxy-7-oxohept-2-en-1-yl)cyclopentyl)oxy)-3-oxopropyl)histidine

[0234] ##STR00068## [0235] 66.

Na-alanyl-Nt-(3-(((2R,3 S,4S,5R,6S)-4-amino-6-(((1S,2S,3R,4S,6R)-4,6-diamino-3-(((2S,3R,5 S,6S)-3,6-diamino-5-hydroxytetrahydro-2H-pyran-2-yl)oxy)-2-hydroxycyclohexyl)oxy)-3,5-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)-3-oxopropyl)histidine

[0236] ##STR00069## [0237] 67.

Na-alanyl-Nt-(3-(3-(ethyl(pyridin-4-ylmethyl)amino)-3-oxo-2-phenylpropoxy)-3-oxopropyl)histidine

[0238] ##STR00070## [0239] 68.

Na-alanyl-Nt-(3-(((1R,2R,E)-1-((2S,5S,11S,14S,17S,20S,23R,26S,29S,32S)-5-ethyl-11,17,26,29-tetraisobutyl-14,32-diisopropyl-1,7,10,16,20,23,25,28,31-nonamethyl-3,6,9,12,15,18,21,24,27,30,33-undecaoxo-1,4,7,10,13,16,19,22,25,28,31-undecaazacyclotritriacontan-2-yl)-2-methylhex-4-en-1-yl)oxy)-3-oxopropyl)histidine

[0240] ##STR00071## [0241] 69.

2-(4-(1-((3-(alanyl-Nt-histidino)propanoyl)oxy)-4-(4-(hydroxydiphenylmethyl)piperidin-1-yl)butyl)phenyl)-2-methylpropanoic acid

[0242] ##STR00072## [0243] 70.

Na-alanyl-Nt-(3-(1-(4-(tert-butyl)phenyl)-4-(4-(hydroxydiphenylmethyl)piperidin-1-yl)butoxy)-3-oxoprpyl)histidine

[0244] ##STR00073## [0245] 71.

Na-alanyl-Nt-(3-(2-(2-(4-((4-chlorophenyl)(phenyl)methyl)piperazin-1-yl)ethoxy)ethoxy)-3-oxopropyl)histidine

[0246] In the alternative, the peptide may be conjugated to a protein therapeutic agent or a nucleotide molecule coding for the protein drug. Examples of protein drugs and nucleic acid molecules which may be used in the practice of the invention include, but are not limited to, those having the SEQ ID NOS: listed in the following table:

TABLE-US-00001 TABLE 1 Sequence ID Therapeutic Agent Sequence Type Seq ID NO: 14 VEGF DECOY Homo sapien Nucleotide Sequence Seq ID NO: 15 VEGF DECOY Protein-Ala-His Amino Acid Sequence Seq ID NO: 16 VEGF DECOY Ala-His-Protein Amino Acid Sequence Seq ID NO: 17 VEGF DECOY Homo sapien Amino Acid Sequence Seq ID NO: 18 Pigment Derived Growth Factor Homo sapien Nucleotide Sequence Seq ID NO: 19 Pigment Derived Growth Factor Protein-Ala-his Amino Acid Sequence Seq ID NO: 20 Pigment Derived Growth Factor Ala-His-Protein Amino Acid Sequence Seq ID NO: 21 Pigment Derived Growth Factor Homo sapien Amino Acid Sequence Seq ID NO: 22 Pigment Derived Growth Factor Homo sapien Nucleotide Sequence Seq ID NO: 23 Pigment Derived Growth Factor Protein-Ala-His Amino Acid Sequence Seq ID NO: 24 Pigment Derived Growth Factor Ala-His-Protein Amino Acid Sequence Seq ID NO: 25 Pigment Derived Growth Factor Homo sapien Amino Acid Sequence Seq ID NO: 26 Pigment Derived Growth Factor Homo sapien Nucleotide Sequence Seq ID NO: 27 Pigment Derived Growth Factor Protein-Ala-His Amino Acid Sequence Seq ID NO: 28 Pigment Derived Growth Factor-Ala-His-Protein Amino Acid Sequence Seq ID NO: 29 Pigment Derived Growth Factor Homo sapien Amino Acid Sequence Seq ID NO: 30 Pigment Derived Growth Factor Homo sapien Nucleotide Sequence Seq ID NO: 31 Pigment Derived Growth Factor Protein-Ala-His Amino Acid Sequence Seq ID NO: 32 Pigment Derived Growth Factor Ala-His-Protein Amino Acid Sequence Seq ID NO: 33 Pigment Derived Growth Factor Homo sapien Amino Acid Sequence Seq ID NO: 34 Endostatin(HumanRecombinant) Homo sapien Nucleotide Sequence Seq ID NO: 35 Endostatin(HumanRecombinant)Protein-Ala-His Amino Acid Sequence Seq ID NO: 36 Endostatin(HumanRecombinant)Ala-His-Protein Amino Acid Sequence Seq ID NO: 37 Endostatin(HumanRecombinant)Homo sapien Amino Acid Sequence Seq ID NO: 38 Type XVIII Collagen Homo sapien Nucleotide Sequence Seq ID NO: 39 Type XVIII Collagen Protein-Ala-His Amino Acid Sequence Seq ID NO: 40 Type XVIII Collagen Ala-His-Protein Amino Acid Sequence Seq ID NO: 41 Type XVIII Collagen Homo sapien Amino Acid Sequence Seq ID NO: 42 Angiostatin Homo sapien Nucleotide Sequence Seq ID NO: 43 Angiostatin Protein-Ala-His Amino Acid Sequence Seq ID NO: 44 Angiostatin Ala-His-Protein Amino Acid Sequence Seq ID NO: 45 Angiostatin Homo sapien Amino Acid Sequence Seq ID NO: 46 Plasminogen Homo sapien Nucleotide Sequence Seq ID NO: 47 Plasminogen Protein-Ala-His Amino Acid Sequence Seq ID NO: 48 Plasminogen Ala-His-Protein Amino Acid Sequence Seq ID NO: 49 Plasminogen Homo sapien Amino Acid Sequence Seq ID NO: 50 Angiopoietin-1 Homo sapien Nucleotide Sequence Seq ID NO: 51 Angiopoietin-1 Protein-Ala-His Amino Acid Sequence Seq ID NO: 52 Angiopoietin-1 Ala-His-Protein Amino Acid Sequence Seq ID NO: 53 Angiopoietin-1 Homo sapien Amino Acid Sequence Seq ID NO: 54 Angiopoietin-1 Homo sapien Nucleotide Sequence Seq ID NO: 55 Angiopoietin-1 Protein-Ala-His Amino Acid Sequence Seq ID NO: 56 Angiopoietin-1 Ala-His-Protein Amino Acid Sequence Seq ID NO: 57 Angiopoietin-1 Homo sapien Amino Acid Sequence Seq ID NO: 58 Angiopoietin-1 Homo sapien Nucleotide Sequence Seq ID NO: 59 Angiopoietin-1 Protein-Ala-His Amino Acid Sequence Seq ID NO: 60 Angiopoietin-1 Ala-His-Protein Amino Acid Sequence Seq ID NO: 61 Angiopoietin-1 Homo sapien Amino Acid Sequence Seq ID NO: 62 ABCA4 Homo sapien Nucleotide Sequence Seq ID NO: 63 ABCA4 Protein-Ala-His Amino Acid Sequence Seq ID NO: 64 ABCA4 Ala-His-Protein Amino Acid Sequence Seq ID NO: 65 ABCA4 Homo sapien Amino Acid Sequence Seq ID NO: 66 NADH Dehydrogenase Unit4 Homo sapien Nucleotide Sequence Seq ID NO: 67 NADH Dehydrogenase Unit4 Protein-Ala-His Amino Acid Sequence Seq ID NO: 68 NADH Dehydrogenase Unit4 Protein-Ala-His Amino Acid Sequence Seq ID NO: 69 NADH Dehydrogenase Unit4 Homo sapien Amino Acid Sequence Seq ID NO: 70 GDNF Homo sapien Nucleotide Sequence Seq ID NO: 71 GDNF Protein-Ala-His Amino Acid Sequence Seq ID NO: 72 GDNF Ala-His-Protein Amino Acid Sequence Seq ID NO: 73 GDNF Homo sapien Amino Acid Sequence Seq ID NO: 74 GDNF Homo sapien Nucleotide Sequence Seq ID NO: 75 GDNF Protein-Ala-His Amino Acid Sequence Seq ID NO: 76 GDNF Ala-His-Protein Amino Acid Sequence Seq ID NO: 77 GDNF Homo sapien Amino Acid Sequence Seq ID NO: 78 GDNF Homo sapien Nucleotide Sequence Seq ID NO: 79 GDNF Protein-Ala-His Amino Acid Sequence Seq ID NO: 80 GDNF Ala-His-Protein Amino Acid Sequence Seq ID NO: 81 GDNF Homo sapien Amino Acid Sequence Seq ID NO: 82 GDNF Homo sapien Nucleotide Sequence Seq ID NO: 83 GDNF Protein-Ala-HisProtein-Ala-His Amino Acid Sequence Seq ID NO: 84 GDNF Protein-Ala-HisAla-His-Protein Amino Acid Sequence Seq ID NO: 85 GDNF Homo sapien Amino Acid Sequence Seq ID NO: 86 Peripherin-2 Homo sapien Nucleotide Sequence Seq ID NO: 87 Peripherin-2 Protein-Ala-His Amino Acid Sequence Seq ID NO: 88 Peripherin-2 Ala-His-Protein Amino Acid Sequence Seq ID NO: 89 Peripherin-2 Homo sapien Amino Acid Sequence Seq ID NO: 90 RPE65 Homo sapien Nucleotide Sequence Seq ID NO: 91 RPE65 Protein-Ala-His Amino Acid Sequence Seq ID NO: 92 RPE65 Ala-His-Protein Amino Acid Sequence Seq ID NO: 93 RPE65 Homo sapien Amino Acid Sequence

[0247] In one embodiment, the therapeutic agent is an antisense oligonucleotide that inhibits viral replication. In another embodiment, the antisense oligonucleotide inhibits cytomegalovirus (CMV) replication. Antisense oligonucleotides that are useful for the treatment of cytomegalovirus are disclosed in Henry et al., (2001).

[0248] When the therapeutic agent is a nucleotide molecule, it may be contained by a vector including plasmids, cosmids, artificial chromosomes, and modified viruses, as are known in the art. See, for example, Current Protocols in Molecular Biology (eds. Ausubel et al., Wiley, 2004 edition) and Molecular Cloning: A Laboratory Manual (Sambrook and Russell (Cold Spring Harbor Laboratory Press, 2001, third edition).

[0249] In one embodiment, the therapeutic agent is an antibody. In another embodiment, the antibody is bevacizumab (Avastin) or ranibizumab (Lucentis). In another embodiment, the ocular disorder is macular degeneration.

[0250] The nanoparticle composition may further comprise a targeting agent such as an antibody. The term antibody is used to refer to any antibody-like molecule that has an antigen binding region, and includes antibody fragments such as Fab, Fab, F(ab).sub.2, single domain antibodies (DABs), Fv, scFv (single chain Fv), and the like. The techniques for preparing and using various antibody-based constructs and fragments are well known in the art. Means for preparing and characterizing antibodies are also well known in the art (See, e.g., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, 1988). Antibody targeting agents which are expected to be useful in the eye include growth factors (e.g., VEGF and PDGF), growth factor receptors (e.g., VEGF and PDGF), receptors of inflammatory mediators, and integrin receptors.

[0251] Monoclonal antibodies (MAbs) are recognized to have certain advantages, e.g., reproducibility and large-scale production. The antibodies may be of human, murine, monkey, rat, hamster, rabbit and chicken origin.

[0252] Humanized antibodies are also contemplated, as are chimeric antibodies from mouse, rat, or other species, bearing human constant and/or variable region domains, bispecific antibodies, recombinant and engineered antibodies and fragments thereof. Methods for the development of antibodies that are custom-tailored to the patient's disease are likewise known and such custom-tailored antibodies are also contemplated.

[0253] Antibodies may be further purified, if desired, using filtration, centrifugation and various chromatographic methods such as HPLC or affinity chromatography. Fragments of the antibodies can be obtained from the antibodies so produced by methods which include digestion with enzymes such as pepsin or papain, and/or by cleavage of disulfide bonds by chemical reduction. Alternatively, antibody fragments encompassed by the present invention can be synthesized using an automated peptide synthesizer or by expression of full-length gene or gene fragments in E. coli.

[0254] A molecular cloning approach may be used to generate monoclonal antibodies. In one embodiment, combinatorial immunoglobulin phagemid libraries are prepared from RNA isolated from the spleen of the immunized animal, and phagemids expressing appropriate antibodies are selected by panning using cells expressing the antigen and control cells. The advantages of this approach over conventional hybridoma techniques is that many more antibodies can be produced and screened in a single round, and that new specificities are generated by H and L chain combination which further increases the chance of finding appropriate antibodies.

[0255] The peptide-therapeutic agent conjugates may be made by methods of solid phase synthesis exemplified by FIG. 3.

[0256] The nanoparticle compositions may be formulated with a pharmaceutically acceptable carrier for ophthalmic use. Particular carriers include saline, buffered saline, together with optional ingredients such as reduced glutathione, vitamin A, vitamin E. See U.S. Pat. No. 6,194,457.

[0257] The compositions may be administered by any means that achieves contact to the eye. In some embodiments, the composition is administered by intravitreal injection, eye drops, and the like. The location of the nanoparticle composition within the vitreous may be determine by ophthalmoscopy.

[0258] In one embodiment, the nanoparticle composition is exposed to light. In one embodiment, the method further comprises exposing the nanoparticle to light sufficient to induce the quantum dot to emit energy, wherein the energy cleaves the linkage and the therapeutic agent is released. In another embodiment, the wavelength of the light is in the range of 600-2000 nm. In another embodiment, the wavelength of the light is in the range of 700-1200 nm. In another embodiment, the wavelength of the light is in the range of 750-1100 nm. In a further embodiment, a laser provides the light to the nanoparticle.

[0259] The invention provides a method of treating an ocular disorder, comprising: (a) intravitreal administration to an eye of a subject in need thereof with an effective amount of a therapeutic nanoparticle composition, the therapeutic nanoparticle composition comprising (i) at least one population of nanostructures and (ii) at least one peptide attached to the at least one population of nanostructures. When in the eye, the nanostructures are exposed to light thereby electrostimulating the eye and treating the ocular disorder.

[0260] It has been discovered that therapeutic nanparticle compositions comprising a peptide bound to the surface thereof have a longer residence time compared to drugs which are administered by intravitreal means. Thus, the nanoparticle compositions have much longer half-lifes within the eye, e.g., from 5-15 days. In another embodiment, the half-life is 7-9 days.

[0261] The therapeutic nanoparticle compositions provide electrical stimulation to the eye and are useful for treating a number of disorders which are treatable by electrostimulation. Examples of such disorders include loss of vision resulting from non-arteritic anterior ischemic optic neuropathy, multiple sclerosis and clinically isolated syndrome (see, the web at willseye.org/transcorneal-electrical-stimulation-tes); retinitis pigmentosa (see, Adam et al., Exp. Eye Res. 149:75-83 (2016)); to treat longstanding retinal artery occlusion (see, Inomata et al., Clin. Invest. 245:1773-80 (2007)); partial atrophy of the optic nerve in neurological patients (see, Shandurina et al., Neurosci. Behav. Physiol. 26:137 (1996)); fibromyalgia (see, Hargrove et al., Pain Med. 13:115-124 (2012)); light-induced photoreceptor degeneration (see, Ying-qin Ni et al., Exp. Neurol. 219:439-52 (2009)); progressive myopia (see, Okovitov, Vestn Oftalmol. 113:24-6 (1997)); amblyopia (see, S. B. Slobodyanik and V. S. Ponomarchuk, Electrical stimulation of the visual pathway based on phosphen phenomenon in amblyopia therapy, in: XI Congress of the European Society of Ophthalmology (Hungary, Budapest, Jun. 1-5, 1997), Budapest (1997), p. 193), and acute ocular hypertension related injury (see, Fu et al., on the web at hub.hku.hk/handle/10722/207462).

[0262] The therapeutic nanoparticle compositions also provide increased circulation and restore optimal eye function by increasing blood flow and metabolism. The therapeutic nanoparticle compositions also increase blood vessel permeability and achieve a more normal cellular electrical potential, increase ATP levels, and restore normal cell metabolism. In addition, the therapeutic nanoparticle compositions have a healing effect on the small blood vessels in the retina, providing a more efficient delivery of nutrients to the retinal cells and a more efficient uptake of proteins that can accumulate on the retina, thus rejuvenating the cells in the eye (see, U.S. Pat. 6,275,735).

[0263] The therapeutic nanoparticle compositions may be used for the rehabilitation of unilateral neglect syndrome in stroke patients (see, Yang et al., Frontiers in Human Neurosci. 7:187 (2013)). In another embodiment, the therapeutic nanoparticle compositions may be used for the treatment of degeneration of the retina. In another embodiment, the therapeutic nanoparticle compositions may be used for the treatment of loss of vision resulting from non-arteritic anterior ischemic optic neuropathy, multiple sclerosis, clinically isolated syndrome, retinitis pigmentosa, longstanding retinal artery occlusion, partial atrophy of the optic nerve in neurological patients, fibromyalgia, light-induced photoreceptor degeneration, progressive myopia, amblyopia, and acute ocular hypertension related injury, and for the rehabilitation of unilateral neglect syndrome in stroke patients. In another embodiment, the therapeutic nanoparticle compositions may be used for the treatment of loss of vision resulting from glaucoma, ischemic neuropathy or retinal vascular occlusion. In another embodiment, the therapeutic nanoparticle compositions may be used for the treatment of glaucoma including Open Angle Glaucoma, Angle Closure Glaucoma, Aniridic Glaucoma, Congenital Glaucoma, Juvenile Glaucoma, Lens-Induced Glaucoma, Neovascular Glaucoma, Post-Traumatic Glaucoma, Steroid-Induced Glaucoma, Sturge-Weber Syndrome Glaucoma, and Uveitis-Induced Glaucoma.

[0264] Also provided is a method to tailor the physical/chemical properties of the nanoparticle to capitalize or the biological ocular environment to enhance contact time of therapeutic agents in different anatomical areas of the eye. In this embodiment, nanoparticles of different sizes and compositions are administered to different anatomical areas of the eye and the residence time within the eye is measured. For example, the nanoparticle compositions may be administered by injection into the vitreous body just outside of the lens, into the center of the vitreous body, on top of the retina, in the subconjunctival space, in subretinal space, or on top of the optic nerve, and the residence time measured to determine which compositions have the longest residence time when injected into a particular location.

EXAMPLE

[0265] Intravitreal administration has been an effective way of delivering agents including drugs into the posterior chamber of the eye for the treatment of diseases such as macular degeneration (Kuppermann et al., 2007), diabetic retinopathy (Martidis et al., 2002) or viral infections (Henry et al., 2001). In most cases the agents have to be administered periodically in part due to the clearance from the vitreous and retina or due to enzymatic inactivation. Thus compounds that have slow clearance out of the vitreous or retina or longer residence in these tissues have the benefit of fewer administrations. In this experiment the clearance of SeeQ Cd/Se 655 Alt (a 6.5 nm quantum dot coated with the dipeptides His-Leu and Gly-His in a weight ratio of about 8:1; obtained from ThermoFisher Scientific) and its duration in the vitreous and retina was evaluated after intravitreal injection in rabbit eyes.

Methods

[0266] New Zealand white rabbits weighing 2 to 2.5 kg were used in the study. Rabbits were anesthetized with intramuscular injection of 50 mg/kg of ketamine and 10 mg/kg xylazine. The eyes were topically anesthetized with proparacaine (0.5%) and the ocular surface was cleaned with providone iodine 0.5% before injection. Intravitreal injection was made 2 to 3 mm from the limbus in the superior quadrant of the globe. Twelve rabbits received single injection of 40 l of SeeQ Cd/Se 655 Alt (4.2 M aqueous solution, at pH 8) into both eyes, using 0.5 ml tuberculin syringe. Vitreous and retinal samples were collected at times 0, 4 hours, 1, 3, 7 and 14 days after injection. Two rabbits (4 eyes) were used per time point. Eyes were routinely examined for inflammation and toxicity using indirect ophthalmoscope and slit lamp. The localization of the drug in the vitreous was also assessed with indirect ophthalmoscope before the rabbits were sacrificed. Rabbits were euthanized with intravenous injection of ketamine and xylazine and eyes were enucleated. Animals that were not injected served as blank control. The vitreous and the retina samples were collected into pre-weighed tubes, weighed and frozen until they were analyzed. SeeQ Cd/Se 655 Alt concentration in the retina and vitreous were determined by measuring fluorescence (excitation 410 and emission 660). Samples were prepared, diluted and values measured from an external standard curve with concentrations of 40, 30, 20, 10, 5, 4, 3, 2, 1, and 0.5 nM. The limit of detection and limit of quantitation were 0.1 nM and 0.5 nM respectively.

Results

[0267] Examination of the eyes during the two-week period showed no inflammation or any toxic effect of the drug. Examination of the posterior part of the eye using indirect ophthalmoscope and slit lamp showed the presence of the therapeutic in the vitreous. At time 0 it was located at the site of injection. Four hours after injection the drug was distributed in most of the vitreous humor and moved towards the retina. During the rest of the experimental periods (1 to 14 days), the presence of the drug was evident as seen by the orange color in the vitreous.

Clearance of SeeQ Cd/Se 655 Alt

[0268] The concentration of SeeQ Cd/Se 655 Alt in the vitreous humor is shown in FIG. 1. The maximum concentration was constant in the first three days after injection with average of 80.910.7 nM, indicating very little clearance during this period. After day 7 and 14 vitreal concentration decreased to 56.4 nM (by 25%) and 25.6 nM (67%) respectively. The half-life of the clearance from the vitreous was 9 days. It appears to follow first order process.

[0269] In the retina the drug increased in the first three days reaching maximum in 1 day and remained high after 3 days. After day 7 and 14 retinal concentrations decreased to 0.152 nmoles/g (by 47%) and 0.0399 nmoles/g (88%) respectively. The half-life of the clearance from the retina was 7.5 days. The rate of clearance from the retina was also similar to that of the vitreous.

Discussion

[0270] Intravitreal injection of drugs is a very effective way of targeted drug delivery to the posterior portion of the eye. In this experiment we showed the distribution and clearance of SeeQ Cd/Se 655 Alt in the vitreous and retina. After a single intravitreal injection, SeeQ Cd/Se 655 Alt was cleared from these tissues slowly with half-life of 7.5 days in the retina and 9 days in the vitreous. The concentration gradient created between the vitreous and retina allowed the retina to reach Cmax at 3 days after injection.

[0271] The rate of clearance of intravitreally administered material depends on the physicochemical properties of the material. These properties include lipophilicity, molecular size, structure, and surface charge of the material. In addition there are also active transport mechanisms, enzymatic degradation that can affect the residency and clearance of the drug. For example small molecule dexamethasone in a solution form disappears quickly (half-life of 3 days) (Berthe et al., 1992). On the other hand when it is prepared in a sustained release form (embedded in a lactic co-glycolic copolymer) it maintained a constant concentration for longer than one month (Chang-Lin et al., 2011). In this case a much higher concentration was accumulated in the retina. Unlike dexamethasone solution a suspension of triamcinolone acetonide had a different profile of clearance (Kim et al., 2006). The half-life of triamcinolone acetonide was 24 days for 4 mg and 39 days for 16 mg and the drug lasted for up to 4 to 6 months for the two doses administered (Kim et al., 2006). This long duration of triamcinolone is due to a very low solubility of the compound, and therefore the dissolution rate contributes to the steady state concentration in the retina. Other small molecules such as the hydrophilic antiviral foscarnet have a half-life of 12 hours (Kwak et al., 1994) Large protein molecules like Avastin, an antibody that are used for the treatment of macular edema, had short half-life of 6 days but can be detected for longer than 30 days (Sinapis et al., 2011). This study also demonstrated that Avastin is delivered systemically as bevacizumab was found in the untreated contralateral eye.

[0272] In this experiment, SeeQ Cd/Se 655 Alt (168 pmoles/eye) was prepared in aqueous solution. In the retina and vitreous, the half-life was 7.5 and 9 days respectively. These values are 2.5 to three times longer than that reported for aqueous solution of dexamethasone sodium. Bioavailability of a material depends on the concentration present at the site of action. At present the physicochemical property of SeeQ Cd/Se 655 Alt in the rabbit vitreous is not known. However the observation that there was higher concentration of drug in the vitreous than in the retina at the end of two weeks suggests that the vitreous may act as a slow release depot.

REFERENCES

[0273] Berthe P, Baudouin C, Garraffo R, et al. Toxicologic and pharmacokinetic analysis of intravitreal injections of foscarnet, either alone or in combination with ganciclovir. Invest. Ophthalmol. Vis. Sci 1994:35:1038-1045.

[0274] Chang-Lin J-E, Burke JA, Peng Q. Pharmacokinetics of a Sustained-release Desamethasone Intravitreal Implant in Vitrectomized and Nonvitrectomized Eyes. Invest. Ophthalmol. Vis. Sci. 52:4605-4609, 2011

[0275] Henry SP, Miner RC, Drew WL, et al. Antiviral activity and ocular kinetics of antisense oligonucleotides designed to inhibit CMV replication. Invest Ophthalmol Vis Sci. 42:2646-51, 2001

[0276] Kim H, Csaky KG, Gravlin L. Safety and pharmacokinetics of a Preservative-free Triamcinolone Acetonide Formulation for Intravitreal Administration. Retina, 26:523530, 2006

[0277] Kuppermann BD, Blumenkranz MS, Haller JA, et al. Randomized controlled study of an intravitreous dexamethasone drug delivery system in patients with persistent macular edema. Arch Ophthalmol. 125:309-317, 2007

[0278] Kwak HW, D'Amico DJ. Evaluation of the retinal toxicity and pharmacokinetics of dexamethasone after intravitreal injection. Arch. Ophthalmol. 110:259-266, 1992

[0279] Martidis A, Duker JS, Greenberg PB, et al. Intravitreal triamcinolone for refractory diabetic macular edema. Ophthalmology,109:920-7, 2002.

[0280] Sinapis, C I, Routsias, JG, et al. Pharmacokinetics of intravitreal bevacizumab (Avastin) in rabbits. Clin. Pharmacol. 5:697-704, 2011.

[0281] Having now fully described this invention, it will be understood by those of ordinary skill in the art that the same can be performed within a wide and equivalent range of conditions, formulations, and other parameters without affecting the scope of the invention or any embodiment thereof.

[0282] Other embodiments of the invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the invention being indicated by the following claims. All patents, patent applications and publications cited herein are fully incorporated by reference.