Stable indocyanine green formulation

Abstract

This invention relates to indocyanine green compositions with improved stability, particularly improved storage stability. In particular, this invention relates to compositions comprising indocyanine green and at least one stabilizer selected from ascorbic acid and its salts.

Description

[0001] This invention relates to the fields of medicine and pharmaceuticals. Specifically, it relates to indocyanine green compositions with improved stability, particularly improved storage stability. More particularly, this invention relates to indocyanine green compositions comprising at least one stabilizer selected from ascorbic acid and its salts. The invention also relates to methods for preparing such compositions and methods of using them. Background Technology

[0002] Indocyanine green (ICG) is a water-soluble tricyanine fluorescent dye approved by the U.S. Food and Drug Administration for medical diagnostics. ICG is used to measure liver function, cardiac output, hepatic blood flow, flap microcirculation, and ocular angiography. These medical uses of ICG are based on its high optical absorbance in a relatively transparent spectral region (750 nm to 800 nm) in human tissues. These medical uses are made possible by its extremely low toxicity. After intravenous administration, ICG is rapidly extracted from the liver into bile, and its effective half-life is approximately 3 to 4 minutes, depending on liver function.

[0003] For the applications described above, ICG must be administered in aqueous solution. It is commercially available as a lyophilized powder soluble in sterile water for injection and is typically supplied as a kit. However, ICG exhibits poor stability and rapid degradation in aqueous environments, limiting its effectiveness in time-sensitive applications such as surgical procedures. Instructions from ICG suppliers recommend immediate use of the reconstituted ICG solution within 10 hours, and that any unused portion should be discarded.

[0004] Studies have shown that ICG molecules undergo physicochemical transformations such as aggregation in aqueous media, especially at high concentrations. ICG readily forms dimers and oligomers in water, while remaining monomeric in human plasma, methanol, and dimethyl sulfoxide (DMSO). [1] Furthermore, singlet oxygen can be generated via ICG, resulting in dioxane, which thermally decomposes into several carbonyl compounds. [2] These breakdown products reduce cell viability in vitro.

[0005] Under light, high temperature, and high concentrations of ICG, the physicochemical transformation of ICG is more likely to occur. [3,4,5] The type and intensity of light that ICG is exposed to also affect degradation. Physicochemical transformations lead to rapid degradation of ICG's optical properties, resulting in discoloration, decreased fluorescence intensity, and a shift in peak absorbance. It has been reported that after 10 hours of storage in water and under direct light, the absorbance of ICG's spectral peaks decreased by 10%. [4] .

[0006] To overcome these shortcomings, many studies have been conducted to improve the stability of indocyanine green in aqueous solutions.

[0007] Engel et al. reported that the addition of the singlet oxygen quencher sodium azide (NaN3) can inhibit the chemical degradation of ICG. [2] .

[0008] Mindt et al. investigated the stability of ICG samples stored in dark or clear vials at different temperatures. Absorbance and fluorescence measurements showed that ICG remained stable for three days when stored in the dark at 4°C, with a 20% loss in fluorescence intensity during this period. Based on these experiments, they suggested that if ICG solutions are stored in the dark at 4°C, ICG can be used for more than one day after preparation from the raw material. [6] .

[0009] Other researchers have suggested encapsulating ICG in nanoscale carriers to protect ICG molecules from degradation. For example, CN103301482 discloses micelles loaded with a triblock peptide ICG, comprising a polyleucine core, a polylysine intermediate layer, and a polyethylene glycol shell, with indocyanine green dispersed in the core. The core-shell structure encapsulating indocyanine green effectively prevents ICG molecule aggregation, thereby enhancing the stability of ICG.

[0010] Kirchherr et al. disclosed the encapsulation of ICG in a Solutol HS15 (polyoxyethylene ester of 12-hydroxystearic acid) micelle system. Compared with free ICG in aqueous solution, this formulation exhibited lower aggregation behavior and higher water stability over 4 weeks. [7] .

[0011] US6944493 discloses indocyanine green compositions exhibiting enhanced stability and increased indocyanine green concentration. This invention is based on indocyanine green liposome formulations that are stable for at least 24 hours after reconstitution. All formulations disclosed in US6944493 are reconstituted with a diluent solution containing ethanol and polysorbate 80, etc.

[0012] WO2016 / 123864 discloses a composition comprising ICG in a lipid membrane embedded with lipid nanoparticles. The fluorescence intensity of the nanoparticles is 4 to 5 times higher than that of free ICG. After storage at 4°C for 0 to 300 days, the fluorescence intensity of the nanoparticles is 4 to 100,000 times higher than that of free ICG in aqueous solution.

[0013] Dedora et al. disclosed the reaction of ICG in aqueous solution with highly soluble methyl β-CyD and FDA-approved sulfobutyl ether β-CyD. The complexation effect of these complexes can reduce ICG aggregation and exhibit a more than two-fold sustained increase in fluorescence over 24 hours compared to free ICG in water. However, ICG complexation with methyl β-CyD has been found to severely reduce the viability of some fibroblasts. [8] .

[0014] ICG may have defects in its encapsulation or complexation with the aforementioned systems, such as incompatibility with cells, or it may limit the ability of ICG molecules to interact with plasma proteins after being injected into the bloodstream, thus potentially slowing down the excretion dynamics of the vascular system.

[0015] WO2020 / 240514 discloses an indocyanine green formulation comprising disodium hydrogen phosphate, sodium dihydrogen phosphate, and sodium chloride. After reconstitution with an aqueous diluent, the solution is stable for up to 24 hours.

[0016] US2010 / 0181535 discloses a method for enhancing the fluorescence intensity of a fluorescent dye, wherein the fluorescent dye is mixed with a redox buffer. The fluorescent dye may be selected from rhodamine, carborhodamine, etc. Azine dyes and / or cyanine dyes including indocyanine green. The redox buffer contains at least one reducing agent and / or at least one oxidizing agent and / or at least one reductive-oxidizing agent. Suitable reducing agents may be selected from thiophene, thionaphthol, phenol sulfides, uric acid, urea, bilirubin, ascorbic acid, and / or flavin. Single-molecule fluorescence measurements showed that adding 1 mM ascorbic acid to cyanine dyes immobilized on DNA prolonged the dark state length of the cyanine dye (i.e., the time during which the fluorescent dye remains non-fluorescent). On the other hand, compared to solutions of fluorescent dyes in phosphate-buffered saline, the reducing agent 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid... The combined use of the oxidizing agent methyl viologen with cyanine dyes in solution showed a maximum improvement in photostability of 2.6 times. These experiments were conducted by irradiating the dye with a laser-activated signal in a fluorescence detection instrument. The authors mention that another advantage of using the redox buffer is that the shelf life of the fluorescent dye solution containing the redox buffer can be increased by 10% to 20% compared to the same fluorescent dye solution without the redox buffer. However, the literature does not mention the long-term storage stability of the composition.

[0017] WO9423646 discloses a storage-stable composition comprising a voltage-sensitive dye, particularly indocyanine green, used as a contrast agent in optical imaging. The composition contains an air-protective agent, such as an antioxidant and a surfactant, to inhibit ICG decomposition. The air-protective agent may be selected from sodium sulfite, sodium ascorbate, glutathione, dithiothreitol, EDTA, polysorbate 80, and carboxymethyl cellulose. ICG decomposition, measured by the decrease in maximum absorbance in the UV-Vis spectrum from 600 nm to 900 nm, was inhibited in an airless composition stored in the dark. The best results reported in the literature show a 30% reduction in ICG content within one hour. When the experimental conditions of this literature were reproduced, rapid degradation of the composition as shown in the experimental section of this application was observed.

[0018] All of these existing technologies have reported improvements in the water stability of ICG, but none have provided aqueous ICG compositions that are stable for longer periods, ranging from 1 to 3 days. Therefore, there is a need for aqueous ICG compositions that exhibit stability exceeding that of existing formulations.

[0019] Some existing formulations are prepared in expensive equipment and / or require long manufacturing processes and / or specialized excipients. Some formulations require excipients such as alcohol and polysorbate, which can make the injection of ICG compositions painful and challenging.

[0020] In particular, an aqueous ICG composition that can be stable at room temperature for one or several months is required.

[0021] There is also a need for ICG compositions for intravenous administration that are compatible with target tissues, non-toxic, and rapidly eliminated by metabolism.

[0022] The present invention aims to provide a storage-stable ICG composition that is easy to formulate and based on readily available materials, and that has an ICG concentration sufficient for use in the same manner as prior art short-shelf-life ICG formulations.

[0023] Surprisingly, ICG formulations containing compounds selected from ascorbic acid or its salts showed improved stability compared to prior art compositions.

[0024] The formulations of this invention contain ICG at concentrations comparable to or better than those in the prior art. They can be formulated as lyophilized powders or aqueous solutions. Compared to the prior art, they offer improved storage stability and shelf life.

[0025] Another object of the present invention is to provide a stable, cost-effective, and easy-to-manufacture formulation. An object of the present invention is to provide a formulation that does not cause any discomfort to the patient when administered. Summary of the Invention

[0026] The present invention relates to a composition comprising indocyanine green (a) and a compound selected from ascorbic acid, its salts, and mixtures thereof (b), wherein when the composition is stored at ambient temperature for at least 1 month, the content of indocyanine green in the composition, as measured by HPLC at 240 nm in percentage area, decreases by less than 10% or equal to 10%.

[0027] Advantageously, prior to storage, the ICG content in the composition of the present invention is greater than or equal to 90%, and the indocyanine green content is measured by HPLC at 240 nm as a percentage area.

[0028] Preferably, prior to storage, the total amount of impurities in the ICG composition, measured by HPLC at 240 nm as area %, is less than 10% or equal to 10%.

[0029] Preferably, when the composition is stored at ambient temperature for at least one month, the increase in impurities in the composition according to the invention, as measured by HPLC at 240 nm as area % change, is less than 10% or equal to 10%.

[0030] The present invention also relates to an aqueous composition comprising at least indocyanine green (a) and a compound (b) selected from ascorbic acid, its salts, and mixtures thereof, wherein the weight ratio of compound (b) to indocyanine green (a) is 0.15 to 30, and the concentration of indocyanine green in the aqueous composition is 0.1 mg / ml to 50 mg / ml, preferably 0.1 mg / ml to 30 mg / ml.

[0031] Advantageously, the weight ratio of compound (b) to indocyanine green (a) is 0.15 to 25, preferably 0.15 to 20.

[0032] Advantageously, the weight ratio of compound (b) to indocyanine green (a) is 0.15 to 15.

[0033] According to the implementation plan, compound (b) is ascorbic acid.

[0034] According to another embodiment, compound (b) is ascorbate.

[0035] Preferably, according to this embodiment, compound (b) is selected from sodium ascorbate, potassium ascorbate, calcium ascorbate, magnesium ascorbate, and mixtures thereof. Preferably, according to this embodiment, compound (b) is sodium ascorbate.

[0036] Optionally, the indocyanine green composition according to the invention further comprises at least one compound selected from ethylenediaminetetraacetic acid (EDTA), D-α-tocopherol polyethylene glycol 1000 succinate (vitamin E-TPGS), thiomalic acid, isosorbide, PEG400, PEG 1000, thioglycolic acid, thiourea, α-cyclodextrin and β-cyclodextrin, thiolactic acid, reduced glutathione, thioglycerol, sorbitol, methionine, sodium chloride, histidine, dithiothreitol (DTT), and mixtures thereof (c).

[0037] Preferably, compound (c) is selected from reduced glutathione, thioglycerol, sorbitol, methionine, sodium chloride, histidine, dithiothreitol (DTT), and mixtures thereof.

[0038] Preferably, the weight ratio of compound (c) to indocyanine green (a) is 1:500 to 100:1, more preferably 1:100 to 10:1, and even more preferably 1:100 to 4:1.

[0039] According to an embodiment, the composition according to the invention comprises up to 5% by weight of sodium iodide based on the weight of indocyanine green.

[0040] Preferably, the concentration of indocyanine green in the aqueous composition is from 1 mg / ml to 25 mg / ml.

[0041] According to a preferred embodiment, the aqueous indocyanine green composition according to the present invention is packaged in vials or ampoules.

[0042] The present invention also relates to an aqueous composition comprising indocyanine green (a) and a compound (b) selected from ascorbic acid, its salts, and mixtures thereof, wherein the weight ratio of compound (b) to indocyanine green (a) is 0.15 to 30, and the concentration of indocyanine green in the aqueous composition is 0.1 mg / ml to 50 mg / ml, preferably 0.1 mg / ml to 30 mg / ml, said aqueous composition being used as a pharmaceutical or diagnostic agent.

[0043] The present invention also relates to a kit comprising, in a separate compartment, at least 1 / lyophilized powder of the composition as described above and defined in detail below and 2 / aqueous diluent.

[0044] The present invention also relates to a kit capable of preparing an aqueous composition according to the invention, the kit comprising at least 1 / indocyanine green (a) and 2 / an aqueous solution in a separate compartment, the aqueous solution comprising at least a compound selected from ascorbic acid, its salts, and mixtures thereof (b).

[0045] The present invention also relates to a method for diagnosing and / or treating a patient, the method comprising administering, in an effective amount, particularly via an intravenous route, an aqueous composition according to the invention. Detailed Implementation

[0046] The term “substantially constitutes” followed by one or more features means that components and steps that do not substantially affect the nature and features of the invention, other than those expressly listed, may also be included in the methods or materials of the invention.

[0047] Unless otherwise explicitly stated, the expression "inclusive of X to Y" includes the boundary. This expression means that the target range includes the values ​​of X and Y, as well as all values ​​from X to Y.

[0048] References to treatment methods in the specification may be interpreted as references to the compounds, pharmaceutical compositions, and drugs of the present invention, used in methods of treating humans (or animals) or diagnostic methods by means of therapy.

[0049] Stability of the indocyanine green composition according to the present invention

[0050] The present invention aims to provide an indocyanine green composition with improved stability compared to prior art compositions.

[0051] The terms "stable composition" and "stability" refer to compositions in which the indocyanine green content remains stable during storage for a certain period of time (e.g., 1 week, 1 month, 2 months, 6 months, 1 year, 2 years, etc.).

[0052] The stability of the composition was assessed by measuring the change in indocyanine green content in the composition after prolonged storage.

[0053] Preferably, the compositions according to the invention have the advantage that, after prolonged storage, they show virtually no signs of aggregation, degradation, or precipitation, and are substantially free of or minimally free of impurities resulting from the chemical modification of indocyanine green. Furthermore, the compositions of the invention have the advantage that, after prolonged storage, they do not exhibit significant discoloration or a decrease in fluorescence intensity or peak absorbance.

[0054] In particular, the present invention provides indocyanine green-containing compositions that exhibit improved stability compared to currently available indocyanine green formulations.

[0055] Advantageously, the compositions of the present invention exhibit at least 10 times, more preferably at least 30 times, more preferably at least 50 times, and most preferably 100 times, stability compared to currently approved indocyanine green formulations.

[0056] In one aspect, the composition according to the invention is stable when stored at ambient temperature for at least one month, preferably at least three months, more preferably at least six months, even more preferably at least 12 months, and most preferably at least two years.

[0057] The term “ambient temperature” refers to a controlled room temperature of 20°C to 25°C as defined in the United States Pharmacopeia, or 15°C to 30°C for short periods, or 15°C to 25°C as defined in the European Pharmacopoeia.

[0058] Preferably, the composition according to the invention is stable at ambient temperature for 1 to 3 months, more preferably 3 months to 3 years, more preferably 1 to 3 years, and even more preferably 1 to 2 years.

[0059] Advantageously, the compositions of the present invention are stable when stored in a container protected from natural light and / or ultraviolet light and / or fluorescence for at least one month, preferably at least three months, more preferably at least six months, and most preferably at least two years.

[0060] A container capable of protecting the composition from natural light and / or ultraviolet light and / or fluorescence is a container whose walls have a maximum transmittance of at least 50% at any wavelength. Typically, the container may be an opaque or amber container and / or an opaque secondary packaging component (e.g., a carton or outer packaging).

[0061] Advantageously, the compositions according to the invention are stable when stored in a tightly closed or sealed container for at least one month, preferably at least three months, more preferably at least six months, and most preferably at least two years.

[0062] The stability of the compositions according to the invention can be assessed by visual inspection of color and / or transparency and / or other analytical methods.

[0063] Analytical methods used to assess the stability of a composition are known to those skilled in the art. These may include nuclear magnetic resonance (NMR), high-performance liquid chromatography (HPLC), size exclusion chromatography (SEC), liquid chromatography-mass spectrometry (LC-MS), dynamic light scattering (DLS), differential scanning calorimetry (DSC), ultraviolet spectroscopy, and Fourier transform infrared spectroscopy (FTIR), or combinations of these methods.

[0064] Advantageously, the stability of the composition is assessed by measuring the change in indocyanine green content in the composition by HPLC before and after prolonged storage. More advantageously, the ICG content in the composition and the change in that content are assessed by HPLC at 240 nm as a percentage area.

[0065] Preferably, when the composition is stored at ambient temperature for at least 1 month, preferably 3 months, more preferably 6 months, even more preferably 12 months, and most preferably at least 2 years, the stability of the composition of the present invention corresponds to a decrease of less than 10% or equal to 10% in the content of indocyanine green in the composition as measured by HPLC at 240 nm in percentage area.

[0066] Preferably, when the composition is stored at ambient temperature for at least 1 month, preferably 3 months, more preferably 6 months, even more preferably 12 months, and most preferably at least 2 years, evidence of the stability of the composition of the present invention corresponds to a decrease of less than 5% or equal to 5% in the content of indocyanine green in the composition as a percentage area measured by HPLC at 240 nm.

[0067] Even more preferably, when the composition is stored at ambient temperature for at least 1 month, preferably 3 months, more preferably 6 months, even more preferably 12 months, and most preferably at least 2 years, the evidence of the stability of the composition of the present invention corresponds to a decrease of less than 2% or equal to 2% in the content of indocyanine green in the composition as a percentage area measured by HPLC at 240 nm.

[0068] Within the meaning of this application, "content decrease" should be understood as the percentage difference between the ICG content in the composition measured before storage and the ICG content in the composition measured after a defined storage time, measured by HPLC at 240 nm as % area.

[0069] Preferably, prior to storage, the ICG content in the measured composition is greater than 90% or equal to 90%, more preferably greater than 95% or equal to 95%, more preferably greater than 97% or equal to 97%, and most preferably greater than 99% or equal to 99%, and the indocyanine green content is measured by HPLC at 240 nm as a percentage area.

[0070] The stability of the indocyanine green composition of the present invention can also correspond to the fact that the total amount of impurities contained in the composition of the present invention does not increase during storage, in particular not to the level that reduces the ability of the indocyanine green composition to be used in medical and / or diagnostic applications.

[0071] For example, when the composition is stored at ambient temperature for at least one month and then these impurities are measured by HPLC, only a small amount of impurities are observed in the composition, and this amount does not increase significantly over time.

[0072] In the context of this invention, the term "impurity" should be understood as a chemical entity that is not an excipient or other additive of indocyanine green or the indocyanine green composition of this invention.

[0073] Impurities can be process-related, such as byproducts or intermediates that may form during the manufacture of indocyanine green, or degradation-related impurities arising from the chemical transformation of indocyanine green during storage. Degradation-related impurities are products that may form during storage, such as in response to light exposure, temperature and humidity, or reactions with oxygen.

[0074] In an embodiment, when the composition is stored at ambient temperature for at least 1 month, preferably 2 months, more preferably 6 months, and most preferably at least 2 years, the increase in impurities in the composition of the present invention, as measured by HPLC at 240 nm in area % change, is less than 10% or equal to 10%, preferably less than 5% or equal to 5%, and more preferably less than 2% or equal to 2%.

[0075] In an embodiment, prior to storage, the total amount of impurities in the ICG composition, measured by HPLC at 240 nm as area %, is less than 10% or equal to 10%, preferably less than 5% or equal to 5%, more preferably less than 1% or equal to 1%.

[0076] The term "total amount of impurities" includes both process-related impurities and degradation-related impurities.

[0077] Indocyanine Green Composition

[0078] The present invention provides a composition comprising indocyanine green (a) and at least one stable compound selected from ascorbic acid and its salts (b).

[0079] Optionally, the composition according to the invention may contain at least one additional compound (c) different from compound (b).

[0080] Indocyanine green (a)

[0081] The term indocyanine green refers to indocyanine green and its pharmaceutically acceptable salts, solvates, hydrates, acids, anhydrous forms and free bases, preferably indocyanine green.

[0082]

[0083] Indocyanine green can be prepared by any method known to those skilled in the art. For example, indocyanine green can be prepared according to the methods described in WO95 / 07888 or WO2017093889. Preferably, the method for preparing indocyanine green includes the step of purifying the obtained product.

[0084] Advantageously, the indocyanine green used in the compositions according to the invention is substantially pure. Preferably, the indocyanine green used in the compositions according to the invention is provided in the form of a composition having a purity greater than 90.0%, preferably greater than 95.0%, more preferably greater than 97.0%, and even more preferably greater than 99.7%, as measured by HPLC.

[0085] The term "purity" as used in this article refers to the degree to which the raw material indocyanine green is free of unwanted or adulterated chemical entities.

[0086] According to the present invention, the purity and impurities in the indocyanine green material are evaluated by HPLC at 240 nm as a percentage area.

[0087] Preferably, the indocyanine green used in the composition according to the invention is provided in the form of a composition containing less than 5.0%, preferably less than 1.0%, more preferably less than 0.5% of impurities as measured by HPLC.

[0088] Preferably, the indocyanine green used in the composition according to the invention is provided in the form of a composition comprising less than 0.50%, preferably less than 0.40%, more preferably less than 0.20%, and most preferably less than 0.15% of each impurity as measured by HPLC.

[0089] Impurities contained in indocyanine green can be any reaction byproducts or intermediates generated during the manufacturing process. For example, as described in document US2019 / 0337896, impurities can include, but are not limited to, N-phenylacetamide, 4-(1,1,2-trimethyl-1H-benzo[e]indol-3-yl)butane-1-sulfonate, and 4-(1,1,2-dimethyl-2-((1E,3E,5E)-6-(N-phenylacetamido)hexa-1,3,5-trienyl)-1H-benzo[e]indol-3-yl)butane-1-sulfonate.

[0090] Preferably, indocyanine green is provided in the form of a composition comprising a metallic contaminant in an amount of less than 200 ppm, preferably less than 100 ppm, more preferably less than 50 ppm, and even more preferably less than 20 ppm based on the weight of indocyanine green. The content of the metallic contaminant in indocyanine green can be measured by inductively coupled plasma mass spectrometry (ICP-MS).

[0091] The term "metal pollutants" refers to so-called "heavy" metals, especially: Al, As, Cd, Cr, Cu, Fe, Sn, Mn, Hg, Mo, Ni, Pb, Zn, and their organic and inorganic derivatives.

[0092] More preferably, indocyanine green contains no or less than 2 ppm of lead and arsenic.

[0093] The indocyanine green used in the compositions of this invention is provided as a composition that may also contain sodium iodide. Sodium iodide is an additive commonly found in commercial indocyanine green compositions. When present, the iodide content is ideally limited to less than 5% by weight based on the weight of indocyanine green. The sodium iodide content in indocyanine green can be determined by potentiometric analysis.

[0094] The indocyanine green used in the compositions of this invention can be provided in any suitable form, such as lyophilized or crystalline powder.

[0095] Stable compound (b)

[0096] Compound (b) may be selected from ascorbic acid, ascorbate salts, and mixtures thereof, wherein ascorbic acid is also known as (5R)-5-[(1S)-1,2-dihydroxyethyl]-3,4-dihydroxyfuran-2(5H)-one, 3-oxo-L-gulononic acid furanolactone, L-ascorbic acid, or vitamin C.

[0097] "The mixture thereof" refers to a mixture of ascorbic acid and ascorbate in various proportions.

[0098] Ascorbates may include, but are not limited to, sodium ascorbate, potassium ascorbate, calcium ascorbate, and magnesium ascorbate.

[0099] According to a first embodiment of the present invention, compound (b) is ascorbic acid.

[0100] According to a second embodiment of the invention, compound (b) is ascorbate.

[0101] Preferably, according to this embodiment, compound (b) is sodium ascorbate.

[0102] According to a third embodiment, the composition comprises indocyanine green, ascorbic acid, and at least one ascorbate salt, preferably sodium ascorbate.

[0103] When compound (b) associates with indocyanine green (a), the inventors unexpectedly observed an increase in the stability of the indocyanine green composition in an aqueous medium.

[0104] In a preferred embodiment, the weight ratio of compound (b) to indocyanine green (a) is 0.15 to 30, preferably 0.15 to 25, preferably 0.15 to 20, preferably 0.15 to 15, more preferably 0.20 to 15, and even more preferably 0.20 to 13.

[0105] In a preferred embodiment, the molar ratio of compound (b) to indocyanine green (a) is 0.30 to 120, preferably 0.30 to 60, more preferably 0.40 to 60, and even more preferably 0.40 to 55.

[0106] In a preferred embodiment of the invention, the composition consists essentially of indocyanine green (a) and at least one compound (b) selected from ascorbic acid, its salts, and mixtures thereof.

[0107] In another preferred embodiment of the invention, the composition consists essentially of indocyanine green (a) and at least one compound (b) selected from ascorbic acid, its salts, and mixtures thereof, and also contains an iodide.

[0108] According to another aspect, the present invention relates to the use of ascorbic acid and / or its salts to improve the storage stability of ICG compositions.

[0109] Compound (c)

[0110] In another embodiment of the invention, the composition may further comprise one or more compounds (c), said compound (c) being selected from compounds known to those skilled in the art as suitable for medical and / or diagnostic injectable solutions.

[0111] When present, compound (c) may be selected, for example, from ethylenediaminetetraacetic acid (EDTA), D-α-tocopherol polyethylene glycol 1000 succinate (vitamin E-TPGS), thiomalic acid, isosorbide, PEG 400, PEG 1000, thioglycolic acid, thiourea, α-cyclodextrin and β-cyclodextrin, thiolactic acid, reduced glutathione, thioglycerol, sorbitol, methionine, sodium chloride, histidine, dithiothreitol (DTT), and mixtures thereof.

[0112] Preferably, compound (c) may be selected from reduced glutathione, thioglycerol, sorbitol, methionine, sodium chloride, histidine, dithiothreitol (DTT), and mixtures thereof.

[0113] "The mixture thereof" means a mixture of two or more of the following in all proportions: reduced glutathione, thioglycerol, sorbitol, methionine, sodium chloride, histidine, and dithiothreitol (DTT).

[0114] Preferably, the weight ratio of compound (c) to indocyanine green (a) is 1:500 to 100:1, more preferably 1:500 to 10:1, more preferably 1:100 to 10:1, and most preferably 1:100 to 4:1.

[0115] Preferably, the weight ratio of compound (c) to compound (b) is 1:100 to 100:1, more preferably 1:10 to 10:1.

[0116] When present, compound (c) may be provided in any form, such as solid, liquid, or semi-solid. Compound (c) may also be in the form of an aqueous solution.

[0117] In a preferred embodiment of the invention, the composition is substantially composed of indocyanine green (a), a stable compound selected from ascorbic acid, its salts, and mixtures thereof (b), and a compound selected from reduced glutathione, thioglycerol, sorbitol, methionine, sodium chloride, histidine, dithiothreitol (DTT), and mixtures thereof (c).

[0118] In another preferred embodiment of the invention, the composition is substantially composed of indocyanine green (a), a stable compound selected from ascorbic acid, its salts, and mixtures thereof (b), an iodide, and a compound selected from reduced glutathione, thioglycerol, sorbitol, methionine, sodium chloride, histidine, dithiothreitol (DTT), and mixtures thereof (c).

[0119] Other additives

[0120] The compositions according to the invention may also contain at least one pharmaceutically acceptable additive.

[0121] As used herein, the term “pharmaceutically acceptable” means a compound, material, or composition suitable for contact with tissues in humans and animals without excessive toxicity, irritation, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0122] The term "additive" includes components in a composition other than those described above (a), (b), and (c) or sodium iodide. Examples of such additives include buffers such as phosphate buffers, pH adjusters, isotonic agents, surfactants, preservatives, penetration enhancers, antimicrobial agents, wetting agents, and emulsifiers.

[0123] For example, the compositions of the present invention may contain one or more additives selected from sodium hydroxide, potassium hydroxide, methylparaben, propylparaben, sodium acetate, sodium citrate, sodium carbonate, ammonium carbonate, sodium bicarbonate, benzoate / ester, acetate / ester, boric acid, lactic acid, glutaric acid, malic acid, succinic acid, and carbonic acid, as well as alkali metal or alkaline earth metal salts of these acids, meglumine, hydrochloric acid, cyclodextrin derivatives, sorbitan monolaurate, triethanolamine acetate, triethanolamine oleate, citric acid, lactic acid, phosphoric acid, sulfuric acid, arginine, alanine, glycine, and lysine.

[0124] These additives are generally available to those skilled in the art and can be in any form, such as solid, liquid or semi-solid.

[0125] Preferably, the amount of additive in the composition of the present invention is within a range that will not substantially adversely affect the activity of indocyanine green. Preferably, the composition of the present invention contains the minimum amount and quantity of additive required to provide a stable and effective composition.

[0126] The composition according to the invention

[0127] The compositions according to the invention may be provided in any suitable form, but are preferably provided as aqueous compositions or lyophilized solid compositions.

[0128] Aqueous composition

[0129] According to a first aspect, the composition according to the invention is in the form of an aqueous composition.

[0130] "Aqueous composition" should be understood as aqueous solution, aqueous suspension, colloidal aqueous suspension, aqueous dispersion, preferably aqueous solution.

[0131] The term "aqueous solution" refers to the composition of indocyanine green as described above, the components of which are soluble in an aqueous medium.

[0132] Advantageously, according to this variant, the composition comprises an aqueous diluent, indocyanine green (a), one or more compounds as defined above (b), optionally one or more compounds as defined above (c), and optionally one or more pharmaceutically acceptable additives.

[0133] Aqueous diluents are pharmaceutically acceptable solvents that are safe and non-toxic when administered to humans or animals and can be used to prepare pharmaceutical formulations.

[0134] The aqueous diluent contains water and may contain one or more pharmaceutically acceptable additives. In some variations of the methods disclosed herein, one or more of the compounds (b) and optionally one or more of the compounds (c) as defined above are introduced into the aqueous diluent prior to mixing with ICG. In such embodiments, one or more of the compounds (b) and optionally one or more of the compounds (c) may be considered as part of the aqueous diluent.

[0135] A suitable aqueous diluent can be any biologically acceptable liquid in which the compositions of the present invention are completely soluble. Water, particularly sterile water for injection (SWFI) and / or bacteriostatic water for injection (BWFI), is a preferred diluent because it does not contain salts or other compounds that could affect the stability of ICG. However, other aqueous diluents, such as sterile saline solutions, Ringer's solution, dextran solution, phosphate-buffered saline (PBS) solution, glucose solution, etc., may also be used. The aqueous diluent may also contain buffers, bacteriostatic agents, and solutes that make the composition isotonic with the blood of the intended recipient.

[0136] According to the implementation plan, the aqueous diluent is selected from water for injection, antibacterial water for injection, sterile saline solution, Ringer's solution, dextran solution, phosphate buffered saline solution, and glucose solution.

[0137] Those skilled in the art can select a diluent based on the intended use and relative to other compounds in the composition. They can also adjust the amount of excipient based on its solubility in aqueous solution.

[0138] In a preferred embodiment, the aqueous diluent is water for injection, particularly sterile water for injection (SWFI) and / or bacteriostatic water for injection (BWFI).

[0139] Advantageously, the osmotic pressure of the aqueous solution is at least 1 mOsM. More advantageously, the osmotic pressure of the aqueous solution is from 1 mOsM to 300 mOsM, and even more preferably from 1 mOsM to 200 mOsM.

[0140] The concentration of indocyanine green in the aqueous composition of the present invention can be any concentration suitable for medical or diagnostic purposes, particularly for producing satisfactory quality angiographic images.

[0141] According to a preferred embodiment, the concentration of indocyanine green in the aqueous composition according to the present invention is from 0.1 mg / ml to 50 mg / ml.

[0142] When the concentration of the ICG solution is less than 0.1 mg / ml, the fluorescence is insufficient for satisfactory detection by the analytical instrument.

[0143] According to a preferred embodiment, the concentration of indocyanine green in the aqueous composition according to the invention is from 0.1 mg / ml to 30 mg / ml, preferably from 1 mg / ml to 25 mg / ml, and more preferably from 1 mg / ml to 10 mg / ml.

[0144] When one or more of the components of the composition of the present invention, such as compound (b) or possibly compound (c), are added as an aqueous solution, the volume of the aqueous diluent can be adjusted to obtain the desired ICG concentration.

[0145] In embodiments of the present invention, the aqueous composition consists essentially of an aqueous diluent, indocyanine green (a), and a stable compound selected from ascorbic acid, its salts, and mixtures thereof (b).

[0146] In embodiments of the present invention, the aqueous composition consists essentially of an aqueous diluent, indocyanine green (a), a stable compound selected from ascorbic acid, its salts, and mixtures thereof (b), and an iodide.

[0147] In another embodiment, the aqueous composition comprises essentially an aqueous diluent, indocyanine green (a), a stable compound selected from ascorbic acid, its salts, and mixtures thereof (b), and a compound selected from reduced glutathione, thioglycerol, sorbitol, methionine, sodium chloride, histidine, dithiothreitol (DTT), and mixtures thereof (c).

[0148] In another embodiment, the aqueous composition consists essentially of an aqueous diluent, indocyanine green (a), a stable compound selected from ascorbic acid, its salts, and mixtures thereof (b), and a compound selected from reduced glutathione, thioglycerol, sorbitol, methionine, sodium chloride, histidine, dithiothreitol (DTT), and mixtures thereof (c) and an iodide.

[0149] The most preferred percentages of compounds (ICG(a), stable compound(b) and compound(c)) in the aqueous composition are the same as those expressed in the chapter “Indocyanine Green Compositions” above.

[0150] lyophilized solid compositions

[0151] According to another preferred embodiment, the composition according to the invention is in the form of a lyophilized composition or a lyophilized powder.

[0152] The terms “lyophilized composition,” “lyophilized powder,” or “lyophilized product” refer to the solid composition according to the invention obtained by lyophilization of an aqueous solution.

[0153] The term "freeze-drying" or "lyophilization" refers to the process of first freezing a composition in an aqueous solvent and then removing the frozen solvent by sublimation in a vacuum environment.

[0154] When provided as a lyophilized product, the composition according to the invention can be converted into a solution by using an aqueous diluent therein before application. The reconstituted aqueous solution, also a composition according to the invention, is then stable at ambient temperature for at least 1 month, preferably 2 months, more preferably 6 months, and most preferably at least 2 years.

[0155] All the above-mentioned limitations regarding aqueous diluents, ICG concentration, pH, osmotic pressure, and stability also apply to aqueous solutions reconstituted from the lyophilized composition according to the invention.

[0156] Furthermore, in embodiments, the lyophilized composition according to the invention has a short reconstitution time of less than 2 minutes, preferably less than 1 minute, and is suitable for intravenous administration after dilution with an aqueous diluent.

[0157] The term "reconstitution time" refers to the time required to rehydrate the lyophilized composition with an aqueous solution to form a clear, particle-free solution.

[0158] After the composition is reconstituted into an aqueous solution for medical or diagnostic purposes, the solution is preferably stored in a container that protects it from light and oxygen.

[0159] Reagent test kit

[0160] According to a preferred embodiment, the present invention relates to a kit comprising, in a first portion, the composition of the invention in the form of a lyophilized powder, and in a second portion, an aqueous diluent.

[0161] According to another preferred embodiment, the present invention relates to a kit comprising indocyanine green (a) in a first portion and an aqueous solution (b) comprising at least a stable compound selected from ascorbic acid and its salts in a second portion. Optionally, the aqueous solution may comprise a compound (c) as defined above, and optionally one or more additives as described above.

[0162] According to this implementation scheme, indocyanine green is preferably in the form of lyophilized powder.

[0163] All preferred embodiments of the above-described ICG composition and aqueous diluent are also applicable to any kit variant.

[0164] According to the embodiments, the kit according to the invention comprises a portion (powder and diluent) in an amount such that the ICG concentration in the aqueous solution reconstituted from the kit is from 0.1 mg / ml to 50 mg / ml, preferably from 0.1 mg / ml to 30 mg / ml, more preferably from 1 mg / ml to 25 mg / ml, and most preferably from 1 mg / ml to 10 mg / ml.

[0165] According to these preferred embodiments, the kit includes at least two separate compartments in which the lyophilized powder and the aqueous diluent are stored separately.

[0166] The kit may include two separate vials, packaged with or without syringes, or the kit may include a pre-filled dual-chamber syringe, such as a dual-chamber bypass syringe.

[0167] This kit allows the powder and aqueous solution to be mixed before application.

[0168] Kits of one or another variant are designed for the temporary reconstitution of stable aqueous solutions of indocyanine green as described herein. Their advantage lies in providing the essential components of the composition for injection under optimized storage conditions, delivering the desired concentration in appropriate amounts. Therefore, aqueous compositions can be reconstituted without dilution, in addition to the diluent contained in the kit. This kit packaging avoids the risk of dosage errors or the introduction of contaminants.

[0169] Preparation method of the composition according to the present invention

[0170] The compositions according to the invention can be prepared using any known method, and are not limited to any particular method.

[0171] For example, a composition can be prepared by mixing indocyanine green (a) with a stable compound (b), optionally one or more of the above-mentioned compounds (c), and optionally one or more pharmaceutically acceptable excipients.

[0172] Preferably, the mixing of the composition components according to the invention is carried out in the above-mentioned aqueous solvent. The order of addition of the components constituting the composition according to the invention can be changed. For example, indocyanine green can be dissolved in an aqueous diluent before adding the stabilizing compound (b) and optionally compound (c) and the additive. Compound (b) and compound (c) can be mixed before adding the dissolved indocyanine green. Alternatively, indocyanine green can be introduced into an aqueous solution, wherein compound (b) and optionally compound (c) have been pre-dissolved.

[0173] Preferably, mixing is carried out under an inert atmosphere, such as argon or nitrogen. Preferably, the aqueous diluent and / or liquid components are degassed before use.

[0174] When provided as a lyophilized product, the lyophilization of the compositions according to the invention can be carried out using standard lyophilization or vacuum drying equipment. The lyophilization cycle can vary depending on the equipment and facilities used, and can be adjusted by those skilled in the art.

[0175] Preferably, the pre-lyophilized aqueous solution is an aqueous solution of indocyanine green as described above. This pre-lyophilized aqueous solution according to the invention can be sterilized before lyophilization. Sterilization is typically achieved by using a suitable membrane filtration solution.

[0176] Preferably, the aqueous solution is freeze-dried shortly after preparation to prevent any degradation of the composition. The freeze-dried product can be stored for several months before use. Preferably, the freeze-dried product is stored under conditions that protect it from air and / or humidity and / or light.

[0177] When provided as an aqueous composition, the indocyanine green composition according to the invention can also be obtained by reconstitution using a lyophilized product from the kit described above and an aqueous diluent. For reconstitution, the lyophilized composition can be introduced into a vial, and then the aqueous diluent can be added.

[0178] Advantageously, the composition prepared in lyophilized or aqueous form is immediately placed in a light-protected, airtight container after preparation. The container can be, for example, a bottle, ampoule, vial, syringe, or tube. The container can be made of, for example, glass, polymer, metal, etc. The container can be a single-dose or multi-dose container.

[0179] In a preferred embodiment of the invention, the container is preferably a colored glass vial or a colored glass ampoule. Advantageously, the container is selected from colored glass vials or colored glass ampoules having a vacuum headspace. Preferably, the composition according to the invention is stored in a colored glass ampoule.

[0180] Advantageously, the compositions according to the invention are stored at ambient temperature.

[0181] Use of the composition according to the invention

[0182] The compositions of the present invention can be used in diagnostic and / or therapeutic methods for patients. The methods include administering an effective amount of the aqueous solution according to the invention. This administration can be carried out via enteral route, parenteral route, particularly intravenous injection, or local application. Preferably, administration is carried out via intravenous injection.

[0183] The term "effective amount" refers to, for example, the amount of indocyanine green that researchers or clinicians are seeking to elicit a biological or medical response in tissues, systems, animals, or humans.

[0184] Advantageously, the compositions of the present invention can be used for the diagnostic and / or therapeutic methods described above, wherein the aqueous ICG composition is administered to a patient at least 1 month after preparation or reconstitution from one of the above-described kits, preferably at least 6 months after preparation, more preferably at least 12 months after preparation, and most preferably at least 2 years after preparation.

[0185] In particular, the compositions according to the invention can be used in thermotherapy, ICG-enhanced selective photocoagulation, photodynamic therapy (PDT), and photodynamic and thermotherapy (PHT). More particularly, as a PDT application, the compositions according to the invention can be used in the treatment of infections, acne, macular surgery, cancer treatment, etc. The use of the compositions according to the invention can be associated with other therapies such as immunotherapy, radiotherapy, ultrasound, and chemotherapy.

[0186] The compositions of the present invention can also be used to obtain angiographic images of patient tissues, to measure cardiac output, to measure liver function and hepatic blood flow, and to diagnose and treat age-related macular degeneration, related choroidal neovascularization, and tumors.

[0187] The compositions according to the invention are based on components that have been clinically tested and approved for use in humans and / or animals.

[0188] For diagnostic applications, the amount of ICG composition administered to the patient should be sufficient to allow the dye to fluoresce upon irradiation at an appropriate wavelength, with peak absorption and emission of ICG known to be between 800 nm and 850 nm. The same standard applies to therapeutic applications using ICG solutions; sufficient dye should be administered to make the treatment effective. Those skilled in the art can readily determine the amount of ICG composition to be administered, and it should be at least at an acceptable concentration currently used for ophthalmic angiography, e.g., 2 ml of 20 mg / ml ICG solution for diagnostic purposes. Higher dye concentrations can be advantageously used for any of these diagnostic and therapeutic methods, as is recognized by those skilled in the art.

[0189] Compared to existing ICG compositions, the stability of ICG in solution according to the present invention is improved, thus allowing for better diagnosis / treatment of patients when the same amount of compound is injected. This is because the improved stability of ICG will provide a stronger response to fluorescence irradiation.

[0190] Example

[0191] I-Materials and Methods :

[0192] 1. Reagents and chemicals:

[0193] Indocyanine green was supplied by Biophore India Pharmaceuticals and was ready for use without further purification. Ascorbic acid was supplied by Fisher Scientific. Sodium ascorbate was supplied by Aldrich. Other reagents were purchased from Aldrich and were ready for use without further purification.

[0194] 2. Equipment:

[0195] The stability of the formulations in the above examples was determined by high-performance liquid chromatography (HPLC). The analytical apparatus used was an Agilent Infinity 1260 HPLC / MS system equipped with a diode array detector (UV-Vis), a Waters Xbridge Shield RP18-4.6x100-5μm column, and an MSESI spectrometer. The injection volume was 5μL. The flow rate was 1ml / min, the column temperature was 30℃, and the detection wavelength was 240nm.

[0196] Elution was performed using a gradient method. The mobile phase consisted of a mixture of 10 mM ammonium acetate / acetic acid buffer (elution buffer A) and acetonitrile (elution buffer B) at pH 5.5. The composition of the mobile phase changed continuously during elution, as detailed in Table I.

[0197] Table I

[0198] Time (minutes) Eluent A% Eluent B% 0.0 85 15 3.0 85 15 40.0 30 70 40.1 85 15 47.0 85 15

[0199] 3. Sample preparation:

[0200] To prepare the following ICG formulation, a certain amount of the stable compound (b) and optionally one or more compounds (c) are weighed into a Type I clear glass vial. The vial is then tare and 25 mg of indocyanine green is weighed into it. The vial is sealed with a bromobutyl rubber stopper (from VWS) and then purged with a stream of nitrogen.

[0201] Then add 10 ml of pre-degassed WFI using a syringe. Gently shake the vial manually until the solute dissolves. Then store the vial at room temperature, protected from light.

[0202] II - Compositions comprising indocyanine green (a) and a stabilizing compound (b) according to the invention:

[0203] Table II

[0204]

[0205] * Composition C3 was prepared in 10 ml of phosphate buffered saline solution according to the same procedure as described above.

[0206] Table III below shows the stability changes of an ICG composition containing 25 mg ICG and 10 mg sodium ascorbate (ICG concentration = 2.5 mg / ml, sodium ascorbate / ICG weight ratio = 0.4) over several months.

[0207] Table III

[0208]

[0209] The results shown in Tables II and III indicate that ICG does not degrade in aqueous solution in the presence of ascorbic acid or sodium ascorbate and remains stable over time.

[0210] III - Compositions comprising indocyanine green (a), a stabilizing compound (b), and other compounds (c) according to the present invention:

[0211] Table IV

[0212]

[0213] To prepare a solution containing sodium chloride, a certain amount of ICG, compound (b), and methionine or histidine (compositions C8 and C10) were weighed into a vial, sealed with a bromobutyl rubber stopper, and then purged with a stream of nitrogen. Then, 5 ml of degassed WFI and 5 ml of 0.9% degassed NaCl solution were added. Composition C9, without methionine or histidine, was prepared according to the same procedure.

[0214] The results reported in Table IV indicate that aqueous indocyanine green compositions containing sodium ascorbate and one or more additional compounds (c) selected from reduced glutathione, thioglycerol, sorbitol, methionine, sodium chloride, histidine, or DTT are also stable over time. Compound (c) in Table IV does not significantly affect the stability of the ICG compositions of the present invention containing stabilizing compound (b).

[0215] IV. Effect of changes in the ratio of compound (b) to indocyanine green (a):

[0216] Table V

[0217]

[0218] The solution is not clear; a precipitate of the solute is observed.

[0219] V-Contrast Composition

[0220] Composition C18 was prepared according to the scheme in §I-3.

[0221] Composition C19 was prepared according to the following procedure: 1.0 mg of ICG was weighed and added to 250 ml of phosphate buffer solution (pH 7.41). Then, 3 ml of 0.0002 M sodium ascorbate solution was added. The solution used to prepare this composition was degassed before use. The resulting ICG solution was stored at room temperature protected from light. This composition is according to Example 7 of prior art WO9423646.

[0222] Composition C20 was prepared according to the following procedure: 2.5 mg of ICG was weighed and added to 250 ml of phosphate buffer solution (pH 7.41). 1.5 ml of this ICG / phosphate buffer solution was combined with 1.5 ml of 0.10 M ascorbate solution. The solution used to prepare this composition was degassed before use. The resulting ICG solution was stored at room temperature protected from light. This composition is according to Example 8 of prior art WO9423646.

[0223] Table VI

[0224]

[0225] *** ICG content cannot be measured by HPLC because the sodium ascorbate peak is much higher than the ICG peak. After 10 days of storage, the solution color changes from light green to yellow.

[0226] The results in Table VI indicate that even at very low ICG concentrations, ICG solutions are not storage stable, even with a sodium ascorbate / ICG weight ratio of 0.15 to 30.

[0227] Table VII

[0228]

[0229] The solution is not clear, and Trolox (6-hydroxy-2,5,7,8-tetramethylchromo-2-carboxylic acid) is insoluble at this concentration.

[0230] Composition C25 was prepared by weighing ICG in a vial, sealing the vial with a bromobutyl rubber stopper, and then purging with a stream of nitrogen. Then, 5 ml of degassed WFI and 5 ml of 0.9% degassed NaCl solution were added.

[0231] Compositions C26 and C27 were prepared by weighing ICG and other compounds (c) in vials, sealing the vials with bromobutyl rubber stoppers, and then purging with a stream of nitrogen. 10 ml of phosphate buffered saline solution was then added.

[0232] The results in Table VII indicate that the ICG content in the aqueous solution varies considerably over a short time in the absence of the stabilizer compound (b), and the solution is unstable in the presence of compound (c) alone.

[0233] References

[0234] [1] Holzer W, Mauerer M, Penzkofer A, Szeimies RM, Abels C, Landthaler M, W.Photostability and thermal stability of indocyanine green.JPhotochem Photobiol B.1998Dec:47(2-3):155-64.

[0235] [2]Engel E,Schraml R,Maisch T,Kobuch K, B,Szeimies RM,HillenkampJ, W,Vasold R.Light-induced decomposition of indocyanine green.InvestOphthalmol Vis Sci.2008May:49(5):1777-83.

[0236] [3]Saxena V,Sadoqi M,Shao J.Degradation kinetics of indocyanine greenin aqueous solution.J Pharm Sci.2003Oct:92(10):2090-7.

[0237] [4]Gathje J,Steuer RR,Nicholes KR.Stability studies on indocyanine green dyes.J Appl Physiol.1970Aug:29(2):181-5.

[0238] [5]Barbier f,Deweerdt ga.Chromatography and IRspectrography ofindocyanine green.Clin Chim Acta.1964Dec:10:549-5

[0239] [6]Mindt S,Karampinis I,John M,Neumaier M,Nowak K.Stability and degradation of indocyanine green in plasma,aqueous solution and whole blood.Photochem Photobiol Sci.2018Sep 12:17(9):1189-1

[0240] [7]Kirchherr AK,Briel A, K.Stabilization of indocyanine green byencapsulation within micellar systems.Mol Pharm.2009Mar-Apr;6(2):480-91.

[0241] [8]DeDora DJ,Suhrland C,Goenka S,Mullick Chowdhury S,Lalwani G,Mujica-Parodi LR,Sitharaman B.Sulfobutyl etherβ-cyclodextrin( )andmethylβ-cyclodextrin enhance and stabilize fluorescence of aqueousindocyanine green.J Biomed Mater Res B Appl Biomater.2016 Oct;104(7):1457-64.

Claims

1. An aqueous composition comprising at least indocyanine green (a) and a compound (b) selected from ascorbic acid, its salts, and mixtures thereof, wherein the weight ratio of compound (b) to indocyanine green (a) is 0.15 to 15, and the concentration of indocyanine green in the aqueous composition is 0.1 mg / ml to 30 mg / ml.

2. The indocyanine green composition according to claim 1, wherein the weight ratio of compound (b) to indocyanine green (a) is 0.20 to 13.

3. The indocyanine green composition according to claim 1 or 2, wherein compound (b) is ascorbic acid.

4. The indocyanine green composition according to claim 1, wherein compound (b) is ascorbate.

5. The indocyanine green composition according to claim 4, wherein compound (b) is selected from sodium ascorbate, potassium ascorbate, calcium ascorbate, magnesium ascorbate, and mixtures thereof.

6. The indocyanine green composition according to claim 5, wherein compound (b) is sodium ascorbate.

7. The indocyanine green composition according to claim 1, further comprising at least one compound selected from reduced glutathione, thioglycerol, sorbitol, methionine, sodium chloride, histidine, dithiothreitol (DTT), and mixtures thereof (c).

8. The indocyanine green composition according to claim 7, wherein the weight ratio of compound (c) to indocyanine green (a) is 1:500 to 100:

1.

9. The indocyanine green composition according to claim 8, wherein the weight ratio of compound (c) to indocyanine green (a) is from 1:100 to 10:

1.

10. The indocyanine green composition according to claim 9, wherein the weight ratio of compound (c) to indocyanine green (a) is from 1:100 to 4:

1.

11. The indocyanine green composition of claim 1, wherein the composition comprises up to 5% by weight of sodium iodide based on the weight of indocyanine green.

12. The indocyanine green composition according to claim 1, wherein the concentration of indocyanine green in the aqueous composition is from 1 mg / ml to 25 mg / ml.

13. The indocyanine green composition according to claim 1, wherein, prior to storage, the content of indocyanine green in the composition is greater than or equal to 90%, and the content of indocyanine green is measured by HPLC at 240 nm as a percentage area.

14. The indocyanine green composition according to claim 1, which is used as a pharmaceutical or diagnostic agent.

15. A kit comprising a lyophilized powder of the composition as defined in claim 1 in a first portion and an aqueous diluent in a second portion.

16. A kit for preparing the composition according to claim 1, comprising indocyanine green (a) in a first portion and an aqueous solution in a second portion, said aqueous solution comprising at least a compound selected from ascorbic acid, its salts, and mixtures thereof (b).

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