Stable indocyanine green formulation

Abstract

The present invention relates to indocyanine green compositions with improved stability, in particular with improved storage stability. In particular, the present invention relates to compositions comprising a combination of indocyanine green and stabilizing compounds, said combination comprising ethylenediaminetetraacetic acid (EDTA) and / or one or more than one salt thereof, histidine and sodium chloride.

Description

[0001] The present invention relates to the field of medicine and pharmaceuticals. The present invention relates to indocyanine green compositions with improved stability, in particular improved storage stability. In particular, the present invention also relates to indocyanine green compositions comprising ethylenediaminetetraacetic acid (EDTA) and / or one or more than one salt thereof, histidine and sodium chloride. The present invention also relates to methods of preparing such compositions and methods of using them. BACKGROUND

[0002] Indocyanine green (ICG) is a water-soluble tricarbocyanine fluorescent dye approved by the U.S. Food and Drug Administration for medical diagnostics. Indocyanine green is used for the determination of 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 the relatively transparent spectral region of human tissue (750 nm to 800 nm). These medical uses are possible due to its extremely low toxicity. After intravenous administration, ICG is rapidly extracted by the liver into the bile with an effective half-life of about 3 to 4 minutes depending on liver function.

[0003] For the above-mentioned applications, ICG has to be administered in the form of an aqueous solution. It is marketed as a lyophilized powder soluble in sterile water for injection and is usually supplied as a kit. However, ICG has poor stability in aqueous environments and degrades rapidly, which limits its effectiveness in time-sensitive applications such as surgical procedures. The instructions of the ICG supplier include immediate use of the reconstituted ICG solution up to 10 hours, any unused portion should be discarded.

[0004] Studies have shown that ICG molecules undergo physical-chemical transformations, such as aggregation, in aqueous media, especially at high concentrations. ICG readily forms dimers and oligomers in water, while it remains monomeric in human plasma, methanol and dimethyl sulfoxide (DMSO) [1] In addition, singlet oxygen can be generated by ICG, which generates dioxetanes that thermally decompose into several carbonyl compounds [2] These decomposition products reduce the viability of cells in vitro.

[0005] The physical-chemical transformations of ICG are more likely to occur under light, high temperatures, in the presence of oxygen and at high concentrations of ICG [3,4,5] The type and intensity of light to which ICG is exposed also affects degradation. Physical-chemical transformations cause rapid degradation of the optical properties of ICG, resulting in discoloration, reduced fluorescence intensity and shift in peak absorbance. It has been reported that the absorbance of the spectral peak of ICG decreases by 10% after 10 hours of storage in water and direct light [4] .

[0006] To overcome these drawbacks, many studies have been carried out to improve the stability of indocyanine green in aqueous solution.

[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 at different temperatures in either dark or clear bottles. Absorbance and fluorescence measurements indicated that ICG remained stable for three days when stored in the dark at 4°C, during which time the fluorescence intensity was lost by 20%. On the basis of these experiments, they suggested that ICG can be used for more than one day after preparation from the raw material if the ICG solution is stored in the dark at 4°C [6] .

[0009] Other researchers have suggested encapsulating ICG into nanoscale carriers to protect the ICG molecules from degradation. For example, CN103301482 discloses a triblock polypeptide ICG-loaded micelle comprising a poly-leucine core, a poly-lysine middle layer, and a poly-ethylene glycol shell, wherein the indocyanine green is dispersed in the core. The core-shell structure encapsulating the indocyanine green can effectively prevent the aggregation of ICG molecules, thereby enhancing the stability of ICG.

[0010] Kirchherr et al. disclose the encapsulation of ICG in Solutol HS15 (polyoxyethylene alkyl ester of 12-hydroxystearic acid) micellar systems. The formulation exhibits lower aggregation behavior and higher aqueous stability over 4 weeks compared to free ICG in aqueous solution [7] .

[0011] US 6944493 discloses indocyanine green compositions exhibiting enhanced stability and enhanced indocyanine green concentration. The invention is based on indocyanine green liposome formulations that are stable for at least 24 hours after reconstitution. All formulations disclosed in US 6944493 are reconstituted with a diluent solution comprising ethanol and polysorbate 80, among others.

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

[0013] Dedora et al. disclose ICG in aqueous solution with highly soluble methyl beta-CyD and FDA-approved sulfobutyl ether beta-CyD complexation. These complexes can reduce aggregation of ICG and exhibit greater than two-fold increase in sustained fluorescence over 24 hours compared to free ICG in water. However, it has been found that complexation of ICG with methyl beta-CyD severely reduces the viability of some fibroblasts [8] .

[0014] ICG encapsulation or complexation with the above systems suffers from drawbacks such as incompatibility with cells or can limit the ability of ICG molecules to interact with plasma proteins after injection into the blood, thus potentially slowing down the excretion kinetics from the vasculature.

[0015] 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 can be selected from rhodamine, carborhodamine, xanthene dyes and / or cyanine dyes including indocyanine green. The redox buffer can comprise carotenoids, in particular tocopherols, thiols, in particular glutathione, N-acetyl cysteine, dihydrolipoic acid, amino acids, in particular tryptophan, tyrosine, histidine, cysteine, methionine and / or peptides and proteins containing them. 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, this document does not mention the long-term storage stability of the composition.

[0016] WO2020 / 240514 discloses an indocyanine green formulation comprising disodium hydrogen phosphate, sodium dihydrogen phosphate and sodium chloride. The composition can optionally include other pharmaceutically acceptable excipients such as sodium carbonate, sodium bicarbonate, boric acid, lactic acid, glutaric acid, malic acid, succinic acid and carbonic acid, buffers of amino acid buffers such as arginine, alanine, histidine, glycine and lysine. Upon reconstitution with an aqueous diluent, the solution is stable for up to 24 hours.

[0017] All these prior arts report increasing the aqueous stability of ICG, but none of them provide an ICG aqueous solution that is stable for more than 1 to 3 days. Therefore, there is a need for an ICG aqueous composition having a stability that exceeds the prior art formulations.

[0018] WO9423646 discloses storage stable compositions comprising a voltage sensitive dye, in particular indocyanine green, for use as an optical imaging contrast agent. The compositions comprise an air protection agent, such as an antioxidant and a surfactant, which inhibits the decomposition of ICG. The air protection agent can be selected from sodium sulfite, sodium ascorbate, glutathione, dithiothreitol, EDTA, polysorbate 80 and carboxymethylcellulose. The decomposition of ICG, measured by a decrease in the maximum absorbance in the UV-visible spectrum between 600 nm and 900 nm, is inhibited in air-free storage. The experimental section of WO9423646 shows that compositions containing ICG and EDTA do not have storage stability.

[0019] Some formulations of the prior art are prepared in expensive equipment and / or require long manufacturing process times and / or special excipients. Some formulations require excipients such as alcohol and polysorbate, which make the injection of the ICG composition painful and challenging.

[0020] In particular, there is a need for ICG aqueous compositions that are stable at room temperature for one month or several months.

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

[0022] The present invention aims to provide ICG compositions that are easy to formulate and based on readily available materials, and that have an ICG concentration sufficient to be used in the same way as short shelf-life ICG formulations of the prior art.

[0023] It has surprisingly been found that ICG compositions containing specific compositions of stabilizing compounds have better stability compared to prior art compositions.

[0024] The formulations of the present invention contain ICG at concentrations comparable to or superior to the prior art. They can be formulated as lyophilized powders or aqueous solutions. They have improved storage stability and shelf life compared to the prior art.

[0025] Another object of the present invention is to provide formulations that are stable, cost-effective and easy to manufacture. An object of the present invention is to provide formulations that do not cause any discomfort to the patient at the time of administration. SUMMARY

[0026] The present invention relates to a composition comprising indocyanine green (a), ethylenediaminetetraacetic acid (EDTA) and / or one or more than one salt thereof, histidine and sodium chloride, wherein the decrease in the content of indocyanine green in the composition is less than 10% or equal to 10% measured by HPLC at 240 nm as % area when the composition is stored at ambient temperature for at least 1 month.

[0027] Advantageously, the content of ICG in the composition of the application is greater than 90% or equal to 90% before storage, the content of indocyanine green being measured by HPLC at 240 nm in % area.

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

[0029] Preferably, the increase in impurities in the composition of the application, expressed in area % variation measured by HPLC at 240 nm, is less than 10% or equal to 10% when the composition is stored at ambient temperature for at least 1 month.

[0030] The application also relates to a composition comprising indocyanine green, ethylenediaminetetraacetic acid (EDTA) and / or one or more than one salt thereof, histidine and sodium chloride.

[0031] The application also relates to a kit comprising, in separate compartments, at least 1 / a composition comprising indocyanine green, ethylenediaminetetraacetic acid (EDTA) and / or one or more than one salt thereof, histidine and sodium chloride, and 2 / an aqueous diluent.

[0032] The application also relates to a kit comprising, in separate compartments, at least 1 / indocyanine green (a) and 2 / an aqueous solution comprising at least ethylenediaminetetraacetic acid (EDTA) and / or one or more than one salt thereof, histidine and sodium chloride.

[0033] The application also relates to a composition comprising indocyanine green, ethylenediaminetetraacetic acid (EDTA) and / or one or more than one salt thereof, histidine and sodium chloride for use as a medicament or diagnostic agent.

[0034] The application also relates to a method for diagnosing and / or treating a patient, comprising administering an effective amount of a composition comprising indocyanine green, ethylenediaminetetraacetic acid (EDTA) and / or one or more than one salt thereof, histidine and sodium chloride, in particular by intravenous route.

[0035] According to an embodiment, the indocyanine green composition of the application comprises indocyanine green, one or more than one ethylenediaminetetraacetic acid salt, histidine and sodium chloride.

[0036] Advantageously, according to this embodiment, the ethylenediaminetetraacetic acid salt is chosen from disodium EDTA, sodium calcium EDTA, tetrasodium EDTA, dicalcium EDTA, or a mixture thereof.

[0037] Preferably, the indocyanine green composition according to the application comprises, relative to the total weight of the four compounds:

[0038] - 10 to 70% by weight of indocyanine green, - 0.1 to 10% by weight of ethylenediaminetetraacetic acid (EDTA) and / or one or more than one salt thereof,

[0039] - 0.1 to 15% by weight of EDTA or a salt thereof,

[0040] - 1 to 20% by weight of histidine, and

[0041] - 20 to 80% by weight of sodium chloride.

[0042] Preferably, the indocyanine green composition comprises, relative to the total weight of the four compounds:

[0043] - 10 to 70% by weight of indocyanine green,

[0044] - 0.1 to 15% by weight of one or more ethylenediaminetetraacetic acid salts,

[0045] - 1 to 20% by weight of histidine, and

[0046] - 20 to 80% by weight of sodium chloride. According to an embodiment, the composition comprises at most 5% by weight of sodium iodide, based on the weight of indocyanine green.

[0047] According to a preferred embodiment, the indocyanine green composition according to the application is an aqueous composition.

[0048] According to a more preferred embodiment, the indocyanine green composition according to the application is an aqueous composition contained in a vial or ampoule.

[0049] According to an embodiment, the aqueous composition is reconstituted from a kit comprising, in separate compartments, at least 1 / a composition comprising indocyanine green, ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, histidine and sodium chloride, in particular in the amounts as described above, and 2 / an aqueous diluent.

[0050] According to another embodiment, the aqueous composition is reconstituted from a kit comprising, in separate compartments, at least 1 / indocyanine green and 2 / an aqueous solution comprising at least ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, histidine and sodium chloride.

[0051] According to another embodiment, the aqueous composition is reconstituted from a kit comprising, in a first part, indocyanine green, and in a second part, an aqueous solution, the kit comprising at least ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, histidine and sodium chloride (stabilizing compounds), wherein one of the stabilizing compounds is in the first part and the other two stabilizing compounds are in the other part.

[0052] Advantageously, the concentration of indocyanine green in the aqueous composition is between 0.1 mg / ml and 50 mg / ml, preferably between 0.1 mg / ml and 30 mg / ml.

[0053] According to another embodiment, the composition is in the form of a lyophilized powder. DETAILED DESCRIPTION

[0054] The term "consisting essentially of' followed by one or more characteristics, means that the components and steps, in addition to those explicitly recited, can be included in the process or material of the present application, as long as the nature and features of the invention are not materially affected.

[0055] The expression "comprising X to Y" includes the boundaries, unless otherwise explicitly stated. This expression indicates that the target range includes the X and Y values, as well as all values from X to Y.

[0056] References in the specification to treatment methods can be interpreted as references to the compounds, pharmaceutical compositions and medicaments of the present application, for use in methods of treatment of the human body (or animals) by therapy or diagnostic methods.

[0057] Stability of indocyanine green compositions according to the invention

[0058] The present application aims to provide indocyanine green compositions having improved stability compared to prior art compositions.

[0059] The terms "stable composition" and "stability" refer to a composition in which the indocyanine green content remains stable over a certain period of time (e.g. 1 week, 1 month, 2 months, 6 months, 1 year, 2 years, etc.).

[0060] The stability of the composition is evaluated by measuring the change in indocyanine green content in the composition over long-term storage.

[0061] Preferably, the advantage of the compositions according to the present application is that, after storage over a long period of time, they show substantially no signs of aggregation, degradation, precipitation, and substantially no or minimal impurities formed as a result of chemical modification of indocyanine green. Furthermore, the advantage of the compositions of the present application is that, after long-term storage, they show no significant discoloration or decrease in fluorescence intensity or peak absorbance.

[0062] In particular, the present application provides compositions comprising indocyanine green that exhibit improved stability compared to currently available indocyanine green formulations.

[0063] Advantageously, the compositions of the present application exhibit at least 10-fold, more preferably at least 30-fold, still more preferably at least 50-fold, most preferably 100-fold stability with respect to currently approved indocyanine green formulations.

[0064] In one aspect, the composition according to the application is stable when stored at ambient temperature for at least one month, preferably for at least 3 months, more preferably for at least 6 months, still more preferably for at least 12 months, most preferably for at least 2 years.

[0065] The term "ambient temperature" means 20°C to 25°C according to the definition of the US Pharmacopeia, controlled room temperature from 15°C to 30°C for a short time, or 15°C to 25°C according to the definition of the European Pharmacopeia.

[0066] Preferably, the composition according to the application is stable at ambient temperature for 1 month to 3 years, preferably for 3 months to 3 years, more preferably for 1 year to 3 years, still more preferably for 1 year to 2 years.

[0067] Advantageously, the composition of the application is stable when stored in a container protecting it from natural light and / or ultraviolet and / or fluorescent light for at least one month, preferably for at least 3 months, more preferably for at least 6 months, most preferably for at least 2 years.

[0068] The container capable of protecting the composition from natural light and / or ultraviolet and / or fluorescent light is a container whose walls have a maximum percentage of light transmission of at least 50% at any wavelength. Typically, the container can be an opaque or amber container and / or an opaque secondary packaging component (e.g. a carton or an overwrap).

[0069] Advantageously, the composition according to the application is stable when stored in a container that is tightly closed or sealed for at least one month, preferably for at least 3 months, more preferably for at least 6 months, most preferably for at least 2 years.

[0070] The stability of the composition according to the application can be evaluated by visual inspection of the color and / or the transparency and / or other analytical methods.

[0071] The analytical methods for evaluating the stability of the composition are known to the person skilled in the art. They can include nuclear magnetic resonance (NMR), high-performance liquid chromatography (HPLC), size exclusion chromatography (SEC), liquid chromatography coupled with mass spectrometry analysis (LC-MS), dynamic light scattering (DLS), differential scanning calorimetry (DSC), ultraviolet spectroscopy and Fourier transform infrared spectroscopy (FTIR) or a combination of these methods.

[0072] Advantageously, the stability of the composition is evaluated by the change in the content of indocyanine green in the composition measured by HPLC before and after long-term storage. More advantageously, the content of ICG in the composition and the change in this content are evaluated by HPLC at 240 nm in % area.

[0073] Preferably, the stability of the composition of the application corresponds to a decrease of less than 10% or equal to 10% of the content of indocyanine green in the composition measured by HPLC at 240 nm in area % when the composition is stored at ambient temperature for at least 1 month, preferably 3 months, more preferably 6 months, still more preferably 12 months, most preferably at least 2 years.

[0074] Preferably, the stability of the composition of the application corresponds to a decrease of less than 5% or equal to 5% of the content of indocyanine green in the composition measured by HPLC at 240 nm in area % when the composition is stored at ambient temperature for at least 1 month, preferably 3 months, more preferably 6 months, still more preferably 12 months, most preferably at least 2 years.

[0075] Even more preferably, the stability of the composition of the application corresponds to a decrease of less than 2% or equal to 2% of the content of indocyanine green in the composition measured by HPLC at 240 nm in area % when the composition is stored at ambient temperature for at least 1 month, preferably 3 months, more preferably 6 months, still more preferably 12 months, most preferably at least 2 years.

[0076] Within the meaning of the present application, "decrease in content" is understood to be the percentage difference between the content of ICG in the composition measured before storage and the content of ICG in the composition measured after a defined storage time, the content being measured by HPLC at 240 nm in area %.

[0077] Preferably, the content of indocyanine green in the composition measured before storage is greater than 90% or equal to 90%, preferably greater than 95% or equal to 95%, more preferably greater than 97% or equal to 97%, most preferably greater than 99% or equal to 99%, the content of indocyanine green being measured by HPLC at 240 nm in area %.

[0078] The stability of indocyanine green of the composition of the application can also correspond to the fact that the total amount of impurities contained in the composition of the application does not increase during storage, in particular not to a level that reduces the ability of the indocyanine green composition for medical and / or diagnostic applications.

[0079] For example, only a small amount of impurities is observed in the composition when it is stored at ambient temperature for at least 1 month, and this amount does not increase significantly over time, when these impurities are measured by HPLC.

[0080] In the context of the present application, the term "impurities" is understood to be chemical entities that are not indocyanine green or excipients or other additives of the indocyanine green composition of the application.

[0081] The impurities can be process-related impurities, such as by-products or intermediates that can form during the manufacturing process of indocyanine green, or degradation-related impurities that result from chemical transformation of indocyanine green during storage. Degradation-related impurities are products that can form during storage, for example in response to light exposure, temperature and humidity, or reaction with oxygen.

[0082] In an embodiment, the increase of impurities in the composition of the application, measured by HPLC at 240 nm as % area change, is less than 10% or equal to 10%, preferably less than 5% or equal to 5%, more preferably less than 2% or equal to 2%, when the composition is stored at ambient temperature for at least 1 month, preferably 2 months, more preferably 6 months, most preferably at least 2 years.

[0083] In an embodiment, 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%, before storage.

[0084] The term "total amount of impurities" includes the above-mentioned process-related impurities and degradation-related impurities.

[0085] Compositions of indocyanine green according to the invention

[0086] The present application provides a composition comprising indocyanine green and a combination of compounds comprising ethylenediaminetetraacetic acid (EDTA) and / or one or more than one salt thereof, histidine and sodium chloride. Surprisingly, the inventors have noticed that this combination of compounds is able to stabilize and / or avoid, and / or reduce, and / or prevent degradation of the ICG composition in an aqueous diluent, and / or improve the shelf life, and / or extend the shelf life. This combination will be named "stabilizing compounds" hereinafter.

[0087] Optionally, the composition according to the present application can comprise at least one pharmaceutically acceptable additive different from the stabilizing compounds described above and hereinafter in details.

[0088] Indocyanine green

[0089] The term indocyanine green refers to indocyanine green and its pharmaceutically acceptable salts, solvates, hydrates, acids, anhydrous forms and free base forms, preferably to indocyanine green.

[0090]

[0091] Indocyanine green can be prepared by any method known to the person skilled in the art. For example, indocyanine green can be prepared according to the method described in WO95 / 07888 or WO2017093889. Preferably, the method of preparation of indocyanine green comprises a step of purification of the obtained product.

[0092] Advantageously, the indocyanine green used in the composition according to the application is essentially pure. Preferably, the indocyanine green used in the composition according to the application 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%, still more preferably greater than 99.7%, measured by HPLC.

[0093] The term "purity" as used herein refers to the extent to which the raw material indocyanine green is free of unwanted or adulterating chemical entities.

[0094] According to the application, the purity and impurities in the indocyanine green material are assessed by HPLC at 240 nm in % area.

[0095] Preferably, the indocyanine green used in the composition according to the application is provided in the form of a composition comprising less than 5.0%, preferably less than 1.0%, more preferably less than 0.5% of impurities, measured by HPLC.

[0096] Preferably, the indocyanine green used in the composition according to the application is provided in the form of a composition comprising less than 0.50%, preferably less than 0.40%, more preferably less than 0.15% of each impurity, measured by HPLC.

[0097] The impurities contained in the indocyanine green can be any reaction by-products or intermediates produced during the manufacturing process. For example, the impurities can include N-phenylacetamide, 4-(1,1,2-trimethyl-1 H-benzo[e]indol-3-yl)butane-1 -sulfonate, 4-(1,1,2-dimethyl-2-((1 E,3E,5E)-6-(N-phenylacetamido)hexa-1,3,5-trienyl)-1 H- benzo[e]indol-3-yl)butane-1 -sulfonate, as described in document US2019 / 0337896, but are not limited thereto.

[0098] Preferably, the indocyanine green is provided in the form of a composition comprising a content of metal contaminants less than 200 ppm, preferably less than 100 ppm, more preferably less than 50 ppm, even more preferably less than 20 ppm, based on the weight of the indocyanine green. The content of metal contaminants in the indocyanine green can be measured by inductively coupled plasma mass spectrometry (ICP-MS).

[0099] By "metal contaminants" is meant so-called "heavy" metals, in particular: Al, As, Cd, Cr, Cu, Fe, Sn, Mn, Hg, Mo, Ni, Pb, Zn, and organic and inorganic derivatives thereof.

[0100] Still more preferably, the indocyanine green does not comprise or comprises less than 2 ppm of lead and arsenic.

[0101] The indocyanine green used in the composition of the present application is provided as a composition which can also comprise sodium iodide. Sodium iodide is an additive which is usually present in commercial compositions of indocyanine green. When present, the iodide content is desirably limited to less than 5% by weight, based on the weight of the indocyanine green. The sodium iodide content in indocyanine green can be determined by potentiometric analysis.

[0102] The indocyanine green used in the composition of the present application can be provided in any suitable form, such as a lyophilisate or a crystalline powder.

[0103] Stabilizing compounds

[0104] The composition according to the present application comprises a combination of indocyanine green and stabilizing compounds, said combination comprising ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, histidine and sodium chloride.

[0105] Advantageously, the ethylenediaminetetraacetic acid salt can be selected from disodium EDTA, calcium sodium EDTA, tetrasodium EDTA, disodium calcium EDTA, or a mixture thereof. These ethylenediaminetetraacetic acid salts can also be in the form of a hydrated salt.

[0106] By "a mixture thereof" is meant a mixture of different EDTA salts in various proportions.

[0107] For the purposes of the present application, "histidine" means the L- or D- enantiomer of the amino acid histidine. Advantageously, histidine means the L- enantiomer of the amino acid histidine.

[0108] According to a preferred embodiment, the composition comprises indocyanine green, one or more EDTA salts, histidine and sodium chloride.

[0109] In a preferred embodiment, the EDTA salt is selected from tetrasodium EDTA and calcium sodium EDTA, or a mixture thereof.

[0110] When using the combination of the components ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, histidine and sodium chloride, the inventors have surprisingly observed an enhanced stability of the indocyanine green composition in aqueous media.

[0111] In a preferred embodiment of the present application, the composition consists essentially of indocyanine green, ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, histidine and sodium chloride.

[0112] In another preferred embodiment of the present application, the composition consists essentially of indocyanine green, ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, histidine, sodium chloride and iodide.

[0113] Advantageously, the composition according to the application comprises, or preferably consists essentially of, relative to the total weight of the four following compounds:

[0114] - 10% to 70% by weight of indocyanine green,

[0115] - 0.1% to 15% by weight of EDTA or a salt thereof,

[0116] - 1% to 20% by weight of histidine, and

[0117] - 20% to 80% by weight of sodium chloride.

[0118] Even more preferentially, the composition according to the application comprises, or preferably consists essentially of, relative to the total weight of the four following compounds:

[0119] - 15% to 60% by weight of indocyanine green,

[0120] - 1% to 10% by weight of EDTA or a salt thereof,

[0121] - 2% to 15% by weight of histidine, and

[0122] - 30% to 70% by weight of sodium chloride.

[0123] According to a second embodiment, advantageously, the composition according to the application comprises, or preferably consists essentially of, relative to the total weight of the five following compounds:

[0124] - 10% to 70% by weight of indocyanine green,

[0125] - 0.1% to 10% by weight of EDTA or a salt thereof,

[0126] - 1% to 20% by weight of histidine,

[0127] - 20% to 80% by weight of sodium chloride, and

[0128] - 0% to 5% by weight of sodium iodide.

[0129] Even more preferentially, according to this embodiment, the composition according to the application comprises, or preferably consists essentially of, relative to the total weight of the five following compounds:

[0130] - 15% to 60% by weight of indocyanine green,

[0131] - 1% to 10% by weight of EDTA or a salt thereof,

[0132] - 2% to 15% by weight of histidine,

[0133] - 0.1 % to 15% by weight of one or more than one pharmaceutically acceptable excipient.

[0134] - 0.1 % to 15% by weight of one or more than one pharmaceutically acceptable excipient.

[0135] According to a third embodiment, advantageously, the composition according to the application comprises, or preferably consists essentially of, relative to the total weight of the six compounds:

[0136] - 10% to 70% by weight of indocyanine green,

[0137] - 0.1 % to 10% by weight of EDTA or a salt thereof,

[0138] - 1 % to 20% by weight of histidine,

[0139] - 20% to 80% by weight of sodium chloride,

[0140] - 0% to 5% by weight of sodium iodide, and

[0141] - 0.1 % to 15% by weight of one or more than one pharmaceutically acceptable excipient.

[0142] Even more preferably, according to this embodiment, the composition according to the application comprises, or preferably consists essentially of, relative to the total weight of the six compounds:

[0143] - 15% to 60% by weight of indocyanine green,

[0144] - 1 % to 10% by weight of EDTA or a salt thereof,

[0145] - 2% to 15% by weight of histidine,

[0146] - 30% to 70% by weight of sodium chloride,

[0147] - 0% to 5% by weight of sodium iodide, and

[0148] - 0.1 % to 15% by weight of one or more than one pharmaceutically acceptable excipient.

[0149] According to another aspect, the application relates to the use of the association of the stabilizing compounds described above to improve the storage stability of ICG compositions.

[0150] Further additives

[0151] The composition according to the application can also comprise at least one pharmaceutically acceptable additive.

[0152] As used herein, the term "pharmaceutically acceptable" means a compound, material, composition or an article that is suitable for use with humans and animals without undue toxicity, irritation, or other problem or complication, commensurate with a reasonable benefit / risk ratio, and is acceptable to the dental community.

[0153] The term "additive" includes ingredients contained in the composition other than ICG, ethylenediaminetetraacetic acid (EDTA) or a salt thereof, histidine, sodium chloride or sodium iodide. Examples of such additives include buffers, pH adjusting agents, isotonic agents, surfactants, preservatives, tonicity agents, antimicrobial agents, wetting agents and emulsifying agents.

[0154] For example, the composition of the present application can comprise one or more additives selected from sodium hydroxide, potassium hydroxide, methyl paraben, propyl paraben, sodium acetate, sodium citrate, sodium carbonate, ammonium carbonate, sodium bicarbonate, benzoate, acetate, boric acid, lactic acid, glutaric acid, malic acid, succinic acid and carbonic acid, and alkali or alkaline earth metal salts of these acids, glutamine, hydrochloric acid, cyclodextrin derivatives, sorbitan monolaurate, triethanolamine acetate, triethanolamine oleate, citric acid, lactic acid, phosphoric acid, sulfuric acid, arginine, alanine, glycine and lysine.

[0155] These additives are generally available to those of ordinary skill in the art and can be in any form, for example, solid, liquid or semi-solid.

[0156] Preferably, the amount of additive in the composition of the present application is in a range that does not substantially adversely affect the activity of indocyanine green. Preferably, the composition of the present application contains the minimum number and amount of additives required to provide a stable and effective composition.

[0157] Form of the composition according to the invention

[0158] The composition according to the present application can be provided in any suitable form, but is preferably provided in the form of an aqueous composition or a lyophilised solid composition.

[0159] Aqueous composition

[0160] According to a first aspect, the composition according to the present application is in the form of an aqueous composition.

[0161] An "aqueous composition" is understood to mean an aqueous solution, an aqueous suspension, a colloidal aqueous suspension, an aqueous dispersion, preferably an aqueous solution.

[0162] The term "aqueous solution" means a composition of indocyanine green as described above, the ingredients of which are soluble in an aqueous medium.

[0163] Advantageously, according to this variant, the composition comprises, in particular in the amounts as defined above, an aqueous diluent, indocyanine green, ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, histidine, sodium chloride, and optionally one or more pharmaceutically acceptable additives.

[0164] The aqueous diluent is a pharmaceutically acceptable solvent, safe and non-toxic when administered to a human or an animal, and useful for the preparation of pharmaceutical formulations.

[0165] The aqueous diluent comprises water and can comprise one or more pharmaceutically acceptable additives. In some variants of the method disclosed herein, the components ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, histidine and sodium chloride as defined above are introduced into the aqueous diluent before mixing with the ICG. In such embodiments, one or more of these compounds can be considered as part of the aqueous diluent.

[0166] A suitable aqueous diluent can be any liquid that is biologically acceptable and in which the composition of the application is completely soluble. Water, in particular sterile water for injection (SWFI) and / or bacteriostatic water for injection (BWFI), is the preferred diluent as it does not contain salts or other compounds that can affect the stability of the ICG. However, another aqueous diluent can also be used, such as a sterile saline solution, a Ringer's solution, a dextrose solution, a glucose solution, etc.

[0167] According to embodiments, the aqueous diluent is selected from the group consisting of water for injection, bacteriostatic water for injection, a sterile saline solution, a Ringer's solution, a dextrose solution and a glucose solution.

[0168] The person skilled in the art is able to select the diluent according to the intended use and in relation to the other compounds of the composition. He is also able to adjust the amount of excipient according to the solubility of the excipient in aqueous solution.

[0169] In preferred embodiments, the aqueous diluent is water for injection, in particular sterile water for injection (SWFI) and / or bacteriostatic water for injection (BWFI).

[0170] The aqueous diluent can also include buffers, bacteriostatic agents and solutes that make the composition isotonic with the blood of the intended recipient.

[0171] According to embodiments, the aqueous diluent is not a buffered solution or does not contain a buffer.

[0172] Advantageously, the osmolarity of the aqueous solution is at least 1 mOsM. More advantageously, the osmolarity of the aqueous solution is between 1 mOsM and 300 mOsM, more preferably between 1 mOsM and 200 mOsM.

[0173] The concentration of indocyanine green in the aqueous composition of the present application can be any concentration suitable for medical or diagnostic use, in particular for producing angiographic images of satisfactory quality.

[0174] According to a preferred embodiment, the concentration of indocyanine green of the aqueous composition according to the present application is comprised between 0.1 mg / ml and 50 mg / ml, most preferably between 0.1 mg / ml and 30 mg / ml.

[0175] When the concentration of the ICG solution is less than 0.1 mg / ml, the fluorescence is not sufficient for a satisfactory detection by analytical instruments.

[0176] According to a preferred embodiment, the concentration of indocyanine green of the aqueous composition according to the present application is comprised between 0.1 mg / ml and 25 mg / ml, preferably between 1 mg / ml and 25 mg / ml, more preferably between 1 mg / ml and 10 mg / ml.

[0177] In case one or more of the components of the composition of the present application are added as aqueous solutions, the volume of the aqueous diluent can be adjusted to obtain the desired concentration of ICG.

[0178] In an embodiment of the present application, the aqueous composition consists essentially of an aqueous diluent, indocyanine green, ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, histidine and sodium chloride.

[0179] In another embodiment, the aqueous composition consists essentially of an aqueous diluent, indocyanine green, ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, histidine, sodium chloride and sodium iodide.

[0180] The preferred percentages of the compounds (ICG and stabilizing compounds) in the aqueous composition are the same as expressed in the chapter "Stabilizing compounds" above.

[0181] Lyophilized solid composition

[0182] According to another preferred embodiment, the composition according to the present application is in the form of a lyophilized composition or a lyophilized powder.

[0183] The term "lyophilized composition", "lyophilized powder" or "lyophilizate" refers to a solid composition according to the present application obtained by lyophilization of an aqueous solution.

[0184] The term "lyophilization" or "freeze-drying" 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.

[0185] When provided as a lyophilizate, the composition according to the application can be converted into a solution before administration by using the aqueous diluent contained therein. The reconstituted aqueous solution, also a composition according to the application, is then stable at ambient temperature for at least 1 month, preferably 2 months, more preferably 6 months, most preferably at least 2 years.

[0186] All the above limitations regarding the aqueous diluent, the ICG concentration, the pH, the osmolarity and the stability also apply to the aqueous solution reconstituted from the lyophilized composition according to the application.

[0187] Furthermore, in embodiments, the lyophilized composition according to the application has a short reconstitution time of less than 2 minutes, preferably less than 1 minute, and is suitable for intravenous administration after dilution with the aqueous diluent.

[0188] By "reconstitution time" is meant the time required to rehydrate the lyophilized composition with an aqueous solution into a clear solution free of particles.

[0189] After the composition has been reconstituted into an aqueous solution for medical or diagnostic use, the solution is preferably stored in a container that protects it from light and oxygen.

[0190] Kit

[0191] According to a preferred embodiment, the present application relates to a kit comprising in a first part the composition of the application in the form of a lyophilized powder and in a second part the aqueous diluent.

[0192] According to another preferred embodiment, the present application relates to a kit comprising in a first part indocyanine green and in a second part at least ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, histidine and sodium chloride.

[0193] According to another embodiment, the present application relates to a kit comprising in a first part indocyanine green and in a second part an aqueous solution, the kit comprising at least ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, histidine and sodium chloride (stabilizing compounds), wherein one stabilizing compound is in the first part and the other two stabilizing compounds are in the other part.

[0194] In all kit embodiments, preferably, the solid part comprising indocyanine green is in the form of a lyophilized powder.

[0195] In all kit embodiments, optionally, the aqueous solution can comprise one or more additives as described above.

[0196] All the preferred embodiments of the ICG composition and the aqueous diluent described above also apply to any kit variant.

[0197] According to an embodiment, the kit according to the application consists of parts (powder and diluent) in amounts such that the concentration of ICG in the aqueous solution reconstituted from the kit is between 0.1 mg / ml and 50 mg / ml, preferably between 0.1 mg / ml and 30 mg / ml, preferably between 0.1 mg / ml and 25 mg / ml, more preferably between 1 mg / ml and 25 mg / ml, most preferably between 1 mg / ml and 10 mg / ml.

[0198] According to these preferred embodiments, the kit comprises at least two separate compartments in which the lyophilized powder and the aqueous diluent are stored separately.

[0199] The kit can comprise two separate vials, packaged with or without a syringe, or the kit can comprise a pre-filled dual chamber syringe, such as a dual chamber bypass syringe.

[0200] Such a kit allows the mixing of the powder and the aqueous solution before administration.

[0201] The kits according to one or the other variant are intended for the extemporaneous reconstitution of a stable aqueous solution of indocyanine green as described herein. Their advantage is to provide the essential components of the composition for injection under optimized storage conditions, in appropriate amounts to provide the ideal concentration. Thus, in addition to the diluent contained in the kit, it is possible to reconstitute the aqueous composition without dilution with the diluent. Thanks to this packaging as a kit, it is possible to avoid dosage errors or the introduction of contaminants.

[0202] Process for the preparation of the composition according to the invention

[0203] The composition according to the application can be prepared using any known method and is not limited to a particular method.

[0204] For example, the composition can be prepared by mixing indocyanine green with ethylenediaminetetraacetic acid (EDTA) and / or one or more than one salt thereof, histidine and sodium chloride, and optionally one or more than one pharmaceutically acceptable excipient.

[0205] Preferably, the mixing of the components of the composition according to the application is carried out in the aqueous solvent described above. The order of addition of the components constituting the composition according to the application can vary. For example, indocyanine green can be dissolved in the aqueous diluent, then ethylenediaminetetraacetic acid (EDTA) and / or one or more than one salt thereof, histidine and sodium chloride, and the additives can be added. The stabilizing compounds can be mixed before being added to the dissolved indocyanine green. Alternatively, indocyanine green can be introduced into an aqueous solution in which the stabilizing compounds and the optional pharmaceutically acceptable additives have been previously dissolved.

[0206] 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.

[0207] 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.

[0208] 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.

[0209] 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.

[0210] When provided as an aqueous composition, the indocyanine green composition according to the invention can also be reconstituted using the lyophilized product from the kit described above and an aqueous diluent. To reconstitute, the lyophilized composition can be introduced into a vial, and then the aqueous diluent can be added.

[0211] 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.

[0212] 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.

[0213] Advantageously, the compositions according to the invention are stored at ambient temperature.

[0214] Use of the composition according to the invention

[0215] 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.

[0216] 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.

[0217] Advantageously, the composition of the application can be used in a diagnostic and / or therapeutic method as described above, wherein the aqueous ICG composition is administered to the patient after reconstitution or preparation from one of the above-mentioned kits, at least 1 month later, preferably at least 6 months later, more preferably at least 12 months later, most preferably at least 2 years later.

[0218] In particular, the composition according to the application can be used in thermotherapy, ICG-enhanced selective photocoagulation, photodynamic therapy (PDT), photodynamic and thermotherapy (PHT) therapy. More particularly, as PDT applications, the composition according to the application can be used in infection treatment, acne treatment, macular surgery, cancer treatment, etc. The use of the composition according to the application can be associated with other therapies such as immunotherapy, radiotherapy, ultrasound and chemotherapy.

[0219] The composition of the application can also be used to obtain angiographic images of the patient's tissues, for the determination of cardiac output, for the determination of liver function and liver blood flow, for the diagnosis and treatment of age-related macular degeneration, associated choroidal neovascularization and tumors.

[0220] The composition according to the application is based on components that have already been tested clinically and approved for administration to humans and / or animals.

[0221] For diagnostic applications, the amount of ICG composition administered to the patient should be sufficient to allow the dye to emit fluorescence when irradiated at the appropriate wavelength, the peak absorption and emission of ICG being known to be located at 800-850 nm. The same criterion applies to therapeutic methods using ICG solutions; sufficient dye should be administered for the treatment to be effective. The person skilled in the art can easily determine the amount of ICG composition to be administered and should be at least the acceptable concentration currently used for ophthalmic angiography, for example, 2 ml of a 20 mg / ml ICG solution for diagnosis. Higher dye concentrations can advantageously be used in any of these diagnostic and therapeutic methods, as recognized by the person skilled in the art.

[0222] Compared to prior art ICG compositions, the stability of ICG in the solution according to the application is improved, thus allowing a better diagnosis / treatment of the patient when the same amount of compound is injected. This is because the improvement in the stability of ICG will provide a more intense response to fluorescence irradiation.

[0223] Example

[0224] I - Materials and methods:

[0225] 1. Reagents and chemicals:

[0226] Indocyanine green was supplied by Biophore India Pharmaceuticals and used without further purification. EDTA tetrasodium salt hydrate (EDTA 4Na) (CAS number: 194491-31-1), EDTA 2Na Ca (CAS number: 62-33-9) and Histidine (CAS number: 71-00-1) were all purchased from Aldrich and used without purification. NaCI solution (CAS number: 7647-14-5) was purchased from VWR.

[0227] 2. Equipment:

[0228] The stability of the formulations in the above examples was determined by high pressure liquid chromatography (HPLC). The equipment used to perform the analysis was an Agilent Infinity 1260 HPLC / MS system equipped with a diode array detector (UV-Vis), a Waters Xbridge Shield RP18-4.6x100-5pm column and a MS ESI spectrometer. The injection volume of the solution was 5 pL. The flow rate was 1 ml / min, the column temperature was 30 °C and the detection wavelength was 240 nm.

[0229] The elution was performed with a gradient method. The mobile phase consisted of a mixture of 10 mM ammonium acetate / acetic acid buffer at pH 5.5 (eluent A) and acetonitrile (eluent B). The composition of the mobile phase was constantly changing during the elution, as detailed in Table I:

[0230] Table I

[0231] Time (min) Eluent A % Eluent B % 0.0 85 15 3.0 85 15 40.0 30 70 40.1 85 15 47.0 85 15

[0232] II - Sample preparation:

[0233] The ICG compositions were prepared as follows: the required amount of EDTA salt and / or histidine was weighed into a Type I clear glass vial. The vial was then de-capped and 25 mg of indocyanine green was weighed in. The vial was closed with a bromobutyl rubber stopper (from VWS) and then purged with a nitrogen gas stream.

[0234] For compositions C3, C4, C7 and C8, 10 ml of WFI previously degassed with nitrogen was then added using a syringe.

[0235] For compositions C1 and C5, 5 ml of WFI previously degassed and 5 ml of a 0.9% NaCI degassed solution were added.

[0236] For compositions C2 and C6, 5 ml of WFI previously degassed and 5 ml of a 0.45% NaCI degassed solution were added.

[0237] Composition C9 was prepared by weighing 25 mg ICG into a vial, sealing the vial with a bromobutyl rubber stopper and purging it with a nitrogen stream, and then adding 5 ml of degassed WFI and 5 ml of 0.9% NaCl degassed solution.

[0238] Gently shake the vial manually until the solute dissolves. Then store the vial at room temperature, away from light.

[0239] III - Results:

[0240] Table II

[0241]

[0242] *: Tetrasodium ethylenediaminetetraacetate hydrate

[0243] These results indicate that the storage stability of ICG compositions containing EDTA salts and histidine can be improved when used alone. A synergistic effect was also observed when the salts EDTA2NaCa or EDTA4Na were combined with histidine and NaCl: ICG solutions C1 and C2 were significantly more stable than compositions containing only one or two components of the conjugate containing the desired stable compounds.

[0244] References

[0245] [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.

[0246] [2]Engel E, Schraml R, Maisch T, Kobuch K, B, Szeimies RM, Hillenkamp J, Baumler W, Vasold R. Light-induced decomposition of indocyanine green. InvestOphthalmol Vis Sci. 2008 May; 49(5): 1 777-83.

[0247] [3] Saxena V, Sadoqi M, Shao J. Degradation kinetics of indocyanine green in aqueous solution. J Pharm Sci. 2003 Oct;92(10):2090-7.

[0248] [4] Gathje J, Steuer RR, Nicholes KR. Stability studies on indocyanine green dye. J Appl Physiol. 1970 Aug;29(2):181-5.

[0249] [5] Barbier f, Deweerdt ga. Chromatography and I.R. spectrography of indocyanine green. Clin Chim Acta. 1964 Dec;10:549-54.

[0250] [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. 2018 Sep 12;17(9):1189-1196.

[0251] [7] Kirchherr AK, Briel A, K. Stabilization of indocyanine green by encapsulation within micellar systems. Mol Pharm. 2009 Mar-Apr;6(2):480-91.

[0252] [8] DeDora DJ, Suhrland C, Goenka S, Mullick Chowdhury S, Lalwani G, Mujica-Parodi LR, Sitharaman B. Sulfobutyl ether p-cyclodextrin (Captisol )and methyl beta-cyclodextrin enhance and stabilize fluorescence of aqueous indocyanine green. J Biomed Mater Res B Appl Biomater. 2016 Oct; 104(7): 1457-64.

Claims

1. An indocyanine green composition comprising at least: - from 10% to 70% by weight of indocyanine green, - from 0.1% to 15% by weight of ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, - from 1% to 20% by weight of histidine, and - from 20% to 80% by weight of sodium chloride, the percentages by weight being expressed with respect to the total weight of the four compounds.

2. The indocyanine green composition according to claim 1, wherein the composition comprises - from 10% to 70% by weight of indocyanine green, - from 0.1% to 15% by weight of one or more ethylenediaminetetraacetic acid salts, - from 1% to 20% by weight of histidine, and - from 20% to 80% by weight of sodium chloride, the percentages by weight being expressed with respect to the total weight of the four compounds.

3. The indocyanine green composition according to claim 1 or 2, wherein the ethylenediaminetetraacetic acid salt is selected from disodium EDTA, sodium calcium EDTA, tetrasodium EDTA, dicalcium EDTA, or a mixture thereof.

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

5. The indocyanine green composition according to claim 1, which is an aqueous composition.

6. The indocyanine green composition according to claim 5, wherein the concentration of indocyanine green in the aqueous composition is from 0.1 mg / ml to 50 mg / ml.

7. The indocyanine green composition according to claim 5 or 6, wherein the content of indocyanine green in the composition is greater than 90% or equal to 90% before storage, the content of indocyanine green being measured by HPLC at 240 nm in % area.

8. The indocyanine green composition according to claim 1, wherein the composition is in the form of a lyophilized powder.

9. The indocyanine green composition according to claim 1, for use as a medicament or diagnostic agent.

10. A kit comprising in separate compartments at least 1 / a composition as defined in claim 8 and 2 / an aqueous diluent.

11. A kit capable of being used to prepare a composition according to claim 5, comprising in separate compartments at least 1 / indocyanine green and 2 / an aqueous solution comprising at least ethylenediaminetetraacetic acid (EDTA) and / or one or more salts thereof, histidine and sodium chloride.

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