WO2020120970A1 - Procédés pour améliorer l'imagerie médicale basée sur le vert d'indocyanine et la photothérapie - Google Patents

Procédés pour améliorer l'imagerie médicale basée sur le vert d'indocyanine et la photothérapie Download PDF

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WO2020120970A1
WO2020120970A1 PCT/GB2019/053522 GB2019053522W WO2020120970A1 WO 2020120970 A1 WO2020120970 A1 WO 2020120970A1 GB 2019053522 W GB2019053522 W GB 2019053522W WO 2020120970 A1 WO2020120970 A1 WO 2020120970A1
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icg
icgs
tissue
qds
conjugated
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Joe BROUGHTON
Imad Naasani
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Nanoco Technologies Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
    • A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6851—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell
    • A61K47/6855—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell the tumour determinant being from breast cancer cell
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
    • A61K47/6923—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being an inorganic particle, e.g. ceramic particles, silica particles, ferrite or synsorb
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
    • A61K49/0032—Methine dyes, e.g. cyanine dyes
    • A61K49/0034—Indocyanine green, i.e. ICG, cardiogreen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0063—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres
    • A61K49/0069—Preparation for luminescence or biological staining characterised by a special physical or galenical form, e.g. emulsions, microspheres the agent being in a particular physical galenical form
    • A61K49/0089—Particulate, powder, adsorbate, bead, sphere
    • A61K49/0091—Microparticle, microcapsule, microbubble, microsphere, microbead, i.e. having a size or diameter higher or equal to 1 micrometer
    • A61K49/0093—Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle

Definitions

  • This invention relates generally to methods for enhancing the performance of indocyanine green imaging and photodynamic therapy.
  • Photodynamic therapy is a treatment that uses a photosensitive drug, called a photosensitizer (PS), along with light to kill undesirable cells including precancerous and cancer cells. PDT drugs only work after they have been activated by light. Upon irradiation with appropriate light, the photosensitizer produces reactive oxygen species (ROS) for the destruction of the undesired tissue such as, in particular, neoplastic tissue.
  • ROS reactive oxygen species
  • ICG Indocyanine green
  • NIR near-infrared
  • ICG has been employed clinically to locate sentinel lymph node metastases in breast cancer as well as in ophthalmic angiography and coronary artery blood flow evaluation.
  • ICG-PDT has been FDA approved in the US as a contrast agent in retinal and choroidal vascular imaging, and in liver function tests since 1956 because of its low toxicity and low incidence of side effects.
  • ICG-PDT is used to evaluate liver function and for intraoperative local diagnosis of hepatocellular carcinoma (HCC) by fluorescence imaging.
  • HCC hepatocellular carcinoma
  • ICG-PDT methicillin-resistant Staphylococcus aureus
  • MRSA methicillin-resistant Staphylococcus aureus
  • ICG photodynamic therapy
  • PDT photodynamic therapy
  • pancreatic adenocarcinoma cells pancreatic adenocarcinoma cells.
  • Cytotoxicity was hypothesized to be primarily due to an oxidative reaction mechanism, and secondarily because of production of heat as a by-product.
  • antitumor activity of ICG-PDT has been variously reported as involving the generation of singlet oxygen species the yield of l 02 has been found to be extremely low due to a low yield of excited triplet states ( ⁇ 10 -5 ). This, coupled with a short blood half-life of 2-4 min, poor photo- and thermal-stability, non-specific binding with proteins, and susceptibility to aggregation, has limited the application of ICG-PDT.
  • Embodiments disclosed describe methods for enhancing the performance of ICG-PDT in which quantum dot nanoparticles (QDs) are conjugated to ICG or an ICG derivative (for example, tethered to the QD surface).
  • QDs quantum dot nanoparticles
  • Embodiments disclosed include quantum dot nanoparticles, wherein each QD is bonded (e.g., covalently bonded or physically bonded by, for example, ion pairing or van der Waals interactions) to ICG, e.g., by aliphatic chains, p-p stacking, p interactions, an amide, ester, thioester, or thiol anchoring group directly on an inorganic surface of the QD, or on an organic corona layer that is used to render the QD water-soluble and biocompatible.
  • Water- soluble QDs include a core of one semiconductor material and at least one shell of a different semiconductor material (core/shell particle).
  • the water-soluble QD includes an alloyed semiconductor material having a bandgap value that increases outwardly by compositionally graded alloying.
  • Such embodiments are useful, for example, to improve and enhance existing uses of ICG-PDT and for ex vivo and in vivo visualization and treatment of cancer, for visualization and treatment of bacterial infections, and visualization and treatment of retinal vascular proliferation among others.
  • each QD comprises: a core semiconductor material, and an outer layer, wherein the outer layer comprises a corona of organic coating (a functionalization organic coating) to render the particles water soluble and biocompatible, and ICG or an ICG derivative.
  • each QD comprises one or more shells of semiconductor material, the outer shell comprising an outer layer, wherein the outer layer comprises a corona of an organic coating (a functionalization organic coating) to render the particles water-soluble and biocompatible, and ICG or an ICG derivative.
  • each QD comprises: an alloyed quantum dot and ICG or an ICG derivative. In at least one embodiment, each QD comprises: a doped quantum dot and ICG or an ICG derivative. In at least one embodiment of any of the QDs described herein, the nanoparticle comprises a II- VI material, a III-V material, or I-III-IV material, or any alloy or doped derivative thereof.
  • any of the QDs described herein are associated with an emission spectrum ranging from about 600 nm to about 900 nm and further from about 750 nm to about 850 nm.
  • any of the QDs described herein may further comprise a cellular uptake enhancer, a tissue penetration enhancer, or any combination thereof.
  • cellular uptake enhancers include, for example, trans-activating transcriptional activators (TAT), Arg-Gly-Asp (RGD) tri-peptides, linear and cyclic peptides including the RGD motif, or poly arginine peptides.
  • tissue penetration enhancers include but are not limited to saponins, cationic lipids, and Streptolysin O (SLO).
  • At least one target specific ligand is conjugated to a water- soluble non-toxic QD together with ICG.
  • targets to which the target specific ligands are specific include EGFR, PD-L1, PD-L2, HER2, CEA, CA19-9, CA125, telomerase proteins and subunits, CD20, CD25, CD30, CD33, CD52, CD73, CD109, VEGF-A, CTLA-4, and RANK ligand.
  • a method of inducing cell death is provided. In another embodiment, a method of inducing cell death and imaging affected tissues is provided.
  • a method of visualizing and treating tumors (both malignant and benign), inflammatory tissue, and/or undesired cells is provided.
  • the tumor is soft or solid.
  • the method of visualizing tumors and/or inflammatory tissue can be used for intraoperative imaging and fluorescence guided surgery of the tumors and/or inflammatory tissue.
  • Inflammatory tissues may include, for example, arthritis, Crohn’s disease, Inflammatory Bowel Disease, psoriasis, acne, multiple sclerosis, Alzheimer’s, and Parkinson’s Disease.
  • QD-ICG conjugates are used to detect atherosclerosis plaques, atheromatous lesions and stenosis levels, particularly those of the carotid artery.
  • the QD-ICG conjugates are used to treat atherosclerosis plaques, atheromatous lesions and stenosis levels, particularly those of the carotid artery.
  • a method of visualizing and treating circulating cells in blood or body fluids is provided.
  • a method of visualizing sentinel lymph nodes (SLNs) is provided.
  • a method of visualizing interstitial fluids of tissues is provided.
  • any of the methods described herein comprises i) contacting QD- ICG conjugates (e.g., a plurality or a panel of QD conjugates) according to any of the embodiments described herein with a cell, tumor or unwanted tissue, and (ii) exciting the QD- ICG conjugate to emit light and induce formation of reactive oxygen species (ROS).
  • a polymerizable ligand is affixed to the QD- ICG wherein the ligand is polymerized by excitation of the QDs with an energy source (e.g., a light source, such as a UV or visible light source).
  • the QD-ICG are excited using multi-photon excitation (e.g., a two-photon excitation).
  • multi-photon excitation e.g., a two-photon excitation
  • the combined energy of two or more light beams is used to excite a particular QD-ICG.
  • any of the methods described herein are performed in bodily fluids (e.g., blood, pancreatic secretions, plasma, fine needle aspirate) and/or tissues samples in vivo via administration of QD-ICG followed by administration of radiation at a wavelength the generates ICG fluorescence directly and/or at a wavelength that excites the QD to emit at a wavelength at induced ICG fluorescence.
  • bodily fluids e.g., blood, pancreatic secretions, plasma, fine needle aspirate
  • tissues samples in vivo via administration of QD-ICG followed by administration of radiation at a wavelength the generates ICG fluorescence directly and/or at a wavelength that excites the QD to emit at a wavelength at induced ICG fluorescence.
  • any of the methods described herein are performed in bodily fluids and/or tissues samples taken and examined ex vivo.
  • the detection of an emission signal can be performed on biological samples removed and tested ex vivo using fluorescence microscopy, flow cytometry or fluorimeters.
  • Fig. 1 depicts one embodiment of a scheme for derivatization of a water-soluble QD with ICG or an ICG derivative.
  • Fig. 2 graphs the fluorescence signal intensity (mV) versus retention time through the column of an original unmodified water-soluble cadmium-free QD versus QD-ICG at a fluorescence of 820 nm, and the absorbance of QD-ICG at approximately 278 nm versus retention time.
  • Fig. 3A - Fig. 3D depict the quantum yield (QY) of original unmodified water-soluble cadmium-free QD versus QD-ICG at different excitation wavelengths.
  • Fig. 3A depicts ICG at an excitation wavelength lec at 388 nm and an emission wavelength lah at 820 nm.
  • Fig. 3B depicts a QD-ICG conjugate at an excitation wavelength lec at 388 nm and an emission wavelength lah at 820 nm.
  • Fig. 3C depicts ICG at an excitation wavelength lec at 750 nm and an emission wavelength lah at 820 nm.
  • Fig. 3D depicts an QD-ICG conjugate at an excitation wavelength lec at 750 nm and an emission wavelength lah at 820 nm.
  • Fig. 4A and Fig. 4B depict the absorption and fluorescence spectra of ICG and QD- ICG.
  • Fig. 4A depicts the absorption spectrum of QD-ICG versus dilutions of ICG.
  • Fig. 4B shows the fluorescence spectra of the samples at different wavelengths, overlaid for direct comparison.
  • Fig. 5A - Fig. 5C depict characterization data for near-IR emitting AgInS2 (AIS) QDs.
  • Fig. 5A depicts dynamic light scattering (DLS) data showing a single peak with the greatest size distribution at 16.24 nm and a polydispersity index of 0.104.
  • Fig. 5B presents raw correlation data revealing a single drop, which means one particle size due to uniform particle movement.
  • Fig. 5C presents fluorescence emission peak data of the near-IR emitting AIS QD.
  • QD-ICG conjugates features high safety and biocompatibility profiles and methods for enhancing ICG-based medical imaging and phototherapy with the use of QDs.
  • biocompatible, non-toxic, fluorescent QDs are conjugated with ICG.
  • the QD-ICG conjugates are formed of semiconductor materials that are themselves toxic and contribute to the cytotoxicity of the system.
  • Embodiments disclosed herein relate to QDs conjugated to ICG, and in particular methods for enhancing ICG based medical imaging and phototherapy with the use of QDs.
  • Provided herein are methods for enhancing the performance of ICG by conjugation to QD, and using the fluorescent emission of the QD to excite the ICG dye in vitro and in vivo.
  • Certain attributes of this combination include that ICG tethered to QD enhances carcinoma specific uptake of ICG via antibodies.
  • emission by the ICG at 820 nm to 830 nm is maximized when QD and ICG are conjugated.
  • QD tunability allows for a range of excitation wavelengths beyond that of ICG alone.
  • CA125 Cancer Antigen-125 is also known as mucin 16 (MUC16)
  • CA19-9 Carbohydrate antigen 19-9 (CA19-9), a.k.a. cancer antigen 19-9 or sialylated Lewis (a) antigen
  • CD 109 Cluster of Differentiation 109 is a glycosyl phosphatidylinositol (GPI) - linked glycoprotein that is involved in transforming growth factor beta binding
  • CD20 CD20 is an activated-glycosylated phosphoprotein encoded by the MS4A1 gene and expressed on the surface of B-cells but not early pro-B cells or plasma cells.
  • CD25 CD25 is the interleukin-2 receptor alpha chain (IL2RA) protein
  • CD30 a.k.a. TNFRSF8 is a tumor necrosis factor receptor family protein
  • CD33 a.k.a. sialic acid binding Ig-like lectin 3 (SIGLEC-3)
  • CD73 Cluster of Differentiation 73 a.k.a. ecto-5 '-nucleotidase (NT5E)
  • CTLA-4 Cytotoxic T-lymphocyte-associated protein 4
  • EGFR HER1
  • HER2 Human Epidermal Growth Factor Receptor 2
  • PD-L1 Programmed death ligand- 1 (PD-L1), also known as cluster of differentiation 274 (CD274) or B7 homolog 1 (B7-H1).
  • CD274 cluster of differentiation 274
  • B7-H1 B7 homolog 1
  • the phrase“at least one of A, B and C,” means“at least one from the group A, B, C, or any combination of A, B and C.”
  • the phrase requires one or more, and not necessarily not all, of the listed items.
  • Indocyanine Green and Derivatives thereof.
  • Indocyanine green is a tricarbocyanine dye having a molecular weight of 751.4 Da. It is a negatively charged ion that belongs to the large family of cyanine dyes.
  • ICG Indocyanine Green
  • ICG indocyanine Green
  • cyanine derivatives thereof that fluoresce in response to red to near-infrared light (650-900 nm).
  • ICG has the CAS number 3599-32-4 and the IUPAC name sodium 4-[2-[( 1 A,3/y5/y7Z)-7-[ l , l -dimethyl-3-(4-sulfonatobutyl)benzo[T]indol-2-ylidene] hepta- 1 ,3 , 5-trienyl]- 1 , 1 -dimethylbenzo[e]indol-3 -ium-3 -yl]butane- 1 -sulfonate.
  • ICG The chemical formula of ICG is C 43 H 47 N 2 Na0 6 S 2 and the molecule has a molecular mass of 774.96 g/mol in sodium salt form. ICG has the following chemical structure in its sodium salt form:
  • ICG-Der-02-/V-hydroxysuccinimide ester which contains one carboxyl functional group for covalent conjugation to amino groups.
  • ICG derivative having an available COOH group for coupling is ICG bis-carboxylic acid, which has the following structure:
  • ICG derivatives include ICG derivatives including pendant alkyl chains, ICG with folate-polyethylene glycol, and hydrophilic derivatives such as l, l '-bis-(4-sulfobutyl) indotricarbocyanine-5,5'-dicarboxylic acid diglucamide monosodium salt (SIDAG).
  • SIDAG has the following structure:
  • ICG derivatives including various functional groups are commercially available from Iris Biotech GMBH including indocyanine green maleimide (ICG-Mal), indocyanine green NHS active ester (ICG-NHS), ICG-azide, indocyanine green 2- cyanobenzothiazole (ICG-CBT), indocyanine green tetrazine (ICG-Tz), indocyanine green dib enzoazacy cl oocty ne (ICG-DBCO), and indocyanine green alkyne (ICG-alkyne).
  • ICG-Mal indocyanine green maleimide
  • ICG-NHS indocyanine green NHS active ester
  • ICG-azide ICG-azide
  • ICG-CBT indocyanine green 2- cyanobenzothiazole
  • ICG-Tz indocyanine green tetrazine
  • ICG-DBCO indo
  • QDs are fluorescent semiconductor nanoparticles with unique optical properties. QDs represent a particular very small size form of semiconductor material in which the size and shape of the particle results in quantum mechanical effects upon light excitation. Generally, larger QDs such as having a radius of 5-6 nm will emit longer wavelengths in orange or red emission colors and smaller QDs such as having a radius of 2-3 nm emit shorter wavelengths in blue and green colors, although the specific colors and sizes depend on the composition of the QD. In certain embodiments disclosed herein the QDs emit in the red to near-infrared (near-IR) range of 700 850 nm. In particular embodiments the QDs emit in an ICG excitation wavelength range of 750 nm - 800 nm.
  • QDs shine around 20 times brighter and are many times more photo-stable than any of the conventional organic fluorescent dyes including ICG. Importantly, QD residence times are longer due to their chemical nature and nano-size. QDs can absorb and emit much stronger light intensities. In certain embodiments, the QD can be equipped with more than one binding tag, forming bi- or tri-specific nano-devices. The unique properties of QDs enable several medical applications that serve unmet needs.
  • QD precursors are provided in the presence of a molecular cluster compound under conditions whereby the integrity of the molecular cluster is maintained and acts as a well-defined prefabricated seed or template to provide nucleation centers that react with the chemical precursors to produce high quality nanoparticles on a sufficiently large scale for industrial application.
  • Suitable types of QDs useful in the present invention include, but are not limited to, core materials comprising the following types (including any combination or alloys or doped derivatives thereof):
  • IIA-VIA (2-16) material, incorporating a first element from group 2 of the periodic table and a second element from group 16 of the periodic table and also including ternary and quaternary materials and doped materials.
  • Suitable nanoparticle materials include, but are not limited to: MgS, MgSe, MgTe, CaS, CaSe, CaTe, SrS, SrSe, SrTe, BaS, BaSe, BaTe.
  • IIB-VA (12-15) material incorporating a first element from group 12 of the periodic table and a second element from group 15 of the periodic table and also including ternary and quaternary materials and doped materials.
  • Suitable nanoparticle materials include, but are not limited to: ZmP2, Zm As2, CM3P2, Cd3 As2, Cd3N2, ZmN2.
  • heavy metal-free IIB-VA materials are selected from ZmP2 and ZmN2.
  • Suitable nanoparticle materials include, but are not limited to: CdSe, CdTe, ZnS, ZnSe, ZnTe, ZnO, HgS, HgSe, HgTe, CdSeS, CdSeTe, CdSTe, ZnSeS, ZnSeTe, ZnSTe, HgSeS, HgSeTe, HgSTe, CdZnS, CdZnSe, CdZnTe, CdHgS, CdHgSe, CdHgTe, HgZnS, HgZnSe, HgZnTe, CdZnSeS, CdZnSeTe, CdHgSeS, CdHgS, C
  • IIIA-VA (13-15) material incorporating a first element from group 13 of the periodic table and a second element from group 15 of the periodic table and also including ternary and quaternary materials and doped materials.
  • Suitable nanoparticle materials include, but are not limited to: BP, A1P, AlSb; GaN, GaP, GaSb; InN, InP, InSb, AIN, and BN.
  • IIIA-IVA 13-14 material incorporating a first element from group 13 of the periodic table and a second element from group 14 of the periodic table and also including ternary and quaternary materials and doped materials.
  • Suitable nanoparticle materials include, but are not limited to: B 4 C, AI4C3, Ga 4 C, Si, SiC.
  • IIIA-VIA (13-16) material incorporating a first element from group 13 of the periodic table and a second element from group 16 of the periodic table and also including ternary and quaternary materials.
  • Suitable nanoparticle materials include, but are not limited to: AI 2 S 3 , AhSe3, AfTei, Ga2S3, Ga2Se3, GeTe; fii2S3, hi2Se3, Ga2Te3, fii2Te3, InTe.
  • IV A- VIA (14-16) material incorporating a first element from group 14 of the periodic table and a second element from group 16 of the periodic table, and also including ternary and quaternary materials and doped materials.
  • Suitable nanoparticle materials include, but are not limited to: PbS, PbSe, PbTe, Sb 2 Te 3 , SnS, SnSe, SnTe.
  • heavy metal-free the IV A- VI material is selected from SnS, SnSe, and SnTe.
  • VA-VIA (15-16) material incorporating a first element from group 15 of the periodic table and a second element from group 16 of the periodic table, and also including ternary and quaternary materials and doped materials.
  • a suitable nanoparticle material includes but is not limited to SbiC , Sb2S3, Sb2Se3, and Sb2Te3.
  • a IB-IIIA-VIA (11-13-16) material incorporating a first element from group 1 1 of the periodic table, a second element from group 13 of the periodic table, and a third element of group 16 of the periodic table including ternary and quaternary materials and doped materials.
  • An example of a suitable nanoparticle materials includes, but is not limited to: AgInS2, CuInS2, CuGaS 2 , CuInSe 2 , CuGaSe 2 .
  • the QDs is substantially free of heavy metals such as cadmium (e.g., contains less than 5 wt. %, such as less than 4 wt. %, less than 3 wt. %, less than 2 wt. %, less than 1 wt. %, less than 0.5 wt. %, less than 0.1 wt. %, less than 0.05 wt. %, or less than 0.01 wt.
  • heavy metals such as cadmium (e.g., contains less than 5 wt. %, such as less than 4 wt. %, less than 3 wt. %, less than 2 wt. %, less than 1 wt. %, less than 0.5 wt. %, less than 0.1 wt. %, less than 0.05 wt. %, or less than 0.01 wt.
  • Examples of cadmium-, lead- and arsenic-free nanoparticles include nanoparticles comprising semiconductor materials, e.g., ZnS, ZnSe, ZnTe, InP, InSb, A1P, A1S, AlSb, GaN, GaP, GaSb, AgInS2, AgInS2/ZnS (referring to a core of AgInS2 with a ZnS shell: this molecule may alternatively be referred to as ZnS-AgInS2), CuInS2, Si, Ge, and alloys and doped derivatives thereof, particularly, nanoparticles comprising cores of one of these materials and one or more shells of another of these materials.
  • semiconductor materials e.g., ZnS, ZnSe, ZnTe, InP, InSb, A1P, A1S, AlSb, GaN, GaP, GaSb, AgInS2, AgInS2/ZnS (referring to a core of AgInS2 with a
  • nanoparticles that include a single semiconductor material, e.g., CdS, CdSe, ZnS, ZnSe, InP, GaN, etc. may have relatively low quantum efficiencies because of non- radiative electron-hole recombination that occurs at defects and dangling bonds at the surface of the nanoparticles.
  • the nanoparticle cores may be at least partially coated with one or more layers (also referred to herein as“shells”) of a material different than that of the core, for example a different semiconductor material than that of the“core.”
  • the material included in the one or more shells may incorporate ions from any one or more of groups 2 to 16 of the periodic table.
  • each shell may be formed of a different material.
  • the core is formed from one of the materials specified above and the shell includes a semiconductor material of larger band-gap energy and similar lattice dimensions as the core material.
  • Exemplary shell materials include, but are not limited to, ZnS, ZnO, ZnSe, MgS, MgSe, MgTe and GaN.
  • a multi-shell nanoparticle is InP/ZnS/ZnO. The confinement of charge carriers within the core and away from surface states provides nanoparticles of greater stability and higher quantum yield.
  • doped nanoparticle for the purposes of specifications and claims refers to nanoparticles of the above and a dopant comprising one or more main group or rare earth elements, this most often is a transition metal or rare earth element, such as but not limited to zinc sulfide with manganese, such as ZnS nanoparticles doped with Mn + .
  • a transition metal or rare earth element such as but not limited to zinc sulfide with manganese, such as ZnS nanoparticles doped with Mn + .
  • a suitable nanoparticle material can incorporate a first element from any group in the transition metal of the periodic table and a second element from group 16 of the periodic table and also including ternary and quaternary materials and doped materials.
  • a I-III-VI material incorporates a first element from group 11 of the periodic table, a second element from group 13 of the periodic table and a third element from group 16 of the periodic table, and including quaternary, higher order and doped materials.
  • suitable nanoparticle materials include, but are not limited to: CuInS2, CuInSe2, CuGaS2, CuGaSe2, CuIn x Gai- x SySe2-y (0 ⁇ x ⁇ 1 and 0 ⁇ y ⁇ 2), AgInS2, AgInSe2, NiS, CrS and AgS.
  • light responsive QD-ICG conjugates utilize a water-soluble QD nanoparticle that is considered a“core only” nanoparticle formed of a semiconductor material but lacking an inorganic shell of a different semiconductor material.
  • Core only QDs are capable of light absorption but in some cases do not exert strong fluorescence emission and thus have been disfavored for purposes where light emission is the purpose of the QD.
  • a core/shell particle having a central region or“core” of at least one semiconductor composition buried in or coated by one or more outer layers or “shells” of distinctly different semiconductor compositions.
  • the core may be comprised of an alloy of In, P, Zn and S, optionally involving molecular seeding of indium- based QDs over a ZnS molecular cluster, followed by formation of a shell of ZnS.
  • light responsive QD-ICG conjugates utilize a water-soluble QD nanoparticle comprising an alloyed semiconductor material having a band gap value or energy (E g ) that increases outwardly by graded alloying in lieu of production of a core/shell QD.
  • the band gap energy (E g ) is the minimum energy required to excite an electron from the ground state valence energy band into the vacant conduction energy band.
  • the graded alloy QD composition is considered“graded” in elemental composition from at or near the center of the QD to the outermost surface of the QD, rather than formed as a discrete core overlaid by one or more discrete shell layers.
  • An example would be an Im- x Pi- y Zn x S y graded alloy QD wherein the x and y increase gradually from 0 to 1 from the center of the QD to the surface.
  • the band gap of the QD would gradually change from that of pure InP towards the center to that of a larger band gap value of pure ZnS at the surface.
  • the band gap of a nanoparticle is dependent on particle size, the bulk band gap of ZnS is wider than that of InP such that the band gap of the graded alloy would gradually increase from an inner aspect of the QD to the surface.
  • the coordination about the final inorganic surface atoms in any core, core-shell or core-multi shell nanoparticle may be incomplete, with highly reactive“dangling bonds” on the surface, which can lead to particle agglomeration. This problem is overcome by passivating (capping) the“bare” surface atoms with protecting organic groups, referred to herein as capping ligands or a capping agent.
  • the capping or passivating of particles prevents particle agglomeration from occurring but also protects the particle from its surrounding chemical environment and provides electronic stabilization (passivation) to the particles, in the case of core material.
  • the capping ligands may be but are not limited to a Lewis base bound to surface metal atoms of the outermost inorganic layer of the particle. The nature of the capping ligand largely determines the compatibility of the nanoparticle with a particular medium.
  • the capping ligand may be selected depending on desired characteristics.
  • capping ligands that may be employed include, but are not restricted to, mono- or polythiols, mono- or polycarboxyls, mono- or poly- primary, secondary or tertiary amines, phosphines, phosphine oxides, phosphonic acids, phosphinic acids, imidazoles, mono- or polyalcohols, thioethers, and calixarene groups.
  • calixarenes all of these capping ligands have head groups that can form anchoring centers for the capping ligands on the surface of the particle.
  • the body of the capping ligand can be aliphatic or aromatic.
  • the body of the capping ligand can be a linear or branched chain, cyclic, or a combination thereof.
  • the body of capping ligand can be made up of only carbon and hydrogen or can be substituted with one or more of, for example O, S, N, and P.
  • the capping ligand itself can be large, small, oligomeric or polydentate. The nature of the body of the ligand and the protruding side that is not bound onto the particle, together determine if the ligand is hydrophilic, hydrophobic, amphiphilic, negative, positive or zwitterionic.
  • the capping ligands are hydrophobic (for example, alkyl thiols, alkyl amines fatty acids, alkyl phosphines, alkyl phosphine oxides, and the like).
  • the nanoparticles are typically dispersed in hydrophobic solvents, such as toluene, following synthesis and isolation of the nanoparticles.
  • Such capped nanoparticles are typically not dispersible in more polar media. If surface modification of the QD is desired, the most widely used procedure is known as ligand exchange.
  • Lipophilic ligand molecules that coordinate to the surface of the nanoparticle during core synthesis and/or shelling procedures may subsequently be exchanged with a polar/charged ligand compound.
  • An alternative surface modification strategy intercalates polar/charged molecules or polymer molecules with the ligand molecules that are already coordinated to the surface of the nanoparticle.
  • QY quantum yield
  • a one-pot synthesis process may be employed to generate a ligand-capped QD as a modification of the molecular seeding process described in Example 1 herein. This may be achieved by gradually decreasing the amounts of an indium fatty acid such as indium myristate (In(MA)3) and a tris(trialkylsilyl) phosphine, such as tris(trimethylsilyl) phosphine ((TMS)3P) or a tris(triarlysilyl) phosphine, such as tris(triphenylsilyl) phosphine, added to the reaction solution to maintain particle growth, while adding increasing amounts of zinc and sulfur precursors during a process such as is described for generation of the“core” particle of Example 1.
  • an indium fatty acid such as indium myristate (In(MA)3)
  • a tris(trialkylsilyl) phosphine such as tris(trimethylsilyl) phosphine ((TMS
  • a dibutyl ester and a saturated fatty acid are placed into a reaction flask and degassed with heating. Nitrogen is introduced and the temperature is increased.
  • a molecular cluster compound such as for example a ZnS molecular cluster [Et3 H]4[ZnioS4(SPh)i6], is added with stirring.
  • the temperature is increased as graded alloy precursor solutions are added according to a ramping protocol that involves addition of gradually decreasing concentrations of a first semiconductor material and gradually increasing concentrations of a second semiconductor material.
  • the ramping protocol may begin with additions of In(MA)3 and (TMS) 3 P dissolved in a dicarboxylic acid ester (such as for example di-n- butylsebacate ester) wherein the amounts of added In(MA)3 and (TMS) 3 P gradually decrease over time to be replaced with gradually increasing concentration of sulfur and zinc compounds such as (TMS)2S and zinc acetate.
  • a dicarboxylic acid ester such as for example di-n- butylsebacate ester
  • QD particles of a desired size with an emission maximum gradually increasing in wavelength are formed wherein the concentrations of InP and ZnS are graded with the highest concentrations of InP towards a center of the QD particle and the highest concentrations of ZnS on an outer layer of the QD particle. Further additions to the reaction are stopped when the desired emission maximum is obtained and the resultant graded alloy particles are left to anneal followed by isolation of the particles by precipitation and washing.
  • the QDs are functionalized to present a hydrophilic outer layer or corona that permits use of the QDs in the aqueous environment, such as, for example, in vivo and ex vivo applications in living cells.
  • Such QDs are termed water-soluble QDs.
  • the capped QDs can be further modified by one of the standard methods for water-solubilization and surface functionalization using ligand exchange with thiol-containing ligands, amphiphilic ligands or imidazole-containing ligands as described in US8906699B2, Chan et al. “Quantum Dot Bioconjugates for Ultrasensitive Nonisotopic Detection” Science 281 (1998) 2016-2018, Walling et al. “Quantum Dots for Lice Cell and In Vivo Imaging” Int. J. Mol. Sci.
  • the surface modified particles according to the methods mentioned in the art can be equipped with carboxyl or amine groups to enable the use of standard linking agents (e.g., carbodiimides) to link ICG molecules to the surface of the particle.
  • standard linking agents e.g., carbodiimides
  • the fatty acid-capped QDs can also be modified by a multi-step reaction scheme including, but not necessarily limited to, 1) intercalating a hydrocarbon chain of a ligand interactive agent with alkyl chains of the fatty acid-capped QDs, 2) reacting a functional group (such as a carboxylic acid, carboxylic acid ester, alcohol, thiol, amine, or amide) of the ligand interactive agent with a linking/crosslinking compound such as, for example, a melamine or, more specifically, hexamethoxymethylmelamine (HMMM) together with a crosslinking agent that is able to contribute a reactive group such as for example a -COOH group to form covalent bonds therebetween, and 3) reacting previously unreacted portions of the linking/crosslinking compound with ICG or an ICG derivative and, optionally a surface modifying ligand such as, for example HO-PEG 2000 -OCH 3 .
  • a functional group such as a carboxylic acid,
  • the ligand interactive agent can be a C8-C20 fatty acid such as, but not limited to, carprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoelaidic acid, a-linoleic acid, arachidonic acid, and eicosapentaenoic acid.
  • carprylic acid capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoelaidic acid, a-linoleic acid, arachidonic acid
  • the ligand interactive agent can be a C8-C20 fatty acid ester such as, but not limited to, methyl-, ethyl-, propyl-, isopropyl-, butyl-, secbutyl-, and tertbutyl- esters forms of the listed C8-C20 fatty acids.
  • the ligand interactive agent can be a sterol or stenol such as, but not limited to nat-cholesterol, ent-cholesterol, ergosterol, campesterol, campestanol, brassicasterol, D5- or A7-avenasterol, and cycloartenol.
  • the ligand interactive agent can be a C8-C20 alcohol such as, but not limited to, 1-octanol, 1-decanol, 1- dodecanol, 1-teradecanol, 1-pentadecanol, 1-hexadecanol, and 1-octadecanol.
  • the ligand interactive agent can be a C8-C20 thiol such as, but not limited to, 1-octanethiol, 1-nonanethiol, 1-decanethiol, 1-undecanethiol, 1-dodecanethiol, 1- teradecanethiol, 1-pentadecanethiol, 1-hexadecanethiol, and 1-octadecanethiol.
  • the ligand interactive agent can be a C8-C20 amine such as, but not limited to, octylamine, decylamine, dodecylamine, tretradecylamine, pentadecylamine, hexadecylamine, and octadecylamine.
  • the ligand interactive agent can be a C8-C20 amide such as, but not limited to, octadecanamide, oleamide, 11-bromoundecanamide, 11- mercaptoundecanamide, 16-mercaptoundecanamide, and linoleamide.
  • the QDs may be surface equipped with a conjugation capable functionality (for example, COOH, OH, NH2, SH, azide, alkyne).
  • a water-soluble non-toxic QD is or becomes carboxyl functionalized.
  • the QD-COOH may be linked to ICG using a carbodiimide linking technology employing water-soluble 1 -ethyl-3 -(-3 -dimethylaminopropyl) carbodiimide hydrochloride (EDC).
  • EDC water-soluble 1 -ethyl-3 -(-3 -dimethylaminopropyl) carbodiimide hydrochloride
  • sulfo derivative of N-hydroxysuccinimide is added during the reaction with the primary amine bearing molecule.
  • the EDC couples NHS to carboxyls, forming an NHS ester that is more stable than the O-acylisourea intermediate while allowing for efficient conjugation to primary amines at physiologic pH. In either event, the result is a covalent bond between the QD and the amine bearing molecule.
  • Other chemistries like Suzuki-Miyaura cross-coupling, (succinimidyl 4-(N-maleimidomethyl) cyclohexane- 1-carboxylate) (SMCC), or aldehyde based reactions may alternatively be used.
  • the QDs may non-covalently bound to the surface by physisorption.
  • QD-ICG conjugates are prepared by incubating overnight in an aqueous solution including water-soluble cadmium-free QD prepared in accordance with Examples 1 and 2 together with ICG. After incubation excess ICG was removed by ultrafiltration.
  • the QD is“non-toxic” as defined as substantially free of toxic heavy metals such as cadmium, lead and arsenic (e.g., contains less than 5 wt. %, such as less than 4 wt. %, less than 3 wt. %, less than 2 wt. %, less than 1 wt. %, less than 0.5 wt. %, less than 0.1 wt. %, less than 0.05 wt. %, or less than 0.01 wt.
  • toxic heavy metals such as cadmium, lead and arsenic
  • % of heavy metals such as cadmium, lead and arsenic or is free of heavy metals such as cadmium, lead and arsenic.
  • reduced toxicity QDs that lack heavy metals such as cadmium, lead and arsenic are provided.
  • QDs enable several potential medical applications including unmet in vivo and ex vivo diagnostics in living cells.
  • One of the major concerns regarding the medical applications of QDs has been that the majority of research has focused on QDs containing toxic heavy metals such as cadmium, lead or arsenic.
  • the biologically compatible and water-soluble heavy metal-free QDs described in certain embodiments herein can safely be used in medical applications both in vivo and ex vivo.
  • in vivo compatible water-dispersible cadmium-free QDs are provided that have a hydrodynamic diameter of 10-30 nm (for comparison, within the range of the dimensional size of a full IgG2 antibody).
  • the in vivo compatible water-dispersible cadmium-free QDs are produced.
  • the in vivo compatible water-dispersible cadmium-free QDs are carboxyl functionalized and further derivatized with a ligand or a ligand binding moiety.
  • the ligands are selected from one or more of the group consisting of antibodies, streptavidin, nucleic acids, lipids, saccharides, drug molecules, proteins, peptides, and amino acids.
  • the detecting is used for imaging and detecting one or more of angiogenesis, tumor demarcation, tumor metastasis, diagnostics in vivo , and lymph node progression while in other aspects the detecting is used in one or more of immunochemistry, immunofluorescence, DNA sequence analysis, fluorescence resonance energy transfer (FRET), flow cytometry (FC), fluorescence activated cell sorting (FACS), and high- throughput screening.
  • FRET fluorescence resonance energy transfer
  • FC flow cytometry
  • FACS fluorescence activated cell sorting
  • At least one of the ligands has specificity for a target selected from the group consisting of EGFR, PD-L1, PD-L2, HER2, CEA, CA19-9, CA125, telomerase proteins and subunits, CD20, CD25, CD30, CD33, CD52, CD73, CD109, VEGF- A, CTLA-4, and RANK ligand.
  • a multi-ligand nano-device is provided having at least ICG and at least one target specific ligand.
  • the targeting ligand is an antibody.
  • antibody includes both intact immunoglobulin molecules as well as portions, fragments, and derivatives thereof, such as, for example, Fab, Fab', F(ab')2, Fv, Fsc, CDR regions, or any portion of an antibody that is capable of binding an antigen or epitope including chimeric antibodies that are bi-specific or that combine an antigen binding domain originating with an antibody with another type of polypeptide.
  • antibody thus includes monoclonal antibodies (mAh), chimeric antibodies, humanized antibodies, as well as fragments, portions, regions, or derivatives thereof, provided by any known technique including but not limited to, enzymatic cleavage and recombinant techniques.
  • antibody as used herein also includes single-domain antibodies (sdAb) and fragments thereof that have a single monomeric variable antibody domain (V H ) of a heavy-chain antibody.
  • sdAb which lack variable light (V L ) and constant light (C L ) chain domains are natively found in camelids (V H H) and cartilaginous fish (VNAR) and are sometimes referred to as“Nanobodies” by the pharmaceutical company Ablynx who originally developed specific antigen binding sdAb in llamas.
  • the modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method.
  • Cadmium- free nanoparticles readily degrade when methods such as the aforementioned ligand exchange methods are used to modify the surface of such cadmium-free nanoparticles. For example, attempts to modify the surface of cadmium-free nanoparticles have been observed to cause a significant decrease in the luminescence QY of such nanoparticles.
  • surface-modified cadmium-free nanoparticles with high QY are required.
  • the high QY cadmium-free water dispersible nanoparticles disclosed herein have a QY greater than about 20%, greater than about 25%, greater than about 30%, greater than about 35%, or greater than about 40%.
  • heavy metal-free semiconductor indium-based nanoparticles or nanoparticles containing indium and/or phosphorus are preferred.
  • non-toxic QD nanoparticles are surface modified to enable them to be water soluble and to have surface moieties that allow derivatization by exposing them to a ligand interactive agent to effect the association of the ligand interactive agent and the surface of the QD.
  • the ligand interactive agent can comprise a chain portion and a functional group having a specific affinity for, or reactivity with, a linking/crosslinking agent, as described below.
  • the chain portion may be, for example, an alkane chain.
  • functional groups include nucleophiles such as thiol groups, hydroxyl groups, carboxamide groups, ester groups, and a carboxyl groups.
  • the ligand interactive agent may, or may not, also comprise a moiety having an affinity for the surface of a QD.
  • moieties include thiols, amines, carboxylic acids or esters thereof, and phosphines. If the ligand interactive group does not comprise such a moiety, the ligand interactive group can associate with the surface of the nanoparticle by intercalating with capping ligands.
  • ligand interactive agents include Cx-20 fatty acids and esters thereof, such as for example myristic acid or isopropyl myristate, Cs-2o amines, Cs-2o amides, Cs-2o alcohols, and Cs-2o thiols.
  • the ligand interactive agent may be associated with a QD nanoparticle simply as a result of the processes used for the synthesis of the nanoparticle, obviating the need to expose nanoparticle to additional amounts of ligand interactive agents. In such case, there may be no need to associate further ligand interactive agents with the nanoparticle.
  • a QD nanoparticle may be exposed to a ligand interactive agent after the nanoparticle is synthesized and isolated. For example, the nanoparticle may be incubated in a solution containing the ligand interactive agent for a period of time.
  • Such incubation, or a portion of the incubation period, may be at an elevated temperature to facilitate association of the ligand interactive agent with the surface of the nanoparticle.
  • the QD nanoparticle is exposed to a linking/crosslinking agent and a surface modifying ligand.
  • the linking/crosslinking agent includes functional groups having specific affinity for groups of the ligand interactive agent and with the surface modifying ligand.
  • the ligand interactive agent-nanoparticle association complex can be exposed to a linking/crosslinking agent and surface modifying ligand sequentially.
  • the nanoparticle might be exposed to the linking/crosslinking agent for a period of time to effect crosslinking, and then subsequently exposed to the surface modifying ligand to incorporate it into the ligand shell of the nanoparticle.
  • the nanoparticle may be exposed to a mixture of the linking/crosslinking agent and the surface-modifying ligand thus effecting crosslinking and incorporating surface modifying ligand in a single step.
  • the kem of the water soluble QD is tuned to emit at red to near- infrared (near-IR) wavelengths that maximally excite the ICG, that is, excitation wavelengths of 600 to 900 nm with peak ICG absorption around 750 nm - 800 nm. Exciting the QD induces it to emit light at a peak ICG absorption wavelength that in turn excites the ICG.
  • near-IR near- infrared
  • the QD is an AgInS2 (AIS) QD or a AgInS2/ZnS (ZAIS) QD that is rendered water-soluble as described in Examples 1 - 2 (in reference to InPZnS QD) by including one or more Cx to C20 saturated or unsaturated fatty acids during QD synthesis, which serve as capping ligands on the surface of the QD.
  • AIS AgInS2
  • ZAIS AgInS2/ZnS
  • Suitable fatty acids include but are not limited to lauric acid (C12), myristic acid (C14), palmitic (Ci6), stearic acid (Cis), oleic acid (Cis), and linoleic acid (Ci 8 ) together with indium salts such as for example indium laurate, indium myristate, indium palmitate, indium stearate, indium oleate, etc.
  • AIS and ZAIS QDs are characterized by a very broad emission peak of 750 nm and full width at half maximum (FWHM) of 200 nm, thus encompassing the excitation wavelengths of ICG of 600 to 900 nm with peak ICG absorption around 760 nm - 800 nm.
  • ZAIS QD are used because the excitation wavelengths in the near-infrared have increased tissue permeation (2-3 mm) of excitation and emission light leading to more effective detection.
  • One non-limiting preparation scheme for ZAIS QD precursor molecules would be as described in T. Torimoto, et al. “Facile Synthesis of ZnS-AgInS2 Solid Solution Nanoparticles for a Color-Adjustable Luminophore” J. Am. Chem. Soc., 2007, 129 (41), pp 12388-12389.
  • near-infrared emitting QD are core shell QD that may include toxic heavy metals coated in the ZnS shell to render it non-toxic.
  • One non-limiting example would be an A-acetyl-L-cysteine (NAC) capped CdHgTe/CdS core/shell QDs with an additional ZnS shell to form a CdHgTe/CdS/ZnS core/shell/shell QD. See e.g.
  • any of the QDs described herein include a first layer including a first semiconductor material provided on the nanoparticle core.
  • a second layer including a second semiconductor material may be provided on the first layer.
  • the ICG is non-covalently bound to the QD as described in Example 5.
  • linkers are used to connect ICG or an ICG derivative to the QD.
  • Known linkers such as a thiol anchoring groups directly on the inorganic surface of the QDs may be used. Standard coupling conditions may be employed and will be known to a person of ordinary skill in the art.
  • suitable coupling agents include, but are not limited to, carbodiimides, such as dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), and l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC).
  • DCC dicyclohexylcarbodiimide
  • DIC diisopropylcarbodiimide
  • EDC l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
  • the coupling agent is EDC.
  • QDs bearing a carboxyl end group, a diamine, and an ICG derivative having, for example a reactive -COOH or -OH group may be mixed in a solvent.
  • a coupling agent such as EDC, may be added to the mixture.
  • the reaction mixture may be incubated. Standard incubation conditions for coupling can be employed.
  • the coupling conditions may be a solution in the range of 0.5 to 4 hours.
  • the temperature range of the coupling conditions may be in the range of 0° C to 200° C.
  • the coupling conditions may be constant or varied during the reaction.
  • the reaction conditions may be 130° C for one hour then raised to 140° C for three hours.
  • the crude polymerizable ligand nanoparticle conjugate may be subject to purification and/or isolation. Standard solid-state purification methods may be used. Several cycles of filtering and washing with a suitable solvent may be necessary to remove excess unreacted functionalized ligand and/or coupling agents.
  • Fig. 1 depicts one embodiment of a scheme for derivatization of water-soluble QD with ICG wherein the ICG is covalently bound to ligand interactive agents that intercalate between C8-C20 capping ligands of the QD.
  • cholesterol is the ligand interactive agent having a HMMM reactive -OH group that is reacted with HMMM and salicylic acid to provide -COOH groups available for EDC conjugation.
  • EDC conjugation is employed by reacting the -COOH group located on the QD surface at room temperature with approximately 4% EDC in alkaline buffer for approximately 2 hours to produce the unstable intermediate o- acylisourea active ester.
  • a diamine is added and a stable amide bond is formed between one of the -ME groups of the diamine and the -COOH group located on the QD surface.
  • the second, remaining, -ME can then be reacted with an ICG derivative having a reactive -COOH or -OH group, forming an amide bond therebetween, and isourea side product.
  • Suitable diamines include methanediamine, ethylenediamine, propylenediamine, butylenediamine, pentylenediamine, hexylenediamine, heptylenediamine, octylenediamine, meta-, ortho- or para-phenylene diamine, and ME-PEG-ME and derivatives thereof including ME-COMEPEG-ME . If ICG derivative having a reactive -COOH or -OH group is utilized, it is preferred to use a non-aqueous solvent such as, for example, dichloromethane.
  • EDC conjugation between the ICG-COOH and - COOH groups on a water-soluble QD is conducted by incubation at room temperature with approximately 4% EDC in alkaline buffer for approximately 2 hours to produce the unstable intermediate o-acylisourea active ester, refluxing for 2 hours in the presence of H + or OH to convert the ester into the carboxylic acid.
  • a diamine is added and a stable amide bond is formed between one of the -ME groups of the diamine and the -COOH group located on the QD surface.
  • the second, remaining, -ME can then be reacted with an ICG derivative having a reactive -COOH or -OH group, forming an amide bond therebetween, and isourea side product.
  • Suitable diamines again include methanediamine, ethylenediamine, propylenediamine, butylenediamine, pentylenediamine, hexylenediamine, heptylenediamine, octylenediamine, meta-, ortho- or para-phenylene diamine, and ME-PEG-ME and derivatives thereof including ME-COMEPEG-ME .
  • EDC conjugation is employed by reacting the a -COOH group located on the QD surface at room temperature with approximately 4% EDC in alkaline buffer for approximately 2 hours to produce the unstable intermediate o-acylisourea active ester.
  • a compound having a terminal amine and a carboxylic acid, such as an amino acid or NEE-PEG- COOH is added and a stable amide bond is formed between the -NEE group of the amine/carboxylic acid compound and the -COOH group located on the QD surface.
  • the carboxylic acid of the amine/carboxylic acid compound can then be reacted with an ICG derivative having a reactive -COOH or -OH group, forming an ester bond therebetween, and isourea side product.
  • Any of the above processes can further comprise isolating the QD-ICG conjugate, purifying the QD-ICG conjugate or a combination thereof.
  • QD-ICG Therapeutics The delivery of QD-ICG conjugates is applied to multiple purposes.
  • the QD-ICG conjugates are applicable to photodynamic therapy (PDT).
  • PDT photodynamic therapy
  • the QD-ICG When physically excited, for example, by photons, the QD-ICG are expected to produce reactive oxygen species (ROS), which lead to cell death.
  • ROS reactive oxygen species
  • ICG was prone to photobleaching and signal disappearance such that the ROS pathway would be insufficiently activated for use in ablative procedures. For example, as discussed above, certain tumor types could not be treated by PDT using ICG.
  • the presently disclosed invention provides for treatment and imaging of various abnormal proliferative tissue growths, cancers and precancerous conditions, such as, for example, cancers like gliomas, bladder cancer, melanomas, esophageal cancer, endobronchial cancer, cancers of the lung, bladder, prostate, bile duct, stomach, mouth, throat, larynx, cervix, vagina, and vulva.
  • cancers like gliomas, bladder cancer, melanomas, esophageal cancer, endobronchial cancer, cancers of the lung, bladder, prostate, bile duct, stomach, mouth, throat, larynx, cervix, vagina, and vulva.
  • the PDT with QD-ICG conjugates disclosed herein is utilized in treatment of cancerous skin cancers including melanoma, basal and squamous cell carcinoma and in treatment of precancerous lesions of the skin (including actinic keratosis).
  • proliferative inflammatory diseases of the skin or other tissues including the gastrointestinal tract are treated using PDT with QD-ICG conjugates as disclosed herein.
  • the presently disclosed improvements in ICG-PDT provide therapy using treatment with QD-ICG conjugates for various indications where ICG alone is not effective or marginally effective.
  • the QDs disclosed herein are conjugated to tumor-specific ligands in addition to ICG for tumor-specific targeting of the QD-ICG conjugates.
  • QD-ICG conjugates result in greater generation of detectable fluorescence and are able to sustain a higher emission for a longer period of time, they can be used to as a marker (or label) in fluorescence guided surgery, treatment and imaging of various abnormal proliferative tissue growths, cancers and precancerous conditions, imaging of sentinel lymph nodes (SLNs), the imaging of interstitial fluids of tissues, as well as in treatment of antibiotic resistant bacteria.
  • SSNs sentinel lymph nodes
  • QD-ICG conjugates according the disclosure have been found to sustain emission for a period in excess of three months in aqueous solution, where ICG alone is capable of sustained emission for only about 24 hours in an aqueous solution.
  • the administration of the QD-ICG in embodiments disclosed herein can be enteral or parenteral.
  • the QD-ICG can be administered subcutaneously, intravenously, intramuscular, topically, and orally in various embodiments. Examples include bolus injection or IV infusions.
  • the QD-ICG conjugates will be applied to the tissue being treated and allowed to become absorbed onto or internalized into cells or concentrated in target tissues prior to irradiation. This period will be determined empirically depending on the tissue being treated.
  • the QD-ICG conjugates will be administered 10 to 20 hours prior to irradiation.
  • QD-ICG conjugates will be administered 14 to 18 hours prior to irradiation.
  • QD-ICG conjugates are formulated and packaged in a unit dose volume to be administered in a single procedure, such as through the skin and into a tumor tissue.
  • the volume per unit dose is determined on the basis of the anatomy of the administration site as well as the desired distribution area.
  • the QD-ICG includes a targeting moiety such as for example a tumor marker specific monoclonal antibody to target the tumor, as an ICG delivery system (QD+ICG+mAb).
  • a targeting moiety such as for example a tumor marker specific monoclonal antibody to target the tumor, as an ICG delivery system (QD+ICG+mAb).
  • QD+ICG+mAb an ICG delivery system
  • QD residence times are longer due to their chemical nature and nano-size.
  • QDs can absorb and emit much stronger light intensities thus improving ease of detection.
  • QDs also offer tunability of emission so that spatial imaging is enabled with minimum background from autofluorescence interference.
  • each QD can be equipped with more than one binding tag, forming multi- specific nano-devices such as, without limitation, bi- or tri- specific nano-devices with maximum binding probability, and thus highest detection efficiency.
  • the unique properties of QDs enable several medical applications that serve unmet needs in in vitro and in vivo diagnostics, clinical imaging, targeted drug delivery, and photodynamic therapy.
  • the conjugates are thus“theranostic” nano devices with multimodal properties useful for the imaging and treatment of cancer.
  • the disclosed theranostic nano-devices have imaging and therapeutic capabilities to be used for pre-, intra-, and post-operative detection and therapy of cancer.
  • the QD+ICG+mAb is manufactured for use as a medicament for detecting and treating cancer, wherein the medicament is utilized in preoperative or intraoperative diagnosis with minimally invasive endoscopy or laparoscopy.
  • the QD+ICG+mAb is adapted for injection into the circulation or into the abdomen and after administration is allowed to concentrate in tumor cells relative to normal cells.
  • the QD+ICG+mAb is adapted for concentration in tumor cells relative to normal cells by surface modification with a monomethoxy polyethylene oxide.
  • the concentrated QD+ICG+mAb can be induced to fluoresce by exposure to a QD excitatory light source applied endoscopically or laparoscopically, said fluorescence detectable by an imaging camera.
  • the fluorescence may be utilized to provide therapeutic modulation by the spectral emission from the ICG and the QD including through the generation of singlet oxygen and/or heat.
  • the included ICG can be excited with 800 nm (tissue penetrating) radiation, for cases deemed inoperable due to proximity to blood vessels or other delicate structures.
  • the embodiments disclosed herein provide enhanced ICG based labeling and imaging of tissues, enhanced photodynamic therapy approaches using ICG and enhanced deeper access to tissues treated with ICG.
  • Forms of administration may include preparations for parenteral administration by subcutaneous, intradermal, intramuscular or intravenous administration (e.g., injectable administration) such as sterile suspensions or emulsions. Gastrointestinal routes of administration may also be employed such as for gastrointestinal cancerous and precancerous conditions such as for example Barrett’s Esophagus.
  • Compositions may be topically administered in an admixture with a suitable carrier, diluent, or excipient such as sterile water, physiological saline, glucose, or the like.
  • compositions can contain auxiliary substances such as wetting or emulsifying agents, pH buffering agents, adjuvants, gelling or viscosity enhancing additives, preservatives, flavoring agents, colorants, and the like, depending upon the route of administration and the preparation desired.
  • auxiliary substances such as wetting or emulsifying agents, pH buffering agents, adjuvants, gelling or viscosity enhancing additives, preservatives, flavoring agents, colorants, and the like, depending upon the route of administration and the preparation desired.
  • Standard pharmaceutical texts such as “Remington’s Pharmaceutical Sciences,” 1990 may be consulted to prepare suitable preparations, without undue experimentation.
  • Suitable pharmaceutically acceptable carriers include, but are not limited to, sterile water, saline, glucose, dextrose, or buffered solutions. Carriers may include auxiliary agents including, but not limited to, diluents, stabilizers, sugars and amino acids, preservatives, wetting agents, emulsifying agents, pH buffering agents, viscosity enhancing additives, colorants and the like.
  • the QD-ICG conjugate is administered parenterally and allowed to circulate until the QD-ICG has concentrated in the tumor.
  • Particulates such as QDs are expected to accumulate in the vasculature of tumors after repeated passes through the circulation because the spongey vasculature of tumors is known to trap particulates in circulation to levels higher than those existing systemically. This phenomenon is known as Enhanced Permeability and Retention effect (EPR).
  • the QDs include polyethylene glycol (PEG) moieties that reduce removal of the QDs by the reticuloendothelial system as they circulate such that the QDs are allowed to accumulate in the tumor.
  • PEG polyethylene glycol
  • the loaded drug is released by administering light into the local environment of the target tissue either by open or closed procedures.
  • the tumor is an intra-abdominal tumor and the light source is introduced into the abdomen endoscopically.
  • the presently disclosed invention provides for treatment and imaging of various abnormal proliferative tissue growths, cancers and precancerous conditions, such as, for example, cancers like gliomas, bladder cancer, melanomas, esophageal cancer, endobronchial cancer, cancers of the lung, bladder, prostate, bile duct, stomach, mouth, throat, larynx, cervix, vagina, and vulva.
  • the PDT with QD-ICG conjugates disclosed herein is utilized in treatment of cancerous skin cancers including melanoma, basal and squamous cell carcinoma and in treatment of precancerous lesions of the skin (including actinic keratosis).
  • the QD-ICG conjugate is injected directly into the tumor tissue and ICG is excited by administering light into the local environment of the target tumor either by open or closed procedures.
  • a molecular seeding process was used to generate non-toxic QDs. Briefly, the preparation of non-functionalized indium-based quantum dots with emission in the range of 500 - 720 nm was carried out as follows: Dibutyl ester (approximately 100 ml) and myristic acid (MA) (10.06 g) were placed in a three-neck flask and degassed at ⁇ 70°C under vacuum for 1 h. After this period, nitrogen was introduced and the temperature was increased to ⁇ 90°C. Approximately 4.7 g of a ZnS molecular cluster [Et3 H]4[ZnioS4(SPh)i6] was added, and the mixture was stirred for approximately 45 min.
  • Dibutyl ester approximately 100 ml
  • MA myristic acid
  • the particles were isolated by the addition of dried degassed methanol (approximately 200 ml) via cannula techniques. The precipitate was allowed to settle and then methanol was removed via cannula with the aid of a filter stick. Dried degassed chloroform (approximately 10 ml) was added to wash the solid. The solid was left to dry under vacuum for 1 day. This procedure resulted in the formation of indium-based nanoparticles on ZnS molecular clusters. In further treatments, the quantum yields of the resulting indium-based nanoparticles were further increased by washing in dilute hydrofluoric acid (HF).
  • HF dilute hydrofluoric acid
  • the resulting particles were isolated by adding 40 ml of anhydrous degassed methanol and centrifuging. The supernatant liquid was discarded, and 30 ml of anhydrous degassed hexane was added to the remaining solid. The solution was allowed to settle for 5 h and then centrifuged again. The supernatant liquid was collected and the remaining solid was discarded.
  • the QYs of the final fatty acid-capped indium-based nanoparticle material ranged from approximately 60%-90% in organic solvents.
  • HMMM hexamethoxymethylmelamine
  • One example of preparation of a suitable water-soluble nanoparticle is provided as follows: 200 mg of cadmium-free QDs with red emission at 625 nm having as a core material an alloy comprising indium and phosphorus with Zn-containing shells, made using a procedure substantially as described in Example 1, were dispersed in toluene (1 ml) with isopropyl myristate (100 microliters). The isopropyl myristate is included as the ligand interactive agent. The mixture was heated at 50°C for about 1-2 minutes then slowly shaken for 15 hours at room temperature.
  • HMMM CYMEL 303, available from Cytec Industries, Inc., West Paterson, NJ
  • CH3O- PEG2000-OH 400 mg
  • salicylic acid 50 mg
  • the salicylic acid that is included in the functionalization reaction plays three roles, as a catalyst, a crosslinker, and a source for COOH. Due in part to the preference of HMMM for OH groups, many COOH groups provided by the salicylic acid remain available on the QD after crosslinking.
  • HMMM is a melamine-based linking/crosslinking agent having the following structure:
  • HMMM can react in an acid-catalyzed reaction to crosslink various functional groups, such as amides, carboxyl groups, hydroxyl groups, and thiols.
  • the mixture was degassed and refluxed at 130°C for the first hour followed by 140°C for 3 hours while stirring at 300 rpm with a magnetic stirrer. During the first hour a stream of nitrogen was passed through the flask to ensure the removal of volatile byproducts generated by the reaction of HMMM with nucleophiles. The mixture was allowed to cool to room temperature and stored under inert gas. The surface-modified nanoparticles showed little or no loss in fluorescence quantum yield and no change in the emission peak or full width at half maximum (FWHM) value, compared to unmodified nanoparticles. An aliquot of the surface-modified nanoparticles was dried under vacuum and deionized water was added to the residue.
  • FWHM full width at half maximum
  • the surface-modified nanoparticles dispersed well in the aqueous media and remained dispersed permanently. In contrast, unmodified nanoparticles could not be suspended in the aqueous medium.
  • the fluorescence QY of the surface-modified nanoparticles according to the above procedure is 40 - 50 %. In typical batches, a quantum yield of 47% ⁇ 5% is obtained.
  • cadmium- free QDs 200 mg
  • red emission at 625 nm were dispersed in toluene (1 ml) with cholesterol (71.5 mg).
  • the mixture was heated at 50° C. for about 1-2 minutes then slowly shaken for 15 hours at room temperature.
  • the monomethoxy polyethylene oxide (CTEO-PEG x -OH) and guaifenesin are optional as shown in Fig. 1.
  • the compound“guaifenesin” has the following chemical structure:
  • the mixture was degassed and refluxed at 140° C. for 4 hours while stirring at 300 rpm with a magnetic stirrer.
  • a stream of nitrogen was passed through the flask to ensure the removal of volatile byproducts generated by the reaction of HMMM with nucleophiles.
  • the mixture was allowed to cool to room temperature and stored under inert gas.
  • An aliquot of the surface-modified nanoparticles was dried under vacuum and deionized water was added to the residue.
  • the pH of the solution was adjusted to 6.5 using a 100 mM KOH solution and the excess non reacted material was removed by three cycles of ultrafiltration using Amicon filters (30 kD). The final aqueous solution was kept refrigerated until use.
  • the synthesis example provided here and as originally tested for proof of principal generates a water soluble QD with an emission wavelength (kem) of 630 nm due to a red shift from the original QD lah of 625 nm as a consequence of surface modification.
  • Different quantum dots/dye-doped nanomaterials are tunable to different excitation and emission bands.
  • a QD is utilized that emits in a red to near-infrared wavelength range of 700 - 800 nm.
  • an example of such an AIS QD having peak emission at around 750 nm and having an average dynamic light scattering (DLS) determined size of 18.63 nm with a standard deviation of 4.26 nm wherein about 10 % of the particles have a diameter of 16.24 nm.
  • the water-soluble fatty acid-capped QDs capped with an intercalated ligand interactive agent will have a DLS size of 14 - 30 nm and the QD will emit at 750 - 800 nm.
  • the surface-modified nanoparticles prepared as in this example also disperse well and remain permanently dispersed in other polar solvents, including ethanol, propanol, acetone, methylethylketone, butanol, tripropylmethylmethacrylate, or methylmethacrylate.
  • NanoSep 300K filters PALL NanoSep 300K Omega ultrafilters
  • the MES/EDC/Sulfo-NHS/QD solution was added to the NanoSep 300K filter and topped up 500 m ⁇ with MES. The filter was centrifuged at 5000 rpm/15 min.
  • the retained dots were re-dispersed in 50 m ⁇ activation buffer and transferred to an Eppendorf tube containing 10 m ⁇ of trastuzumab (HERCEPTIN ® , 100 mg/ml stock in a 25 mM solution of HEPES buffer, pH 8.5) + 40m1 HEPES, pH 8.5.
  • the solution was mixed well and incubated at RT overnight (around 16 - 18 hours).
  • the solution was quenched with 16 m ⁇ of 6-amino caproic acid (6AC) (19.7 mg/100 mM). Note that quenching could be alternatively conducted with other compounds having a primary amine, but 6AC was selected for this embodiment because it has a -COOH group and can maintain the colloidal stability of the product.
  • a polymerizable ligand is affixed to the QD-ICG wherein the polymerizable ligand is polymerized by excitation of the quantum dot nanoparticles with an energy source (e.g., a light source, such as a UV or visible light source).
  • an energy source e.g., a light source, such as a UV or visible light source.
  • Suitable polymerizable ligands include, but are not limited to, acrylates, methacrylates, diacetylene, cyanoacrylates, azide/alkyne pairs (click chemistry) and any combination thereof.
  • the polymerizable ligand is a methacrylate (e.g., 2- aminoethyl methacrylate) or a salt thereof, such as a hydrochloride salt.
  • Suitable acryl-based polymerizable ligands include, for example, methacryloyl-L-lysine, 4-methacryloxy-2- hydroxybenzophenone, and salts thereof, and any combination thereof.
  • the polymerizable ligand comprises acrylate and methacrylate ligands.
  • quantum dot nanoparticles comprising methacrylate ligands may be polymerized and crosslinked using excitation light to induce exciton formation that can in turn initiate acrylate polymerization.
  • light active monomers such as, e.g., methacryloyl-L- lysine, 4-methacryloxy-2-hydroxybenzophenone
  • the polymerizable ligand is a cyanoacrylate.
  • the polymerizable ligand is glycidyl cinnamate, or a derivative thereof.
  • the polymerizable ligand is a diacetylene, e.g., tricosa-10,12-diynoic acid.
  • Carboxy functionalized QDs are linked to 2-aminoethyl methacrylate hydrochloride using standard coupling chemistry with l-Ethyl-3-(3- dimethylaminopropyl)carbodiimide (EDC).
  • EDC l-Ethyl-3-(3- dimethylaminopropyl)carbodiimide
  • the resulting dots have pendant methacrylate groups that are delivered to the targeted tissue and polymerized by the excitations of the QDs with an energy source.
  • carboxy functionalized red QDs were linked to methacryloyl-L-lysine using standard EDC coupling chemistry.
  • the resulting QDs have pendant methacrylate groups that are polymerizable by UV/visible excitation at 300-500 nm. Fluorescence microscopy imaging at 1000 x magnification showed that when exposed to 320 nm UV, the nanoparticles aggregated, unlike the ones that were not irradiated.
  • the QDs can be delivered to the targeted tissue and polymerized by the excitations of the QDs with an energy source.
  • carboxy functionalized QDs were surface loaded with 4- methacryloxy-2-hydroxybenzophenone (Formula I) using hydrophobic interaction forces as follows. To an amount of 100 mg water-soluble QDs made in accordance with Examples 1 and 2 were dispersed in 1 mL H2O, a lOOpL solution of 4-methacryloxy-2-hydroxybenzophenone dissolved in DMSO at lOOmg/mL was added with vigorous mixing.
  • a small drop of the polymerizable QD preparation was mounted on a microscope slide, covered with a glass coverslip, and then irradiated for 5 minutes using a 6 Watt handheld UV lamp (UVP, LLC) at 365 nm wavelength.
  • a control slide was prepared in the same manner but was not irradiated. The slides were then examined using a fluorescence microscope. The irradiated sample showed significant aggregation (data not shown).
  • QD-ICG conjugates were prepared by incubating overnight a solution including 300 pL of water-soluble cadmium-free QDs, prepared substantially in accordance with Examples 1 and 2, at a concentration of 121.8 mg/mL in water together with 30 pL ICG (20 mg/mL in PBS) and 2 mL distilled H2O. After overnight, excess ICG was removed by adding 500 pL PBS 10 x and centrifuging @ 3500 rpm.
  • GPC gel permeation chromatography
  • QD-ICG graphs the fluorescence signal intensity (mV) and absorbance (at 278 nm) versus retention time through the column of the original unmodified water-soluble cadmium-free QDs.
  • QD-ICG at a fluorescence of 820 nm is compared with QD- ICG at an absorbance at approximately 278 nm.
  • the original unmodified QD (data shown in FIG. 2 as a dash-dot line) has an excitation wavelength kex at 450 nm and an emission wavelength kem at 630 nm with a fluorescence peak at 29 minutes.
  • the QD-ICG (data shown in Fig.
  • Quantum Yield The washed QD-ICG were analyzed for QY by using a 500 pL deionised water blank scan in a 1 mL vial, and measured between 550 - 850 nm. The sample was then spiked into that vial using a volume approximately 5 - 20 pL of sample and a measurement scan was run. Attenuator and gain settings were constant throughout. The results are shown in Fig. 3A -Fig. 3D. The result depicted in Fig. 3A with ICG at an excitation wavelength kex at 388 nm and an emission wavelength kem at 820 nm shows mostly noise at 820 nm. The result depicted in Fig.
  • Fig. 4A and Fig. 4B depict the absorbance and fluorescence spectra of ICG and QD-ICG at different wavelengths, overlaid for direct comparison.
  • Fig. 4A depicts the absorbance spectrum of QD-ICG versus ICG.
  • QD-ICG was measured using an ultraviolet-visible light spectrometer to determine the concentration of ICG in the sample.
  • the increasing absorbance peak height of ICG shows the increasing concentrations of ICG that were being detected by the instrument.
  • Fig. 4B shows the fluorescence spectra of the samples at different wavelengths, overlaid for direct comparison.
  • the sample prepared above in was also used to measure fluorescence, as the concentration of free ICG was matched to the QD-ICG sample, this means that a fair comparison could be made of the fluorescent emissions at different wavelengths.
  • the ICG absorbance peak, and hence concentration, is actually slightly higher than the QD-ICG peak.
  • the QD-ICG and the ICG samples were measured at two different wavelengths, 388 nm, which should favour the QD excitation, and 750 nm, which should favour the ICG excitation. At both wavelengths the QD-ICG sample gave higher ICG associated emission peaks.
  • the QD- ICG sample excited at 388 nm gave a significantly better signal than its free ICG counterpart when measured at 388 nm.
  • the 750 nm excitation source is longer than the excitation peak for the QD, but is the optimum for ICG.
  • the conjugated QD-ICG sample still yielded a higher emission peak, even though the there was a marginally higher concentration and the excitation favoured ICG. This suggests that the presence of QD has given a boost in performance to the ICG.
  • the fluorescence spectra where concentrations were matched by UV-Vis absorbance; the largest difference was between the QD-ICG and the ICG when tested at 388 nm, the signal peak was enhanced by 250%, from 400,000 to 1,000,000 cps.
  • the comparison at 750 nm were the highest, at 1.35 million and 1.54 million cps; with the QD-ICG giving the greatest emission, despite the ICG samples being more concentrated.
  • AIS near-IR emitting QD were also generated that have a PLmax at 750nm, which is a peak wavelength for ICG excitation.
  • Fig. 5A - Fig. 5C depict characterization data for such near-IR emitting AIS QD.
  • Fig. 5A depicts dynamic light scattering (DLS) data showing a single peak with the greatest size distribution at 16.24 nm and a polydispersity index of 0.104. This is indicative of little polydispersity as a polydispersity of ⁇ 0.05 is considered 100% homogenous while a polydispersity of 0.7 suggests a wide range of sized particles within the sample.
  • the very low polydispersity is also reflected in Fig. 5B presenting raw correlation data.
  • Fig. 5C presents fluorescence emission peak data of the near-IR emitting AIS QDs, showing a PLmax at the 750 nm peak of ICG excitation.
  • Statement 1 A method for the enhancement of indocyanine green (ICG)-based imaging, angiography and detection efficiency comprising administering ICG or ICG derivative-conjugated quantum dots (QD-ICGs) to a tissue; allowing the QD-ICGs to be concentrated within the tissue or absorbed onto or internalized into cells within the tissue; and energetically exciting the QD-ICGs to induce fluorescence.
  • the method may be provided as a method of diagnosis or therapy performed on a tissue comprising at least a part of the human or animal body. Alternatively, the method may be provided on the proviso that it is not a method of diagnosis or therapy performed on a tissue comprising at least part of the human or animal body.
  • Statement 2 A method for the enhancement of indocyanine green (ICG) photodynamic therapy (PDT) comprising administering ICG or ICG derivative-conjugated quantum dots (QD-ICGs) to a tissue; allowing the QD-ICGs to be concentrated within the tissue or absorbed onto or internalized into cells within the tissue; and energetically exciting the QD-ICGs to induce fluorescence.
  • the method may be provided as a method of diagnosis or therapy performed on a tissue comprising at least a part of the human or animal body. Alternatively, the method may be provided on the proviso that it is not a method of diagnosis or therapy performed on a tissue comprising at least a part of the human or animal body.
  • Statement 3 A method according to Statement 1 or 2, wherein the quantum dots (QDs) are water-soluble heavy metal-free QDs.
  • Statement 4 A method according to any one of Statements 1-3, wherein the step of energetically exciting the QD-ICGs comprises irradiation.
  • Statement 5 A method according to any one of Statements 1-4, wherein the induced fluorescence is utilized for photodynamic therapy of abnormal tissue proliferation, pre neoplastic tissues, neoplastic tissues, reduction in tumor size or labelling of tumors, and reduction of bacterial virulence.
  • Statement 6 A method according to any one of Statements 1-5, wherein the induced fluorescence is utilized for labelling and visualization of abnormal tissue proliferation, pre neoplastic tissues, and neoplastic tissues.
  • Statement 7 A method according to Statement 6, wherein the labeling and visualization of neoplastic tissues is used in conjunction with surgery to remove the neoplastic tissue.
  • Statement 8 A method according to any one of Statements 1-4, wherein the administration is used to screen tissues that may be susceptible to QD-ICGs treatment.
  • Statement 9 A method according to any one of Statements 1-4, wherein the administration is used to visualize lymphatic vessels, and nodes, and body interstitial fluids.
  • Statement 10 A method according to any one of Statements 1-9, wherein QD-ICGs are further conjugated to at least one tissue-specific ligand.
  • Statement 11 A method according to Statement 10, wherein the at least one tissue specific ligand originates from any one of a protein, a small peptide, a small molecule, a glycoprotein, a lipoprotein, a nucleic acid, an aptamer and a synthetic polymer.
  • Statement 12 A method according to any one of Statements 1-8, wherein QD-ICGs are further conjugated to at least one tumor-specific ligand.
  • Statement 13 A method according to Statement 12, wherein the tumor specific ligand is specific for EGFR, PD-L1, PD-L2, HER2, CEA, CA19-9, CA125, telomerase proteins and subunits, CD20, CD25, CD30, CD33, CD52, CD73, CD109, VEGF-A, CTLA-4, or RANK ligand.
  • Statement 14 A method according to any one of Statements 1-13, wherein the QD- ICGs further comprise a polymerizable ligand that results in QD aggregation upon physical excitation of the QDs.
  • Statement 15 A method according to any one of Statements 1-14, wherein the QD- ICGs further include a cellular uptake enhancer, a tissue penetration enhancer, or any combination thereof.
  • Statement 16 A method according to Statement 15, wherein the cellular uptake enhancer is selected from one or more of trans-activating transcriptional activators (TAT), Arg- Gly-Asp (RGD) tri-peptides, linear and cyclic peptides including the RGD motif, or poly arginine peptides.
  • TAT trans-activating transcriptional activators
  • RGD Arg- Gly-Asp
  • Statement 17 A method according to Statement 15, wherein the tissue penetration enhancer is selected from one or more of saponins, cationic lipids, and Streptolysin O (SLO).
  • the tissue penetration enhancer is selected from one or more of saponins, cationic lipids, and Streptolysin O (SLO).
  • Statement 18 A method for facilitating cell death comprising administering quantum dots (QDs) conjugated to indocyanine green (ICG) to undesired cells and energetically exciting the QDs conjugated to ICG to induce fluorescence and generation of reactive oxygen species (ROS) that facilitate cell death.
  • QDs quantum dots conjugated to indocyanine green
  • ROS reactive oxygen species
  • the method is not a method of treatment of the human or animal body.
  • the method may be an in vitro method.
  • Statement 19 A method according to Statement 18, wherein the undesired cells are precancerous cells, tumor cells, or inflammatory tissue.
  • Statement 20 A method according to Statement 18 or 19, wherein the method is performed in vivo. Alternatively, the method may be performed in vitro.
  • Statement 21 A composition comprising quantum dots (QDs) conjugated to ICG or an ICG derivative in a pharmaceutically acceptable formulation.
  • QDs quantum dots
  • Statement 22 A composition according to Statement 21, wherein the QDs are water- soluble non-toxic QD nanoparticles.
  • Statement 23 A composition according to Statement 21 or 22, wherein the QD-ICGs are further conjugated to at least one tumor-specific ligand.
  • Statement 24 A composition according to Statement 23, wherein the tumor specific ligand is specific for EGFR, PD-L1, PD-L2, HER2, CEA, CA19-9, CA125, telomerase proteins and subunits, CD20, CD25, CD30, CD33, CD52, CD73, CD109, VEGF-A, CTLA-4, or RANK ligand.
  • Statement 25 A composition according to any one of Statements 21-24, wherein the QD-ICGs further include a polymerizable ligand that results in QD-ICG aggregation upon physical excitation of the QD.
  • Statement 26 A composition according to any one of Statements 21-25, wherein the QDs further include a cellular uptake enhancer, a tissue penetration enhancer, or any combination thereof.
  • Statement 27 A composition according to Statement 26, wherein the cellular uptake enhancer is selected from one or more of trans-activating transcriptional activators (TAT), Arg- Gly-Asp (RGD) tri-peptides, linear and cyclic peptides including the RGD motif, or poly arginine peptides.
  • TAT trans-activating transcriptional activators
  • RGD Arg- Gly-Asp
  • Statement 28 A composition according to Statement 26, wherein the tissue penetration enhancer is selected from one or more of saponins, cationic lipids, and Streptolysin O (SLO).
  • the tissue penetration enhancer is selected from one or more of saponins, cationic lipids, and Streptolysin O (SLO).
  • Statement 29 ICG or ICG derivative conjugated quantum dots (QD-ICGs) for use in therapy, or compositions according to Statements 21 to 28 for use in therapy.
  • QD-ICGs ICG or ICG derivative conjugated quantum dots
  • Statement 30 ICG or ICG derivative conjugated quantum dots (QD-ICGs), or a composition comprising quantum dots (QDs) conjugated to ICG or an ICG derivative in a pharmaceutically acceptable formulation according to any one of Statements 21 to 28, for use in a method (i.e. an in vivo method) for the enhancement of indocyanine green (ICG)-based imaging, angiography and detection efficiency comprising: administering the ICG or ICG derivative conjugated quantum dots (QD-ICGs) to a tissue (i.e. in a patient); allowing the QD- ICGs to be concentrated within the tissue or absorbed onto or internalized into cells within the tissue; and energetically exciting the QD-ICGs to induce fluorescence.
  • the method may additionally comprise intraoperative imaging and fluorescence guided surgery of abnormal proliferative tissue growths and/or inflammatory tissue.
  • Statement 31 ICG or ICG derivative conjugated quantum dots (QD-ICGs), or a composition comprising quantum dots (QDs) conjugated to ICG or an ICG derivative in a pharmaceutically acceptable formulation according to any one of Statements 21 to 28, for use in a method of diagnosing one or more of:
  • abnormal proliferative tissue growths including cancers (such as, for example, cancers like gliomas, bladder cancer, cancerous skin cancers (including melanoma, basal and squamous cell carcinoma), esophageal cancer, endobronchial cancer, gastrointestinal cancer, intra abdominal tumor, cancers of the lung, bladder, prostate, bile duct, stomach, mouth, throat, larynx, cervix, vagina, and vulva), precancerous conditions (such as, for example, precancerous lesions of the skin (including actinic keratosis), and Barrett’s Esophagus) and tumors (both malignant and benign; soft or solid);
  • cancers such as, for example, cancers like gliomas, bladder cancer, cancerous skin cancers (including melanoma, basal and squamous cell carcinoma), esophageal cancer, endobronchial cancer, gastrointestinal cancer, intra abdominal tumor, cancers of the lung, bladder, prostate, bile
  • the method comprises: administering the ICG or ICG derivative conjugated quantum dots (QD- ICGs) to a patient; allowing the QD-ICGs to be concentrated within the patient tissue or absorbed onto or internalized into cells within the patient tissue; and energetically exciting the QD-ICGs to induce fluorescence.
  • ICG or ICG derivative conjugated quantum dots QD- ICGs
  • QD-ICGs indocyanine green (ICG)-based imaging, angiography and detection efficiency (i.e. relative to methods which do not utilize the QD-ICGs of the invention).
  • Statement 32 ICG or ICG derivative conjugated quantum dots (QD-ICGs), or a composition comprising quantum dots (QDs) conjugated to ICG or an ICG derivative in a pharmaceutically acceptable formulation according to any one of Statements 21 to 28, for use in a method (i.e.
  • an in vivo method for the enhancement of indocyanine green (ICG) photodynamic therapy (PDT) comprising: administering the ICG or ICG derivative conjugated quantum dots (QD-ICGs) to a tissue (i.e. in a patient); allowing the QD-ICGs to be concentrated within the tissue or absorbed onto or internalized into cells within the tissue; and energetically exciting the QD-ICGs to induce fluorescence.
  • ICG indocyanine green
  • PDT photodynamic therapy
  • Statement 33 ICG or ICG derivative conjugated quantum dots (QD-ICGs), or a composition comprising quantum dots (QDs) conjugated to ICG or an ICG derivative in a pharmaceutically acceptable formulation according to any one of Statements 21 to 28, for use in a method of treating one or more of:
  • abnormal proliferative tissue growths including cancers (such as, for example, cancers like gliomas, bladder cancer, cancerous skin cancers (including melanoma, basal and squamous cell carcinoma), esophageal cancer, endobronchial cancer, gastrointestinal cancer, intra abdominal tumor, cancers of the lung, bladder, prostate, bile duct, stomach, mouth, throat, larynx, cervix, vagina, and vulva), precancerous conditions (such as, for example, precancerous lesions of the skin (including actinic keratosis), Barrett’s Esophagus) and tumors (both malignant and benign; soft or solid);
  • cancers such as, for example, cancers like gliomas, bladder cancer, cancerous skin cancers (including melanoma, basal and squamous cell carcinoma), esophageal cancer, endobronchial cancer, gastrointestinal cancer, intra abdominal tumor, cancers of the lung, bladder, prostate, bile duct
  • ICG indocyanine green
  • PDT photodynamic therapy
  • the method comprises: administering the ICG or ICG derivative conjugated quantum dots (QD-ICGs) to a patient; allowing the QD- ICGs to be concentrated within the patient tissue or absorbed onto or internalized into cells within the patient tissue; and energetically exciting the QD-ICGs to induce fluorescence.
  • ICG indocyanine green
  • QD-ICGs provides enhancement of indocyanine green photodynamic therapy (PDT).
  • Statement 34 The QD-ICGs for use according to any one of Statements 30 to 33, or a composition for use according to Statement 30 or 31, wherein the step of energetically exciting the QD-ICGs comprises irradiation.
  • Statement 35 The QD-ICGs for use according to any one of Statements 30 to 34, or a composition for use according to any one of Statements 30 to 34, wherein the induced fluorescence is utilized for at least one of photodynamic therapy of abnormal tissue proliferation, photodynamic therapy of pre-neoplastic tissues, photodynamic therapy of neoplastic tissues, reduction in tumor size or labelling of tumors, and reduction of bacterial virulence.
  • Statement 36 The QD-ICGs for use according to any one of Statements 30 to 34, or a composition for use according to any one of Statements 30 to 34, wherein the induced fluorescence is utilized for labelling and visualization of abnormal tissue proliferation, pre neoplastic tissues, and neoplastic tissues.
  • Statement 37 The QD-ICGs for use according to Statement 36, or a composition for use according to Statement 36, wherein the labeling and visualization of neoplastic tissues is used in conjunction with surgery to remove the neoplastic tissue.
  • Statement 38 The QD-ICGs for use according to any one of Statements 30 to 34 or a composition for use according to any one of Statements 30 to 34, wherein the administration is used to screen tissues that may be susceptible to QD-ICGs treatment.
  • Statement 39 The QD-ICGs for use according to any one of Statements 30 to 34 or a composition for use according to any one of Statements 30 to 34, wherein the administration is used to visualize lymphatic vessels, and nodes, and body interstitial fluids.
  • Statement 40 The QD-ICGs for use according to any one of Statements 30 to 39 or a composition for use according to any one of Statements 30 to 39, wherein the QD-ICGs arefurther conjugated to at least one tissue-specific ligand originating from any one of a protein, a small peptide, a small molecules, a glycoprotein, a lipoprotein, a nucleic acid, an aptamer and a synthetic polymer.
  • Statement 41 The QD-ICGs for use according to any one of Statements 30 to 38 or a composition for use according to any one of Statements 30 to 38, wherein the QD-ICGs are further conjugated to at least one tumor-specific ligand.
  • Statement 42 The QD-ICGs for use according to any one of Statements 30 to 41 or a composition for use according to any one of Statements 30 to 41, wherein the QD-ICGs further comprise a polymerizable ligand that results in QD aggregation upon physical excitation of the QDs.
  • Statement 43 The QD-ICGs for use according to any one of Statements 30 to 42 or a composition for use according to any one of Statements 30 to 42, wherein the QD-ICGs further include a cellular uptake enhancer, a tissue penetration enhancer, or any combination thereof.
  • Statement 43 ICG or ICG derivative conjugated quantum dots (QD-ICGs) or a composition according to Statements 21 to 28 for use in a method for facilitating cell death in photodynamic therapy comprising administering the composition to undesired cells in vivo and energetically exciting the QDs conjugated to ICG to induce fluorescence and generation of reactive oxygen species (ROS) that facilitate cell death.
  • QD-ICGs ICG or ICG derivative conjugated quantum dots
  • Statement 44 ICG or ICG derivative conjugated quantum dots (QD-ICGs), or a composition comprising quantum dots (QDs) conjugated to ICG or an ICG derivative in a pharmaceutically acceptable formulation according to any one of Statements 21 to 28, for use in a method of treating one or more of:
  • abnormal proliferative tissue growths including cancers (such as, for example, cancers like gliomas, bladder cancer, cancerous skin cancers (including melanoma, basal and squamous cell carcinoma), esophageal cancer, endobronchial cancer, gastrointestinal cancer, intra abdominal tumor, cancers of the lung, bladder, prostate, bile duct, stomach, mouth, throat, larynx, cervix, vagina, and vulva), precancerous conditions (such as, for example, precancerous lesions of the skin (including actinic keratosis), Barrett’s Esophagus) and tumors (both malignant and benign; soft or solid);
  • cancers such as, for example, cancers like gliomas, bladder cancer, cancerous skin cancers (including melanoma, basal and squamous cell carcinoma), esophageal cancer, endobronchial cancer, gastrointestinal cancer, intra abdominal tumor, cancers of the lung, bladder, prostate, bile duct
  • inflammatory tissue for example, arthritis, Crohn’s disease, Inflammatory Bowel Disease, psoriasis, acne, multiple sclerosis, Alzheimer’s, and Parkinson’s Disease
  • proliferative inflammatory diseases of the skin or other tissues including the gastrointestinal tract and atherosclerosis plaques, atheromatous lesions and stenosis levels in a patient by photodynamic therapy, wherein the method comprises administering the composition to undesired cells in vivo and energetically exciting the QDs conjugated to ICG to induce fluorescence and generation of reactive oxygen species (ROS) that facilitate cell death (i.e. of the undesired cells).
  • ROS reactive oxygen species
  • Statement 46 Use of ICG or ICG derivative conjugated quantum dots (QD-ICGs) or a composition according to any one of Statements 21 to 28 for the manufacture of a medicament.
  • QD-ICGs ICG derivative conjugated quantum dots
  • Statement 47 Use of ICG or ICG derivative conjugated quantum dots (QD-ICGs) or a composition according to Statements 21 to 28 for the manufacture of a medicament for use in a method for the enhancement of indocyanine green (ICG)-based imaging, angiography and detection efficiency, wherein the method comprises: administering the ICG or ICG derivative conjugated quantum dots (QD-ICGs) to a tissue; allowing the QD-ICGs to be concentrated within the tissue or absorbed onto or internalized into cells within the tissue; and energetically exciting the QD-ICGs to induce fluorescence.
  • ICG indocyanine green
  • Statement 48 Use of ICG or ICG derivative conjugated quantum dots (QD-ICGs) or a composition according to Statements 21 to 28 for the manufacture of a medicament for use in a method of diagnosing one or more of:
  • abnormal proliferative tissue growths including cancers (such as, for example, cancers like gliomas, bladder cancer, cancerous skin cancers (including melanoma, basal and squamous cell carcinoma), esophageal cancer, endobronchial cancer, gastrointestinal cancer, intra abdominal tumor, cancers of the lung, bladder, prostate, bile duct, stomach, mouth, throat, larynx, cervix, vagina, and vulva), precancerous conditions (such as, for example, precancerous lesions of the skin (including actinic keratosis), and Barrett’s Esophagus) and tumors (both malignant and benign; soft or solid);
  • cancers such as, for example, cancers like gliomas, bladder cancer, cancerous skin cancers (including melanoma, basal and squamous cell carcinoma), esophageal cancer, endobronchial cancer, gastrointestinal cancer, intra abdominal tumor, cancers of the lung, bladder, prostate, bile
  • the method comprises: administering the ICG or ICG derivative conjugated quantum dots (QD- ICGs) to a patient; allowing the QD-ICGs to be concentrated within the patient tissue or absorbed onto or internalized into cells within the patient tissue; and energetically exciting the QD-ICGs to induce fluorescence.
  • QD- ICGs in such methods can provide enhancement of indocyanine green (ICG)-based imaging, angiography and detection efficiency (i.e. relative to methods which do not utilize the QD-ICGs of the invention).
  • Statement 49 Use of ICG or ICG derivative conjugated quantum dots (QD-ICGs) or a composition according to Statements 21 to 28 for the manufacture of a medicament for the enhancement of indocyanine green (ICG) photodynamic therapy (PDT), wherein the therapy comprises: administering the ICG or ICG derivative conjugated quantum dots (QD- ICGs) to a tissue; allowing the QD-ICGs to be concentrated within the tissue or absorbed onto or internalized into cells within the tissue; and energetically exciting the QD-ICGs to induce fluorescence.
  • ICG indocyanine green
  • PDT photodynamic therapy
  • Statement 50 Use of ICG or ICG derivative conjugated quantum dots (QD-ICGs) or a composition according to Statements 21 to 28 for the manufacture of a medicament for treating one or more of:
  • abnormal proliferative tissue growths including cancers (such as, for example, cancers like gliomas, bladder cancer, cancerous skin cancers (including melanoma, basal and squamous cell carcinoma), esophageal cancer, endobronchial cancer, gastrointestinal cancer, intra abdominal tumor, cancers of the lung, bladder, prostate, bile duct, stomach, mouth, throat, larynx, cervix, vagina, and vulva), precancerous conditions (such as, for example, precancerous lesions of the skin (including actinic keratosis), Barrett’s Esophagus) and tumors (both malignant and benign; soft or solid);
  • cancers such as, for example, cancers like gliomas, bladder cancer, cancerous skin cancers (including melanoma, basal and squamous cell carcinoma), esophageal cancer, endobronchial cancer, gastrointestinal cancer, intra abdominal tumor, cancers of the lung, bladder, prostate, bile duct
  • ICG indocyanine green
  • PDT photodynamic therapy
  • the therapy comprises: administering the ICG or ICG derivative conjugated quantum dots (QD-ICGs) to a patient; allowing the QD- ICGs to be concentrated within the patient tissue or absorbed onto or internalized into cells within the patient tissue; and energetically exciting the QD-ICGs to induce fluorescence.
  • ICG indocyanine green
  • Statement 51 The use according to any one of Statements 47 to 50, wherein the step of energetically exciting the QD-ICGs comprises irradiation.
  • Statement 52 The use according to any one of Statements 47 to 51, wherein the induced fluorescence is utilized for at least one of photodynamic therapy of abnormal tissue proliferation, photodynamic therapy of pre-neoplastic tissues, photodynamic therapy of neoplastic tissues, reduction in tumor size or labelling of tumors, and reduction of bacterial virulence.
  • Statement 53 The use according to any one of Statements 47 to 52, wherein the induced fluorescence is utilized for labelling and visualization of abnormal tissue proliferation, pre-neoplastic tissues, and neoplastic tissues.
  • Statement 54 The use according to Statement 53, wherein the labeling and visualization of neoplastic tissues is used in conjunction with surgery to remove the neoplastic tissue.
  • Statement 55 The use according to any one of Statements 47to 54, wherein the administration is used to screen tissues that may be susceptible to QD-ICGs treatment.
  • Statement 56 The use according to any one of Statements 47 to 51, wherein the administration is used to visualize lymphatic vessels, and nodes, and body interstitial fluids.
  • Statement 57 The use according to any one of Statements 47 to 56, wherein the QD-ICGs are further conjugated to at least one tissue-specific ligand originating from any one of a protein, a small peptide, a small molecules, a glycoprotein, a lipoprotein, a nucleic acid, an aptamer and a synthetic polymer.
  • Statement 58 The use according to any one of Statements 47 to 57, wherein QD- ICGs are further conjugated to at least one tumor-specific ligand.
  • Statement 59 The use according to any one of Statements 47 to 58, wherein the QD-ICGs further comprise a polymerizable ligand that results in QD aggregation upon physical excitation of the QDs.
  • Statement 60 The use according to any one of Statements 47 to 59, wherein the QD-ICGs further include a cellular uptake enhancer, a tissue penetration enhancer, or any combination thereof.
  • Statement 61 Use of ICG or ICG derivative conjugated quantum dots (QD-ICGs) or a composition according to Statements 21 to 28 for the manufacture of a medicament facilitating cell death in photodynamic therapy, the therapy comprising administering the composition to undesired cells in vivo and energetically exciting the QDs conjugated to ICG to induce fluorescence and generation of reactive oxygen species (ROS) that facilitate cell death.
  • QD-ICGs ICG derivative conjugated quantum dots
  • ROS reactive oxygen species
  • Statement 62 Use of ICG or ICG derivative conjugated quantum dots (QD-ICGs), or a composition comprising quantum dots (QDs) conjugated to ICG or an ICG derivative in a pharmaceutically acceptable formulation according to any one of Statements 21 to 28, for the manufacture of a medicament for use in a method of treating one or more of:
  • abnormal proliferative tissue growths including cancers (such as, for example, cancers like gliomas, bladder cancer, cancerous skin cancers (including melanoma, basal and squamous cell carcinoma), esophageal cancer, endobronchial cancer, gastrointestinal cancer, intra abdominal tumor, cancers of the lung, bladder, prostate, bile duct, stomach, mouth, throat, larynx, cervix, vagina, and vulva), precancerous conditions (such as, for example, precancerous lesions of the skin (including actinic keratosis), Barrett’s Esophagus) and tumors (both malignant and benign; soft or solid);
  • cancers such as, for example, cancers like gliomas, bladder cancer, cancerous skin cancers (including melanoma, basal and squamous cell carcinoma), esophageal cancer, endobronchial cancer, gastrointestinal cancer, intra abdominal tumor, cancers of the lung, bladder, prostate, bile duct
  • inflammatory tissue for example, arthritis, Crohn’s disease, Inflammatory Bowel Disease, psoriasis, acne, multiple sclerosis, Alzheimer’s, and Parkinson’s Disease
  • proliferative inflammatory diseases of the skin or other tissues including the gastrointestinal tract and atherosclerosis plaques, atheromatous lesions and stenosis levels in a patient by photodynamic therapy
  • the method comprises administering the composition to undesired cells in vivo and energetically exciting the QDs conjugated to ICG to induce fluorescence and generation of reactive oxygen species (ROS) that facilitate cell death (i.e. of the undesired cells).
  • ROS reactive oxygen species

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Abstract

L'invention concerne des compositions et des procédés pour améliorer les performances de l'imagerie basée sur le vert d'indocyanine (ICG), de l'efficacité de l'angiographie et de la détection ainsi que de l'ICG-PDT dans lesquelles des nanoparticules à points quantiques (QD) sont conjuguées au ICG ou à un dérivé de l'ICG.
PCT/GB2019/053522 2018-12-13 2019-12-12 Procédés pour améliorer l'imagerie médicale basée sur le vert d'indocyanine et la photothérapie WO2020120970A1 (fr)

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RU2818094C1 (ru) * 2023-06-19 2024-04-24 Государственное бюджетное учреждение здравоохранения города Москвы городская клиническая больница имени С.П. Боткина департамента здравоохранения города Москвы (ГБУЗ ГКБ им. С.П. Боткина ДЗМ) Способ проведения биопсии сторожевых лимфатических узлов при хирургическом лечении локальной меланомы кожи туловища и конечностей

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112891557A (zh) * 2021-02-03 2021-06-04 柯阳 一种ICG-β-环糊精载药系统及其制备方法和应用
RU2818094C1 (ru) * 2023-06-19 2024-04-24 Государственное бюджетное учреждение здравоохранения города Москвы городская клиническая больница имени С.П. Боткина департамента здравоохранения города Москвы (ГБУЗ ГКБ им. С.П. Боткина ДЗМ) Способ проведения биопсии сторожевых лимфатических узлов при хирургическом лечении локальной меланомы кожи туловища и конечностей

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