Composition containing a fluorophore-labeled UPAR-targeted peptide conjugate

Abstract

The present invention describes a composition comprising a fluorophore-labeled uPAR targeting component, a buffer and a surfactant, wherein the fluorophore-labeled uPAR targeting component is solubilized in the composition by the surfactant present, and the composition comprises up to 10% water by weight, preferably up to 5% water by weight.

Description

[Technical Field]

[0001] The present invention relates to a composition comprising a fluorophore-labeled receptor targeting component, preferably a fluorophore-labeled receptor-targeting peptide conjugate, and more preferably a fluorophore-labeled uPAR (urokinase-type plasminogen activator receptor)-targeting peptide conjugate. [Background technology]

[0002] There are existing compositions comprising one or more fluorophore-labeled receptor-targeting peptide conjugates, the receptor of which is uPAR. For example, International Publication No. 2016 / 041558 discloses a conjugate that binds to the cell surface receptor uPA (uPAR). The conjugate is based on a fluorescently labeled peptide useful as a diagnostic probe for the surface of cells expressing uPAR. The conjugate can possess appropriate detectable and imageable labeling, which also enables qualitative detection and quantification of uPAR levels in vitro and in vivo. This makes surgical resection of tumors more optimal. Furthermore, various alternatives to the conjugate, e.g., a conjugate containing ICG-Glu-Glu-AE105 (where ICG is indocyanine green, Glu-Glu are two glutamic acids acting as a linker, and AE105 is a uPAR-targeting peptide), are described in International Publication No. 2016 / 041558.

[0003] The present invention relates to providing formulations comprising a fluorophore-labeled uPAR-targeted peptide conjugate, such as ICG-Glu-Glu-AE105 or other alternatives. The formulations according to the present invention result in improved solubility of the fluorophore-labeled uPAR-targeted peptide conjugate, as well as increased stability of the formulation as a whole and the fluorophore-labeled uPAR-targeted peptide conjugate contained therein. [Overview of the project] [Problems that the invention aims to solve]

[0004] The latter objective described above is achieved by a composition comprising a fluorophore-labeled receptor-targeting component, a buffer, and a surfactant, wherein the fluorophore-labeled receptor-targeting component is solubilized in the composition by the present surfactant, and the composition contains up to 10% by weight of water, preferably up to 5% by weight of water. As should be understood from the above, the present invention relates in particular to a composition comprising a fluorophore-labeled receptor-targeting peptide conjugate, more specifically, a fluorophore-labeled uPAR-targeting peptide conjugate, for example, a peptide conjugate having 80% sequence homology to the peptides shown in Table 4 of Biochemstry 2001, 40, 12157-12168, Michael Ploug et al., "Peptide-Derived Antagonists of the Urokinase Receptor. Affinity Maturation by Combinatorial Chemistry, Identification of Functional Epitopes, and Inhibitory Effect on Cancer Cell Intravasation". However, it should be noted that receptor-targeting components may include small molecules, proteins, antibodies, frabs, or, of course, peptides, or any combination thereof. [Means for solving the problem]

[0005] The compositions according to the present invention exhibit several advantages. The two most important are the complete solubilization of the fluorophore-labeled receptor-targeted peptide conjugate and the high stability of the overall composition. Stability is primarily facilitated by the absence of water. For example, the lyophilized product, one type of target product according to the present invention, is an example of a low water content. However, to obtain a suitable formulation, good solubilization is necessary. Freeze dryingThe product should also have the cake situation, and thus solubilization of the fluorophore-labeled receptor-targeted peptide conjugate is very important. Further, it should be noted that the compositions according to the invention can have a water content in different dosage forms well below 5% by weight, for example in the range of 1 to 3% by weight. In this regard, it should also be noted that the water content can also increase, for example, during a storage life of 3 years.

[0006] It should be noted above that the compositions according to the invention also contain a buffer and a surfactant. The surfactant is a component that ensures that the fluorophore-labeled receptor-targeted peptide conjugate is solubilized in the composition. Different types of surfactants and their combinations can be incorporated into the compositions according to the invention.

[0007] Furthermore, the buffer components can be of different types according to the invention. Some possible examples are provided below.

Brief Description of the Drawings

[0008] [Figure AD] None [Figure E] None

Modes for Carrying Out the Invention

[0009] [[ID= 27]]Various aspects of the invention are further described and dealt with below. Further, some specific embodiments of the invention are also further described.

[0010] The characteristics of the fluorophore-labeled receptor-targeting component solubilized in the composition can be viewed or shown in different ways in the composition according to the present invention. One such indicator is when looking at the absorbance spectrum of the composition according to the present invention. This is further explained in the examples. Further, according to one specific embodiment of the present invention, the fluorophore-labeled receptor-targeting component is solubilized in the composition at a level corresponding to a single peak having an absorption maximum near 800 nm when measuring the absorbance spectrum of the composition in the wavelength range of 700 to 825 nm. The absorbance spectrum of the composition according to the present invention does not exclude peak areas exceeding one, such as double peaks, but it should be noted that an important feature in this regard is that the area level of one main peak having an absorption maximum near 800 nm is high. To make this more clear in this regard, an absorbance spectrum having distinct double peaks is an indicator that the fluorophore-labeled receptor-targeting component in the composition is not completely or substantially completely solubilized, and such a composition can be said not to be part of the scope of the present invention.

[0011] Further, according to one specific embodiment of the present invention, the fluorophore-labeled receptor-targeting component is solubilized in the composition at a level corresponding to having an absorbance spectrum peak with a maximum near 800 nm, and the area of the absorbance spectrum peak is at least 50%, preferably at least 60%, more preferably at least 65% of the total area of the absorption spectrum in a given wavelength range of from 600 to 900 nm.

[0012] This relative area value of a single peak to the total area is calculated by dividing the area into a quadrilateral, and the AUC (Area Under Curve) in the wavelength range of 600-900 nm can be determined. Next, a line is drawn in the center, i.e., at 750 nm. Then, the AUC to the left and right of the line at 750 nm is calculated, respectively. Next, the AUC to the right, which represents a single peak near 800 nm, is divided by the total AUC calculated according to the above. As described above, according to one embodiment, this single peak near 800 nm accounts for at least 50%, preferably at least 60%, and more preferably more than 65% of the total area in the wavelength range of 600-900 nm.

[0013] The present invention preferably includes other components. For example, cryoprotectants are such components. Cryoprotectants can be combined with small molecules, peptides, or proteins and are molecules that significantly prevent or reduce the chemical and / or physical instability of small molecules, peptides, or proteins during drying, particularly during freeze-drying and subsequent storage. Some examples of cryoprotectants include sugars, e.g., sucrose or trehalose; amino acids, e.g., monosodium glutamate, histidine, or arginine; methylamines, e.g., betaine; lyotropic salts, e.g., magnesium sulfate. Other examples include polyols, e.g., trivalent or higher sugar alcohols, e.g., glycerin, erythritol, glycerol, arabitol, xylitol, sorbitol, and mannitol. Some further examples include ethylene glycol; propylene glycol; polyethylene glycol; pluronic acid; or hydroxyalkyl starch, e.g., hydroxyethyl starch (HES). Naturally, combinations are also entirely possible. According to one specific embodiment of the present invention, the composition comprises a cryoprotectant, preferably selected from the group consisting of sucrose, trehalose, mannitol, glycine, or combinations thereof, more preferably mannitol or mannitol in combination with one or more other components, more preferably a combination of mannitol and glycine or a combination of mannitol and sucrose. Some of these preferred alternatives according to the present invention are further presented in examples. Furthermore, in this context, it can be said that it is usually appropriate to use enough sugar to prepare an isotonic formulation to avoid a reaction (stinging pain) at the injection site.

[0014] As described above, the compositions according to the present invention include surfactants. Many different forms of surfactants can be used in accordance with the present invention. There are no limitations on possible substitutes according to the present invention, and the following examples can be given. According to one embodiment of the present invention, the compositions include nonionic surfactants. One example is ethoxylates, e.g., fatty alcohol ethoxylates, alkylphenol ethoxylates (APEs), fatty acid ethoxylates, ethoxylated fatty acid esters and oils, or ethoxylated amines or fatty acid amines. Another type is fatty acid esters, e.g., fatty acid esters of polyhydroxy compounds, e.g., fatty acid esters of glycerol, sorbitol, sucrose or alkyl polyglucosides. Other examples are various types of amine oxides, polyoxomers, sulfoxides or phosphine oxides.

[0015] Furthermore, various forms of tween surfactants are interesting examples of the present invention. For example, Tween 20 can be used, which is used in the substitutes presented in the examples. Tween 80 and other polysorbates are also perfectly usable instead. In addition, hydroxyl-β-cyclodextrin is another possible example. Naturally, combinations are also usable.

[0016] It should be noted that according to the present invention, many surfactant substitutes are possible, including ionic surfactants. Furthermore, surfactants at various concentrations are also quite possible according to the present invention, and are naturally not limited to the levels presented as examples.

[0017] Another component included in the composition according to the present invention is at least one buffer / buffer. In this case as well, many substitutes are perfectly possible. Non-limiting examples include various forms of borates, carbonates, citrates, and phosphates, e.g., PBS. Furthermore, glycine and Tris and formulated Tris solutions are also perfectly possible substitutes. Further, some other more specific examples are imidazole and succinic acid formulations. Also possible are bis-tris formulations, e.g., bis-tris or N,N-bis(2-hydroxyethyl)glycine. Further specific examples include sulfonic acid preparations, such as 2-(N-morpholino)ethanesulfonic acid or 4-morpholineethanesulfonic acid, N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid, 3-(cyclohexylamino)-1-propanesulfonic acid, 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid, and N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid).

[0018] According to one general embodiment of the present invention, the buffer solution is supplied so that the composition has a physiological pH or substantially physiological pH. As shown in the example, sodium phosphate can be used according to the present invention to set the pH to, for example, around 7.4.

[0019] Furthermore, the fluorophore may be modified according to the present invention. Firstly, according to one embodiment, the fluorophore-labeled receptor targeting component comprises a fluorophore, a peptide that binds to the receptor, and a linker group, wherein the fluorophore, the peptide that binds to the receptor, and the linker group are linked by covalent bonds. For example, the linker group may include other short oligomers such as oligoethylene glycol or oligoglycerol, oligolactic acid, or a carbohydrate (optionally linked to at least one amino acid by covalent bonds).

[0020] With regard to fluorophore substitutes, many such are possible according to the present invention. According to one embodiment of the present invention, the fluorophore is preferably selected from indocyanine green (ICG), methylene blue, 5-ALA, protoporphyrin IX, IRDye800CW, ZW800-1, Cy5, Cy7, Cy5.5, Cy7.5, IRDye700DX, Alexa fluor 488, fluorescein isothiocyanate, Flav7, CH1055, Q1, Q4, H1, IR-FEP, IR-BBEP, IR-E1, IR-FGP, or IR-FTAP, and the fluorophore is preferably indocyanine green (ICG).

[0021] Furthermore, the peptides involved may also be of different types. According to one specific embodiment, the peptide may be selected from any of the following: -Asp-Cha-Phe-(D)Ser-(D)Arg-Tyr-Leu-Trp-Ser(-); -Asp-Cha-Phe-(D)Ser-(D)Arg-Tyr-Leu-Trp-Ser-OH; or -Asp-Cha-Phe-(D)Ser-(D)Arg-Tyr-Leu-Trp-Ser-NH2.

[0022] Furthermore, amino acids may be selected from proteinogenic and non-proteinogenic amino acids, and include native and synthetic amino acids. In this regard, it can be further noted that native amino acids may include C-α-alkylated amino acids such as aminoisobutyric acid (Aib), N-alkylated amino acids such as sarcosine, and naturally occurring β-amino acids such as β-alanine. Furthermore, synthetic amino acids may include amino acids with non-proteinogenic side chains, such as cyclohexylalanine (Cha), γ-amino acids, and dipeptide mimics. The term dipeptide mimic can be interpreted as an organic molecule that mimics a dipeptide by presenting two amino acid side chains, for example by reducing the amide bond that links two residues together. Amino acids with non-proteinogenic side chains may also include amino acids with side chains whose movement in chi space is restricted. The term restricted movement in chi space can be interpreted as restricted flexibility in the rotation of the side chain group. Oligopeptides may consist of up to 50 amino acids and may include dipeptides, tripeptides, tetrapeptides, and pentapeptides, and may further consist of proteinogenic and non-proteinogenic amino acids.

[0023] In relation to the present invention, it should be noted that when peptides are included, it is appropriate that AE105 is not the only one. Many others are also possible, for example, those disclosed in Table 4 of "Peptide-Derived Antagonists of the Urokinase Receptor. Affinity Maturation by Combinatorial Chemistry, Identification of Functional Epitopes, and Inhibitory Effect on Cancer Cell Intravasation" by Michael Ploug et al., Biochemstry 2001, 40, 12157-12168. Accordingly, according to one specific embodiment of the present invention, the peptides may be AE101, AE105, AE106, AE110, AE112, AE113, AE116, AE133, AE133 *, AE134, AE135, AE136, AE137, AE138, AE139, AE145, AE140, AE141, AE142, AE143, AE144, AE164, AE164 * AE120, AE120 * Alternatively, it is AE151, and the following applies: AE101 is d-Cha-FsrYLWS, AE105 is D-Cha-FsrYLWS, AE106 is D-Cha-FSrYLWS, AE110 is D-Cha-FsRYLWS, AE112 is DFFsrYLWS, AE113 is DNFsrYLWS, AE116 is D-Cha-FsrGYLWS, AE133 is KGSGG-D-Cha-FsrYLWS, and AE133 * AE134 is KGSGG-D-Cha-FsrYLWS, AE134 is KGSGG-D-Cha-FsrYLWA, AE135 is KGSGG-D-Cha-FsrYLAS, AE136 is KGSGG-D-Cha-FsrYAWS, AE137 is KGSGG-D-Cha-FsrALWS, AE138 is KGSGG-D-Cha-FsaYLWS, AE139 is KGSGG-D-Cha-FarLWS, AE145 is KGSGG-D-Cha-FArYLWS, AE140 is KGSGG-D-Cha-AsrYLWS, AE141 is KGSGG-DAFsrYLWS, AE142 is KGSGG-A-Cha-FsrYLWS, and AE143 is KGSGG-D-Chp-FsrYLWS c Therefore, AE144 is KGSGG-D-Cpa-FsrYLWS c Therefore, AE164 is KGSGG-DFFsrYLWS, and AE164 * It is KGSGG-DFFsrYLWS, and AE120 is [D-Cha-FsrYLWS]2- / 3A-K cand AE120 * is [D-Cha-F-s-r-Y-L-W-S]2- / 3A-K c and AE151 is [r-W-D-Cha-S-L-s-F-Y]2- / 3A-K c (), or is selected from combinations thereof, or is a peptide having at least 80% sequence homology to any of these peptides. According to a preferred embodiment, the selected peptide is AE105.

[0024] Furthermore, a "linker group" is a molecule that connects two parts to create a conjugate, and most of their respective functions are maintained. For example, the peptide binds to the receptor and the fluorophore emits light. The linker connects the two (peptide and fluorophore) together, and their desired properties are preserved in whole or in part. This means, for example, that the peptide still binds to the receptor and the fluorophore preserves its properties. The linker can be at least part of either of the two linked molecules. According to one embodiment, the linker is Glu-Glu.

[0025] According to a preferred embodiment of the present invention, the receptor is uPAR (urokinase-type plasminogen activator receptor). Furthermore, according to yet another preferred embodiment, the fluorophore-labeled receptor targeting component is ICG-Glu-Glu-AE105: [Chemical formula] or a pharmaceutically acceptable salt thereof.

[0026] According to one embodiment, the concentration of ICG-Glu-Glu-AE105 is in the range of 0.1 to 10.0 mg / ml, for example in the range of 0.1 to 8.0 mg / ml, for example in the range of 0.1 to 5.0 mg / ml, for example in the range of 0.2 to 3.0 mg / ml, preferably in the range of 0.5 to 2.0 mg / ml.

[0027] In accordance with the above, according to one specific embodiment of the present invention, the fluorophore-labeled receptor targeting component is ICG-Glu-Glu-AE105: [ka] or a pharmaceutically acceptable salt thereof, The concentration of ICG-Glu-Glu-AE105 is in the range of 0.1 to 10.0 mg / ml. The composition also includes a buffer in the form of sodium phosphate at a concentration of 5-50 mM. The composition includes a combination of cryoprotective agents: mannitol at a concentration of 10-50 mg / ml and glycine at a concentration of 1-30 mg / ml.

[0028] Furthermore, preferably, the composition according to the present invention contains at least one polysorbate, preferably polysorbate 20, and more preferably polysorbate 20 at a concentration of more than 0.01% by weight.

[0029] The present invention also provides other interesting aspects of compositions comprising fluorophore-labeled receptor-targeting components. One such aspect is the pharmacokinetic profile. Accordingly, according to one specific embodiment of the present invention, the fluorophore-labeled receptor-targeting component is - Receptors, preferably molecules that bind to uPAR; and - A linker group that covalently attaches to a molecule that binds a fluorophore to a receptor, The conjugate contains and is adapted for systemic administration into the body of a human or animal.

[0030] Furthermore, according to one embodiment, the fluorophore-labeled receptor-targeting component has a pharmacokinetic profile in which a TBR (tumor-to-background ratio) of at least 2.5 is reached within 3.5 hours after administration, and the TBR level of at least 2.5 is maintained for at least 30 minutes before decreasing again, and the fluorophore-labeled receptor-targeting component is a human uPAR-targeting conjugate.

[0031] Furthermore, according to yet another embodiment, the fluorophore-labeled receptor-targeting component has a pharmacokinetic profile in which a TBR (tumor-to-background ratio) of at least 2.5 is reached within 3.5 hours after administration, and the level of TBR of at least 2.5 is maintained for at least 30 minutes before decreasing again, and preferably the fluorophore-labeled receptor-targeting component is a human uPAR-targeting conjugate.

[0032] Furthermore, according to yet another embodiment, The fluorophore-labeled receptor-targeting component has a pharmacokinetic profile in which the plasma half-life is up to 75 hours, preferably up to 20 hours, more preferably up to 15 hours, more preferably in the range of 6 to 15 hours, and most preferably in the range of 6 to 10 hours.

[0033] While "blood" and "plasma" are sometimes used synonymously in relation to some of the expressions above and below, it should be noted that, strictly speaking, plasma is the yellowish liquid component of blood that usually holds blood cells in suspension. Plasma is the liquid part of blood that carries cells and proteins throughout the body.

[0034] In this regard, the target receptor can be of various types according to the present invention. Targeted receptors may be urokinase-type plasminogen-activating factor receptor (uPAR), tissue factor (TF), epidermal growth factor receptor (EGFR), prostate-specific membrane antigen (PSMA), vascular endothelial growth factor (VEGF), folate receptor, matrix metalloproteinase-2 (MMP-2), membrane type I MMP, transmembrane inhibitor of metalloproteinase-2 (TIMP2), ClC-3 chloride ion channel, disaccharides and other glycans, or glycophosphatidylinositol (GPI)-immobilized cell membrane receptors. Furthermore, the type of receptor may have proteolytic activity or other enzymatic activity on the cell surface, such as urokinase (uPA). Moreover, the receptor is expressed in such a way in human cancer and correlates, for example, with poor prognosis, local invasiveness, or metastasis. In this regard, it can be further noted that the conjugate product according to the present invention is mainly / partially immobilized outside the cell expressing the specific receptor, i.e., unlike the case in which the conjugate product is internalized into the cell expressing the receptor.

[0035] In relation to receptors, when we say that the receptor (uPAR) is expressed on cancer cells, it can also be noted that this may mean that it is expressed on “normal” stromal cells of the body that are affected by the cancer cells, with which they are in contact or in very close proximity (e.g., 2-5 cells between each). This is also true when “normal” cells assist cancer cells in invading normal tissue. For clarity, such closely located “normal” stromal cells are also included as cancer cells. Cells that are under the influence of cancer cells and expressing uPAR can be argued to no longer be called normal, hence the term "normal in quotation marks."

[0036] As suggested above, the receptor-targeted conjugate according to the present invention, and more appropriately the uPAR-targeted conjugate, may also have an optimal receptor binding profile and pharmacokinetic profile. A systemically injected uPAR-targeted conjugate is distributed through the circulatory system to hemoperfused tissues and organs in the body. In hemoperfused tissues, the uPAR-targeted conjugate accumulates. When such tissue is exposed to light having a wavelength (color) absorbed by the fluorophores contained in the uPAR-targeted conjugate, the fluorophores emit light. The uPAR-targeted conjugate ("L" in the following formula) binds to the receptor ("R" in the following formula) according to first-order kinetics: R+L⇔RL, the reaction involves onate bonding (K on ), off-rate coupling (Koff) and the resulting equilibrium coupling constant K D (K D Characterized by (=Koff / Kon). Preferably, as will be described later, K on >1×10 3 M -1 s -1 and / or K off <1×10 -1 s -1 , comfortable K on ≥7.3 × 10 5 M -1 s -1 That is the case.

[0037] The product conjugate is distributed via the blood (which is then eliminated via excretion by the liver and / or kidneys) or redistributed to compartments other than the bloodstream. This causes tissue containing cells expressing the targeted receptor to which the uPAR-targeted conjugate binds to become brighter than the background, resulting in a so-called "TBR" (tumor-to-background ratio). The conjugate product is illuminated using a light source that produces light of a specific wavelength, and the light emitted from the fluorophores is then detected using a specific filter for specific emission light. In a conceivable ideal scenario, cancer cells would light up with a sufficiently high relative light intensity immediately after injection, with no background light, and the resulting desired TBR of at least 2.5 would last for several hours. In the real world, it is acceptable if the desired TBR of 2.5 is reached within 3.5 hours after injection and lasts for at least 30 minutes. Further description is given below with reference to this aspect and others in relation to the present invention.

[0038] In accordance with the above, according to one embodiment of the present invention, the fluorophore-labeled receptor-targeting component has a pharmacokinetic profile in which a TBR (tumor-to-background ratio) of at least 2.8 is reached within 3.5 hours after administration, and the level of TBR of at least 2.8 is maintained for at least 30 minutes before decreasing again.

[0039] Furthermore, according to yet another embodiment, the peak TBR of the fluorophore-labeled receptor-targeting component after administration is at least 3.

[0040] The conjugate products according to this embodiment of the present invention exhibit several features, including those relating to their pharmacokinetics and receptor binding affinity. Receptor-targeted conjugates provide a specific combination of plasma half-life and receptor binding affinity.

[0041] In accordance with the above, according to one specific embodiment of the present invention, the rate at which a protein (P)-ligand (L) complex is formed can be defined as follows:

number

[0042] Furthermore, according to yet another embodiment, the K of the fluorophore-labeled receptor targeting component on The K of uPA, a natural ligand. on That's all, K on ≥4.6 × 10 6 M -1 s -1 That's how it works.

[0043] Furthermore, according to yet another specific embodiment of the present invention, IC 50 Receptor binding affinity, as defined, is a measure of ligand / receptor binding affinity. According to one embodiment, the fluorophore-labeled receptor targeting component binds the native ligand (uPA) to uPAR at a maximum IC of 1,000 nM, preferably up to 200 nM, more preferably up to 50 nM, and most preferably up to 25 nM. 50 Replace with a value. Furthermore, according to yet another embodiment of the present invention, the receptor binding affinity of the fluorophore-labeled receptor targeting component to uPAR, defined as Kd, is in the range of up to 2,500 nM, preferably up to 2,000 nM, more preferably up to 500 nM, and most preferably 2,000 to 300 nM.

[0044] Hereinafter, several important aspects and features related to this aspect of the present invention will be further described. The conjugate composition should be administered systemically at a sufficiently high dose to allow sufficient distribution in the body to reach the targeted specific receptor present on cancer cells. This is to ensure that the conjugate rapidly binds to the target receptor, and combined with a short plasma half-life, enables rapid TBR production, which lasts for a sufficiently long time to be useful. In short, high receptor binding affinity combined with a short plasma half-life creates a fast, high, and long-lasting TBR.

[0045] TBR can be calculated from the relative intensity of light from the tumor and the background. Light intensity can be measured using a simple, commercially available camera with physical filters, such as, but not limited to, the clinically approved NIR camera system Fluobeam® 800 (Fluoptics, Grenoble, France) or EleVision® (Medtronic, USA). To enhance TBR, post-image recording optimization of the image may be applied using software.

[0046] Higher concentrations push the equilibrium toward more conjugates binding to the target receptor. However, concentrations should not be too high, as this increases the risk of toxic effects for the patient and increases the cost of administration beyond what is actually acceptable. According to the present invention, administration may be systemic, preferably intravenously, so that the plasma concentration reaches its peak rapidly. The plasma concentration then decreases as the conjugate product is metabolized, excreted (by the liver), eliminated (by the kidneys), and / or distributed to different distribution compartments than the blood. According to the present invention, the concentration should be sufficiently high and maintained for a sufficiently long period so that the conjugate product reaches a certain concentration in the compartment associated with the targeted receptor (e.g., plasma, tissue stoma, cerebrospinal fluid, urine) for a sufficiently long time, so that the conjugate product can bind to the receptor. According to one specific embodiment of the present invention, the drug is administered in the range of 0.1 to 2,000 mg per drug unit, preferably in the range of 1 to 1,000 mg per human drug unit.

[0047] Furthermore, another important feature is the binding affinity to the target receptor, including onset and offset, as discussed above. This marks the target tumor cells as quickly as possible, with the highest possible relative light intensity, and with the highest possible contrast over the longest possible time. Fast onset binding and slow offset are preferred. In relation to the above, it can be mentioned that TBR is a feature measured in vivo and is created by a combination of several other features such as plasma half-life, but receptor binding affinity is one important feature.

[0048] Furthermore, selectivity for cancer tissue is also of interest in relation to the present invention. According to one specific embodiment of the present invention, the receptor-targeted conjugate has selectivity for cancer tissue of at least 60%, preferably more than 70%, more preferably more than 80%, and most preferably more than 90%. Thus, the conjugate product is characterized by having selectivity for at least 60%, or 70%, or 80%, or 90% of cancer tissue. Selectivity is understood as the relative number of tissue samples that a surgeon considers to be cancerous and removes based on their light intensity / contrast, and the number of tissue samples that are subsequently confirmed to be cancerous. For example, if a surgeon removes 10 tissue samples that he / she considers to be cancerous, and 7 of them are histologically confirmed to be cancerous, the selectivity is 7 / 10 (70%). If 100% of the tissue samples removed by a surgeon are proven to be cancerous, the selectivity is 100%. If only half of the tissue samples removed by a surgeon are cancerous and the other half are normal tissue, the selectivity is 50%.

[0049] Another aspect related to the above aspects is the site of discharge and elimination. Various types of conjugates according to the present invention are discharged and / or eliminated in various organs and are therefore unsuitable for types of cancer localized in these organs. Furthermore, according to the present invention, the indication and type of target receptor are also important. The preference here is that the receptor needs to be expressed in cancer where the patient must be in the greatest benefit to the operator (e.g., surgeon). Furthermore, it is of particular interest that the receptor be expressed in the right portion of the cancer. This is interesting because the central part of the cancer is usually easier for the surgeon to see and remove. However, the border and locally invasive growth from the cancer are more difficult for the surgeon to see and separate from normal tissue, and therefore more difficult for the surgeon to remove and / or preserve normal tissue.

[0050] Ideally, the conjugate product would be administered to the patient when the surgeon is examining the removed tissue or planning postoperative procedures, for example, when the surgeon is examining the removed cancerous tissue and when the surgeon is examining whether cancer cells remain in the patient immediately after the removal of the cancerous tissue, thereby investigating the integrity of the surgical procedure. This could be, for example, immediately before or during surgery, or immediately after, during, or before anesthesia. This represents a significant improvement over other known alternatives, which must be administered 6, 12, 18 hours, or even 1-2 days before surgery to be ready for use during surgery, and do not always provide satisfactory TBR, nor do they have satisfactory specificity to cancer, meaning that normal tissue is not mistaken for cancerous tissue and removed. The combination of receptor binding and plasma clearance features according to the present invention would enable such improved use. In other words, the conjugate product according to the present invention has a pharmacokinetic profile that allows it to be administered as close to the time of use as possible for the surgeon, for example, around the time of anesthesia. Furthermore, it can be said that the conjugate product according to the present invention makes it possible for the time from administration to the time suitable for first use to be at least 1200 minutes, for example, 600 minutes, for example, 300 minutes, or 120 minutes, for example, preferably 60 minutes, or still 30 minutes, for example, 15 minutes.

[0051] To summarize several different aspects related to the pharmacokinetic aspects of the present invention, the following can be stated. The conjugate according to the present invention preferably exhibits the following features: TBR is rapidly generated and sustained for a long period (during surgery), which is achieved through the following combinations: - Reaching a sufficiently high concentration in plasma; - Reaching a sufficiently high concentration in cancer tissue; - Possesses an appropriate coupling rate; - Has an appropriate plasma exclusion half-life; - It can be administered at safe dose levels and does not cause serious adverse events; -In a subset of the target cancer, it exhibits high selectivity for the target receptor which is widely expressed in the target cancer, and has high selectivity for the cancer compared to normal tissue adjacent to the target cancer; and - Detectable with available / existing equipment

[0052] The above-mentioned given characteristics may be measured and analyzed by various means and instruments.

[0053] In addition to an optimal pharmacokinetic profile, there are certain other interesting aspects of the present invention. Two such aspects are solubility and protein-binding ability. Compositions according to the present invention offer a preferred combination of beneficial features. According to one specific embodiment, the composition is soluble in less than 10 minutes, preferably less than 5 minutes, more preferably less than 2 minutes, and most preferably less than 1 minute. Furthermore, according to yet another specific embodiment, the composition has in vivo protein binding of more than 50%, preferably more than 75%, more preferably more than 85%, more preferably more than 90%, more preferably more than 95%, and most preferably more than 99%.

[0054] As is clear from the above, the conjugates and compositions according to the present invention are intended for use in cancer surgery, cancer treatment and / or cancer diagnosis. Accordingly, according to one embodiment of the present invention, receptor-targeted conjugates and compositions according to the present invention are provided for use in cancer surgery, cancer treatment or diagnosis, including use in optical imaging / fluorescence imaging (FLI) of cancer. It should be said that the conjugates and compositions according to the present invention find use in several different types of indications. Some examples are gliablastoma, glioma, lung, colorectal, breast, prostate, stomach, gastric, liver, thyroid, bladder, esophagus, pancreas, kidney, uterine body, cervix, melanoma, brain (including central and peripheral nervous systems), ovary, gallbladder, head and neck (e.g., lips, oral cavity, larynx, nasopharynx, oropharynx, hypopharynx), multiple myeloma, testis, vulva, salivary gland, mesothelioma, penis, Kaposi's sarcoma, vagina, neuroendocrine tumors, neuroendocrine carcinomas.

[0055] Imaging naturally involves equipment. The conjugate products and compositions according to the present invention preferably contain a fluorescent chemical element that can re-emit light upon photoexcitation. The excitation light and synchrotron radiation are specific to the fluorophore used. The excitation light typically comes from a laser having wavelengths between, for example, 600 nm and 900 nm. The synchrotron radiation from the fluorophore is typically detected by a camera using a mechanical or software-based filter that detects light between, for example, 750 nm and 950 nm. The equipment used may be a surgical robot, surgical microscope, endoscope, or handheld device. The specifications of the light source (e.g., laser) and photodetector (e.g., camera with filter) depend on the selected fluorophore.

[0056] According to the present invention, several different types of procedures can be used. Non-limiting examples of surgical procedures include day care surgery, open surgery, minimally invasive surgery, and robot-assisted surgery.

[0057] Furthermore, surgeries may have different purposes. Non-limiting examples of surgical purposes include: curative surgery (aimed at removing all cancerous tumors from the body - this is included in marked size calculations), prophylactic surgery (used to remove tissue that does not contain cancerous cells but may develop into malignant tumors, such as polyps in the colon), diagnostic surgery (helping to determine whether cells are cancerous, e.g., performing a biopsy for diagnostic or screening purposes, e.g., using a colorectal scope to look for malignant colorectal polyps), staging surgery (work to reveal the extent of cancer, e.g., laparoscopy (inserting an observation tube equipped with a lens or camera through small incisions to examine the inside of the body)), and debulking surgery (removing a portion of a cancerous tumor). While these may be used in certain situations where removing the entire tumor could cause damage to an organ or body, palliative or supportive surgery (used to treat advanced cancer; it does not cure the cancer but works to alleviate discomfort or correct other problems that may have been caused by the cancer or cancer treatment; an example of supportive surgery is the insertion of a catheter to assist chemotherapy), restorative surgery (may be used as a follow-up to curative surgery or other surgery to change or restore the appearance or function of a person's body part; for example, after breast cancer in women), or corrective surgery (reoperation to resolve problems after surgery (or other procedure), such as bleeding or infection).

[0058] Another area of ​​interest related to the present invention is photodynamic therapy. Photodynamic therapy (PDT) is increasingly being used as an attractive alternative treatment modality for superficial cancers. The treatment involves two relatively simple steps: administration of a photosensitive drug and irradiation of the tumor to activate or heat the drug. Compositions according to the present invention can be used in PDT treatment.

[0059] The present invention also relates to, for example, methods intended for the treatment, staging, or diagnosis of cancer in optical imaging / fluorescence imaging (FLI) of cancer.

[0060] The methods may relate to methods including diagnosing anatomical structures, guiding surgeons / robots, assisting surgeons / robots, increasing survival rates, increasing the amount of cancerous tissue removed under surgery, improving quality of life, decreasing the amount of normal tissue removed, increasing certainty, shortening surgical time, improving the quality of surgery, improving surgical quality assurance, reducing the cost of surgery, improving surgeon performance, and / or improving surgical outcomes in any other way.

[0061] Furthermore, the compositions according to the present invention may be provided in different forms. According to one specific embodiment of the present invention, a freeze-dried composition comprising the composition according to the present invention is provided. In this case, the freeze-protectant can be easily reconstituted. Freeze drying It is suitable for making cakes.

[0062] Furthermore, the compositions according to the present invention may be provided in different forms. According to one specific embodiment of the present invention, a freeze-dried composition comprising the composition according to the present invention is provided. In this case, the freeze-protectant is suitable for producing a Rio cake that can be easily reconstituted.

[0063] Furthermore, dried formulations containing the composition according to the present invention are also of interest.

[0064] Furthermore, the production method is relevant. According to one specific embodiment, the present invention relates to a method for producing a composition in accordance with the above, the method comprising mixing a fluorophore-labeled receptor-targeting component, a buffer, and a surfactant to solubilize the fluorophore-labeled receptor-targeting component in the composition. As described above, preferably, the receptor-targeting component is a uPAR-targeted peptide conjugate. Furthermore, according to one specific embodiment, a cryoprotectant is mixed with the composition, thereby, Freeze drying A cake is produced. Preferably, Freeze drying The cake can be easily reconstituted. One preferred combination of cryoprotectants according to the present invention is mannitol and glycine.

[0065] Furthermore, the production method is relevant. According to one specific embodiment, the present invention relates to a method for producing a composition in accordance with the above, the method comprising mixing a fluorophore-labeled receptor-targeting component, a buffer, and a surfactant to solubilize the fluorophore-labeled receptor-targeting component in the composition. As described above, preferably the receptor-targeting component is a uPAR-targeted peptide conjugate. Furthermore, according to one specific embodiment, a cryoprotectant is mixed with the composition to produce a Rio cake. Preferably, the Rio cake can be easily reconstituted. One preferred combination of cryoprotectants according to the present invention is mannitol and glycine.

[0066] Furthermore, according to yet another specific embodiment of the present invention, the production method is: -(i)ICG-Glu-Glu-AE105: [ka] Alternatively, a pharmaceutically acceptable salt thereof may be mixed with sodium phosphate, mannitol, and glycine. ICG-Glu-Glu-AE105 at concentrations of -0.1 to 10.0 mg / ml; Sodium phosphate in a concentration range of -5 to 50 mM; Mannitol in a concentration range of -10 to 50 mg / ml; Glycine in a concentration range of -1 to 30 mg / ml; Polysorbate 20 with a concentration exceeding -0.01% by weight To obtain a composition containing -(ii) Adjust the pH of the composition from step (i) to a pH in the range of 6.9 to 7.9; -(iii) Transfer an amount of the mixture from step (ii) equal to the desired dosage into a suitable container; -(iv) Drying the mixture; and -(v) sealing a container Includes.

[0067] In relation to the above, the preferred concentration of the fluorophore-labeled uPAR receptor targeting component is in the range of 0.1 to 2,000 mg per drug dose, preferably in the range of 1 to 1,000 mg per human drug dose. [Examples]

[0068] Example 1: Development of an aqueous formulation of the composition according to the present invention A composition containing the fluorophore-labeled receptor-targeted peptide conjugate ICG-Glu-Glu-AE105 (hereinafter referred to as Composition 1) is soluble in DMSO at a maximum concentration of 20 mg / ml, and its peak spectral absorption at 800 nm is similar to that of indocyanine green (ICG), which is currently used in clinics for visual assessment of blood vessels, blood flow, and associated tissue perfusion, Figure A.

[0069] Using excipients suitable for pharmaceutical formulation development, an aqueous formulation of Composition 1 with a concentration of 1.0 mg / ml was developed in accordance with the present invention.

[0070] Composition 1 did not dissolve in phosphate-buffered saline at a test concentration of 1.0 mg / ml, which can be observed from both the spectral absorption with a distinct double peak in the range of 700 nm to 825 nm and the corresponding emission spectrum with a significantly lower fluorescence peak at 800 nm (Figure A). Excitation was performed at 775 nm.

[0071] In a 10 mM sodium phosphate buffer containing 45 mg / ml mannitol at pH 7.4, the peak absorption shifted towards the 800 nm peak absorption, and in a formulation containing 10 mM sodium phosphate, 45 mg / ml mannitol, 1% polysorbate 20, and pH 7.4, the absorption spectrum was similar to that obtained with DMSO. The addition of 0.2 g / ml 2-hydroxypropyl-β-cyclodextrin (HBC) to phosphate-buffered saline was also very efficient in solubilizing composition 1 at 1.0 mg / ml (Figure B).

[0072] The absorbance and emission spectra of the three formulations (1.0 mg / ml) of Composition 1, with complete solubilization, are shown in Figures C and D.

[0073] Further explanation of Figures A-D A. Absorbance and emission spectra of FG001 dissolved in DMSO and PBS at 1.0 mg / ml. Excitation wavelength: 775 nm. The analytical sample for absorbance measurement was further diluted 200-fold and 400-1000-fold for emission spectroscopy. B.1) Absorbance spectra of FG001 dissolved in 10 mM sodium phosphate, 45 mg / ml mannitol, pH 7.4, 2) 10 mM sodium phosphate, 45 mg / ml mannitol, 1% polysorbate 20, pH 7.4, and 3) PBS, 0.2 g / ml HBC. The analytical samples for absorbance measurement were further diluted 200-fold and 400-1000-fold for emission spectra. Overlays of absorption spectra of three formulations (1.0 mg / ml) with complete solubilization of C.FG001: A) DMSO, B) 10 mM sodium phosphate, 45 mg / ml mannitol, 1% polysorbate 20, pH 7.4, and C) phosphate-buffered saline, 0.2 g / ml HBC. The analytical samples for absorbance measurement were further diluted 200-fold and 400-1000-fold for emission spectra. D. Overlay of emission spectra of three formulations (1.0 mg / ml) of Composition 1 with complete solubilization: A) DMSO, B) 10 mM sodium phosphate, 45 mg / ml mannitol, 1% polysorbate 20, pH 7.4, and C) phosphate-buffered saline, 0.2 g / ml HBC. Excitation wavelength: 775 nm. The analytical samples for absorbance measurement were further diluted 200-fold and 400-1000-fold for emission spectroscopy.

[0074] Example 2 Determination of the optimal concentration range of polysorbate 20 for solubilizing composition 1 - See Figure E. The optimal concentration range of polysorbate 20 for solubilizing composition 1 at a concentration of 1.0 mg / ml in 10 mM sodium phosphate, 45 mg / ml mannitol, and pH 7.4 was evaluated by measuring the absorption spectra of formulations containing 0-1.0% polysorbate 20.

[0075] The absorption spectra showed that all formulations containing more than 0.01% polysorbate 20, composition 1, were completely solubilized, with a main peak absorption at 800 nm.

[0076] Example 3: Determination of the stability of composition 1 in three formulations containing polysorbate 20. The stability of composition 1 at a concentration of 1.0 mg / ml was evaluated using three different formulations suitable for the development of lyophilized products. A (mannitol): 10 mM sodium phosphate, 45 mg / ml mannitol, 0.025% polysorbate 20, pH 7.4 B (Mannitol / Glycine): 10 mM sodium phosphate, 26 mg / ml mannitol, 8.7 mg / ml glycine, 0.025% polysorbate 20, pH 7.4 C (mannitol / sucrose): 10 mM sodium phosphate, 26 mg / ml mannitol, 40 mg / ml sucrose, 0.025% polysorbate 20, pH 7.4

[0077] Composition 1 was weighed into a formulation buffer and the pH was adjusted to 7.4 using sodium hydroxide or hydrochloric acid. The bulk formulation was sterile filtered through a suitable sterile filter and filled into 6R vials. The stoppered vials were placed in a lyophilizer and lyophilized using a standard program. After lyophilization, all vials were capped.

[0078] After reconstituting one vial of each formulation with sterile water for injection, the osmolality and purity were measured using the RP-HPLC method described in Example 4. [Table 1]

[0079] All three formulations had the desired osmolality for use as a pharmaceutical formulation for intravenous administration to humans.

[0080] The stability of the recomposed liquid formulation of composition 1 was evaluated for two weeks at room temperature and under daylight (RTL). [Table 2]

[0081] It is well known that fluorophores like ICG are photosensitive and should be protected from light. However, in order to select the most stable formulation, three liquid formulations were exposed to daylight at room temperature for two weeks.

[0082] From the data presented in the table above, it is clear that formulation B: mannitol / glycine is the most stable.

[0083] The stability of the lyophilized products from the three formulations was also evaluated for two weeks under three different storage conditions: I: Room temperature, daylight (RTL) II: Room temperature, dark (RTD) III: 40℃, dark (40D) [Table 3]

[0084] The stability of the lyophilized vial was significantly increased compared to the liquid formulation, with smaller changes observed. The study confirmed that Formulation B: Mannitol / glycine as a lyophilized product exhibited superior stability under both high temperatures (40°C) and exposure to light at room temperature.

[0085] Lyophilized samples stored at room temperature, dark (RTD) and 40°C, dark (40D) for two weeks were reconstituted and stored at room temperature, daylight (RTL) for a further 24 hours. [Table 4]

[0086] The stability of the sample, maintained as a lyophilized product for two weeks and then stored at room temperature under daylight for 24 hours after reconstitution, also supports the selection of formulation B: mannitol / glycine as the preferred formulation.

[0087] It should be noted that another possible method used to investigate characteristic single absorbance spectral peaks is to calculate the area of ​​a single peak located around 800 nm and compare this to the total area of ​​the absorption spectrum in a given wavelength range of 600–900 nm. This is further explained above in the description.

[0088] Example 4 Determination of the long-term storage stability of composition 1 in a formulation containing polysorbate 20. The long-term stability of composition 1 at a concentration of 1.0 mg / ml was evaluated as a lyophilized product in one formulation containing 10 mM sodium phosphate, 26 mg / ml mannitol, 9.0 mg / ml glycine, 0.025% polysorbate 20, and pH 7.4.

[0089] Composition 1 was weighed into a formulation buffer and the pH was adjusted to 7.4 using sodium hydroxide or hydrochloric acid. The bulk formulation was sterile filtered through a suitable sterile filter and filled into 6R vials. The vials were capped and placed in a lyophilizer and lyophilized using a standard program. After lyophilization, all vials were capped, visually inspected, and stored protected from light at 5°C, 25°C / 60%RH, or 40°C / 75%RH for up to 9 months.

[0090] Once selected, the vial is removed from the stability chamber for visual inspection of the lyophilized material and determination of its water content. Before analyzing the corresponding liquid formulation for purity, pH, and visual appearance, another vial is reconstituted with sterile water for injection. [Table 5] [Table 6]

[0091] The formulation of selected composition 1 has been shown to be stable for up to 9 months under storage conditions of 5°C and 25°C / 60%RH, as determined by purity determination. The pH of the formulation, as well as the appearance of the lyophilized product and the clarity of the reconstituted liquid formulation, remain unchanged during storage for up to 9 months. Furthermore, the residual water content is approximately 2% after 9 months of storage at 25°C / 60%RH.

[0092] Analysis method : RP-HPLC for detecting the purity of reconstituted composition 1: Mobile phase A consisted of purified water / acetonitrile (80 / 20 v / v) containing 5 mM ammonium acetate, and mobile phase B consisted of purified water / acetonitrile (10 / 90 v / v) containing 5 mM ammonium acetate. A Waters XBridge BEH Peptide, 3.5 μm, 130 Å, 4.6 × 150 mm column was used. The flow rate was set to 1.0 mL / min, detection was performed at a wavelength of 780 nm, and the column running temperature was 45°C. The sample condenser temperature was set to 5°C, and the sample injection load was 10 μg.

[0093] The water content of the freeze-dried material was determined by Karl Fischer titration according to Ph.Eur.2.5.32.

Claims

1. The component comprises a fluorophore-labeled receptor targeting component, a buffer, and a surfactant. The fluorophore-labeled receptor-targeting component is solubilized in the composition by the present surfactant. It contains up to 10% by weight of water, preferably up to 5% by weight of water. The fluorophore-labeled receptor targeting component is ICG-Glu-Glu-AE105, represented by the following structural formula: 【Chemistry 1】 or a pharmaceutically acceptable salt thereof, It contains a cryoprotective agent which is a combination of mannitol and glycine. The buffer solution is supplied such that the composition has a physiological pH or substantially physiological pH, preferably in the range of 7.3 to 7.

5.

2. The composition according to claim 1, wherein the fluorophore-labeled receptor targeting component is a fluorophore-labeled receptor-targeting peptide conjugate.

3. The composition according to claim 1 or 2, wherein the fluorophore-labeled receptor-targeting component is solubilized in the composition at a level corresponding to a single peak having an absorption maximum around 800 nm when the absorbance spectrum of the composition in the wavelength range of 700 to 825 nm is measured.

4. The composition according to any one of claims 1 to 3, wherein the fluorophore-labeled receptor targeting component is solubilized in the composition to a level corresponding to having an absorbance spectral peak with a maximum value around 800 nm, and the area of ​​the absorbance spectral peak is at least 50%, preferably at least 60%, and more preferably at least 65% of the total area of ​​the absorption spectrum in a given wavelength region of 600 to 900 nm.

5. The composition according to any one of claims 1 to 4, wherein the surfactant is a nonionic surfactant.

6. The fluorophore-labeled receptor-targeting component comprises a fluorophore, a peptide that binds to the receptor, and a linker group. The composition according to any one of claims 1 to 5, wherein the fluorophore, the peptide that binds to the receptor, and the linker group are linked by a covalent bond.

7. The composition according to any one of claims 1 to 6, wherein the receptor is a uPAR.

8. The composition according to claim 1, wherein the concentration of ICG-Glu-Glu-AE105 is in the range of 0.1 to 10.0 mg / ml.

9. The fluorophore-labeled receptor targeting component is ICG-Glu-Glu-AE105, represented by the following structural formula: 【Chemistry 2】 or a pharmaceutically acceptable salt thereof, The concentration of ICG-Glu-Glu-AE105 is in the range of 0.1 to 10.0 mg / ml. The buffer is a buffer in the form of sodium phosphate with a concentration of 5 to 50 mM. The composition according to any one of claims 1 to 8, wherein the cryoprotective agent is a combination of mannitol at a concentration of 10 to 50 mg / ml and glycine at a concentration of 1 to 30 mg / ml.

10. The composition according to any one of claims 1 to 4, wherein the surfactant comprises polysorbate, preferably polysorbate, more preferably polysorbate 20, at a concentration of more than 0.01% by weight.

11. The fluorophore-labeled receptor-targeting component has a pharmacokinetic profile in which a TBR (tumor-to-background ratio) of at least 2.5 is reached within 3.5 hours after administration, and the level of at least 2.5 TBR is maintained for at least 30 minutes before decreasing again. The composition according to any one of claims 1 to 10, wherein the fluorophore-labeled receptor-targeting component is a human uPAR-targeting conjugate.

12. The fluorophore-labeled receptor-targeting component has a pharmacokinetic profile in which a TBR (tumor-to-background ratio) of at least 2.5 is reached within 3.5 hours after administration, and the level of at least 2.5 TBR is maintained for at least 30 minutes before decreasing again. Preferably, the fluorophore-labeled receptor-targeting component is a human uPAR-targeting conjugate, according to any one of claims 1 to 11.

13. The composition according to claim 11 or 12, wherein the fluorophore-labeled receptor-targeting component has a pharmacokinetic profile in which the plasma half-life is up to 75 hours, preferably up to 20 hours, more preferably up to 15 hours, more preferably in the range of 6 to 15 hours, and most preferably in the range of 6 to 10 hours.

14. The composition according to any one of claims 1 to 13, wherein the fluorophore-labeled receptor-targeting component has a pharmacokinetic profile in which a TBR (tumor-to-background ratio) of at least 2.8 is reached within 3.5 hours after administration, and the level of TBR of at least 2.8 is maintained for at least 30 minutes before decreasing again.

15. The composition according to any one of claims 1 to 14, wherein the peak TBR of the fluorophore-labeled receptor-targeting component after administration is at least 3.

16. The composition according to any one of claims 1 to 15, wherein the receptor binding affinity of the fluorophore-labeled receptor targeting component to uPAR, defined as Kd, is in the range of up to 2,500 nM, preferably up to 2,000 nM, more preferably up to 500 nM, and most preferably 2,000 to 300 nM.

17. K of fluorophore-labeled receptor targeting components ｏｎ This is the K of uPA, a natural ligand. ｏｎ That's all, K ｏｎ ≥ 4.6 × 10 ６ M －１ s －１ The composition according to claim 16.

18. The fluorophore-labeled receptor targeting component contains the natural ligand (uPA) that binds to uPAR in an IC50 concentration of up to 1,000 nM, preferably up to 200 nM, more preferably up to 50 nM, and most preferably up to 25 nM. ５０ A composition according to any one of claims 1 to 17, wherein the values ​​are replaced.

19. The composition according to any one of claims 1 to 18, wherein the fluorophore-labeled receptor targeting component has selectivity of at least 60%, preferably more than 70%, more preferably more than 80%, and most preferably more than 90% for the detection of cancer tissue.

20. The composition according to any one of claims 1 to 19, which is soluble in less than 10 minutes, preferably less than 5 minutes, more preferably less than 2 minutes, and most preferably less than 1 minute.

21. A freeze-dried composition comprising the composition according to any one of claims 1 to 20.

22. The process involves mixing a fluorophore-labeled receptor targeting component, a buffer, and a surfactant to solubilize the fluorophore-labeled receptor targeting component in the composition. A freeze-protecting agent is mixed with the composition to produce a freeze-dried cake. A method for producing the composition according to any one of claims 1 to 21.

23. - (i) ICG-Glu-Glu-AE105 represented by the following structural formula: 【Transformation 3】 Alternatively, a pharmaceutically acceptable salt thereof may be mixed with sodium phosphate, mannitol, glycine, and polysorbate 20. ICG-Glu-Glu-AE105 in concentrations of -0.1 to 10.0 mg / ml; Sodium phosphate in a concentration range of -5 to 50 mM; Mannitol in a concentration range of -10 to 50 mg / ml; Glycine in a concentration range of -1 to 30 mg / ml; - Polysorbate 20 with a concentration exceeding -0.01% by weight To obtain a composition containing - (ii) Adjust the pH of the composition in step (i) to a pH in the range of 6.9 to 7.9; - (iii) Transfer an amount of the mixture from step (ii) equal to the desired dosage into a suitable container; - (iv) Drying the mixture; and - (v) sealing a container including, The method according to claim 22.

Images