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Title:
METHOD OF TREATING OR PREVENTING RETINOPATHY
Document Type and Number:
WIPO Patent Application WO/2020/255122
Kind Code:
A1
Abstract:
The invention provides a method of treating or preventing a retinal disease, comprising the steps of administering to a subject in need, or at risk of developing a retinal disease, an effective amount of a composition comprising nanoparticles conjugated to a von Willebrand factor (vWF) targeting agent, and radiating the retina with near infra-red (NIR) radiation, microwave radiation, radio frequency radiation or ultrasound radiation. The invention further provides nanoparticles conjugated to a von Willebrand factor (vWF) targeting agent.

Inventors:
LERNER NOAM (IL)
NAHMIAS YAAKOV (IL)
KAGAN EVGENY (IL)
Application Number:
PCT/IL2020/050662
Publication Date:
December 24, 2020
Filing Date:
June 15, 2020
Export Citation:
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Assignee:
YISSUM RES DEV CO OF HEBREW UNIV JERUSALEM LTD (IL)
International Classes:
A61K47/69; A61K47/66; A61K47/68; A61K49/00; A61K49/04; A61K49/06; A61K49/18; A61N5/02; A61N5/06; A61N5/10; A61N7/02; A61P27/02; A61P35/00; B82Y40/00; C07K7/08; C07K14/755
Domestic Patent References:
WO2012142362A22012-10-18
Foreign References:
US20180311378A12018-11-01
Other References:
GOSWAMI, NIRMAL ET AL.: "Engineering gold-based radiosensitizers for cancer radiotherapy", MATERIALS HORIZONS, vol. 4., 13 July 2017 (2017-07-13), pages 817 - 831, XP055774058, Retrieved from the Internet [retrieved on 20200819], DOI: 10.1039/C7MH00451F
MASHREGHI, MOHAMMAD ET AL.: "Angiogenesis biomarkers and their targeting ligands as potential targets for tumor angiogenesis", JOURNAL OF CELLULAR PHYSIOLOGY, vol. 233, no. 4, April 2018 (2018-04-01), pages 2949 - 2965, XP055774062, Retrieved from the Internet [retrieved on 20200819], DOI: 10.1002/jcp.26049
BANERJEE, DEBOSHRI ET AL.: "Nanotechnology-mediated targeting of tumor angiogenesis", VASCULAR CELL, vol. 3, no. 1, 31 January 2011 (2011-01-31), pages 3, XP021091557, Retrieved from the Internet [retrieved on 20200813], DOI: 10.1186/2045-824X-3-3
YAMASHITA, MARIKO ET AL.: "Intravitreal injection of aflibercept, an anti-VEGF antagonist, down- regulates plasma von Willebrand factor in patients with age-related macular degeneration", SCIENTIFIC REPORTS, vol. 8, no. 1, 24 January 2018 (2018-01-24), pages 1 - 9, XP055774316
ABDALLA, AHMED ME ET AL.: "Current challenges of cancer anti-angiogenic therapy and the promise of nanotherapeutics", THERANOSTICS, vol. 8, 1 January 2018 (2018-01-01), pages 533 - 548, XP055665865, Retrieved from the Internet [retrieved on 20200817], DOI: 10.7150/thno.21674
MUKHERJEE, SUDIP ET AL.: "Therapeutic application of antiangiogenic nanomaterials in cancers", NANOSCALE, vol. 8, no. 25, 2016, pages 12444 - 12470, XP055774314, Retrieved from the Internet [retrieved on 20200817], DOI: 10.1039/C5NR07887C
SUN, HONGGUANG ET AL.: "Oligonucleotide aptamers: new tools for targeted cancer therapy", MOLECULAR THERAPY-NUCLEIC ACIDS, vol. 3, no. el82, 5 August 2014 (2014-08-05), pages 12444 - 12470, XP055537083, Retrieved from the Internet [retrieved on 20200813], DOI: 10.1038/mtna.2014.32
DARWEESH, RUBA S. ET AL.: "Gold nanoparticles and angiogenesis: Molecular mechanisms and biomedical applications", INTERNATIONAL JOURNAL OF NANOMEDICINE, vol. 14, 19 September 2019 (2019-09-19), pages 7643 - 7663, XP055774072, Retrieved from the Internet DOI: 10.2147 /IJN. S22394
Attorney, Agent or Firm:
FRIEDMAN, Nathalie et al. (IL)
Download PDF:
Claims:
What is claimed is:

1 . A method of treating or preventing a retinal disease, comprising the steps of administering to a subject in need, or at risk of developing a retinal disease, an effective amount of a composition comprising nanoparticles conjugated to a von Willebrand factor (vWF) targeting agent, and radiating the retina with near infra-red (NIR) radiation, microwave radiation, radio frequency radiation or ultrasound radiation.

2. The method of claim 1 , wherein the composition is administered systematically or intraocularly.

3. The method of any one of claims 1 or 2, wherein the composition is administered intravenously (IV), parenterally, subcutaneously, nasally, intramuscular injection or orally.

4. The method of claim 1 , wherein the retinal disease is diabetic retinopathy, age-related macular degeneration, retinopathy of prematurity, retinal ischemia, choroidal neovascularization or genetic disease of the retina.

5. The method of any one of claims 1 -4, wherein the retinal disease is associated with an over expression of von Willebrand factor (vWF).

6. The method of any one of claims 1 -5, wherein said method is effective in ablating micro-aneurysm.

7. The method of any one of claims 1 -6, wherein the nanoparticles are gold nanoparticles.

8. The method of claim 7, wherein the nanoparticles are asymmetric.

9. The method of any one of claims 1 -8, wherein the von Willebrand factor (vWF) targeting agent is an antibody, peptide, aptamer, heptamer, oligomer, factor VIII or derivative thereof, targeting vector, a plasmid, a small molecule or nanobody.

10. The method of claim 9, wherein the peptide has a sequence of CGGG TRYLRIHPQSWVHQI (SEQ ID. No. 1 )

1 1 .The method of any one of claims 9 and 10 , wherein the von Willebrand factor (vWF) targeting agent and/or the nanoparticles are bound to a marker for imaging detectable by an OCT, a detector of color, fluorescence, x-ray, CT scan, magnetic resonance imaging (MRI), radio-isotope scan, single photon emission tomography (SPECT) or positron emission tomography (PET).

12. The method of any one of claims 1 -1 1 , wherein the von Willebrand factor (vWF) targeting agent is conjugated to the nanoparticles with a linker.

13. The method of any one of claims 1 -12, wherein the subject is diabetic.

14. The method of any one of claims 1 -13, wherein the subject was diagnosed with a retinal disease or as detected being at risk of developing a retinal disease.

15. Nanoparticles associated with a von Willebrand factor (vWF) targeting agent.

16. The nanoparticles of claim 15, wherein the nanoparticles are gold nanoparticles.

17. The nanoparticles of any one of claims 15 and 16, wherein the nanoparticles are in a rod shape.

18. The nanoparticles of claim 17, wherein the nanoparticles have a length of no more than 1 micron.

19. The nanoparticles of claim 17, wherein the nanoparticles are asymmetric.

20. The nanoparticles of any one of claims 15-19, wherein the von Willebrand factor (vWF) targeting agent is an antibody, peptide, aptamer, heptamer, oligomer, factor VIII or derivative thereof, targeting vector, a plasmid, a small molecule or nanobody.

21 . The nanoparticles of claim 20, wherein the peptide has a sequence of CGGG

TRYLRIHPQSWVHQI (SEQ ID. No. 1 ).

22. The nanoparticles of any one of claims 15-21 , wherein the von Willebrand factor (VWF) targeting agent and/or the nanoparticles are bound to a marker for imaging detectable by an OCT, a detector of color, fluorescence, x-ray, CT scan, magnetic resonance imaging (MRI), radio-isotope scan, single photon emission tomography (SPECT) or positron emission tomography (PET).

23. The nanoparticles of any one of claims 15-22, wherein the von Willebrand factor (vWF) targeting agent is conjugated to the nanoparticles with a linker.

24. The nanoparticles of any one of claims 15-23, wherein the nanoparticles are loaded by a drug.

25. The nanoparticles of claim 24, wherein the drug is an anti-angiogenic drug.

26. A method of treating or preventing a disease or disorder associated with an overexpression of von Willebrand factor (vWF) in tumor cells or in vasculature of a defined organ or area in the body comprising the steps of administering to a subject in need an effective amount of a composition comprising nanoparticles conjugated to von Willebrand factor (vWF) targeting agent, and radiating the defined organ or area in the body.

27. The method of claim 26, wherein the disease is a cancer.

28. The method of claim 27, wherein the disease is a colon cancer, gastric cancer, pancreatic cancer, liver cancer, lung cancer, breast cancer and brain cancer.

29. The method of any one of claims 26-28, wherein the radiation is with high-energy rays (such as x-rays) or with near infra-red radiation.

30. The method of any one of claims 26-29, wherein the nanoparticles are gold nanoparticles.

31 .The method of claim 30, wherein the nanoparticles are asymmetric.

32. The method of any one of claims 26-31 , wherein the von Willebrand factor (vWF) targeting agent is an antibody, peptide, aptamer, heptamer, oligomer, factor V!li or derivative thereof, targeting vector, a plasmid, a small molecule or nanobody.

33. The method of claim 32, wherein the peptide has a sequence of CGGG TRYLRIHPQSWVHQI (SEQ ID. No. 1 ).

34. The method of any one of claims 30 and 31 , wherein the von Willebrand factor (VWF) targeting agent and/or the nanoparticles are bound to a marker for imaging detectable by an OCT, a detector of color, fluorescence, x-ray, CT scan, magnetic resonance imaging (MRI), radio-isotope scan, single photon emission tomography (SPECT) or positron emission tomography (PET).

35. The method of any one of claims 26-34, wherein the von Willebrand factor (vWF) targeting agent is conjugated to the nanoparticles with a linker.

36. The method of claim 28, wherein the disorder is associated with the presence of micro-aneurysm in the brain.

37. The method of claim 36, wherein the method is effective in ablating micro-aneurysm in the brain.

38. The method of any one of claims 26-37, wherein the nanoparticles are in a rod shape.

39. The method of claim 38, wherein the nanoparticles have a length of no more than 1 micron.

40. The method of claim 26, wherein the radiation is delivered adjacent to the target tumor using a surgical device.

41 . The method of any one of claims 1 or 26 or the nanoparticles of claim 15, wherein the nanoparticles have a size of between 5 to 500 nm.

42. The method of any one of claims 1 or 26 or the nanoparticles of claim 15, wherein the nanoparticles have a size of between 5-50 nm and a length of 20-60 nm.

43. The method of any one of claims 1 or 26 or the nanoparticles of claim 15, wherein the nanoparticles are in a rod shape at a size of 10±2 nm x 41 ±5 nm.

44. The method of any one of claims 1 or 26 or the nanoparticles of claim 15, wherein the nanoparticles have a specific absorption peak of 800-900 nm.

45. A pharmaceutical composition comprising the nanoparticles of any one of claims 15- 23 and an acceptable pharmaceutically carrier.

46. A method of treating or preventing a retinal disease, comprising the steps of administering to a subject in need, or at risk of developing a retinal disease, an effective amount of a composition comprising nanoparticles loaded with anti angiogenesis drug, wherein the loaded nanoparticles are conjugated to a von Willebrand factor (vWF) targeting agent.

47. The method of claim 46, wherein said nanoparticles are liposomes, optionally loaded with an anti-angiogenesis drug.

48. The method of claim 47, wherein the anti-angiogenesis drug is avastin.

49. The method of claim 47, when said nanoparticles are a micelle or a liposome.

50. The method of claim 49, wherein the micelle is PEG-PLA micelle conjugated to the anti-angiogenesis drug TNP-470.

Description:
METHOD OF TREATING OR PREVENTING RETINOPATHY

BACKGROUND OF THE INVENTION

[0001 ] Diabetic retinopathy is one of the most common complications associated with chronic hyperglycemia, occurring in 30% of diabetic patients, and is the leading cause of blindness in adults. Diabetic retinopathy is defined by the local expansion of the microvasculature, forming microaneurysms, leading to rupture and blockage resulting in significant vision loss. Diabetic retinopathy affects over 100 million individuals worldwide, a population which is expected to double by 2040, giving rise to medical costs over $1 billion per year.

[0002] In an article published by Ezra et al., the Von Willebrand factor (vWF) expression in diabetic retina blood vessel was shown to correlate with a drop in shear forces required for proper endothelial function. Thus, vWF over-expression was suggested as a marker for the microaneurysm risk in retinopathy (Ezra et al., Integr. Biol., 2013, 5,474-480). The results therein suggest that vWF staining can be used to predict which microaneurysm presents higher risk of leakage.

[0003] Current treatments include preventative intraocular injections of anti-angiogenesis drugs, such as bevacizumab or ranibizumab, blocking blood vessel outgrowth. Intraocular injections are uncomfortable, painful and are only carried out by a qualified physician. Microaneurysm leakage can also be treated after-the-fact by laser photocoagulation in order to limit the damage to the retina.

[0004] Intraocular injection may result in many complications, such as, subconjunctival hemorrhage, foreign body sensation, eye pain, red eye, and blurry vision, inflammation, retinal detachment, ocular hypertension, cataract and the like. Further, a physician is required for such an injection, as well as that an Optical Coherence Tomography (OCT) of the retina is needed prior to the injection. In addition, laser photocoagulation causes significant damage to peripheral healthy tissue.

[0005] Thus, there is a need for a targeted, safe treatment that will avoid the use of intraocular injections and peripheral healthy tissue damage. SUMMARY OF THE INVENTION

[0006] in some embodiments of the invention, there is provided a method of treating or preventing a retinal disease, comprising the steps of administering to a subject in need, or at risk of developing a retinal disease, an effective amount of a composition comprising nanoparticles conjugated to a von Willebrand factor (vWF) targeting agent, and radiating the retina. The radiation may be a near infra-red (NIR) radiation, microwave radiation, radio frequency radiation or ultrasound radiation.

[0007] in some embodiments of the invention, there is provided a method of treating, ameliorating or preventing a retinal disease, comprising the steps of administering to a subject in need, or at risk of developing a retinal disease, an effective amount of a composition comprising nanoparticles conjugated to a von Willebrand factor (vWF) targeting agent, and applying energy to the retina. In some embodiments the energy is applied using laser beam, radiation, thermal heating, and ultrasonic, including focused ultrasound and/or RF (radiofrequency). In some embodiments, the energy applied is near an infra-red (NIR) radiation.

[0008] In some embodiments, the application of the energy on the retina, induces the temperature of the nanoparticles to about 60-100°c, preferably to 70°c.

[0009] In some embodiments of the invention, the composition is administered systematically or intraocular.

[00010] In some embodiments of the invention, the composition is administered one to four times a year. In some embodiments of the invention, the composition is administered two to three times a year.

[0001 1 ] In some embodiments of the invention, the nanoparticles composition can be present in a dry formulation (such as lyophilized composition) or suspended in a biocompatible medium. Suitable biocompatible media include, but are not limited to, water, buffered aqueous media, saline, buffered saline, buffered solutions of amino acids, buffered solutions of proteins, buffered solutions of sugars, buffered solutions of vitamins, buffered solutions of synthetic polymers, lipid-containing emulsions, and the like. [00012] In some embodiments of the invention, the retinal disease is diabetic retinopathy, age-related macular degeneration, retinopathy of prematurity, retinal ischemia, choroidal neovascularization or genetic disease of the retina.

[00013] In some embodiments of the invention, the retinal disease is associated with an over expression of von Willebrand factor (vWF).

[00014] In some embodiments of the invention, the method is effective in ablating micro-aneurysm.

[00015] In some embodiments of the invention, the nanoparticles are metal nanoparticles. In some embodiments of the invention, the nanoparticles are gold nanoparticles.

[00016] In some embodiments of the invention, the nanoparticles are in a rod shape.

[00017] In some embodiments of the invention, the von Willebrand factor (vWF) targeting agent is an antibody, peptide, aptamer, heptamer, oligomer, factor VIII or derivative thereof, targeting vector or nanobody. In some embodiments of the invention, the peptide has a sequence of CGGG TRYLRIHPQSWVHQI (SEQ ID. No. 1 )

[00018] In some embodiments of the invention, the composition is administered as the sole agent or in combination with one or more other therapies. In some embodiments of the invention, the composition is administered in combination with an anti-VEGF agent.

[00019] In some embodiments of the invention, there is provided a method of treating or preventing a disease or disorder associated with an overexpression of von Willebrand factor (vWF) in a defined organ or area in the body comprising the steps of administering to a subject in need an effective amount of a composition comprising nanoparticles conjugated to von Willebrand factor (vWF) targeting agent, and radiating the defined organ or area in the body.

[00020] In some embodiments of the invention, the disease is a cancer.

[00021 ] In some embodiments of the invention, the disease is a colon cancer, gastric cancer pancreatic cancer, liver cancer, lung cancer, breast cancer and brain cancer.

[00022] In some embodiments, the radiation is with high-energy rays (such as x-rays) or with near infra-red radiation.

[00023] In some embodiments of the invention, there are provided nanoparticles conjugated to a von Willebrand factor (vWF) targeting agent. [00024] In some embodiments of the invention, the nanoparticles are gold nanoparticles.

[00025] In some embodiments of the invention, the nanoparticles are in a rod shape.

[00026] In some embodiments of the invention, the nanoparticles have a length of no more than 1 micron.

[00027] In some embodiments of the invention, there is provided a composition comprising a metal nanoparticle conjugated to a von Willebrand factor (vWF) targeting agent and a pharmaceutically acceptable carrier for local or systemic administration.

[00028] In some embodiments of the invention, there is provided a pharmaceutical composition comprising the nanoparticles the invention and an acceptable pharmaceutically carrier, which may be for local or systemic administration.

[00029] In some embodiments of the invention, there is provided a method of treating or preventing a retinal disease, comprising the steps of administering systemically or locally to a subject in need, or at risk of developing a retinal disease, an effective amount of a composition comprising nanoparticles loaded with anti-angiogenesis drug conjugated to a von Willebrand factor (vWF) targeting agent. The nanoparticles may be liposomes or micelles optionally loaded with an anti-angiogenesis drug. The anti-angiogenesis drug may be for example avastin. The micelle may be PEG-PLA micelle conjugated to the anti angiogenesis drug TNP-470.

BRIEF DESCRIPTION OF THE DRAWINGS

[00030] The invention is herein described, by way of example only, with reference to the accompanying drawings. With specific reference now to the drawings in detail, it is stressed that the particulars shown are by way of example and for purposes of illustrative discussion of the preferred embodiments of the present invention only, and are presented in the cause of providing what is believed to be the most useful and readily understood description of the principles and conceptual aspects of the invention. In this regard, no attempt is made to show structural details of the invention in more detail than is necessary for a fundamental understanding of the invention, the description taken with the drawings making apparent to those skilled in the art how the several forms of the invention may be embodied in practice. In the drawings:

[00031 ] Figure 1 is a simulation of Temperature (top) and Velocity (bottom) fields in the aneurysm area.

[00032] Figure 2 is a simulation of Incident radiation (top) and Temperature (bottom) fields in an eye model.

[00033] Figures 3A and 3B show the NIR heating of a 1 pi GNR droplet, using pulsating radiation.

DETAILED DESCRIPTION OF THE INVENTION

[00034] In one embodiment of the invention, there is provided a method of treating retinopathy by retina-wide selective photocoagulation treatment. In another embodiments, there is provided a preventative retina-wide selective photocoagulation treatment.

[00035] As mentioned above, overexpression of von Willebrand factor (vWF) was identified as a biomarker for microaneurysms that are at high risk of bursting. The invention provides a selective delivery of nanoparticles, which may be in an embodiment of the invention, asymmetric gold nanoparticle, to microaneurysms that are at high risk of bursting and that are further radiated with a radiation that may be a near infra-red (NIR) radiation, microwave radiation, radio frequency radiation and ultrasound radiation.

[00036] In an embodiment of the invention, the nanoparticles are conjugated with von Willebrand factor (vWF) targeting agent.

[00037] The targeted treatment provides minimal damage to the healthy retina, is a targeted and localized treatment and enables ablation of micro-aneurysms, which are beyond visual resolution.

[00038] In an embodiment of the invention, the invention is based on the discovery and identification of von Willebrand factor (vWF) as a biomarker for microaneurysms risk in retinopathy.

[00039] In one embodiment of the invention, there is provided a method of treating or preventing a retinal disease, comprising the steps of administering to a subject in need, or at risk of developing a retinal disease, an effective amount of a composition comprising nanoparticles conjugated to a von Willebrand factor (vWF) targeting agent; and radiating the retina with near infra-red (NIR) radiation.

[00040] In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 600-1400.

[00041 ] In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 450-900. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 500-900. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 550-900. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 600-900. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 650-900. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 700-900. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 750-900. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 800-900. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 850-900. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 800-1400. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 900- 1400.

[00042] In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 450-850. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 450-800. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 450-750. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 450-700. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 450-650. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 450-600. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 450-550. In some embodiments, the near infra-red (NIR) radiation is in a wave length of between 450-500. In some embodiments, the near infra-red (NIR) radiation is in a wave length of

[00043] In some embodiments, the composition may be administered systematically or intraocular. According to some embodiments, the systemic administration means that the composition is administered intravenously (IV), parenterally, subcutaneously, nasally, intramuscular injection or orally.

[00044] The retinal disease may be in some embodiments of the invention, a diabetic retinopathy, age-related macular degeneration, retinopathy of prematurity, retinal ischemia, choroidal neovascularization or genetic disease of the retina.

[00045] In some embodiments of the invention, the retinal disease is associated with an over expression of von Willebrand factor (vWF).

[00046] In some embodiments of the invention, the method of treatment described herein is effective in ablating micro-aneurysm.

[00047] In some embodiments of the invention, the nanoparticles are gold nanoparticles.

[00048] In some embodiments of the invention, the nanoparticles are asymmetric.

[00049] In some embodiments of the invention, the nanoparticles are in a rod shape.

[00050] In some embodiments of the invention, the nanoparticles have a length of no more than 1 micron.

[00051 ] In some embodiments of the invention, the nanoparticles are in a rod shape and have a length of no more than 1 micron.

[00052] In some embodiments of the invention, the nanoparticles are in a rod shape and have a size (a length) of between 5 to 500 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 20 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 20 to 50 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 20 to 100 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 20 to 200 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 300 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 200 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 100 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 50 nm. In some embodiments, the length of the nanoparticles is as described above and their width is not more than 1 , 2,3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20. In some embodiments, the length of the nanoparticles is as described above and their width is not more than 1 , 2,3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20.

[00053] In some embodiments, the nanoparticles are in a rod shape at a size of 10±2 nm x 41 ±5 nm.

[00054] In some embodiments of the invention, the nanoparticles, the von Willebrand factor (vWF) targeting agent is an antibody, which maybe monoclonal or polycolonal peptide, aptamer, heptamer, oligomer, factor VIII or derivative thereof, targeting vector, a plasmid, a small molecule or nanobody.

[00055] Examples for a possible antibody is, without limitation, specific antibody directed to the D’D3 region selectively blocked VWF function in vascular inflammation, without interfering with its hemostatic function, a polyclonal vWF-directed antibody that targets the D’D3 region in models of cutaneous inflammation inducing an immediate regression of inflammatory response and a significant reduction in leukocyte recruitment and vascular permeability or monoclonal antibody, such as, GPG-290, 6B4-Fab, h6B4-Fab, and AJW200 that were shown as directly targeting the VWF-Gplb binding as well as 82D6A3 that blocks VWF-collagen interaction or SZ-123.

[00056] In another embodiment of the invention, aptamer may be used. The aptamer is a pharmacological class composed of small RNA/DNA molecules, of 20 to 100 nucleotides, highly specific and nonimmunogenic. Examples for apatmer are without limitation, ARC1779 that targets VWF A1 domain-Gplb interaction and ARC15105 that targets A1 domain on VWF and blocks VWF-collagen binding.

[00057] In another embodiment, nanobody may be used, such as without limitation ALX-0081 (caplacizumab), which is an anti-VWF humanized nanobody that selectively targets the A1 domain, blocking the VWF-Gplb interaction.

[00058] In some embodiments of the invention, the peptide has a sequence of CGGG TRYLRIHPQSWVHQI (SEQ ID. No. 1 ). [00059] In some embodiments of the invention, the von Willebrand factor (vWF) targeting agent and/or the nanoparticles are bound to a marker for imaging detectable by an OCT, a detector of color, fluorescence, x-ray, CT scan, magnetic resonance imaging (MRI), radio-isotope scan, single photon emission tomography (SPECT) or positron emission tomography (PET).

[00060] In some embodiments of the invention, the von Willebrand factor (vWF) binding agent is conjugated to the nanoparticles with a linker. In some embodiments, there is provided a liposome polymeric particle with gold embedded therein and a vWF targeting agent on its surface.

[00061 ] In some embodiments of the invention, the subject is diabetic.

[00062] In some embodiments of the invention, the subject was diagnosed with a retinal disease or as detected being at risk of developing a retinal disease.

[00063] In some embodiments of the invention, there is provided a method of treating or preventing a disease or disorder associated with an overexpression of von Willebrand factor (vWF) or with tumor angiogenesis in a defined organ or area in the body comprising the steps of administering to a subject in need an effective amount of a composition comprising nanoparticles conjugated to von Willebrand factor (vWF) targeting agent, and radiating the defined organ or area in the body.

[00064] In some embodiments of the invention, the disease is a colon cancer, gastric cancer pancreatic cancer, liver cancer, lung cancer, breast cancer and brain cancer.

[00065] In some embodiments, the radiation is with high-energy rays (such as x-rays) or with near infra-red, infra-red or microwave radiation. In such cases, the radiation is delivered adjacent to the target tumor using a surgical device, such as an endoscope.

[00066] In some embodiments of the invention, the nanoparticles are gold nanoparticles.

[00067] In some embodiments of the invention, nanoparticles are asymmetric.

[00068] In some embodiments of the invention, the nanoparticles are in a rod shape and have a length of no more than 1 micron. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size (a length) of between 5 to 500 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 20 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 20 to 50 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 20 to 100 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 20 to 200 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 300 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 200 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 100 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 50 nm. In some embodiments, the length of the nanoparticles is as described above and their width is not more than 1 , 2,3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20. In some embodiments, the length of the nanoparticles is as described above and their width is not more than 1 , 2,3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20.

[00069] In some embodiments, the nanoparticles are in a rod shape at a size of 10±2 nm x 41 ±5 nm.

[00070] In some embodiments of the invention, the nanoparticles, the von Willebrand factor (vWF) targeting agent is an antibody, which maybe monoclonal or polycolonal peptide, aptamer, heptamer, oligomer, factor VIII or derivative thereof, targeting vector, a plasmid, small molecule or nanobody.

[00071 ] In some embodiments of the invention, the peptide has a sequence of CGGG TRYLRIHPQSWVHQI (SEQ ID. No. 1 )

[00072] In some embodiments of the invention, the von Willebrand factor (VWF) targeting agent and/or the nanoparticles are bound to a marker for imaging detectable by an OCT, a detector of color, fluorescence, x-ray, CT scan, magnetic resonance imaging (MRI), radio-isotope scan, single photon emission tomography (SPECT) or positron emission tomography (PET).

[00073] In some embodiments of the invention, the von Willebrand factor (vWF) targeting agent is conjugated to the nanoparticles with a linker. [00074] In some embodiments, the nanoparticles of the invention, are loaded by a drug which may be without being limited a drug is an anti-angiogenic drug.

[00075] In some embodiments of the invention, the nanoparticles of the invention is in a form of a micelle or a liposome that may be loaded with a drug.

[00076] In some embodiments of the invention, the disorder is associated with the presence of micro-aneurysm in the brain, such as for example brain Charcot-Bouchard microaneurysms. In some embodiments of the invention, the method is effective in ablating micro-aneurysm in the brain.

[00077] By one aspect of the invention, the method of treating of the invention requires for detection of an individual with a high risk for developing leakage from a microaneurysm by for example, determining in vivo, the level of von Willebrand factor (vWF) in the microvasculature of the retina to determine which region of the microvasculature has the potential of developing a leak; wherein a significantly higher level of vWF a microaneurysm region as compared to the surrounding microvasculature indicating the specific microaneurysm region is at risk for developing a leakage and said individual is at risk.

[00078] Alternatively, vWF can be detected in the retina, below the resolution needed to identify the microvasculature, and compared to retinal background; this will enable to detect only the areas of high vWF levels (so called “hot spots”) and will indicate a microaneurysm region at risk for developing leakage.

[00079] By another aspect the requires localizing in the retina of an individual a region with a high risk of developing a leakage by localizing in the retina a region with a significantly higher level of vWF as compared to the level in the surrounding microvasculature; the region being at risk to develop leakage.

[00080] By using the above methods it is possible to detect who is at risk to develop rapture or leakage from a retinal microaneurysm, and in addition to detect precisely the suspected region of developing and treat specifically this regions by clinical intervention according to the method described below.

[00081 ] The vWF level is detected by using a vWF binding agent, such as, a vWF antibody, an antigen binding domain of an antibody, a binding peptide (i.e. factor VIII), or a recognizing nucleic acid polymer (i.e. aptamer) linked to a detectable label, such as a fluorescent label for example fluorescein. The labeled vWF targeting agent can be introduced directly into the eye or given systemically.

[00082] The detection is done by direct observation, for example, using intravenous fluorescein angiography (IVFA). This will be done in the framework of a pulse - chase experiment (contrary to standard angiography) where the dye will remain in the microaneurysm after the wash. As indicated above, while this IVFA method may not have the resolution to look at microvasculature, it will be able to detect "hot spots" of elevated vWF expression and compare them to background fluorescence. The "hot spots" will need to be at least 40% above background label and are candidates for treatment.

[00083] In some embodiments of the invention, the invention provides nanoparticles conjugated to a von Willebrand factor (vWF) targeting agent.

[00084] In some embodiments of the invention, the nanoparticles are gold nanoparticles.

[00085] In some embodiments of the invention, the nanoparticles are in a rod shape.

[00086] In some embodiments of the invention, the nanoparticles have a length of no more than 1 micron.

[00087] In some embodiments of the invention, the nanoparticles are in a rod shape and have a size (a length) of between 5 to 500 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 20 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 20 to 50 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 20 to 100 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 20 to 200 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 300 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 200 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 100 nm. In some embodiments of the invention, the nanoparticles are in a rod shape and have a size of between 5 to 50 nm. In some embodiments, the length of the nanoparticles is as described above and their width is not more than 1 , 2,3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20. In some embodiments, the length of the nanoparticles is as described above and their width is not more than 1 , 2,3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20. In some embodiments, the nanoparticles are in a rod shape at a size of 10±2 nm x 41 ±5 nm [00088] In some embodiments of the invention, the nanoparticles are asymmetric.

[00089] In some embodiments of the invention, the von Willebrand factor (vWF) targeting agent is an antibody, peptide, aptamer, factor VIII, a plasmid, a small molecule or nanobody.

[00090] In some embodiments of the invention, the targeting agent is a peptide that has a sequence of CGGG TRYLRIHPQSWVHQI (SEQ ID. No. 1 ).

[00091 ] In some embodiments of the invention, the von Willebrand factor (VWF) targeting agent and/or the nanoparticles are bound to a marker for imaging detectable by an OCT, a detector of color, fluorescence, x-ray, CT scan, magnetic resonance imaging (MRI), radio-isotope scan, single photon emission tomography (SPECT) or positron emission tomography (PET).

[00092] In some embodiments of the invention, the von Willebrand factor (vWF) targeting agent is conjugated to the nanoparticles with a linker.

[00093] In accordance with the present invention, a system for performing photocoagulation treatment is provided. The system comprising, near infrared console generating a near infrared light beam. The near infrared light beam is passed through an optical lens or mask to optically shape the laser light beam.

[00094] In some embodiments of the invention, there is provided a pharmaceutical composition comprising the nanoparticles the invention and an acceptable pharmaceutically carrier, which may be for local or systemic administration.

[00095] In some embodiments of the invention, there is provided a method of treating or preventing a retinal disease, comprising the steps of administering systemically or locally to a subject in need, or at risk of developing a retinal disease, an effective amount of a composition comprising nanoparticles loaded with anti-angiogenesis drug conjugated to a von Willebrand factor (vWF) targeting agent. The nanoparticles may be liposomes or micelles optionally loaded with an anti-angiogenesis drug. The anti-angiogenesis drug, may be for example, avastin or any other drug that is currently used for treating a retinal disease and in particular retinopathy or diabetic retinopathy. In some embodiments, the micelle may be PEG-PLA micelle. In some embodiments it may be conjugated to anti-angiogenesis drug such as for example, TNP-470.

EXAMPLES

Abbreviations:

• GNR - gold nano-rod;

• vWF - von willebrand factor;

• VBP - VWF targeting protein.

• ROI - region of interest.

• DR - diabetic retinopathy.

Example 1

[00096] Bonding efficiency of difference VBP and VBP conjugates is calculated using molecular simulations software YASARA. VBP is selected for experiments based on their bonding efficiency under shear forces. Finite element analysis software, COMSOL is used to calculate the fluid dynamics profile of a simulated microaneurysm. The ability of the VBP to bind and its accumulation with time are modeled to predict optimal dosing regimen. Microfluidic chip is deigned and fabricated using soft lithography based on the size and geometry of common microaneurysms. The device is coated with gelatin and seeding with microvascular endothelial cells. Expansion and loss of shear forces lead specific endothelial region to dysfunction upregulated VWF. Nanoparticles will be perfused into the chip using a syringe pump in physiological shear forces and their ability to selectively bind endothelial cells is experimentally validated. NIR radiation is used to heat the nanoparticles, selectively killing the endothelial cells to which high densities of VBP-nanoparticles bound.

Simulating temperature around a heated micro-aneurysm:

[00097] Given the temperature of the aneurysm wall (generated by heated GNRs that are bound to the VWF), the temperature of the surrounding volume (blood) can be simulated using heat & fluid flow simulations.

[00098] Such a simulation is presented in figure 1 - a model of a 2d blood vessel (30 [pm] diameter) with a half circle aneurysm (30 [pm] radius) is introduced to a 5 mm/s flow at its inlet (left side of model). The model walls are kept at a temperature of 310 [degrees kelvin] as the aneurysm surface is heated to 370 [degrees kelvin]

[00099] The resulting steady-state temperature field is presented and it appears that due to the flow pattern (relatively low velocities inside the aneurysm, as demonstrated at the bottom of figure 1 ) the heating is localized (i.e, the aneurysm wall is mainly heated, as demonstrated at the upper section of figure 1 ) and encompasses the entire aneurysm volume. The simulation predicts targeted heating while maintaining steady blood flow.

Example 2

Simulating radiation inside the eye and temperature of GNRs and surrounding tissue:

[000100] Using the physical properties of GNRs and eye tissue, a simulation of the radiation pattern inside the eye can predict the temperature distribution around the GNRs.

[000101 ] The simulation consists of a 2d eye model with a rod-shaped feature at the back side of the eye (showing at the right side of each part of figure 2), the eye walls are kept at 310 [degrees kelvin] and radiation is entering into the eye from the pupil (on the left side). As demonstrated at the bottom of figure 2, the applied radiation transforms into localized heat, i.e. only the rod-shaped feature was heated. The simulation predicts localized ablation, while keeping the surrounding unheated.

Example 3

NIR heating

[000102] A 1 pi GNR droplet was placed on a rubber surface. A pulsating NIR radiation was applied. The results were measured using a thermal camera for recording the temperature of the surface in time. As can be depicted in figure 3B, the droplet was heated to 100°C within 2 seconds. Figure 3A demonstrates a local heating, while surrounding remained unheated.

Example 4

[000103] In order to determine the radiation profile (power, time, pulsed radiation effects) needed for localized heating, the following experiment is conducted:

1 . Different concentrations of GNRs are placed inside the tissue (pig's retina). 2. ROI is radiated with varying radiation profiles for optimization; a control experiment will be done with GNR-free tissue with the same radiation profiles.

3. The results are measured using a thermal camera for recording the temperature of the ROI in time.

Example 5

[000104] The concluded concentration of GNRs from example 4 is injected into a pathological animal retina and further radiated according to the profile determined in example 4. The experiment is conducted by using an animal model with retinal pathology similar to the one seen in DR. Preferably the experiment is carried out in dog models that appear most similar to human DR. Surprisingly, nonhuman primates are relatively resistant to induced DR. Mice aged 8-10 weeks can be given a single dose of alloxan to induce diabetes. It was previously believed that the alloxan-induced diabetic mouse did not develop cellular or vascular lesions, but a recent study found that alloxan does induce microaneurysms with increased acellular capillaries by 21 days in mice from the FOT_FB strain. Mice are introduced with VBP-bound nanoparticles bound to a fluorescent marker (e.g. FITC) by IV injection. Accumulation of fluorescence marker in the retina is assessed by fluorescence angiography. Heating is introduced using a near infrared diode in wide angle. Coagulation is assessed through OCR as well as histology analysis post mortem.

Example 6

Low temperature ablation of micro-aneurysm

[000105] After the GNRs are localized at the correct location and heated, the ablation of the micro-aneurysm is conducted.

[000106] A micro-fluid channel resembling a typical retinal blood vessel with an aneurysm is created. The model has blood flowing through it and the aneurysm is heated until the desired response is reached (ablation), thus determining the radiation/GNR concentration needed (it is speculated that the temperature will be around.

[000107] While certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes, and equivalents will now occur to those of ordinary skill in the art. It is, therefore, to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention.