Dual-labeled probes for molecular imaging and their uses

Abstract

The present invention provides a compound or a pharmaceutically acceptable salt thereof having the following chemical structure: (A)-x1-(B)-x2-(C), wherein (A) is at least one motif that specifically binds to the cell membrane of a neoplastic cell; (B) is at least one radiometal chelator moiety; (C) is a dye moiety; x1 is a spacer that covalently connects (A) and (B); and x2 is a spacer or a single chemical bond connecting (B) and (C); and wherein (C) is a compound of the formula: [Formula 1] (In the formula, R 1 ~R 4 , R 9 , a, b, Y and X 1 ~X 4 The present invention further relates to compounds of the formula (I) or pharmaceutically acceptable salts thereof, including compounds of the formula (I) or pharmaceutically acceptable salts thereof, having the meanings given in the claims and in the specification.

Description

[Technical Field]

[0001] The present invention relates to a compound having the chemical structure of formula (I): (A)-x1-(B)-x2-(C), where (A) is at least one motif that specifically binds to the cell membrane of a neoplastic cell; (B) is at least one radiometal chelator moiety; (C) is a dye moiety; x1 is a spacer covalently linking (A) and (B); and x2 is a spacer or a single chemical bond linking (B) and (C), or a pharmaceutically acceptable salt thereof. The present invention further relates to compositions comprising the compound, as well as methods for detecting neoplastic cells in a sample in vitro utilizing the compound or composition. [Background technology]

[0002] In recent years, molecular imaging has become increasingly important for the diagnosis of neoplasms, especially cancer, as it allows physicians to obtain useful information about the size, location and shape of neoplastic pitch.

[0003] Various techniques have been developed to visualize neoplastic tissue in vivo, including magnetic resonance imaging (MRI), radiography (especially computed tomography (CT)), fluorescence molecular tomography (FMT), and positron emission tomography (PET), which are commonly used methods for localizing neoplasms today.

[0004] However, these methods still have significant drawbacks. While MRI achieves high-resolution visualization and allows measurements without any staining, MRI makes it relatively difficult, if not impossible, to reliably distinguish between healthy and neoplastic tissue. X-ray imaging techniques such as CT also achieve relatively high resolution, but, like MRI, cannot clearly distinguish between diseased and healthy tissue and often require undesirably high doses of contrast agents. While FMT allows for the detection of specifically stained tissue, it typically has low resolution and only allows for the detection of neoplasms located near the patient's body's outer surface. Furthermore, it cannot visualize surrounding tissues, making it difficult for the examiner to infer diagnostic results and decide on further treatment strategies. PET allows for the detection of neoplasms inside the patient's body, but it only depicts the neoplasm itself and, like FMT, does not provide insight into the neoplasm's localization in relation to its tissues.

[0005] MRI and PET are usually combined with each other (sometimes in a single device) to allow for clear detection of the neoplasm and surrounding tissue. This combination allows for precise localization of the neoplasm in relation to its tissue and further indicates the shape and size of the neoplasm.

[0006] However, MRI and PET devices are quite large, encompassing the entire patient's body, preventing simultaneous surgical intervention. Once molecular imaging with MRI and / or PET is complete, the surgeon attempting to remove the neoplastic tissue must assess the location, size, and shape of the neoplastic tissue within the patient's body by mentally projecting the images obtained from molecular imaging onto the patient's body. In other words, while performing surgery, the surgeon often cannot visually see the neoplastic tissue within the patient's body, as the neoplastic tissue appears no different, or only slightly different, from the surrounding non-neoplastic tissue. For example, lymph nodes containing neoplastic cells are generally indistinguishable from their healthy counterparts. Therefore, the surgeon must either recall the location of each neoplastic tissue previously seen with molecular imaging or occasionally divert their attention from the patient's body to the results of molecular imaging in order to mentally project these results onto the patient's body.

[0007] A major drawback of this procedure is that the surgeon can never be completely sure that the entire neoplastic tissue has been removed, and therefore, often a significant portion of tissue is removed, often containing a large amount of healthy tissue, and alternatively, residual portions of the neoplastic tissue are often still left in the patient.

[0008] Improved radiopharmaceuticals have already been proposed, where compounds with a binding site for neoplastic cells and a chelator moiety are described, where the improvement lies in the combination of dual-targeting radioligands with bispecific PSMA / GRPr for the chemical moiety (C. Liolios et al., Bioconjugate Chemistry, 2016, 27, 737-751) or in the reduction of the uptake of peptidic radioligands in vital organs (Eder et al., The Journal of Nuclear Medicine, 2013, 54(8), 1327-1330). 68An improved imaging contrast agent for Ga-PSMA-11 is described in A.-C. Baranski et al., Bioconjugate Chem. 2017, 28(9), 1485-2492.

[0009] However, there remains a need for improvement by further combining radiochemicals having binding sites and chelators with fluorescent chemical moieties.

[0010] Thus, compounds have been developed that contain a cell-binding moiety (e.g., prostate-specific membrane antigen (PSMA)) that binds to neoplastic cells, a fluorophore, and a chelator (particularly the 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA) chelator) (Banerjee SR, Pullambhatla M, Byun Y, Nimmagadda S, Foss C, Green G, Fox JJ, Lupold SE, Mease R, Pomper MG (2011); Sequential SPECT and optical imaging of experimental models of prostate cancer with a dual modality inhibitor of the prostate-specific membrane antigen. Angew Chem Int Ed Engl. 50(39):9167-9170; and WO 2010 / 108125). Here, both the chelator and the fluorophore are conjugated via independent spacers to a common molecular scaffold that is conjugated to the cell-binding moiety.

[0011] However, this approach has a significant disadvantage in that the flexibility of using various dyes is limited. In fact, it has been found that the use of chelating agents such as DOTA has significant disadvantages, such as weak binding to target structures, unless they are combined with a specific fluorophore structure, such as IRDye800CW, as used by Banerjee et al. Therefore, structures known in the art are not used in a modular manner. In particular, the dyes conjugated thereto cannot be freely selected, and some fluorophores commonly and preferably used in the art cannot be used with this strategy.

[0012] Furthermore, cell staining structures containing motifs that bind to fluorophores, chelators, and cell markers present in various cell populations are known (Seibold U, Waengler B, Schirrmacher R, and Waengler C (2014); Bimodal imaging probes for combined PET and OI: recent developments and future directions for hybrid agent development. Biomed Res Int. 2014:153741. doi: 10.1155 / 2014 / 153741). However, these structures do not contain motifs that specifically bind to the plasma membrane of neoplastic cells, but rather contain motifs that bind to cellular structures present in various cell types, including non-neoplastic physiological cells. Furthermore, in these structures shown by Seibold et al., binding sites cannot be freely selected in a modular manner; rather, they are incorporated into the above structures in a complex manner.

[0013] In view of the above, there remains an unmet need for compounds that are easy to synthesize in a modular manner and have wide flexibility in dye choice, allowing for in vivo molecular imaging in patients as well as intraoperative imaging.

[0014] US 2015 / 110715 A1 describes compounds having a chemical structure comprising at least one motif (A) that specifically binds to the cell membrane of neoplastic cells, at least one radiometal chelator moiety (B), and at least one dye moiety (C). The chemical moieties (A), (B), and (C) may be directly linked to each other or may be linked via a spacer. One of the most important fluorescent chemical structures described is the above-mentioned IRDye800CW and its derivatives.

[0015] Although the radioactive compounds described in US 2015 / 110715 A1 show good performance, there is still a need to further improve the properties of said compounds, especially when IRDye800CW is used.

[0016] One important property is the selectivity of such radioactive compounds, including IRDye800CW, i.e., specific uptake in tumors compared to low uptake in vital organs, as expressed by the respective tumor-to-organ ratios for various different vital organs. [Prior art documents] [Patent documents]

[0017] [Patent Document 1] WO 2010 / 108125 [Patent Document 2] US 2015 / 110715 A1 [Non-patent literature]

[0018] [Non-Patent Document 1] C. Liolios et al., Bioconjugate Chemistry, 2016, 27, 737-751 [Non-patent document 2] Eder et al., The Journal of Nuclear Medicine, 2013, 54(8), 1327-1330 [Non-patent document 3] A.-C. Baranski et al., Bioconjugate Chem. 2017, 28(9), 1485-2492 [Non-patent document 4] Banerjee SR, Pullambhatla M, Byun Y, Nimmagadda S, Foss C, Green G, Fox JJ, Lupold SE, Mease R, Pomper MG(2011);Sequential SPECT and optical imaging of experimental models of prostate cancer with a dual modality inhibitor of the prostate-specific membrane antigen. Angew Chem Int Ed Engl. 50(39):9167-9170 [Non-Patent Document 5] Seibold U, Waengler B, Schirrmacher R and Waengler C(2014);Bimodal imaging probes for combined PET and OI: recent developments and future directions for hybrid agent development. Biomed Res Int. 2014:153741. doi: 10.1155 / 2014 / 153741 Summary of the Invention [Problem to be solved by the invention]

[0019] It is therefore an object of the present invention to provide compounds, including IRDye800CW, that have improved tumor-to-organ ratios. [Means for solving the problem]

[0020] The object is to provide a compound of formula (I): (A)-x1-(B)-x2-(C) (In the formula, (A) is at least one motif that specifically binds to the plasma membrane of neoplastic cells; (B) is at least one radiometal chelator moiety; (C) is a dye moiety; x1 is a spacer covalently linking (A) and (B); x2 is a spacer or a single chemical bond connecting (B) and (C); (C) is a compound of the formula:

[0021] [ka] (In the formula, X 1 and X 4 is -N=, -N(R 5 )=, and -C(R 6 )= independently selected from the group consisting of; X 2 and X 3 are O, S, Se, N(R 5 ), and C(R 6 R 7 ) independently selected from the group consisting of: Y is a linker that connects the two moieties of (C) and allows for electron delocalization between said moieties, and Y is optionally a group (L-) c Z 0 Includes; a and b are independently selected from the group consisting of 1, 2, and 3; Each R 1 and each R 2 are independently (L-) c Z, (L-) c Z 0 or H; two adjacent R 1 and / or two adjacent R 2 may contain one or more (L-) c Z or (L-) c Z 0 It is also possible to form an aromatic ring substituted with; R 3 , R 4 , R 5 , R6 , R 7 , R 9 is (L-) c Z, (L-) c Z 0 independently selected from the group consisting of each c is independently 0 or 1; Each L is independently T 1 , -OT 1 -, -ST 1 -, -C(O)T 1 -, -C(O)OT 1 -, -OC(O)T 1 -, -C(O)NHT 1 -,-NHC(O)T 1 , or optionally -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)O-, and T 1 C that is interrupted and / or terminated by one or more of 1-10 is an alkylene group; T 1 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, decalinyl, adamantyl, C 3-7 cycloalkyl, 3- to 7-membered heterocyclyl, or 7- to 11-membered heterobicyclyl, where T 1 is sometimes halogen, CN, C(O)R 8 , COOR 8 , OR 8 , C(O)N(R 8 R 8a ), S(O)2N(R 8 R 8a ), S(O)N(R 8 R 8a ), S(O)2R 8 , N(R 8 )S(O)2N(R 8a R 8b ), SR 8 , N(R 8 R 8a ), NO2, OC(O)R 8 , N(R 8 )C(O)R 8a , N(R 8 )S(O)2R 8a , N(R 8)S(O)R 8a , N(R 8 )C(O)N(R 8a R 8b ), N(R 8 )C(O)OR 8a , OC(O)N(R 8 R 8a ), oxo(=O) (wherein the ring is at least partially saturated), or C 1-6 Alkyl (C 1-6 alkyl is optionally substituted with one or more of the same or different halogens); Each Z is independently H, halogen, CN, or C(O)R 8 , C(O)OR 8 , C(O)O - , OR 8 , C(O)N(R 8 R 8a ), S(O)2OR8, S(O)2O - , S(O)2N(R 8 R 8a ), S(O)N(R 8 R 8a ), S(O)2R 8 , S(O)R 8 , N(R 8 )S(O)2N(R 8a R 8b ), SR 8 , N(R 8 R 8a ), NO2, P(O)(OR 8 )2, P(O)(OR 8 )O - ,OC(O)R 8 , N(R 8 )C(O)R 8a , N(R 8 )S(O)2R 8a , N(R 8 )S(O)R 8a , N(R 8 )C(O)N(R 8a R 8b ), N(R 8 )C(O)OR 8a , or OC(O)N(R 8 R 8a ) and; R 8 , R 8a , R 8b is H or C 1-6 Alkyl (C 1-6 alkyl is optionally substituted with one or more of the same or different halogens; Z 0 is a chemical bond that connects (C) to x2 or to (B) if x2 is a single chemical bond; However, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 9 One of them is (L-) c Z 0 or Y is (L-) c Z 0 (including and wherein x1 comprises the group -AA- (AA is an amino acid sequence of 4 to 8 (preferably 5 to 7, more preferably 6) naturally occurring amino acids, at least 2 of which are histidines); Any remaining positive or negative charge(s) are compensated by pharmaceutically acceptable negatively or positively charged counterion(s). or a pharmaceutically acceptable salt thereof.

[0022] Surprisingly, it has been found that compounds having moieties (A), (B) and (C) as described in the prior art and comprising a dye derived from dye moiety (C) as described herein, in particular IRDye800CW, reduce the tumor-to-organ ratio compared to compounds having moieties (A) and (B), and that said lower ratio can be improved by the introduction of the amino acid sequence AA as part of the spacer x1 between moieties (A) and (B).

[0023] The compounds of the present invention, or pharmaceutically acceptable salts thereof, enable, by way of example, positron emission tomography (PET) scanning and fluorescence imaging. DETAILED DESCRIPTION OF THE INVENTION

[0024] The term " pharmaceutically acceptable salt " used herein can be understood in the broadest sense as any charged form of the compound of the present invention. Depending on the chemical structure of the compound and the environment in which it is dissolved, the compound may contain one or more charged residues selected from the group consisting of, for example, but not limited to, carboxylate anion residue(s), primary ammonium cation(s), secondary ammonium cation residue(s), tertiary ammonium cation residue(s), primary phosphate anion residue(s), secondary phosphate anion residue(s), sulfate anion residue(s), sulfite anion residue(s), and alkoxide residue(s). The counter ion can be any ion known in the art to be pharmaceutically acceptable, such as, for example, acetate ions, fatty acid carboxylate ions, chloride ions, sodium ions, potassium ions, magnesium ions, calcium ions, aluminum ions, lithium ions, ammonium ions, phosphate ions, hydroxyl ions, proton ions, and fluoride ions.

[0025] The term "motif" as used throughout this invention can be understood in the broadest sense as a molecular structural pattern that allows specific binding to the cell membrane of neoplastic cells.

[0026] The term "neoplastic cells" as used in the context of the present invention can be understood in the broadest sense as any cell that exhibits an abnormal growth and / or division rate, including metaplastic and dysplastic cells. Typically, neoplastic cells tend to form a cell mass known as a neoplasm. Neoplastic cell proliferation is typically less than or not equivalent to the surrounding normal tissue. Neoplastic cell proliferation preferably continues in the same excessive manner even after cessation of stimulation. Neoplastic cells can form benign neoplasms, premalignant neoplasms (carcinoma in situ), or malignant neoplasms (cancer). Neoplasms can also be characterized according to the International Classification of Diseases, Vol. 10 (ICD-10 nomenclature) (2013 edition), i.e., as any pathological condition according to ICD-10 classification C00-D48. In the context of the present invention, cancer also encompasses metastasis.

[0027] Cancer in the sense of the present invention is any malignant neoplasm. By way of example, cancer may be a carcinoma (e.g., prostate carcinoma, breast carcinoma, lung carcinoma, pancreatic carcinoma, liver carcinoma or colon carcinoma), a sarcoma (e.g., a sarcoma in bone, cartilage, fat and / or nervous tissue, or a mesenchymal sarcoma), a lymphoma, a leukemia, a germ cell (e.g., testicular cancer or ovarian cancer (seminoma and dysgerminoma, respectively)) or a blastoma (e.g., hepatoblastoma).

[0028] At least one motif (A) that specifically binds to the cell membrane of a neoplastic cell allows binding of its target structure present on the surface of the neoplastic cell with high affinity compared to other molecular structures.

[0029] The target structure is preferably typical of neoplastic cells. Thus, the target structure can preferably be found on the surface of neoplastic cells exclusively or at a higher local concentration compared to normal cells (i.e., non-neoplastic cells). Thus, the local concentration of target structures recognized by at least one motif (A) according to the present invention at the cell membrane of neoplastic cells is preferably at least 2-fold, more preferably at least 5-fold, even more preferably at least 10-fold, even more preferably at least 100-fold, and even more preferably at least 500-fold higher compared to corresponding normal (i.e., non-neoplastic) cells.

[0030] Preferably, the motif binds to its target structure on a neoplastic cell with an affinity that is at least 5-fold higher, more preferably at least 10-fold higher, even more preferably at least 20-fold higher, even more preferably at least 50-fold higher, and especially at least 100-fold higher than to other molecular structures of similar charge and hydrophobicity in a comparable chemical environment. Preferably, the motif binds to its target structure on the plasma membrane of a neoplastic cell with a dissociation constant of 10 μM or less, more preferably 5 μM or less, even more preferably 1 μM or less, even more preferably 100 nM or less, especially 50 nM or less.

[0031] The compounds of the present invention, or pharmaceutically acceptable salts thereof, comprise a moiety (A) having at least one motif. Thus, the moiety (A) may represent one or more motifs. Illustratively, the moiety (A) represents a chemical structure having one, two, or three motifs. However, preferably, (A) represents one motif.

[0032] Preferably, the motif comprises at least one natural amino acid moiety, more preferably at least two natural amino acid moieties.

[0033] As used throughout the present invention, the terms "moiety," "residue," and "remainder" in the context of chemical structures can be understood interchangeably in the broadest sense as a portion of a molecule that is firmly attached to the rest of the molecule, particularly via a covalent bond. Furthermore, as used herein, the terms "conjugated to" and "bound to" can be understood interchangeably.

[0034] Preferably, the motif comprises at least one non-proteinogenic amide bond, more preferably at least two non-proteinogenic amide bonds, more preferably at least one natural amino acid moiety conjugated via a non-proteinogenic amide bond, even more preferably at least two natural amino acid moieties conjugated via non-proteinogenic amide bonds.

[0035] Preferably, the motif that specifically binds to the cell membrane of a neoplastic cell comprises a moiety of 20 or fewer amino acids, more preferably a moiety of 10 or fewer amino acids, even more preferably a moiety of 5 or fewer amino acids, even more preferably a moiety of 4 or fewer amino acids, and especially a moiety of 3 or fewer amino acids.

[0036] Preferably, the above motif further comprises at least one urea moiety, more preferably at least one urea moiety covalently attached to two amino acids via amide bond formation.

[0037] At least one motif (A) that specifically binds to the cell membrane of neoplastic cells is covalently linked to at least one radiometal chelator moiety (B) via a spacer x1. In this context, the spacer x1 is preferably 5 nm or less in length, more preferably 2 nm or less in length, and particularly preferably 1 nm or less in length. The motif may be conjugated to the radiometal chelator moiety (B) preferably via the epsilon-amino group of a lysine moiety.

[0038] The term "chelator moiety" as used in the context of the present invention may be understood in the broadest sense as any moiety capable of forming a complex with a radiometal under suitable conditions. Herein, the terms "chelator moiety," "chelant," "chelating moiety," "sequestering moiety," and "complexing moiety" may be understood interchangeably. The chelator moiety is preferably an organic moiety. Complexation by chelation preferably involves the formation or presence of two or more separate coordinate bonds between a polydentate (multiple-bonded) ligand and a single central radiometal. The International Union of Pure and Applied Chemistry (IUPAC) general definition of chelation (interpreted in its broadest sense) may also be noted. A "chelator moiety" as used herein typically has at least two heteroatoms that allow interaction with a radiometal. Preferably, the chelator moiety has at least three, especially at least four, heteroatoms that allow interaction with the radiometal.

[0039] A "radiometal" as used in the context of the present invention may be understood in the broadest sense as any radioactive metal or radioactive metal ion (i.e., a metal or metal ion that emits radiation). This may be a typically radioactive metal or metal ion, or may be a radioisotope of a metal that also has a non-radioactive isotope. By way of example, a radiometal may be gallium (Ga) (e.g., 67 Ga, 68 Ga), copper (e.g. 64 Cu, 67 Cu), iron (e.g. 59 Fe), zirconium (e.g. 89 Zr), scandium (e.g. 44 Sc), indium (e.g. 111 In), yttrium (e.g. 90 Y), rubidium (e.g. 82 Rb), cobalt (e.g.60 Co), ruthenium (e.g. 177 Lu), gadolinium (e.g. 153 Gd, 155 Gd, 157 Gd), bismuth (e.g. 213 Bi), strontium (e.g. 90 Sr), actinium (e.g. 225 Ac), or technetium (e.g. 99m Preferably, the radiometal is a radioisotope of 67 Ga, 68 Ga, 111 In, 90 Y, 177 Lu, 64 Cu, 67 Cu, 153 Gd, 155 Gd, 157 Gd, 89 Zr, 44 Sc, 99m Tc, 213 Bi, 225 Ac, 59 Fe or 82 Rb, more preferably 68 Ga, 64 Cu, 89 Zr, 44 Sc, or 82 Rb, even more preferably 68 Ga or 64 Cu, especially 68 A radioactive isotope of Ga. One skilled in the art will recognize several examples of chelator moieties suitable for complexing each of the above radiometals and will be able to select the chelator moiety accordingly. For example: 99m Tc or 82 Suitable chelator moieties for complexing Rb include: 68 Ga or 64 It may or may not be different from the chelating agent suitable for complexing Cu.

[0040] Preferably, the radiometal is one that has a half-life of 4 days or less, more preferably 1 day or less, even more preferably 12 hours or less, even more preferably 6 hours or less, even more preferably 3 hours or less, even more preferably 2.5 hours or less, even more preferably 120 minutes or less, even more preferably 100 minutes or less, even more preferably 80 minutes or less, especially 70 minutes or less.

[0041] The radiometal can be obtained from any source suitable for this purpose. The radiometal may be obtained and isolated from nature, or may be artificially produced (e.g., produced from a gallium-68 generator). 68 Ga, etc.) Those skilled in the art will understand how to obtain each radiometal.

[0042] The compounds of the present invention, or pharmaceutically acceptable salts thereof, comprise a moiety (B) having at least one radiometal chelator moiety. Thus, moiety (B) may represent one or more chelators. Illustratively, moiety (B) represents a chemical structure having one, two, or three chelators. Preferably, however, (B) represents one chelator. As outlined herein, moiety (B) is linked to moiety (A) via spacer x1.

[0043] The compounds of the present invention, or pharmaceutically acceptable salts thereof, comprise a dye moiety (C), which is linked to (B) via x2, which represents a spacer or a single chemical bond.

[0044] The moiety (C) has the formula:

[0045] [ka] (In the formula, X 1 and X 4 is -N=, -N(R 5 )=, and -C(R 6 )= independently selected from the group consisting of; X 2 and X 3are O, S, Se, N(R 5 ), and C(R 6 R 7 ) independently selected from the group consisting of: Y is a linker that connects the two moieties of (C) and allows for electron delocalization between said moieties, and Y is optionally a group (L-) c Z 0 Includes; a and b are independently selected from the group consisting of 1, 2, and 3; Each R 1 and each R 2 are independently (L-) c Z, (L-) c Z 0 or H; two adjacent R 1 and / or two adjacent R 2 may contain one or more (L-) c Z or (L-) c Z 0 It is also possible to form an aromatic ring substituted with; R 3 , R 4 , R 5 , R 6 , R 7 , R 9 is (L-) c Z, (L-) c Z 0 independently selected from the group consisting of each c is independently 0 or 1; Each L is independently T 1 , -OT 1 -, -ST 1 -, -C(O)T 1 -, -C(O)OT 1 -, -OC(O)T 1 -, -C(O)NHT 1 -,-NHC(O)T 1 , or optionally -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)O-, and T 1 C that is interrupted and / or terminated by one or more of 1-10 is an alkylene group; T 1is phenyl, naphthyl, indenyl, indanyl, tetralinyl, decalinyl, adamantyl, C 3-7 cycloalkyl, 3- to 7-membered heterocyclyl, or 7- to 11-membered heterobicyclyl, where T 1 is sometimes halogen, CN, C(O)R 8 , COOR 8 , OR 8 , C(O)N(R 8 R 8a ), S(O)2N(R 8 R 8a ), S(O)N(R 8 R 8a ), S(O)2R 8 , N(R 8 )S(O)2N(R 8a R 8b ), SR 8 , N(R 8 R 8a ), NO2, OC(O)R 8 , N(R 8 )C(O)R 8a , N(R 8 )S(O)2R 8a , N(R 8 )S(O)R 8a , N(R 8 )C(O)N(R 8a R 8b ), N(R 8 )C(O)OR 8a , OC(O)N(R 8 R 8a ), oxo(=O) (wherein the ring is at least partially saturated), or C 1-6 Alkyl (C 1-6 alkyl is optionally substituted with one or more of the same or different halogens); Each Z is independently H, halogen, CN, or C(O)R 8 , C(O)OR 8 , C(O)O - , OR 8 , C(O)N(R 8 R 8a ), S(O)2OR8, S(O)2O - , S(O)2N(R 8R 8a ), S(O)N(R 8 R 8a ), S(O)2R 8 , S(O)R 8 , N(R 8 )S(O)2N(R 8a R 8b ), SR 8 , N(R 8 R 8a ), NO2, P(O)(OR 8 )2, P(O)(OR 8 )O - ,OC(O)R 8 , N(R 8 )C(O)R 8a , N(R 8 )S(O)2R 8a , N(R 8 )S(O)R 8a , N(R 8 )C(O)N(R 8a R 8b ), N(R 8 )C(O)OR 8a , or OC(O)N(R 8 R 8a ) and; R 8 , R 8a , R 8b is H or C 1-6 Alkyl (C 1-6 alkyl is optionally substituted with one or more of the same or different halogens; Z 0 is a chemical bond that connects (C) to x2 or to (B) if x2 is a single chemical bond; However, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 9 One of them is (L-) c Z 0 or Y is (L-) c Z 0 (including It has.

[0046] The term "optionally substituted" means unsubstituted or substituted. Generally (but not exclusively), "one or more substituents" means 1, 2, or 3, preferably 1 or 2, and more preferably 1 substituent. Generally, the substituents may be the same or different.

[0047] "Alkyl" means a straight or branched hydrocarbon chain. Each hydrogen of an alkyl carbon may be optionally replaced with a substituent as further specified herein.

[0048] The terms "alkyl," "alkyl residue," and "alkyl group" and "alkyl moiety" used throughout this application may be understood as a straight-chain or branched saturated hydrocarbon chain. "Straight-chain" may also be referred to as "unbranched" or "linear." Preferably, alkyl is straight-chain.

[0049] The term "alkylene" as used throughout this application means a straight or branched saturated hydrocarbon chain in which two portions of the molecule are connected by an alkylene residue. "Straight-chain" can also be referred to as "unbranched" or "linear." Each hydrogen of an alkylene carbon may or may not be substituted (i.e., substituted or unsubstituted) with a substituent further specified herein.

[0050] "C 1-4 "Alkyl" means, for example, an alkyl chain having 1 to 4 carbon atoms at the end of the molecule, if present: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, or, for example, when two parts of the molecule are linked by an alkyl group, -CH2-, -CH2-CH2-, -CH(CH3)-, -CH2-CH2-CH2-, -CH(C2H5)-, -C(CH3)2-. C 1-4 Each hydrogen of an alkyl carbon may be substituted with a substituent as further specified herein.

[0051] "C 1-6 "Alkyl" refers, for example, to an alkyl chain having 1 to 6 carbon atoms at the end of the molecule, if present: C 1-4 Alkyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, or, for example, when two parts of the molecule are connected by an alkyl group, -CH2-, -CH2-CH2-, -CH(CH3)-, -CH2-CH2-CH2-, -CH(C2H5)-, -C(CH3)2-. C 1-6 Each hydrogen of an alkyl carbon may be substituted with a substituent as further specified herein.

[0052] "C 1-8 The term "alkylene residue" refers to an alkylene chain having 1 to 8 carbon atoms, for example, when two parts of the molecule are linked by an alkylene group, -CH-, -CH-CH-, -CH(CH)-, -CH-CH-CH-, -CH(C2H5)-, -C(CH3)-, -CH-C(CH3)-, -C(CH2-CH3)-, -CH(CH2-CH3)-, -CH-CH(CH3)(CH2-CH3)-, -CH(CH3)(CH2-CH3)-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)7-, -(CH2)8-, etc. 4-8 The terms "C alkylene" and "C alkylene" are defined accordingly. 3-7 "C5 (C4) alkylene" and "C5 (C4) alkylene" are defined accordingly.

[0053] The term “C 1-10 An "alkylene group" means a divalent straight or branched hydrocarbon chain having 1 to 10 carbon atoms. Each hydrogen of an alkyl carbon may be replaced with a substituent as further specified herein. Examples are methylene (-CH-), -CH-CH-, -CH(CH)-, -CH-CH-CH-, -CH(CH)-, -C(CH)-. 1-10Each hydrogen of an alkylene group carbon may be replaced with a substituent as further specified herein.

[0054] Therefore, "C 1-10 "Alkylene residue" means an alkylene chain having 1 to 10 carbon atoms when two parts of the molecule are linked by an alkylene group. Preferably, but not necessarily, C in the context of residue f of spacer y 1-10 The alkylene residue is a straight-chain (i.e., unbranched) C alkylene group optionally having one or more hydrogen(s) replaced and / or optionally having one or more -CH2- moieties replaced with -O- or -NH-. 1-10 It is an alkylene residue.

[0055] The phrase "one or more -CH2- moieties optionally substituted with ..." means that the specified number of CH2 groups may be replaced with an atom or group specified herein. Additionally, one or more hydrogens specified herein may be replaced with a substituent.

[0056] "Possibly blocked and / or terminated" C 1-10 An alkylene group means that the alkylene chain is interrupted between two carbon atoms by an atom or chemical group specified herein, or the alkylene group is terminated by said atom or group following the carbon at at least one end of the alkylene chain, or the alkylene chain is both interrupted and terminated, or the alkylene chain is neither interrupted nor terminated. By way of example, and not by way of limitation, a C alkylene group optionally interrupted and / or terminated by one or more X can have the sequence CCC, CCCX, XCCC, XCCCX, CXCC, CCXC, CXCXC, XCCXC, XCXCXC, XCXCCX, XCXCXCX.

[0057] The term "carbocycle" refers to a partially or fully saturated or aromatic carbocyclic monocyclic, bicyclic or tricyclic fused or non-fused ring system. This term includes phenyl and C 3-7 Preferred carbocycles having 5, 6 or 7 carbon atoms are cyclopentene, cyclohexene, phenyl, cycloheptane, especially cyclohexane.

[0058] "C 3-7 cycloalkyl" or "C 3-7 "Cycloalkyl ring" means a cyclic alkyl chain having 3 to 7 carbon atoms, e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl. Preferably, cycloalkyl refers to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or cycloheptyl. Each hydrogen of a cycloalkyl carbon may be replaced with a substituent as further specified herein. The term "C 3-5 cycloalkyl" or "C 3-5 "Cycloalkyl ring" is defined accordingly.

[0059] "Halogen" means fluoro, chloro, bromo or iodo. Generally, it is preferred that halogen is fluoro or chloro.

[0060] Within the meaning of the present invention, the term "aromatic ring" means a carbocyclic or heterocyclic aromatic ring. Examples are benzene, naphthalene, 5- to 6-membered aromatic heterocycles and 9- to 11-membered aromatic heterobicyclyls.

[0061] A "3- to 7-membered heterocyclyl" or "3- to 7-membered heterocycle" refers to a ring (an aromatic ring or a fully, partially saturated, or unsaturated non-aromatic ring) having 3, 4, 5, 6, or 7 ring atoms, which may include up to a maximum number of double bonds, in which at least one ring atom and up to four ring atoms are replaced with a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)-), oxygen, and nitrogen (including =N(O)-), and the ring is connected to the remainder of the molecule via a carbon or nitrogen atom. Examples of 3- to 7-membered heterocycles are aziridine, azetidine, oxetane, thietane, furan, thiophene, pyrrole, pyrroline, imidazole, imidazoline, pyrazole, pyrazoline, oxazole, oxazoline, isoxazole, isoxazoline, thiazole, thiazoline, isothiazole, isothiazolidine, thiadiazole, thiadiazoline, tetrahydrofuran, tetrahydrothiophene, pyrrolidine, imidazolidine, pyrazolidine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, thiadiazolidine, sulfolane, pyran, dihydropyran, tetrahydropyran, imidazolidine, pyridine, pyridazine, pyrazine, pyrimidine, piperazine, piperidine, morpholine, tetrazole, triazole, triazolidine, tetrazolidine, diazepane, azepine, or homopiperazine. The term "4- to 7-membered heterocyclyl" or "4- to 7-membered heterocycle" is defined accordingly. The term "5- to 6-membered heterocyclyl" or "5- to 6-membered heterocycle" is defined accordingly.

[0062] The term "5- to 6-membered aromatic heterocyclyl" or "5- to 6-membered aromatic heterocycle" refers to a heterocycle derived from cyclopentadienyl or benzene, in which at least one carbon atom is replaced with a heteroatom selected from the group consisting of sulfur (including -S(O)- and -S(O)-), oxygen, and nitrogen (including =N(O)-). Examples of such heterocycles include furan, thiophene, pyrrole, imidazole, pyrazole, oxazole, isoxazole, thiazole, isothiazole, thiadiazole, triazole, tetrazole, pyridine, pyrimidine, pyridazine, pyrazine, and triazine.

[0063] "5-membered aromatic heterocyclyl" or "5-membered aromatic heterocycle" means a heterocycle derived from cyclopentadienyl in which at least one carbon atom is replaced with a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)-), oxygen, and nitrogen (including =N(O)-). Examples of such heterocycles are furan, thiophene, pyrrole, imidazole, pyrazole, oxazole, isoxazole, thiazole, isothiazole, thiadiazole, triazole, and tetrazole.

[0064] A "7- to 11-membered heterobicyclyl" or "7- to 11-membered heterobicycle" refers to a bicyclic heterocyclic system (aromatic ring or fully, partially saturated, or unsaturated non-aromatic ring) having 7 to 11 ring atoms, in which at least one ring atom is shared by both rings and may contain up to a maximum number of double bonds, in which at least one ring atom and up to six ring atoms are replaced with a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)-), oxygen, and nitrogen (including =N(O)-), and in which the ring is connected to the remainder of the molecule via a carbon or nitrogen atom. Examples of 7-11 membered heterobicycles are indole, indoline, benzofuran, benzothiophene, benzoxazole, benzisoxazole, benzothiazole, benzisothiazole, benzimidazole, benzimidazoline, quinoline, quinazoline, dihydroquinazoline, quinoline, dihydroquinoline, tetrahydroquinoline, decahydroquinoline, isoquinoline, decahydroisoquinoline, tetrahydroisoquinoline, dihydroisoquinoline, benzazepine, purine or pteridine. The term "7-11 membered heterobicycle" also encompasses bicyclic spiro structures such as 6-oxa-2-azaspiro[3,4]octane, 2-oxa-6-azaspiro[3.3]heptan-6-yl or 2,6-diazaspiro[3.3]heptan-6-yl, or bridged heterocycles such as 8-aza-bicyclo[3.2.1]octane or 2,5-diazabicyclo[2.2.2]octan-2-yl or 3,8-diazabicyclo[3.2.1]octane.

[0065] A "9- to 11-membered aromatic heterobicyclyl" or "9- to 11-membered aromatic heterobicycle" means a bicyclic heterocyclic ring system in which at least one ring is aromatic and the heterocyclic ring system has 9 to 11 ring atoms, where two ring atoms are shared by both rings and both rings may contain up to a maximum number of double bonds (fully or partially aromatic), where at least one ring atom and up to six ring atoms are replaced with a heteroatom selected from the group consisting of sulfur (including -S(O)-, -S(O)-), oxygen, and nitrogen (including =N(O)-), and where the ring is connected to the remainder of the molecule via a carbon or nitrogen atom. Examples of 9-11 membered aromatic heterobicycles are indole, indoline, benzofuran, benzothiophene, benzoxazole, benzisoxazole, benzothiazole, benzisothiazole, benzimidazole, benzimidazoline, quinoline, quinazoline, dihydroquinazoline, dihydroquinoline, tetrahydroquinoline, isoquinoline, tetrahydroisoquinoline, dihydroisoquinoline, benzazepine, purine or pteridine. The terms "9-10 membered aromatic heterobicyclyl" or "9-10 membered aromatic heterobicycle" are defined accordingly.

[0066] Preferred compounds of formula (I) are those in which one or more of the residues contained therein have the meanings given below, and all combinations of preferred substituent definitions are the subject of the present invention. For all preferred compounds of formula (I), the present invention also includes all tautomeric and stereoisomeric forms, as well as mixtures thereof in all ratios and their pharmaceutically acceptable salts.

[0067] Where tautomerism of the compounds of formula (I) can occur, such as keto-enol tautomerism, the individual forms, such as the keto and enol forms, are included both separately and together as mixtures in any ratio. The same applies to stereoisomers, such as enantiomers, cis / trans isomers, conformers, etc.

[0068] In particular, in the compounds according to formula (I), when enantiomeric or diastereomeric forms are presented, each pure form separately and also any mixture of at least two of said pure forms in any ratio are encompassed by formula (I) and are the subject of the present invention.

[0069] Isotopically labeled compounds of formula (I) are also within the scope of the present invention. Methods for isotopic labeling are known in the art. Preferred isotopes are isotopes of the elements H, C, N, O and S. Solvates of compounds of formula (I) are also within the scope of the present invention.

[0070] If desired, isomers can be separated by methods well known in the art, for example, by liquid chromatography. The same applies to enantiomers, for example, by using a chiral stationary phase. Furthermore, enantiomers can be isolated by converting them to diastereomers (i.e., coupling with an enantiomerically pure auxiliary compound, followed by separation of the resulting diastereomers and cleavage of the auxiliary residue). Alternatively, any enantiomer of a compound of formula (I) can be obtained from stereoselective synthesis using optically pure starting materials, reagents and / or catalysts.

[0071] When a compound according to formula (I) contains one or more acidic or basic groups, the present invention also includes their corresponding pharmaceutically or toxicologically acceptable salts, particularly their pharmaceutically usable salts. Thus, compounds of formula (I) containing acidic groups can be used according to the present invention, for example, as alkali metal salts, alkaline earth metal salts, or ammonium salts. More specific examples of such salts include sodium salts, potassium salts, calcium salts, magnesium salts, or salts with ammonia or organic amines (e.g., ethylamine, ethanolamine, triethanolamine, or amino acids). Compounds of formula (I) containing one or more basic groups (i.e., protonatable groups) can exist and be used according to the present invention in the form of their addition salts with inorganic or organic acids. Examples of suitable acids include hydrogen chloride, hydrogen bromide, phosphoric acid, sulfuric acid, nitric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid, and other acids known to those skilled in the art. When the compound of formula (I) contains both an acidic and a basic group in the molecule, the present invention encompasses inner salts or betaines (zwitterions) in addition to the above salt forms. Salts according to formula (I) can be obtained by conventional methods known to those skilled in the art, for example, by contacting them with organic or inorganic acids or bases in a solvent or dispersant, or by anion or cation exchange with other salts. The present invention also covers all salts of compounds of formula (I) which, because of their low physiological compatibility, are not directly suitable for use in medicines but which may be used, for example, as intermediates for chemical reactions or for the preparation of pharmaceutically acceptable salts.

[0072] It should be understood that the compounds of the present invention or their pharmaceutically acceptable salts exhibit one or more charged functional groups, where each negative charge is compensated by a positive charge, resulting in a neutral molecule. Intramolecular compensation is also possible with other ions in the form of internal and external salts. Suitable counterions are those mentioned above. Thus, in the formula of the present invention, any remaining positive or negative charge(s) are compensated by pharmaceutically acceptable negative or positively charged counterions(s). Similarly, any protonated form can be partially or completely deprotonated with suitable counterions(s).

[0073] Counterions used herein can be any pharmaceutically acceptable ion suitable for neutralizing the charge of a residue or compound of the present invention. It is understood that a single counterion does not necessarily have the same valence as a charged residue or compound of the present invention. Two or more counterions can be used to neutralize a compound having a charge valence higher than +1 or -1. Conversely, a single counterion having a charge valence higher than +1 or -1 can also be used to neutralize two or more compounds. Preferably, the counterion is sufficiently soluble in an aqueous liquid.

[0074] A pharmaceutically acceptable negatively charged counterion X in the context of the present invention - can have any charge valence. - X may have, for example, a charge of -1, -2, -3 or -4, preferably -1 or -2. The charge may be determined by the ionic strength and pH, respectively, as the case may be. - X can be any pharmaceutically acceptable negatively charged ion. Preferably, the ion is thus sufficiently soluble in aqueous liquids. For example, X - is a halide anion (e.g., F - or Cl -), acetate, phosphate, hydrogen phosphate, and pharmaceutically acceptable carboxylates (e.g., fatty acid carboxylates). Furthermore, it is understood that the counterion will typically depend on the surrounding fluid, such as the fluid contained in the buffer in which the compound is dissolved and the body fluid after injection in vivo. In vivo, outside the cell, one of the main (but not the only) negatively charged counterions is Cl. - is.

[0075] The terms "dye moiety," "label," and "stain" as used in the context of the present invention may be understood interchangeably in the broadest sense as any moiety having the above formula that provides a visible stain. Preferably, the dye moiety may be a fluorescent dye moiety and / or a colored moiety, and particularly preferably, the dye moiety is a fluorescent dye moiety.

[0076] As used herein, the term "fluorochrome moiety" can be understood in the broadest sense as any dye moiety that allows for fluorescent detection. Preferably, such fluorescent detection is within the 400-1000 nm range, i.e., within the visible spectrum, and within the near-infrared (NIR) spectrum, particularly within the 400-800 nm range, i.e., within the visible spectrum. Preferably, the fluorescent signal emitted by the fluorescent dye moiety is sufficiently distinguishable from the autofluorescence of the neoplasm and surrounding tissue. Numerous fluorescent dye moieties are known in the art and readily apparent to those skilled in the art. Many fluorescent dyes are commercially available that have activating groups that can be used to react with other compounds, such as protein side chains or precursor compounds for the preparation of the compounds of the present invention.

[0077] Additionally or alternatively, the dye moiety can be colored (i.e., induces a color perception when illuminated by any light). Such a color effect can be induced by absorbing light in one or more specific wavelength range(s) within the visible range (i.e., within the range(s) of about 400 nm to about 800 nm) and / or by emitting light in one or more specific wavelength range(s) within the visible range. Preferably, the color is different from that of the neoplasm and surrounding tissue being examined. Therefore, if the dye moiety is not intended for fluorescence detection, it is preferably not red or brown, but rather blue or green. If the dye moiety is intended for fluorescence detection, color differences typically play only a minor role, as long as the fluorescence is detectable against an autofluorescent background. Preferably, the colored dye moiety in the context of the present invention is a small molecule dye, i.e., a dye moiety having a molecular weight (MW) of 1000 Da or less, preferably 750 Da or less, and particularly 500 Da or less.

[0078] The dye moiety (C) is covalently linked to at least one motif (A) that specifically binds to the cell membrane of a neoplastic cell and at least one radiometal chelator moiety (B) via the structure (A)-x1-(B)-x2-(C). Such covalent conjugation to the radiometal chelator moiety (B) may be a direct covalent bond between the dye moiety (C) and the radiometal chelator moiety (B) or may be a covalent bond via a spacer, preferably a spacer-mediated covalent conjugation. Preferably, the spacer is 5 nm or less in length, preferably 2 nm or less in length, and particularly preferably 1 nm or less in length.

[0079] Therefore, the molecular distance between the motif (A) that specifically binds to the cell membrane of neoplastic cells and the dye moiety (C) is preferably 20 nm or less, more preferably 10 nm or less, and particularly preferably 5 nm or less.

[0080] Depending on the chemical properties of the fluorescent dye moiety(s) in the compounds of the invention and the presence of fluorophores(s) and / or quenchers on the surface of the target cells (i.e., the cell membrane of each neoplastic cell), it may be possible to observe effects such as fluorescence energy transfer (FRET) and / or fluorescence quenching upon binding of the compounds of the invention to said cell membrane. Additionally or alternatively, the presence of the fluorescent dye moiety(s) may also allow for further fluorescence-based testing methods, such as fluorescence recovery after photobleaching (FRAP) and fluorescence decay after photobleaching (FLIP). These methods may provide information about the mobility of the compound or its salt bound to or attached to the cell membrane of neoplastic cells.

[0081] Preferably, the compounds according to the invention, or pharmaceutically acceptable salts thereof, have a molecular weight (MW) of 10 kDa or less, more preferably 5 kDa or less, even more preferably 3.5 kDa or less, even more preferably 3 kDa or less, especially 2.5 kDa or less.

[0082] As described above, motif (A) that specifically binds to the cell membrane of neoplastic cells can bind to any molecular structure typically found in neoplastic cells and can have any molecular structure.

[0083] In a preferred embodiment, the motif (A) that specifically binds to the cell membrane of a neoplastic cell is a motif that specifically binds to the cell membrane of a cancerous cell, and preferably said motif comprises a prostate-specific membrane antigen (PSMA) binding motif.

[0084] As used herein, the terms "prostate-specific membrane antigen," "prostate-specific membrane antigen-binding motif," and "PSMA-binding motif" may be understood interchangeably.

[0085] In a more preferred embodiment, the motif (A) that specifically binds to the cell membrane of neoplastic cells has the following structure:

[0086] [ka] (In the formula, Z 1 , Z 2 and Z 3 is -C(O)OR 1a , -SO2R 1a , -SO3R 1a , -SO4R 1a , -PO2R 1a , -PO3R 1a , and -PO4R 1a R 2a where R 1a and R 2a are, independently of each other, H or C 1-4 - alkyl residues (preferably identical, more preferably H); a' is -[CH2] o -residue, where o is an integer from 1 to 4, preferably o is 3 or 4, in particular o is 4; b' represents a residue selected from the group consisting of -NH-, -C(O)- and -O-, in particular b' is -NH-; The wavy line indicates the conjugation site of the radiometal to the chelator moiety (B), which is conjugated via a spacer molecule x1. The PSMA binding motif is

[0087] Preferably, Z 1 , Z 2 , Z 3 are identical, and more preferably, Z 1 , Z 2 , Z 3 -C(O)OR 1a is.

[0088] In a particularly preferred embodiment, the motif (A) that specifically binds to the cell membrane of neoplastic cells has the following structure:

[0089] [ka] where the wavy line indicates the conjugation site of the radiometal to the chelator moiety (B), which is conjugated via a spacer molecule x1. The PSMA binding motif has the following structure:

[0090] Spacers x1 and x2 may be spacers having a length of 5 nm or less, preferably a length of 2 nm or less, in particular a length of 1 nm or less.

[0091] The spacer x1 comprises a group -AA-, where AA is an amino acid sequence of 4 to 8 (preferably 5 to 7, more preferably 6) naturally occurring amino acids, and at least two amino acids are histidines.

[0092] The term "naturally occurring amino acid" refers to an amino acid selected from the group consisting of:

[0093] [Table 1]

[0094] Natural amino acids may be present in the racemic, D or L form, preferably in the L form.

[0095] Preferably, AA comprises three histidine amino acids. Preferably, AA consists of histidine and glutamic acid. Preferably, AA is represented by the formula -His-Glu-His-Glu-His-Glu-. Thus, the compound or a pharmaceutically acceptable salt thereof may comprise the amino acid sequence -His-Glu-His-Glu-His-Glu-, linked via the C-terminus and N-terminus to the remainder of the molecule, thus giving the following free polypeptide sequence: SEQ ID NO: 1 EHEHEH It comes from.

[0096] Preferably, the histidine contained in the C-terminus or N-terminus of AA, preferably the C-terminus, preferably AA, forms an amide bond with (A).

[0097] Thus, in a preferred embodiment, (A)-x1 has the following substructure with histidine as part of AA and x1 attached to (A):

[0098] [ka] (wherein the dotted line indicates the bond to the remainder of the AA contained in x1) Includes.

[0099] In a preferred embodiment, the spacer x1 has the following structure: -AA-[b''-e-b'''] n -b''''-d 1 - (In the formula, b″ is —C(O)— or —N(H)—, preferably forming an amide bond with AA; e is one or more -CH2- moieties, optionally one or more of -O-, -S-, -C(O)NH-, -C(O)N(C 1-6 C optionally substituted with alkyl, -C(O)O-, succinimide, triazole 1-8 -represents a residue selected from the group consisting of alkylene; b''' is selected from the group consisting of -NH- and -C(O)-; b'''' is selected from the group consisting of -C(O)- and -NH-; b''' and b'''', or the end of AA (preferably the N-terminus of AA) and b'''', together form an amide group; d 1 -[CH2] p where p is 1 or 2, in particular 2; n is 0 or 1, preferably 0. It has.

[0100] Preferably, b'' is C(O).

[0101] Preferably, e does not have any substitution or has 1, 2 or 3 substitutions, preferably 1 substitution, especially with triazole.

[0102] Preferably, e is unsubstituted C 1-8 -Alkylene, one -CH2- replaced by triazole 1-8 Alkylene, one -CH2- replaced by triazole 4-8 Alkylene (especially C6 alkylene), -CH2-(O-CH2-CH2)2-CH2-, -(CH2)2-(O-CH2-CH2)2-, -(CH2) 3-O-CH2-CH2-O-CH2-, -CH2-O-(CH2)6-, -(CH2)2-O-(CH2)5-, -(CH2)3-O-(CH2)4-, - (CH2)4-O-(CH2)3-, -(CH2)5-O-(CH2)2-, -(CH2)6-O-CH2-, -CH2-(O-CH2-CH2)2-, - (CH2)2-O-CH2-CH2-O-CH2-, -CH2-O-(CH2)5-, -(CH2)2-O-(CH2)4-, -(CH2)3-O-(CH2 )3-, -(CH2)4-O-(CH2)2-, -(CH2)5-O-CH2-, -CH2-O-CH2-CH2-O-CH2-, -CH2-O-(CH2)4-, -(CH2)2-O-(CH2)3-, -(CH2)3-O-(CH2)2-, -(CH2)4-O-CH2-, -CH2-O-(CH2)3-, -(CH2)2-O-(CH2)2-, -(CH2)3-O-CH2-, -CH2-O-(CH2)2-, -(CH2)2-O-CH2-, and -CH2-O-CH2-, in particular butylene, pentylene, hexylene or C in which one -CH2- is replaced by triazole. 4-8 is a residue selected from the group consisting of alkylene.

[0103] In particular, e has the following structure:

[0104] [ka] (wherein the dotted line with an asterisk represents a bond to b'', and the dotted line without an asterisk represents a bond to b''') It is expressed by:

[0105] Preferably, b''' is -NH-. Preferably, b''' is -C(O)-. Preferably, n is 1.

[0106] Preferably, b''' and b'''', or when n=0, the end of AA (preferably the N-terminus of AA) and b'''', combine to form an amide group.

[0107] In a more preferred embodiment, the spacer x1 has the following structure: -AA-[C(O)-e-NH] n -C(O)-(CH2) p - (In the formula, e is C in which one -CH2- is replaced by triazole 4-8 alkylene, particularly C alkylene in which one -CH2- is replaced by triazole; n is 0 or 1, in particular 1; p is 1 or 2, especially 2 It has.

[0108] In a particularly preferred embodiment, the spacer x1 has the following structure:

[0109] [ka] It has.

[0110] In a particularly preferred embodiment, (A)-x1 has the following structure:

[0111] [ka] It is expressed by:

[0112] In formula (I), x2 is a spacer or a single chemical bond connecting (B) and (C). Preferably, x2 is a spacer. The spacer x2 is preferably quite hydrophilic.

[0113] In a preferred embodiment, the spacer x2 has the following structure: -d 2 -e-[f-e'] m - (In the formula, d 2 -[CH2] r where r is 1 or 2, in particular 2; e is -C(O)-NH-, -NH-C(O)-, -C(O)-O-, -OC(O)-, -NH-C(O)-NH-, -NH-C(S)-NH-,

[0114] [ka] (In the formula, one of the wavy lines is d 2 The other wavy line indicates the conjugation site for f) In particular, e is -C(O)-NH-; Preferably, e is —C(O)NH—; Each f independently represents a C group in which one or more -CH2- moieties are optionally replaced with -O- or -NH-. 1-10 -alkylene, and f is unsubstituted or is selected from the group consisting of -NH, -COOH and R 3a and is substituted with one or more groups independently selected from the group consisting of: R 3a are -(CH2)2-COOH, -(CH2)4-NH2, -(CH2)4-N + (CH3)3+ X - , -CH2-COOH, -CH2-SH, -CH2-SO3H, and

[0115] [ka] is selected from the group consisting of X - is a pharmaceutically acceptable negatively charged counterion; Preferably, f is -CH2-(O-CH2-CH2)2-CH2-, -(CH2)2-(O-CH2-CH2)2-, -(CH2)2-(CH2-CH2-O)2-(CH2)2-, -(CH2)3-(CH2-CH2-O)2-CH2-, -(CH2-CH2-O)3-CH2-, -(CH2)2-(CH2-CH2-O)2-CH2-, -(CH2)2-(CH2-CH2-O)2-CH2-, -(CH2)2-(CH2-CH2-NH)2-(CH2)2-, -(CH2)3-(CH2-CH2-NH)2-CH2-, -(CH2-CH2-NH)3-CH2-, -(CH2)2-(CH2-CH2- NH)2-CH2-、-(CH2)3-O-CH2-CH2-O-CH2-、-CH2-O-(CH2)6-、-(CH2)2-O-(CH2)5-、-(CH2)3-O-(CH2)4-、-(CH2)4-O-(CH2)3-、-(CH2)5-O-(CH2)2-、-( CH2)6-O-CH2-、-CH2-(O-CH2-CH2)2-、-(CH2)2-O-CH2-CH2-O-CH2-、-CH2-O-(CH2)5-、-(CH2)2-O-(CH2)4-、-(CH2)3-O-(CH2)3-、-(CH2)4-O-(CH2)2 -、-(CH2)5-O-CH2-、-CH2-O-CH2-CH2-O-CH2-、-CH2-O-(CH2)4-、-(CH2)2-O-(CH2)3-、-(CH2)3-O-(CH2)2-、-(CH2)4-O-CH2-、-CH2-O-(CH2)3-、-(CH 2)2-O-(CH2)2-、-(CH2)3-O-CH2-、-CH2-O-(CH2)2-、-(CH2)2-O-CH2-、-CH2-O-CH2-、-(CH2)3-(O-CH2-CH2)2-CH2-、-(CH2)2-(O-CH2-CH2)2-(CH2)2 , -CH2-(O-CH2-CH2)2-(CH2)3, -CH2-(O-CH2-CH2)3-, -(CH2)2-(O-CH2-CH2)2-CH2-, -CH2-(O-CH2-CH2)2-(CH2)2-, and -(CH2)3-(O-CH2-CH2)2-, CH2-(NH-CH2-CH2)2-CH2-, -(CH2)2-(NH-CH2-CH2)2-, -(CH2)3-NH-CH2-CH2-NH-CH2-, -CH2-NH-(CH2)6-, -(CH2)2-NH-(CH2)5-, -(CH2)3-NH-(CH2)4-,-(CH2)4-NH-(CH2)3-、-(CH2)5-NH-(CH2)2-、-(CH2)6-NH-CH2-、-CH2-(NH-CH2-CH2)2-、-(CH2)2-NH-CH2-CH2-NH-CH2-、-CH2-NH-(CH2)5-、-(CH2)2-NH-(CH2)4-、-(CH2)3-NH-(CH2)3-、-(CH2)4-NH-(CH2)2-、-(CH2)5-NH-CH2-、-CH2-NH-CH2-CH2-NH-CH2-、-CH2-NH-(CH2)4-、-(CH2)2-NH-(CH2)3-、 -(CH2)3-NH-(CH2)2-、-(CH2)4-NH-CH2-、-CH2-NH-(CH2)3-、-(CH2)2-NH-(CH2)2-、-(CH2)3-NH-CH2-、-CH2-NH-(CH2)2-、-(CH2)2-NH-CH2-、-CH2-NH-CH2-、-(CH2)3-(NH-CH2-CH2)2-CH2-、-(CH2)2-(NH-CH2-CH2)2-(CH2)2、-CH2-(NH-CH2-CH2)2-(CH2)3、-CH2-(NH-CH2-CH2)3-、-(CH2)2-(NH-CH2-CH2)C H2)2-CH2-、-CH2-(NH-CH2-CH2)2-(CH2)2-、-(CH2)3-(NH-CH2-CH2)2-、-CH2-O-(CH2)8-、-(CH2)2-O-(CH2)7-、-(CH2)3-O-(CH2)6-、-(CH2)4-O-(CH2)5-、-(CH2)5-O-(CH2)4-、-(CH2)6-O-(CH2)3-、-(CH2)7-O-(CH2)2-、-(CH2)8-O-CH2-、-CH2-O-(CH2)7-、-(CH2)2-O-(CH2)6-、-(CH2)3-O-(CH2)5-、 -(CH2)4-O-(CH2)4-、-(CH2)5-O-(CH2)3-、-(CH2)6-O-(CH2)2-、-(CH2)7-O-CH2-、-CH2-NH-(CH2)8-、-(CH2)2-NH-(CH2)7-、-(CH2)3-NH-(CH2)6-、-(CH2)4-NH-(CH2)5-、-(CH2)5-NH-(CH2)4-、-(CH2)6-NH-(CH2)3-、-(CH2)7-NH-(CH2)2-、-(CH2)8-NH-CH2-、-CH2-NH-(CH2)7-、-(CH2)2-NH-(CH2)6-、-(CH2)3-NH-(CH2)5-, -(CH2)4-NH-(CH2)4-, -(CH2)5-NH-(CH2)3-, -(CH2)6-NH-(CH2)2-, -(C H2)7-NH-CH2-, -CH(NH2)-CH2-, -CH2-CH(NH2)-, -CH(COOH)-CH2-, -CH2-CH(COOH)-, and -CH(R, 3 )-, In particular, f is a residue selected from the group consisting of -(CH2)2-(O-CH2-CH2)2-, -CH2-(O-CH2-CH2)2-CH2-, and -(CH2)3-O-CH2-CH2-O-CH2-, especially -(CH2)2-(O-CH2-CH2)2-; Each e'' is a chemical bond, -NH-C(O)-, -C(O)-NH-, -C(O)-O- and -OC(O)-, -NH-C(O)-NH-, -NH-C(S)-NH-, -C(O)-N(CH3)-, -N(CH3)-C(O)-, -NH-C(S)-, -C(S)-NH-,

[0116] [ka] wherein one wavy line indicates a conjugation site for f and the other wavy line indicates a conjugation site for at least one dye moiety (C), in particular e″ is —NH—C(O)—; m represents an integer of 0 to 8, preferably 0 to 4, even more preferably 0 to 2, even more preferably 0 or 1, and particularly preferably 1. It has.

[0117] In a more preferred embodiment, the spacer x2 has the following structure: -(CH2) t -C(O)-NH-(CH2) u -(O-CH2-CH2) v -(CH2) w -e''-, or -(CH2) t -C(O)-NH-(CH2-CH2-O) v -CH2-e''- (In the formula, t is 1 or 2, in particular 2; u is an integer from 1 to 10, preferably from 1 to 3, in particular 2; v is an integer from 0 to 3, in particular 2; w is an integer of 0 to 2, particularly 0. It has one of the following.

[0118] In an even more preferred embodiment, the spacer y has the following structure: -(CH2)2-C(O)-NH-(CH2)2-(O-CH2-CH2)2-e''- -(CH2)2-C(O)-NH-(CH2)2-(O-CH2-CH2)2-NH-C(O)-CH2-(O-CH2-CH2) n' -O-CH2-e''-, -(CH2)2-C(O)-NH-(CH2)2-(O-CH2-CH2)2-NH-[C(O)-CH((CH2)2COOH)-NH] n'' -C(O)-CH((CH2)2COOH)-e''-, -(CH2)2-C(O)-NH-(CH2)2-(O-CH2-CH2)2-NH-[C(O)-CH((CH2)4NH2)-NH] n'' -C(O)-CH((CH2)4NH2)-e''-, or -(CH2)2-C(O)-NH-(CH2)2-(O-CH2-CH2)2-NH-[C(O)-CH((CH2)4N + (CH3)3)-NH] n'' -C(O)-CH((CH2)4N + (CH3)3)-e''- + X - , (In the formula, n' is an integer from 1 to 3; n'' is an integer from 0 to 2; X - is a pharmaceutically acceptable negatively charged counterion; Each e'' is a chemical bond, -NH-C(O)-, -C(O)-NH-, -C(O)-O- and -OC(O)-, -NH-C(O)-NH-, -NH-C(S)-NH-, -C(O)-N(CH3)-, -N(CH3)-C(O)-, -NH-C(S)-, -C(S)-NH-,

[0119] [ka] wherein one wavy line indicates a conjugation site for f and the other wavy line indicates a conjugation site for at least one dye moiety (C), and in particular e″ is —NH—C(O)—. It has one of the following.

[0120] In particular, x2 is represented by the formula -(CH2)2-C(O)-NH-(CH2)2-(O-CH2-CH2)2-e''-.

[0121] The compound according to the present invention or its pharmaceutically acceptable salt is preferably capable of complexing one or more radioactive metal(s), thereby forming a compound-radioactive metal complex, or may be uncomplexed. Obviously, for the detection of radioactive signals, the compound or its salt is preferably complexed with at least one radioactive metal, while for the detection of fluorescent signals, it is optional whether the compound or its salt complexes a radioactive metal. Thus, in the context of detecting radioactive signals, the term "compound" can be understood as meaning that the compound preferably complexes at least one radioactive metal. However, the molecular weight of the compound or salt shown herein is based on the structure without the radioactive metal.

[0122] As noted above, the radiometal chelator moiety (B) is preferably: 68 Ga, 99m Tc or 82 Suitable for complexing Rb (especially in aqueous environments) 68 It is a chelating agent suitable for complexing Ga.

[0123] Gallium-68( 68 Gallium-68 (Ga) has a half-life of about 68 minutes, making it rather inconvenient for long-distance transportation. 68 Ga) can typically be generated near the location where it is complexed with a compound of the invention or a pharmaceutically acceptable salt thereof and administered to a patient in vivo and / or a sample in vitro.

[0124] Thus, in a preferred embodiment, the radiometal chelator moiety (B) is 68 Ga-chelator moieties, preferably 68 Ga-chelator moiety.

[0125] The radiometal chelator moiety (B) is preferably:

[0126] [ka] wherein in each of the above structures, one of the wavy lines indicates a conjugation site to at least one motif (A) that specifically binds to the cell membrane of a neoplastic cell, via a spacer x1, as defined herein, and the other wavy line indicates a conjugation site to a dye moiety (C), preferably via a spacer x2, as defined herein. is.

[0127] In particular, the chelating moiety:

[0128] [ka] is.

[0129] When this chelating agent is used, b'''' is preferably -C(O)- and d 1 and d 2are preferably each -(CH2)2-, and e' is preferably -C(O)-NH or -C(O)-O-, especially -C(O)NH-. Thus, the chelator moiety is a group consisting of two -C(O)-(CH2)2- # The residue is also referred to as "HBED-CC."

[0130] Preferably, (A)-x1-(B)-x2- is of the following formula:

[0131] [ka] It is expressed by:

[0132] These examples given above are 68 Some of the "Ga-chelator moieties" also react with one or more other radiometal(s) (e.g., 64 It will be noted that the cations may serve as complexing structures for the cations (e.g., Cu).

[0133] Preferably, in an aqueous environment of approximately neutral pH (i.e., pH 6-8, particularly 6.5-7.5), neither the motif (A) that specifically binds to the cell membrane of neoplastic cells nor the complexed radiometal or radiometal chelator moiety (B) quenches the intensity of the fluorescent signal obtainable from the dye moiety (C) by more than 50% at its emission maximum.

[0134] Preferably, the dye moiety (C) is suitable for emitting light in an aqueous environment of approximately neutral pH (ie, pH 6-8, particularly 6.5-7.5, especially pH 7.0-7.5).

[0135] In a preferred embodiment, the dye moiety (C) is a fluorescent dye moiety having an emission maximum in the range of 400 nm to 1000 nm.

[0136] The dye moiety (C) has the formula:

[0137] [ka] (In the formula, X 1 and X 4 is -N=, -N(R 5 )=, and -C(R 6 )= independently selected from the group consisting of; X 2 and X 3 are O, S, Se, N(R 5 ), and C(R 6 R 7 ) independently selected from the group consisting of: Y is a linker that connects the two moieties of (C) and allows for electron delocalization between said moieties, and Y is optionally a group (L-) c Z 0 Includes; a and b are independently selected from the group consisting of 1, 2, and 3; Each R 1 and each R 2 are independently (L-) c Z, (L-) c Z 0 or H; two adjacent R 1 and / or two adjacent R 2 may contain one or more (L-) c Z or (L-) c Z 0 It is also possible to form an aromatic ring substituted with; R 3 , R 4 , R 5 , R 6 , R 7 , R 9 is (L-) c Z, (L-) c Z 0 independently selected from the group consisting of each c is independently 0 or 1; Each L is independently T 1 , -OT 1 -, -ST 1-, -C(O)T 1 -, -C(O)OT 1 -, -OC(O)T 1 -, -C(O)NHT 1 -,-NHC(O)T 1 , or optionally -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)O-, and T 1 C that is interrupted and / or terminated by one or more of 1-10 is an alkylene group; T 1 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, decalinyl, adamantyl, C 3-7 cycloalkyl, 3- to 7-membered heterocyclyl, or 7- to 11-membered heterobicyclyl, where T 1 is sometimes halogen, CN, C(O)R 8 , COOR 8 , OR 8 , C(O)N(R 8 R 8a ), S(O)2N(R 8 R 8a ), S(O)N(R 8 R 8a ), S(O)2R 8 , N(R 8 )S(O)2N(R 8a R 8b ), SR 8 , N(R 8 R 8a ), NO2, OC(O)R 8 , N(R 8 )C(O)R 8a , N(R 8 )S(O)2R 8a , N(R 8 )S(O)R 8a , N(R 8 )C(O)N(R 8a R 8b ), N(R 8 )C(O)OR 8a , OC(O)N(R 8 R 8a ), oxo(=O) (wherein the ring is at least partially saturated), or C 1-6 Alkyl (C 1-6alkyl is optionally substituted with one or more of the same or different halogens); Each Z is independently H, halogen, CN, or C(O)R 8 , C(O)OR 8 , C(O)O - , OR 8 , C(O)N(R 8 R 8a ), S(O)2OR8, S(O)2O - , S(O)2N(R 8 R 8a ), S(O)N(R 8 R 8a ), S(O)2R 8 , S(O)R 8 , N(R 8 )S(O)2N(R 8a R 8b ), SR 8 , N(R 8 R 8a ), NO2, (O)(OR 8 )2, P(O)(OR 8 )O - ,OC(O)R 8 , N(R 8 )C(O)R 8a , N(R 8 )S(O)2R 8a , N(R 8 )S(O)R 8a , N(R 8 )C(O)N(R 8a R 8b ), N(R 8 )C(O)OR 8a , or OC(O)N(R 8 R 8a ) and; R 8 , R 8a , R 8b is H or C 1-6 Alkyl (C 1-6 alkyl is optionally substituted with one or more of the same or different halogens; Z 0 is a chemical bond that connects (C) to x2 or to (B) if x2 is a single chemical bond; However, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 9 One of them is (L-) c Z 0 or Y is (L-) c Z 0 (including It has.

[0138] Therefore, residue R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 One of the groups is a linking group, atom or bond (L-) of the dye moiety (C) to x2 or (B). c Z 0 Preferably, R 3 or R 4 is (L-) c Z 0 More preferably, R 3 (L-) c Z 0 is.

[0139] Preferably, in formula (C), X 1 and X 4 are identical, preferably C(R 6 ), more preferably CH.

[0140] Preferably, in formula (C), X 2 and X 3 are identical, preferably C(R 6 R 7 ), more preferably R 6 and R 7 are identical, and even more preferably LZ (L=C 1-10 alkylene, even more preferably L=CH2 and Z=H).

[0141] Preferably, R9 is H.

[0142] Preferably, in formula (C), Y is (L-) c Z 0 and preferably Y does not contain:

[0143] [ka] (In the formula, g is 1, 2, 3, or 4 (preferably 2 or 3, more preferably 3), and each R 9a is (L-) c Z or H; two R 9a can also form a carbocyclic ring having 5, 6, or 7 carbon atoms or a 4- to 7-membered heterocyclic ring; each c is independently 0 or 1; Each L is independently T 1 , -OT 1 -, -ST 1 -, -C(O)T 1 -, -C(O)OT 1 -, -OC(O)T 1 -, -C(O)NHT 1 -,-NHC(O)T 1 , or optionally -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)O- and T 1 C that is interrupted and / or terminated by one or more of 1-10 is an alkylene group; T 1 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, decalinyl, adamantyl, C 3-7 cycloalkyl, 4- to 7-membered heterocyclyl, or 7- to 11-membered heterobicyclyl, where T 1 is sometimes halogen, CN, C(O)R 8 , COOR 8 , OR 8 , C(O)N(R 8 R 8a ), S(O)2N(R 8 R 8a ), S(O)N(R8 R 8a ), S(O)2R 8 , N(R 8 )S(O)2N(R 8a R 8b ), SR 8 , N(R 8 R 8a ), NO2, OC(O)R 8 , N(R 8 )C(O)R 8a , N(R 8 )S(O)2R 8a , N(R 8 )S(O)R 8a , N(R 8 )C(O)N(R 8a R 8b ), N(R 8 )C(O)OR 8a , OC(O)N(R 8 R 8a ), oxo(=O) (wherein the ring is at least partially saturated), or C 1-6 Alkyl (C 1-6 alkyl is optionally substituted with one or more of the same or different halogens); R 8 , R 8a , R 8b is H or C 1-6 Alkyl (C 1-6 alkyl is optionally substituted with one or more of the same or different halogens; Each Z is independently H, halogen, CN, or C(O)R 8 , C(O)OR 8 , C(O)O - , OR 8 , C(O)N(R 8 R 8a ), S(O)2OR8, S(O)2O - , S(O)2N(R 8 R 8a ), S(O)N(R 8 R 8a ), S(O)2R 8 , S(O)R 8 , N(R 8 )S(O)2N(R8a R 8b ), SR 8 , N(R 8 R 8a ), NO2, P(O)(OR 8 )2, P(O)(OR 8 )O - ,OC(O)R 8 , N(R 8 )C(O)R 8a , N(R 8 )S(O)2R 8a , N(R 8 )S(O)R 8a , N(R 8 )C(O)N(R 8a R 8b ), N(R 8 )C(O)OR 8a , or OC(O)N(R 8 R 8a ) is) is.

[0144] Preferably, Y is:

[0145] [ka] or

[0146] [ka] (g = 2 and each R 9a = H) is.

[0147] Preferably, in formula (C), a and b are the same and are preferably 1, more preferably R 1 and R 2 = SO3 - is.

[0148] Preferably, in formula (C), a and b are the same and 2, and preferably, two adjacent R 1 and two adjacent R 2 forms a phenyl ring.

[0149] Preferably, in formula (C), R 3 and R 4 One of them is (L-) c Z Z0 and the other is (L-) c Z(L- = C 1-10 Alkylene, Z = H or SO3 - , preferably c = 1).

[0150] Preferably, (L-) c Z 0 is a C bond that bonds (C) to x2, or to (B) if x2 is a single chemical bond. 1-10 Alkylene (c=1, Z 0 is a chemical bond), preferably C 3-7 It is alkylene, more preferably C5 alkylene.

[0151] Preferably, (L-) c Z is C 1-10 Alkylene, preferably C 3-7 alkylene, more preferably C4 alkylene, c is 1, and Z is SO3 - is.

[0152] In a highly preferred embodiment, the dye moiety (C) has the following structure:

[0153] [ka] (In the formula, X - is a pharmaceutically acceptable negatively charged counterion; Y + is a pharmaceutically acceptable positively charged counterion; The wavy line indicates the site of conjugation to the remainder of the compound of the invention. is a fluorescent dye moiety selected from the group consisting of:

[0154] Most preferred is

[0155] [ka] is.

[0156] a pharmaceutically acceptable negatively charged counterion X - can be understood in the broadest sense set out above.

[0157] Similarly, a pharmaceutically acceptable positively charged counterion Y + can have any valence. + Y may, for example, have a charge of +1, +2, +3 or +4, preferably +1 or +2. + Y can be any pharmaceutically acceptable positively charged ion. Preferably, the ion is one that is sufficiently soluble in aqueous liquids. For example, Y + is the cation of an alkali metal (e.g., Na + , K. + , Li + ), alkaline earth metal cations (e.g., Mg 2+ , Ca 2+ ), Al 3+ , NH4 + , H + and cations of organically bound amines. It is further understood that the counter ion is typically determined by the surrounding fluid, such as the buffer in which the compound is dissolved and that contained in body fluids after injection in vivo. In vivo, outside the cell, the predominant (but not the only) positively charged counter ion is Na + is.

[0158] Preferably, the wavy line indicates the conjugation site for spacer x2. More preferably, the wavy line indicates the conjugation site for e''.

[0159] Thus, in a highly preferred embodiment, the compound of the present invention has the following chemical structure:

[0160] [ka] It has.

[0161] The compounds according to the invention can be obtained by rational chemical synthesis. Exemplary routes for the preparation of the compounds of the invention or pharmaceutically acceptable salts thereof are described in US 2015 / 0110715 A1.

[0162] The compounds according to the invention may be substantially pure or may form part of a composition further comprising a radiometal and one or more pharmaceutically acceptable carriers.

[0163] The present invention further comprises: (a) a compound according to the invention as defined above or a pharmaceutically acceptable salt thereof; (b) a radioactive metal, preferably 89 Zr, 44 Sc, 111 In, 90 Y, 67 Ga, 68 Ga, 177 Lu, 99m Tc, 82 Rb, 64 Cu, 67 Cu, 153 Gd, 155 Gd, 157 Gd, 213 Bi, 225 Ac and 59 Fe, in particular 68 Ga; and optionally (c) one or more pharmaceutically acceptable carriers The present invention relates to a composition comprising:

[0164] The pharmaceutically acceptable carrier may be any agent that is pharmaceutically acceptable and can be added to the compound complexed with a radioactive metal (compound-radioactive metal complex).

[0165] For example, pharmaceutically acceptable carriers may include non-toxic or low-toxicity solvents such as water, aqueous buffers (e.g., Hepes, Tris, or phosphate buffer), pharmaceutically acceptable organic solvents (e.g., dimethyl sulfoxide (DMSO), ethanol, vegetable oils, paraffin oil), or combinations of two or more thereof.

[0166] Additionally, the pharmaceutically acceptable carrier may contain one or more cleaning agents, one or more foaming agents (e.g., sodium lauryl sulfate (SLS) / sodium dodecyl sulfate (SDS)), one or more coloring agents (e.g., TiO2, food coloring), one or more vitamins, one or more salts (e.g., sodium salts, potassium salts, calcium salts, zinc salts), one or more humectants (e.g., sorbitol, glycerol, mannitol, propylene glycol, polydextrose), one or more enzymes, one or more preservatives, preservatives (e.g., benzoic acid, methylparaben), one or more texturing agents (e.g., carboxymethylcellulose (CMC), polyethylene glycol (PEG), sorbitol), one or more emulsifiers, one or more bulking agents, one or more glazing agents, one or more separating agents, one or more antioxidants, one or more herbal and plant extracts, one or more stabilizers, one or more polymers (e.g., hydroxypropyl methacrylamide (HPMA), polyethyleneimine (PEI), carboxymethylcellulose (CMC), polyethylene glycol (PEG), sorbitol), and one or more uptake mediators (e.g., polyethyleneimine (PEI), dimethyl sulfoxide (DMSO), cell penetrating peptides (CPPs), protein transduction domains (PTDs), antimicrobial peptides, etc.), one or more antibodies, one or more sweeteners (e.g., acesulfame, acesulfame salts (e.g., acesulfame potassium (acesulfame K), aspartame, cyclamate, saccharin, saccharin salts ( For example, saccharin sodium (saccharin Na), alitame, neotame, sucralose, dulcin, aspartame-acesulfame salt, sorbitol, stevia, glycerol, inulin, mannitol, isomalt, maltitol, maltooligosaccharide, lactitol, xylitol, glucin, neohesperidin dihydrochalcone, P-4000, brazzein, curculin, erythritol, glycyrrhizin, hydrogenated starch hydrolysate, luo han guo, mabinlin, miraculin, monatin, monellin, osladin, pentadin, tagatose, thaumatin, one or more counterstain dyes (e.g., fluorescein, fluorescein derivatives, Cy dyes, Alexa Fluor 800),The diagnostic composition may comprise one or more homeopathic ingredients, one or more tastants, and / or one or more flavorings (e.g., Fluor dyes, rhodamines, quantum dots, etc.), one or more homeopathic ingredients, one or more tastants, and / or one or more flavorings. The diagnostic composition is typically formed by contacting the above-mentioned ingredients with other ingredients by any method known in the art. Preferably, the diagnostic composition is formed, and thus the above-mentioned ingredients are contacted with other ingredients, before being administered to a patient.

[0167] The radioactive metal may be commercially available, naturally occurring, or obtained from a cyclotron. Preferably, the radioactive metal is obtained from a gallium-68 generator or cyclotron, particularly one located near the site where the composition according to the invention is mixed and, optionally, where in vivo and / or in vitro diagnosis is also performed. Particularly preferably, the radioactive metal is 68 Ga is a gallium-68 generator, which produces a positron-emitting isotope of gallium from a source of decaying germanium-68. 68 It is obtained from the equipment used to extract Ga. 68 Ge is known to have a half-life of 271 days, allowing it to be transported to locations where gallium-68 generators are located.

[0168] Such compositions may be used in vivo in patients and in vitro in tissue culture to diagnose patients for neoplasia, particularly cancerous tissue or tissue at risk of becoming cancerous.

[0169] Thus, a further aspect of the present invention relates to a method for diagnosing a neoplasm in a patient suffering from or at risk of a neoplasm, comprising administering to said patient a sufficient amount of a composition according to the present invention.

[0170] Preferably, the compositions of the present invention are used as diagnostic agents.

[0171] In the context of the diagnostic method, the definitions of the terms specified in the context of the compound, compound-radiometal complex or pharmaceutically acceptable salt thereof above also apply.

[0172] The term "patient" as used in the context of the present invention may be understood in the broadest sense as a subject or individual to whom a compound of the present invention or a compound-radiometal complex or a pharmaceutically acceptable salt thereof is administered, regardless of whether the subject is a human or an animal, and whether or not clinical symptoms occur. Preferably, the patient is a mammal, including a human, more preferably a human, dog, horse, cow, pig, mule, donkey, sheep, goat, or camel. Particularly preferably, the patient is a human patient.

[0173] A patient suffering from a neoplasm may be understood in the broadest sense as any patient with a neoplasm. As used herein, a patient suffering from a neoplasm does not necessarily have any clinical symptoms. The patient may or may not be aware that they have a neoplasm. Similarly, the patient's treating physician may or may not be aware that a neoplasm exists. The patient may optionally suffer from pain, pressure, and / or gastrointestinal and / or urinary dysfunction.

[0174] The term "patient at risk" may be understood in the broadest sense as any patient who may potentially develop a neoplasm. In particular, the patient may be of a certain age, may be living under various conditions, may be exposed to drug therapies that affect gene expression, and / or may have a genetic heredity that carries an increased risk of developing a neoplasm. By way of example, the at-risk patient may be over 40 years of age, preferably over 45 years of age, more preferably over 50 years of age, even more preferably over 60 years of age, even more preferably over 70 years of age, and particularly over 80 years of age. Alternatively or additionally, the patient may be overweight. Alternatively or additionally, the patient may have a family history in which cancer is relatively common, particularly a family history of first-degree family members affected by cancer.

[0175] The above-mentioned compound, compound-radioactive metal complex, or pharmaceutically acceptable salt thereof can be administered to a patient by any means. Preferably, they are injected into a target tissue (i.e., a neoplastic tissue or a tissue at risk of being a neoplastic tissue) or blood vessel via a syringe or infusion. Alternatively, they may be injected intraperitoneally, or administered orally, nasally, respiratoryly, topically, or subcutaneously.

[0176] For example, the compound, compound-radiometal complex, or pharmaceutically acceptable salt thereof may be injected intravenously (iv), intraperitoneally (ip), intraarterially (ia), intramuscularly (im), and / or subcutaneously (sc). Alternatively, the compound, compound-radiometal complex, or pharmaceutically acceptable salt thereof may be taken orally, for example, as a powder, tablet, pill, capsule, chewable capsule, syrup, juice, gel, liquid, or paste. Alternatively, the compound, compound-radiometal complex, or pharmaceutically acceptable salt thereof may be taken nasally (intranasally) (e.g., as a spray or aerosol), transdermally (e.g., as a cream, spray, or ointment, and / or via a coated plaster), and / or respiratoryly (e.g., by inhaling an aerosol or spray). It will be understood that the above-described compounds, compound-radiometal complexes, or pharmaceutically acceptable salts thereof may be administered locally or systemically.

[0177] A sufficient amount of the compound, compound-radiometal complex, or pharmaceutically acceptable salt thereof suitable for treatment can be determined by the physicochemical and pharmacological properties (e.g., bioavailability, charge, lipophilicity, molecular weight, etc.) of the compound or pharmaceutically acceptable salt thereof, the route of administration (e.g., including or excluding the first-pass effect), the patient's weight, the patient's metabolism (e.g., the rate of metabolism and excretion of the compound, compound-radiometal complex, or pharmaceutically acceptable salt thereof), and the accuracy of the analytical device utilized (i.e., a more sensitive analytical device may typically require a lower amount of the compound, compound-radiometal complex, or pharmaceutically acceptable salt thereof).

[0178] The neoplasm detectable by the compositions of the present invention can be any neoplasm known in the art.

[0179] In a preferred embodiment, the neoplasm is cancer, particularly prostate cancer.

[0180] As used herein, prostate cancer can be understood in the broadest sense as any form of cancer that begins in the prostate gland (i.e., a gland in the male reproductive system). Preferably, the prostate cancer is prostate carcinoma.

[0181] Patients at risk for developing prostate cancer may optionally be treated with: significant family history (e.g., having a first-degree relative (father or brother) with prostate cancer); genetic background of risk (e.g., one or more mutations in the BRCA1 and / or BRCA2 genes, one or more mutations in hereditary prostate cancer gene 1 (HPC1), one or more mutations in the androgen receptor, one or more mutations in the vitamin D receptor, TMPRSS2-ETS gene family fusions (particularly TMPRSS2-ERG or TMPRSS2-ETV1 / 49), defects in one or more tumor suppressor genes (e.g., defects in the p53 gene, PTEN, KAI1, E-cadherin, and / or CD44); low blood levels of vitamin D; High blood levels of testosterone; and / or Development of infection or inflammation of the prostate gland (prostatitis) (e.g., infection with chlamydia, gonorrhea, xenotropic MuLV-related virus (XMRV), HPV-16, HPV-18, HSV-2, and / or syphilis) The patient may have one or more risk factors selected from the group consisting of:

[0182] As already mentioned above, the diagnosis of a patient may involve the readout of a radioactive signal arising from a complexed radiometal and / or a fluorescent signal arising from a fluorescent dye, both of which may allow the detection of the localization and / or size of a neoplasm.

[0183] In a preferred embodiment, the method comprises at least the following steps: (i) administering said composition to a patient; (ii) detecting the radioactive signal of the radioactive metal and preferably step (ii) is carried out by three-dimensional imaging, including in particular positron emission tomography (PET).

[0184] The term "three-dimensional imaging" as used throughout the context of the present invention can be understood in the broadest sense as any method that allows determining the localization of a compound-radioactive metal complex in a three-dimensional object. Such methods can be, for example, positron emission tomography (PET), magnetic resonance imaging (MRI), X-ray imaging (especially computed tomography), single-photon emission computed tomography (SPECT), or a combination of two or more of these.

[0185] Most preferably, the method involves positron emission tomography (PET).

[0186] PET can be understood in the broadest sense as a method based on the detection of counter-emitted gamma ray / photon pairs, where the emission of said gamma ray / photon is caused by an annihilation event induced by the liaison between electrons present in the patient's body or sample and protons released during the decay of radioactive metals. A three-dimensional image of the tracer concentration in the body or sample can then be constructed by computer analysis. PET is a nuclear medicine imaging method that allows the generation of three-dimensional image reconstructions, thus enabling visualization of the shape, size, and location of neoplasms.

[0187] Those skilled in the art will understand how to make such measurements and that PET can be very well combined with other imaging techniques, such as three-dimensional fluorescence detection (e.g., by fluorescence molecular tomography (FMT)), CT, and / or MRI. Such combined detection may be performed simultaneously or sequentially, and may be performed on the same device or on different devices.

[0188] The combination of two or more imaging methods may allow for the overlay of data obtained from such methods, thereby allowing for high resolution images (e.g., images obtained from MRI and / or CT) to be combined with methods that allow for the depiction of the concentration of the compound-radiometal complex of the present invention in specific regions. Such results may allow for a particularly accurate determination of neoplastic tissue in a patient.

[0189] Prior to administration to a patient, the compositions of the present invention can typically be produced by mixing the compound-radiometal complex or a pharmaceutically acceptable salt thereof with the radiometal (to form the complex) and optionally a pharmaceutically acceptable carrier. Thus, the method can optionally further comprise the preceding step of mixing the composition according to the present invention.

[0190] As noted above, an advantage of the compounds, compound-radiometal complexes, or pharmaceutically acceptable salts thereof of the present invention is their ability to diagnose neoplasms in vivo and in vitro by detecting two distinct signals: (i) radiation produced by the radiometal, and (ii) fluorescence produced by the dye moiety.

[0191] This method allows for the precise localization of the neoplasm in the entire patient's body, or at least in a large portion thereof, to be determined in one step by detecting a radioactive signal, particularly using PET imaging. In a further step, fluorescence can be detected. This method can be performed in the entire patient's body, or at least in a large portion thereof, as well as when the patient's body is incised (i.e., when the tissue of interest is cut open during surgery).

[0192] Thus, in a further preferred embodiment, the method further comprises: (iii) detecting the dye moiety (C), preferably comprising molecular imaging.

[0193] However, it will be appreciated that alternatively, one detection method or the other may be used in some cases (i.e., one may detect only the radioactive signal from the radiometal, or only the fluorescent signal from the dye moiety). Detection of fluorescent signal only allows the radiometal to be omitted, which may simplify operation and reduce costs, but may typically reduce detection flexibility and data accuracy.

[0194] Throughout this invention, the term "molecular imaging" in the context of detecting dye moieties is understood in its broadest sense as any method that allows for the location of dye moieties and thereby the location of compounds according to the invention (or their radiometal complexes) in an object of interest. Molecular imaging in this context may also be called "fluorescence molecular imaging" and abbreviated as "FMI." As used in the context of detecting dye moieties, detection may be performed visually or in an instrument-assisted manner.

[0195] Preferably, molecular imaging refers to detecting fluorescence in a patient's body or cell culture at a resolution that allows detecting the localization of a compound in the object of interest (i.e., the closest object that can still be distinguished from another object).Therefore, in a patient's body, the closest object that can still be distinguished from another object by molecular imaging can preferably be less than 1 cm, more preferably less than 5 mm, particularly less than 2 mm.In a cell culture, the closest object that can still be distinguished from another object by molecular imaging can preferably be less than 2 mm, more preferably less than 1 mm, particularly less than 0.5 mm, or even within the microscopic range (i.e., less than 0.1 mm).When carried out in a patient's body, molecular imaging can be, for example, fluorescence molecular tomography (FMT), optical imaging, or two-photon fluorescence detection.When carried out in a cell culture, molecular imaging can be, for example, fluorescence microscopy, confocal microscopy (e.g., laser scanning microscopy (LSM)), two-photon fluorescence microscopy, fluorescence energy transfer (FRET)-based methods, fluorescence correlation spectroscopy (FCS), or fluorescence cross-correlation spectroscopy (FCCS). The above imaging can optionally be further combined with other imaging methods, such as, for example, positron emission tomography (PET), magnetic resonance imaging (MRI), radiography (e.g., computed tomography), or ultrasound tomography (UT).

[0196] Step (iii) of detecting the dye moiety (C) (preferably involving molecular imaging) may be performed after, simultaneously with, or before step (ii) of detecting the radioactive signal of the radiometal.

[0197] In a preferred embodiment, step (iii) is carried out after step (ii).

[0198] Prior to detecting the dye moiety, one skilled in the art may administer the compound or compound-radiometal complex or a pharmaceutically acceptable salt thereof of the present invention to the patient for an additional time, or may use a fluorescent signal obtained from the compound, compound-radiometal complex or a salt thereof administered once prior to detection of a preceding radioactive signal, particularly in a PET scan.

[0199] Either strategy may have certain advantages. Administering the compound, compound-radiometal complex, or salt thereof for an additional time before detecting the fluorescent signal allows optimizing the concentration range to obtain a qualitatively good fluorescent signal. Furthermore, local administration to the incised neoplastic tissue and its surrounding tissue may also be possible. Such additional administration may preferably be the administration of the compound, compound-radiometal complex, or salt thereof described above that does not contain a radioactive metal.

[0200] On the other hand, administering the compound, compound-radiometal complex, or salt thereof before detecting only the radioactive signal can prevent the patient from receiving frequent treatment, improving patient compliance, especially when it is administered only once. Furthermore, a lower dose of the compound, compound-radiometal complex, or salt thereof can prevent potentially undesirable side effects. In this case, it may be advantageous for the half-life of the radiometal not to be too long, because if a surgical interaction occurs after the previous molecular imaging by detecting the radioactive signal, the radioactivity of the radiometal may weaken to a low and therefore harmless level when the surgical interaction actually occurs.

[0201] A person skilled in the art of removing neoplastic tissue, typically a surgeon, can optionally emit light that excites the dye moiety during surgery, thereby visualizing the neoplastic tissue to be removed. This can be done at intervals or continuously. The excitation light can come from a standard lamp, a surgical lamp, and / or a headlamp worn by the surgeon and / or any other participant(s). The wavelength of the light can be such that it efficiently excites the dye moiety (particularly a wavelength near its excitation maximum), or it can be twice the wavelength that efficiently excites the dye moiety (for two-photon excitation), particularly twice the wavelength near its excitation maximum. In a highly preferred embodiment, step (iii) is performed during surgery, when the cancerous tissue is at least partially dissected open.

[0202] As previously mentioned, the method for diagnosing a patient in vivo is based on the use of a composition according to the invention, optionally further comprising a pharmaceutically acceptable carrier, and thus on the use of each radiometal-complexed compound, compound-radiometal complex or pharmaceutically acceptable salt thereof, which represents their first and also particular pharmaceutical use.

[0203] Thus, in a further aspect, the present invention relates to a composition according to the invention as specified above for use as a medicament, in particular for use as a diagnostic agent.

[0204] In a second further aspect, the present invention relates to a composition according to the invention as specified above for use in a method for diagnosing a neoplasm in a patient suffering from or at risk of having a neoplasm.

[0205] In the context of the composition for use, the definitions of the terms specified in the context of the compound, compound-radiometal complex or pharmaceutically acceptable salt thereof and the diagnostic method above also apply.

[0206] As noted above, in a preferred embodiment, the neoplasm is cancer, particularly prostate cancer.

[0207] In a preferred embodiment, the method comprises the following steps: (i) administering said composition to a patient; (ii) Radioactive metals, especially 68 detecting a radioactive signal of Ga; Preferably, step (ii) is carried out by three-dimensional imaging, in particular using positron emission tomography (PET).

[0208] As noted above, in a preferred embodiment, the method comprises: (iii) detecting the dye moiety (C), preferably using molecular imaging; In particular, step (iii) is carried out after step (ii).

[0209] As mentioned above, the compound or compound-radiometal complex or pharmaceutically acceptable salt thereof according to the present invention can also be used to detect neoplasms in vitro (e.g., in cell culture or in cells obtained from a patient). In this context, the user of the compound, compound-radiometal complex or pharmaceutically acceptable salt thereof according to the present invention can also be provided with the compound or salt thereof in the form of a kit.

[0210] Thus, further aspects of the present invention include: (a) a compound according to the invention as specified above or a pharmaceutically acceptable salt thereof, or a composition according to the invention as specified above; and (b User Manual The present invention relates to a kit comprising:

[0211] In the context of the kit, the definitions of terms specified in the context of the compound or pharmaceutically acceptable salt thereof, the diagnostic methods and compositions for use listed above also apply.

[0212] In the context of the present invention, the term "kit" can be understood in the broadest sense as a composition of various products that can be used to perform the detection of neoplasms. The kit may include, but is not limited to, the compound or composition according to the present invention, or a pharmaceutically acceptable salt thereof, one or more pharmaceutically acceptable carriers, a user manual, a syringe, needles, etc. Optionally, the compound or pharmaceutically acceptable salt thereof, or the composition according to the present invention, may be dissolved, dried, or lyophilized. Preferably, the kit of the present invention includes a lyophilized, labeled form of the compound or composition according to the present invention, or a pharmaceutically acceptable salt thereof, a buffer for dissolving it, and an injection or infusion device for mixing the compound or pharmaceutically acceptable salt thereof with a buffer and administering the mixture to a patient.

[0213] The user manual may include instructions on how to store the compound or composition according to the invention or a pharmaceutically acceptable salt thereof (e.g., temperature, humidity, storage period, etc.), which radiometals to use and, if applicable, where to obtain them from, how to complex the compound according to the invention or a pharmaceutically acceptable salt thereof with each radiometal (e.g., buffer conditions), and / or how to use the compound or composition according to the invention or a pharmaceutically acceptable salt thereof to detect neoplasms in vivo and / or in vitro (e.g., recommended amounts / concentrations, recommended detection techniques, etc.).

[0214] The kits may also include one or more radiometal(s) suitable for complexing with the compounds of the invention, provided that the radioactive half-life of the radiometal is sufficiently long.

[0215] The kit may also be used for the in vitro detection and surveillance of neoplasms.

[0216] Thus, a still further aspect of the present invention is a method for detecting neoplastic cells in a sample in vitro, comprising the steps of: (i) providing a cell that is neoplastic or at risk of being neoplastic, in particular a cell that is cancerous or at risk of being cancerous; (ii) administering to said cells a compound according to the invention as specified above or a pharmaceutically acceptable salt thereof, or a composition according to the invention as specified above; (iii) detecting a fluorescent signal and / or a radioactive signal from the cells. The present invention relates to the above method, which comprises:

[0217] In the context of the in vitro methods of the present invention, the definitions of terms specified in the context of the above compounds or pharmaceutically acceptable salts thereof, the diagnostic methods, compositions for use and kits listed above also apply.

[0218] Cells that are neoplastic or at risk of being neoplastic may be isolated (i.e., singularized cells), or the cells may still be present in their physiological context (i.e., in their tissue, which may also contain different cell types). Cells may be obtained as tissue samples from patients suffering from or at risk of developing a neoplasm, or may be obtained from tissue culture. In vitro samples used in the context of the present invention may be part of or whole cadavers of patients or experimental animals (e.g., mice, rats, rabbits, etc.).

[0219] When cultured, the cells can be cultured under suitable conditions, i.e., typically at about 37°C, at about pH 6.5 to 7.5, particularly pH 7.0 to 7.5, with suitable nutrients, vitamins, minerals, and optionally growth factors. Those skilled in the art can select suitable standard cell culture techniques suitable for the cells of interest.

[0220] Optionally, the cells may be viable throughout the entire in vitro method, or may be fixed (e.g., in ethanol, acetone, or another fixative) prior to step (ii) of administering to the cells a compound according to the invention or a pharmaceutically acceptable salt thereof or a composition according to the invention, or prior to step (iii) of detecting the fluorescent and / or radioactive signal of said cells.

[0221] The cells can optionally be counterstained with other dyes (eg, DAPI dye or HOECHT dye to stain the nucleus, and / or labeled antibodies to stain another structure of interest).

[0222] In the in vitro method of the present invention, the compound or composition according to the present invention, or a pharmaceutically acceptable salt thereof, can be administered to cells by preparing a buffer containing the compound, composition, or salt thereof and adding it to cells. Alternatively, the compound or pharmaceutically acceptable salt thereof according to the present invention, or a composition thereof, can be administered to a patient in vivo, and then cells and / or tissue samples can be removed from the patient and examined in vitro.

[0223] The in vitro method of the present invention particularly allows for the sequential and simultaneous detection of the radioactive signal from the radiometal and the fluorescent signal from the dye moiety. However, it will be noted that alternatively, one detection method or the other may be used in some cases (i.e., only the radioactive signal from the radiometal or only the fluorescent signal from the dye moiety may be detected). Detection of the fluorescent signal only allows the radiometal to be omitted, which can simplify the operation and reduce costs, but typically can reduce the flexibility of detection and the accuracy of the data.

[0224] Depending on the detection method used, the cells may optionally be washed with fresh buffer or cell culture medium to remove excess unbound compound or compound-radiometal complex before carrying out step (iii).

[0225] According to the present invention, neoplastic cells will typically exhibit a higher staining rate than corresponding non-neoplastic cells of the same cell type.

[0226] The cells are preferably obtained from a patient, in particular a human.

[0227] In a preferred embodiment, the cells are obtained from a patient suffering from or at risk of a neoplasia, preferably cancer, in particular prostate cancer.

[0228] Detection can be performed by any means known in the art.

[0229] In a preferred embodiment, step (iii) comprises detecting fluorescence via microscopic imaging, in particular confocal laser scanning microscopy (LSM) or two-photon microscopy.

[0230] Herein, confocal laser scanning microscopy (LSM) or two-photon microscopy may enable the detection of the compound according to the present invention or the compound-radioactive metal complex or its pharmaceutically acceptable salt at the cell surface of a cell at a microscopic level. Thus, a relatively small local increase at a specific membrane site may be detectable.

[0231] In another preferred embodiment, step (iii) comprises detecting fluorescence by flow cytometry and / or fluorescence activated cell sorting (FACS).

[0232] Detection by flow cytometry may allow for quantification of the fraction of neoplastic cells relative to the fraction of non-neoplastic cells in a sample. Fluorescence-activated cell sorting (FACS) may further allow for isolation of a cell population of interest, such as, for example, a fraction of neoplastic cells, a fraction of a particular cell type (optionally identified by counterstaining with antibodies representative of membrane proteins of said cells), or a fraction of neoplastic cells of a particular cell type.

[0233] In a preferred embodiment, step (iii) comprises detecting radioactivity by gamma counting.

[0234] The term "gamma counting" as used in the context of the present invention can be understood in the broadest sense as any method based on the quantification of gamma irradiation. Those skilled in the art are aware of several methods based on gamma counting and how to carry them out. For example, gamma counting can be used in the context of radioactive binding assays and / or radioimmunoassays (RIA). The readout can be performed by direct gamma counting or indirect gamma counting, such as scintillation counting, particularly liquid scintillation counting. Gamma counting can make it possible to quantify the fraction of a specific cell type. Furthermore, when combined with a previous step of isolating specific cell fractions, for example, using FACS (e.g., as described above), gamma counting can also provide information about the fraction of a specific cell population of interest.

[0235] As mentioned above, the detection of the fluorescence of the cells and the detection of the radioactive signal (particularly gamma irradiation) can be combined with each other, or alternatively, only the fluorescence or only the radioactive signal can be detected.

[0236] In either case, when analyzing the data, cells exhibiting different intensities of fluorescent and / or radioactive signals can be grouped into different fractions. This is typically done by setting a specific threshold. As described above, according to the present invention, neoplastic cells typically exhibit a higher staining rate, and therefore a higher signal intensity, compared to corresponding non-neoplastic cells of the same cell type.

[0237] Thus, in a preferred embodiment, the method further comprises the steps of: (iv) Below: (a) cell counts exceeding fluorescent and / or radioactive signals indicative of neoplastic cells (especially cancerous cells); and (b) Cell counts below the fluorescent and / or radioactive signal, indicating non-neoplastic cells determining the (v) determining the ratio of (a):(b) to assess the severity of the neoplasm in the patient from whom the cells were obtained; Includes.

[0238] The number of cells that exceeds the fluorescent and / or radioactive signal (and thus exceeds a given threshold) indicative of neoplastic cells exhibits a higher signal intensity compared to corresponding non-neoplastic cells of the same cell type. Those skilled in the art will recognize that the signal intensity representing the threshold is determined by the cells being investigated and the detection method. The signal intensity representing this threshold can be calculated as follows: (a) a signal obtained from cells known to be neoplastic; and (b) Signals obtained from known healthy (i.e., non-neoplastic) counterpart cells. can be determined by measuring

[0239] In this specification, (a) typically provides a higher measurement intensity than (b), and a threshold value may be set between the two measurement intensities of (a) and (b).

[0240] This may allow for the determination of the severity of the neoplasm in the patient. A high percentage of cells determined to be neoplastic may indicate a severe neoplasm. Thus, a relatively high percentage of cells in the examined tissue are neoplastic, and thus the neoplasm is relatively widespread. In contrast, a low percentage may indicate a less severe neoplasm in the examined tissue sample, and the absence of neoplastic cells may indicate the absence of a neoplasm in the examined tissue sample.

[0241] Therefore, the compound or compound-radiometal complex according to the present invention or a pharmaceutically acceptable salt thereof can also be used to assess the severity of neoplasms in a sample in vitro.

[0242] Therefore, in a still further aspect, the present invention relates to the use of a compound according to the invention as specified above or a pharmaceutically acceptable salt thereof or a composition according to the invention as specified above, for assessing the severity of a neoplasm in a sample in vitro, wherein said sample contains cells which are neoplastic or at risk of being neoplastic, in particular cells which are cancerous or at risk of being cancerous, and wherein these cells are contacted with said compound or a pharmaceutically acceptable salt thereof or said composition.

[0243] In the context of this usage, the definitions of terms specified throughout the present invention above also apply.

[0244] In a preferred embodiment, the assessment of the severity of the neoplasm is based on the following: (a) a cell count exceeding the fluorescent and / or radioactive signal indicative of neoplastic cells (especially cancerous cells); (b) A cell count below the fluorescent and / or radioactive signal indicative of non-cancerous cells This includes determining the ratio of

[0245] The following examples are intended to illustrate the invention but not to limit the scope of protection conferred by the claims. [Example]

[0246] Abbreviation PSMA-(HE)3-HBED-CC-IRdye800CW (Alternative name: Glu-Urea-Lys-(HE)3-HBED-CC-PEG2-IRDye800CW)

[0247] [ka]

[0248] PSMA-(HE)1-HBED-CC-IRDye800CW (also known as: Glu-urea-Lys-(HE)1-HBED-CC-PEG2-IRDye800CW) (comparative example)

[0249] [ka]

[0250] PSMA-(WE)1-HBED-CC-IRDye800CW (also known as: Glu-urea-Lys-(WE)1-HBED-CC-PEG2-IRDye800CW) (comparative example)

[0251] [ka]

[0252] PSMA-HBED-CC-(HE)3-IRDye800CW (also known as: Glu-urea-Lys-HBED-CC-(HE)3-PEG2-IRDye800CW) (comparative example)

[0253] [ka]

[0254] PSMA-HBED-CC-IRDye800CW (also known as: Glu-urea-Lys-HBED-CC-PEG2-IRDye800CW) (comparative example)

[0255] [ka]

[0256] PSMA-HBED-CC (also known as PSMA-11, Glu-urea-Lys-(Ahx)-HBED-CC) (comparative example)

[0257] [ka]

[0258] Experimental procedure All commercial chemicals were of analytical grade and were used without further purification. 68Ga (half-life 68 min; β + 89%;E β+ (up to 1.9 MeV) is based on a pyrogallol resin support 68 Ge / 68 The data were obtained from a Ga generator (1). Compounds were analyzed using reverse-phase high-performance liquid chromatography (RP-HPLC; Chromolith RP-18e, 100 × 4.6 mm; Merck, Darmstadt, Germany). Analytical HPLC runs were performed using a linear gradient (0.1% aqueous TFA (A) to 100% B (0.1% TFA in CHCN)) at 2 mL / min within 10 min. The system L6200 A (Merck-Hitachi, Darmstadt, Germany) was equipped with a variable UV detector and a gamma detector (Bioscan; Washington, USA).

[0259] For preparative HPLC, a LaPrep P110 system (VWR, Darmstadt, Germany) was equipped with a variable UV detector (P314, VWR, Darmstadt, Germany). Analytical HPLC runs were performed using an Agilent 1100 series system (Agilent Technologies, Santa Clara, CA, USA). UV absorption was measured at 214 nm and 254 nm, respectively. For mass spectrometry, a MALDI-MS (Daltonics Microflex, Bruker Daltonics, Bremen, Germany) was used.

[0260] Glu-urea-Lys-(Ahx)-HBED-CC (PSMA-11) was purchased from ABX (Radeberg, Germany). PSMA-HBED-PEG2-IRDye800CW was synthesized according to the U.S. Patent Publication No. 20150110715 entitled "Dual-Labeled Probes for Molecular Imaging and Their Use."

[0261] Synthesis of Glu-urea-Lys-(HE)3-HBED-CC-PEG2-IRDye800CW Synthesis of Glu-urea-Lys-(HE)3-CO(CH2)4-N3 The synthesis of the pharmacophore Glu-urea-Lys was carried out as previously described (2). Briefly, the synthesis was initiated by the formation of the isocyanate of the glutamyl moiety using triphosgene. Resin-immobilized (2-chloro-trityl resin, Merck, Darmstadt) ε-allyloxycarbonyl-protected lysine was added and allowed to react for 16 h with gentle stirring. The resin was filtered off, and the allyloxy protecting group was removed by reacting twice with Pd(PPh3)4 (0.3 equiv.) and morpholine (15 equiv.) under ambient conditions (1 h, room temperature). The resin was then separated, and the (HE)3 linker was synthesized using a standard Fmoc solid-phase protocol. To obtain the (HE)3-containing molecule, coupling of Fmoc-His(Trt)-OH, Fmoc-Glu(otBu)-OH, and 5-azidopentanoic acid (4 equiv.) was carried out using HBTU (4 equiv.) and DIPEA (4 equiv.) in DMF. The coupling of Fmoc-His(Trt)-OH and Fmoc-Glu(otBu)-OH was repeated to form (HE)3.

[0262] The product was cleaved from the resin using TFA / TIPS / HO (95 / 2.5 / 2.5, v / v / v) at room temperature for 3 h to yield the azide-functionalized intermediate. All products were purified using RP-HPLC and identified by mass spectrometry. Purification of Glu-Urea-Lys-(HE)3-CO-(CH2)4-N3 was performed using a NUCLEODUR® Sphinx RP column (VP250 / 21, 5 μm 250 x 21 mm; Macherey-Nagel, Düren, Germany) with a 20-min gradient starting at 10% B and increasing to 100% B within 20 min. Solvent A consisted of 0.1% aqueous TFA, and solvent B was 0.1% TFA in CH3CN. The flow rate was 20 mL / min.

[0263] Synthesis of Glu-urea-Lys-(HE)3-HBED-CC-PEG2 HBED-CC(TFP)2 was prepared using Fe as previously described. 3+ Protection with [Fe(HBED-CC)] -(3) The bis-TFP ester was isolated by preparative HPLC using a NUCLEODUR® Sphinx RP column with a 20 min gradient starting at 10% B and increasing to 100% B within 20 min. Solvent A consisted of 0.1% aqueous TFA, and solvent B was 0.1% TFA in CH3CN. The flow rate was 20 mL / min. The product was identified by mass spectrometry (MW: 828.7).

[0264] [Fe(HBED-CC)]TFP2 and 0.95 equivalents of propargylamine were dissolved in DMF in the presence of DIPEA. After 4 hours at room temperature, excess 2,2'-(ethylenedioxy)bis(ethylamine) (100 μl) was added and stirred at room temperature for 16 hours. The alkenyl-functionalized chelator was purified via preparative HPLC using a NUCLEODUR® Sphinx RP column with a 20-minute gradient starting at 10% B and increasing to 100% B. Solvent A consisted of 0.1% aqueous TFA, and solvent B was 0.1% TFA in CH3CN. The flow rate was 20 mL / min (MW: 699.8).

[0265] PEG-[Fe(HBED-CC)]-propargylamine (1 equiv.) was then reacted with Glu-urea-Lys-(HE)-CO(CH)-N (1 equiv.) via CuAAC, CuSO (1 equiv.), and Na ascorbate (1 equiv.) in 3 mL THF / HO (1:1, v / v) at room temperature for 16 h. The Fe-protected product was isolated via preparative HPLC using a NUCLEODUR® Sphinx RP column (0–100% B within 20 min, flow rate 20 ml / min) and identified by mass spectrometry (MW: 1943.1).

[0266] Glu-Urea-Lys-(HE)3-HBED-CC-PEG2-IRDye800CW IRDye800CW-NHS ester (1 equiv.) (LI-COR Biosciences) was conjugated to Glu-urea-Lys-(HE)3-[Fe(HBED-CC)]-PEG2 in PBS buffer (pH 8.5) at room temperature for 24 h. The Fe-protected product was isolated via semi-preparative HPLC using a Chromolith RP-18e column (100 × 10 mm; Merck, Darmstadt, Germany) (0–100% B within 10 min, flow rate 5 ml / min) and identified by mass spectrometry (MW: 2929.2).

[0267] Complexed Fe 3+ was removed as previously described (3). Briefly, the Fe-containing product was captured on a C18 cartridge (Waters SepPak-Classic C18; Waters Corp., Milford, MA, USA), which was then flushed with 10 mL of 1 M HCl and washed with 5 mL of HO. The remaining product was eluted with 2 mL of HO / CH3CN (3:1) and evaporated to dryness.

[0268] 68 Ga labeling Precursor peptide [1 nmol (580 mg / ml) in HEPES buffer, 90 μL] was added to 40 μL [ 68 Ga]Ga 3+ The eluate (approximately 40 mBq) was added. The pH was adjusted to 3.8 using 30% NaOH and 10% NaOH, respectively. The reaction mixture was incubated at 98°C for 10 minutes. The radiochemical yield (RCY) was determined by HPLC.

[0269] cell culture PSMA + LNCaP cells (ATCC CRL-1740) were cultured in RPMI medium supplemented with 10% fetal bovine serum and 2 mmol / L L-glutamine (all purchased from PAA). Cells were grown at 37°C in a humidified atmosphere containing 5% CO2 and harvested using trypsin-ethylenediaminetetraacetic acid (trypsin-EDTA; 0.25% trypsin, 0.02% EDTA, Invitrogen).

[0270] Cell Binding and Uptake Competitive cell binding assays and uptake experiments were performed as previously described. Briefly, cells (10 per well) were incubated in the presence of 12 different concentrations of analyte (0–5000 nM, 100 μL / well). 5 )of, 68 Cells were incubated with a 0.2 nM solution of Ga-labeled radioligand [Glu-urea-Lys(Ahx)]-HBED-CC (PSMA-10, precursor ordered from ABX, Radeberg, Germany). After incubation, the mixture was removed, and the wells were washed three times with PBS using a multiscreen vacuum manifold (Millipore, Billerica, MA). Cell-bound radioactivity was measured using a gamma counter (Packard Cobra II, GMI, MN, USA). Median inhibitory concentration (IC50) values ​​were calculated by fitting the data using a nonlinear regression algorithm (GraphPad Software).

[0271] For uptake experiments, 10 cells per well were added 24 hours before incubation. 5 Cells were seeded in 24-well cell culture plates coated with poly-L-lysine. After washing, the cells were incubated with 30 nM of radiolabeled compound for 45 min at 37°C and 4°C, respectively. Cell uptake was terminated by washing three times with 1 mL of ice-cold PBS. To remove surface-bound radioactivity, the cells were incubated twice for 5 min with 0.5 mL of glycine-HCl (50 mM, pH = 2.8) in PBS. The cells were washed with 1 mL of ice-cold PBS and lysed with 0.3 N NaOH (0.5 mL). The surface-bound and internalized fractions were measured in a gamma counter. Cell uptake was measured at 10 5 The percentage of initially added radioactivity bound to cells [%ID / 10 5 cells].

[0272] Distribution in the body For experimental tumor models, 5 × 10 6 LNCaP cells (in 50% Matrigel; Becton Dickinson, Heidelberg, Germany) were inoculated subcutaneously into the right flank of 7-8 week-old male BALB / c nu / nu mice (Charles River). Tumors were grown to approximately 1 cm in size. 3 It grew to. 68 Ga-labeled compounds were injected into the tail vein (1–2 MBq; 60 pmol). One hour after injection, the animals were sacrificed. The organs of interest were dissected, blotted dry, and weighed. Radioactivity was measured using a gamma counter and calculated as % ID / g. All animal experiments were conducted in accordance with the current legislation of the Federal Republic of Germany.

[0273] Statistical Aspects All experiments were performed at least in triplicate and repeated at least three times. Quantitative data were expressed as mean ± SD. Where applicable, means were compared using Student's t-test. A P value of <0.05 was considered statistically significant.

[0274] result To investigate the effect of the linker on binding properties, the uptake efficiency and PSMA binding affinity of the above conjugates were determined. The results are summarized in Table 1. The binding affinities (K) determined for LNCaP cells were: i ) decreased slightly from 9.82 ± 1.26 nM for PSMA-HBED-CC (PSMA-11) to 17.53 ± 4.98 nM for PSMA-HBED-CC-PEG2-IRDye800CW and 36.70 ± 9.77 nM for PSMA-(HE)3-HBED-CC-PEG2-IRDye800CW.

[0275] [Table 2]

[0276] Specific cell surface binding and specific uptake of PSMA-HBED-CC-PEG2-IRDye800CW and PSMA-(HE)3-HBED-CC-PEG2-IRDye800CW were improved compared to the reference structure PSMA-11.

[0277] The higher PSMA-specific uptake of the conjugates observed in these assays resulted in significantly higher tumor accumulation (13.66±3.73%ID / g (PSMA-HBED-CC-PEG2-IRDye800CW) and 7.59±0.95%ID / g (PSMA-(HE)3-HBED-CC-PEG2-IRDye800CW) compared to the reference (4.89±1.34%ID / g (PSMA-11); P<0.05) as shown by organ distribution in LNCaP-tumor-bearing nude mice (Table 2). PSMA-(HE)3-HBED-CC-PEG2-IRDye800CW showed lower uptake in all background organs compared to PSMA-HBED-CC-PEG2-IRDye800CW. Through (HE)3 delivery, spleen uptake significantly decreased from 38.12 ± 14.62%ID / g (PSMA-HBED-CC-PEG2-IRDye800CW) to 3.47 ± 1.39%ID / g (PSMA-(HE)3-HBED-CC-PEG2-IRDye800CW).

[0278] [Table 3]

[0279] [Table 4]

[0280] To assess the impact of (HE)3 linker introduction on uptake in background organs, tumor-to-organ ratios were calculated and summarized in Table 3. Compared with PSMA-HBED-CC-PEG2-IRDye800CW, the compound bearing the linker, PSMA-(HE)3-HBED-CC-PEG2-IRDye800CW, showed higher T / O ratios in all organs except the kidney, clearly indicating an improved pharmacokinetic profile.

[0281] Notably, the placement of (HE)3 as a linker between the binding motif and the chelator resulted in a clear reduction of the spleen signal.

[0282] To compare the impact of introducing the (HE)3 linker at various positions relative to other amino acid linkers, their uptake in background organs was determined and tumor-to-organ ratios were calculated. The data are summarized in Table 4.

[0283] [Table 5]

[0284] PSMA-(HE)3-HBED-CC-PEG2-IRDye800CW (7.59 ± 0.95%ID / g) showed significantly higher tumor uptake than compounds with other linkers (PSMA-HBED-CC-(HE)3-PEG2-IRDye800CW: 3.32 ± 1.51%ID / g, PSMA-(HE)1-HBED-CC-PEG2-IRDye800CW: 3.92 ± 0.31%ID / g, PSMA-(WE)1-HBED-CC-PEG2-IRDye800CW: 3.85 ± 1.10%ID / g). Spleen uptake of PSMA-(HE)3-HBED-CC-PEG2-IRDye800CW was significantly reduced compared to all other compounds shown in Table 4.

[0285] Tumor-to-organ ratios for all background organs (Table 5) were highest for PSMA-(HE)3-HBED-CC-PEG2-IRDye800CW (except for T / heart: 5.57 vs. T / heart: 5.80 for PSMA-HBED-CC-(HE)3-PEG2-IRDye800CW).

[0286] [Table 6]

[0287] To further analyze the pharmacokinetic effects of the (HE)3 linker, 68 The Ga-labeled compound PSMA-(HE)3-HBED-CC-PEG2-IRDye800CW was 68 Organ distribution studies were performed at 2 hours post-injection using Ga-labeled PSMA-11 compared to PSMA-HBED-CC-PEG2-IRDye800CW (Table 6).

[0288] [Table 7]

[0289] The introduction of (HE)3 between the PSMA-binding motif and the chelator resulted in rapid clearance from background organs, particularly reduced kidney and spleen uptake compared to other compounds.

[0290] References (1)Schuhmacher, J., and Maier-Borst, W. (1981) A new Ge-68 / Ga-68 radioisotope generator system for production of Ga-68 in dilute HCl. Int J Appl Radiat Isot 32, 31-36. (2)Schafer, M., Bauder-Wust, U., Leotta, K., Zoller, F., Mier, W., Haberkorn, U., Eisenhut, M., and Eder, M. (2012) A dimerized urea-based inhibitor of the prostate-specific membrane antigen for 68Ga-PET imaging of prostate cancer. EJNMMI Res 2, 23. (3)Eder, M., Wangler, B., Knackmuss, S., Legall, F., Little, M., Haberkorn, U., Mier, W., and Eisenhut, M. (2008) Tetrafluorophenolate of HBED-CC: a versatile conjugation agent for (68)Ga-labeled small recombinant antibodies. Eur J Nucl Med Mol Imaging 35, 1878-86. (4)Eder, M., Schafer, M., Bauder-Wust, U., Hull, W. E., Wangler, C., Mier, W., Haberkorn, U., and Eisenhut, M. (2012) (68)Ga-Complex Lipophilicity and the Targeting Property of a Urea-Based PSMA Inhibitor for PET Imaging. Bioconjug Chem 23, 688-97. The following is one embodiment of the present invention. (1) Formula (I): (A)-x 1 -(B)-x 2 -(C) (In the formula, (A) is at least one motif that specifically binds to the plasma membrane of neoplastic cells; (B) is at least one radiometal chelator moiety; (C) is a dye moiety; x 1 is a spacer that covalently bonds (A) and (B); x 2 is a spacer or a single chemical bond connecting (B) and (C); (C) is a compound of the formula:

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Claims

1. Formula (I): (A)-x 1 -(B)-x 2 -(C) (In the formula, (A) is at least one motif that specifically binds to the plasma membrane of a neoplastic cell; (B) is at least one radiometal chelator moiety; (C) is a dye moiety; x 1 is a spacer that covalently bonds (A) and (B); x 2 is a spacer or a single chemical bond connecting (B) and (C); (C) is a compound of the formula: 【Chemical 1】 (In the formula, X 1 and X 4 is -N=, -N(R 5 )=, and -C(R 6 )=; X 2 and X 3 are O, S, Se, N(R 5 ), and C(R 6 R 7 ) independently selected from the group consisting of: Y is a linker that connects the two moieties of (C) and allows for electron delocalization between said moieties, and Y is optionally a group (L-) c Z 0 Including; a and b are independently selected from the group consisting of 1, 2, and 3; Each R 1 and each R 2 are independently (L-) c Z, (L-) c Z 0 or H, and two adjacent R 1 and / or two adjacent R 2 may contain one or more (L-) c Z or (L-) c Z 0 It is also possible to form an aromatic ring substituted with R 3 , R 4 , R 5 , R 6 , R 7 , R 9 is (L-) c Z, (L-) c Z 0 independently selected from the group consisting of each c is independently 0 or 1; Each L is independently T 1 , -OT 1 -, -ST 1 -, -C(O)T 1 -, -C(O)OT 1 -, -OC(O)T 1 -, -C(O)NHT 1 -,-NHC(O)T 1 , or optionally -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)O-, and T 1 C that is interrupted and / or terminated by one or more of 1-10 an alkylene group; T 1 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, decalinyl, adamantyl, C 3-7 cycloalkyl, 3- to 7-membered heterocyclyl, or 7- to 11-membered heterobicyclyl, where T 1 is sometimes halogen, CN, C(O)R 8 , COOR 8 , OR 8 , C(O)N(R 8 R 8a ), S(O) 2 N(R 8 R 8a ), S(O)N(R 8 R 8a ), S(O) 2 R 8 , N(R 8 )S(O) 2 N(R 8a R 8b ), SR 8 , N(R 8 R 8a ), NO 2 ,OC(O)R 8 , N(R 8 )C(O)R 8a , N(R 8 )S(O) 2 R 8a , N(R 8 )S(O)R 8a , N(R 8 )C(O)N(R 8a R 8b ), N(R 8 )C(O)OR 8a , OC(O)N(R 8 R 8a ), oxo(=O) (wherein the ring is at least partially saturated), or C 1-6 Alkyl (C 1-6 alkyl is optionally substituted with one or more of the same or different halogens; Each Z is independently H, halogen, CN, C(O)R 8 、C(O)OR 8 、C(O)O - 、OR 8 、C(O)N(R 8 R 8a )、S(O) 2 OR 8 、S(O) 2 O - 、S(O) 2 N(R 8 R 8a )、S(O)N(R 8 R 8a )、S(O) 2 R 8 、S(O)R 8 、N(R 8 )S(O) 2 N(R 8a R 8b )、SR 8 、N(R 8 R 8a )、NO 2 、P(O)(OR 8 ) 2 、P(O)(OR 8 )O - 、OC(O)R 8 、N(R 8 )C(O)R 8a 、N(R 8 )S(O) 2 R 8a 、N(R 8 )S(O)R 8a 、N(R 8 )C(O)N(R 8a R 8b )、N(R 8 )C(O)OR 8a 、or OC(O)N(R 8 R 8a ) and; R 8 , R 8a , R 8b is H or C 1-6 Alkyl (C 1-6 alkyl is optionally substituted with one or more of the same or different halogens; Z 0 (C) x 2 or x 2 is a single chemical bond, it is a chemical bond that binds to (B); However, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 9 One of them is (L-) c Z 0 or Y is (L-) c Z 0 (including and Here, (A)-x 1 -has the following structure: 【Chemistry 2】 is represented by Any remaining positive or negative charge(s) are compensated by pharmaceutically acceptable negatively or positively charged counterion(s). or a pharmaceutically acceptable salt thereof.

2. 2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound has a molecular weight of 10 kDa or less, preferably 5 kDa or less.

3. Spacer x 2 but has the following structure: -d 2 -e'-[f-e''] m - (In the formula, d 2 Ha-[CH 2 ] r - in which r is 1 or 2, in particular 2; e' is -C(O)-NH-, -NH-C(O)-, -C(O)-O- and -OC(O)-, -NH-C(O)-NH-, -NH-C(S)-NH-, 【Chemistry 3】 (In the formula, one of the wavy lines is d 2 the other wavy line indicates a conjugation site for f; In particular, e' is -C(O)-NH-; Each f is independently 1-10 - alkylene (wherein one or more -CH 2 - represents a residue selected from the group consisting of - (the moiety is optionally substituted with -O- or -NH-), and f is unsubstituted or -NH 2 , -COOH and R 3a and is substituted with one or more groups independently selected from the group consisting of: R 3a is -(CH 2 ) 2 -COOH, -(CH 2 ) 4 -NH 2 , -(CH 2 ) 4 -N + (CH 3 ) 3 +X - , -CH 2 -COOH, -CH 2 -SH, -CH 2 -SO 3 H, and 【Chemistry 4】 where X is selected from the group consisting of - is a pharmaceutically acceptable negatively charged counterion; Preferably, f is -CH 2 -(O-CH 2 -CH 2 ) 2 -CH 2 -、-(CH 2 ) 2 -(O-CH 2 -CH 2 ) 2 -、-(CH 2 ) 2 -(CH 2 -CH 2 -O) 2 -(CH 2 ) 2 -、-(CH 2 ) 3 -(CH 2 -CH 2 -O) 2 -CH 2 -、-(CH 2 -CH 2 -O) 3 -CH 2 -、-(CH 2 ) 2 -(CH 2 -CH 2 -O) 2 -CH 2 -、-(CH 2 ) 2 -(CH 2 -CH 2 -NH) 2 -(CH 2 ) 2 -、-(CH 2 ) 3 -(CH 2 -CH 2 -NH) 2 -CH 2 -、-(CH 2 -CH 2 -NH) 3 -CH 2 -、-(CH 2 ) 2 -(CH 2 -CH 2 -NH) 2 -CH 2 -、-(CH 2 ) 3 -O-CH 2 -CH 2 -O-CH 2 -、-CH 2 -O-(CH 2 ) 6 -、-(CH 2 ) 2 -O-(CH 2 ) 5 -、-(CH 2 ) 3 -O-(CH 2 ) 4 -、-(CH 2 ) 4 -O-(CH 2 ) 3 -、-(CH 2 ) 5 -O-(CH 2 ) 2 -、-(CH 2 ) 6 -O-CH 2 -、-CH 2 -(O-CH 2 -CH 2 ) 2 -、-(CH 2 ) 2 -O-CH 2 -CH 2 -O-CH 2 -、-CH 2 -O-(CH 2 ) 5 -、-(CH 2 ) 2 -O-(CH 2 ) 4 -、-(CH 2 ) 3 -O-(CH 2 ) 3 -、-(CH 2 ) 4 -O-(CH 2 ) 2 -、-(CH 2 ) 5 -O-CH 2 -、-CH 2 -O-CH 2 -CH 2 -O-CH 2 -、-CH 2 -O-(CH 2 ) 4 -、-(CH 2 ) 2 -O-(CH 2 ) 3 -、-(CH 2 ) 3 -O-(CH 2 ) 2 -、-(CH 2 ) 4 -O-CH 2 -、-CH 2 -O-(CH 2 ) 3 -、-(CH 2 ) 2 -O-(CH 2 ) 2 -、-(CH 2 ) 3 -O-CH 2 -、-CH 2 -O-(CH 2 ) 2 -、-(CH 2 ) 2 -O-CH 2 -、-CH 2 -O-CH 2 -、-(CH 2 ) 3 -(O-CH 2 -CH 2 ) 2 -CH 2 -、-(CH 2 ) 2 -(O-CH 2 -CH 2 ) 2 -(CH 2 ) 2 、-CH 2 -(O-CH 2 -CH 2 ) 2 -(CH 2 ) 3 、-CH 2 -(O-CH 2 -CH 2 ) 3 -、-(CH 2 ) 2 -(O-CH 2 -CH 2 ) 2 -CH 2 -、-CH 2 -(O-CH 2 -CH 2 ) 2 -(CH 2 ) 2 - and -(CH 2 ) 3 -(O-CH 2 -CH 2 ) 2 -、CH 2 -(NH-CH 2 -CH 2 ) 2 -CH 2 -、-(CH 2 ) 2 -(NH-CH 2 -CH 2 ) 2 -、-(CH 2 ) 3 -NH-CH 2 -CH 2 -NH-CH 2 -、-CH 2 -NH-(CH 2 ) 6 -、-(CH 2 ) 2 -NH-(CH 2 ) 5 -、-(CH 2 ) 3 -NH-(CH 2 ) 4 -、-(CH 2 ) 4 -NH-(CH 2 ) 3 -、-(CH 2 ) 5 -NH-(CH 2 ) 2 -、-(CH 2 ) 6 -NH-CH 2 -、-CH 2 -(NH-CH 2 -CH 2 ) 2 -、-(CH 2 ) 2 -NH-CH 2 -CH 2 -NH-CH 2 -、-CH 2 -NH-(CH 2 ) 5 -、-(CH 2 ) 2 -NH-(CH 2 ) 4 -、-(CH 2 ) 3 -NH-(CH 2 ) 3 -、-(CH 2 ) 4 -NH-(CH 2 ) 2 -、-(CH 2 ) 5 -NH-CH 2 -、-CH 2 -NH-CH 2 -CH 2 -NH-CH 2 -、-CH 2 -NH-(CH 2 ) 4 -、-(CH 2 ) 2 -NH-(CH 2 ) 3 -、-(CH 2 ) 3 -NH-(CH 2 ) 2 -、-(CH 2 ) 4 -NH-CH 2 -、-CH 2 -NH-(CH 2 ) 3 -、-(CH 2 ) 2 -NH-(CH 2 ) 2 -、-(CH 2 ) 3 -NH-CH 2 -、-CH 2 -NH-(CH 2 ) 2 -、-(CH 2 ) 2 -NH-CH 2 -、-CH 2 -NH-CH 2 -、-(CH 2 ) 3 -(NH-CH 2 -CH 2 ) 2 -CH 2 -、-(CH 2 ) 2 -(NH-CH 2 -CH 2 ) 2 -(CH 2 ) 2 、-CH 2 -(NH-CH 2 -CH 2 ) 2 -(CH 2 ) 3 、-CH 2 -(NH-CH 2 -CH 2 ) 3 -、-(CH 2 ) 2 -(NH-CH 2 -CH 2 ) 2 -CH 2 -、-CH 2 -(NH-CH 2 -CH 2 ) 2 -(CH 2 ) 2 -、-(CH 2 ) 3 -(NH-CH 2 -CH 2 ) 2 -、-CH 2 -O-(CH 2 ) 8 -、-(CH 2 ) 2 -O-(CH 2 ) 7 -、-(CH 2 ) 3 -O-(CH 2 ) 6 -、-(CH 2 ) 4 -O-(CH 2 ) 5 -、-(CH 2 ) 5 -O-(CH 2 ) 4 -、-(CH 2 ) 6 -O-(CH 2 ) 3 -、-(CH 2 ) 7 -O-(CH 2 ) 2 -、-(CH 2 ) 8 -O-CH 2 -、-CH 2 -O-(CH 2 ) 7 -、-(CH 2 ) 2 -O-(CH 2 ) 6 -、-(CH 2 ) 3 -O-(CH 2 ) 5 -、-(CH 2 ) 4 -O-(CH 2 ) 4 -、-(CH 2 ) 5 -O-(CH 2 ) 3 -、-(CH 2 ) 6 -O-(CH 2 ) 2 -、-(CH 2 ) 7 -O-CH 2 -、-CH 2 -NH-(CH 2 ) 8 -、-(CH 2 ) 2 -NH-(CH 2 ) 7 -、-(CH 2 ) 3 -NH-(CH 2 ) 6 -、-(CH 2 ) 4 -NH-(CH 2 ) 5 -、-(CH 2 ) 5 -NH-(CH 2 ) 4 -、-(CH 2 ) 6 -NH-(CH 2 ) 3 -、-(CH 2 ) 7 -NH-(CH 2 ) 2 -、-(CH 2 ) 8 -NH-CH 2 -、-CH 2 -NH-(CH 2 ) 7 -、-(CH 2 ) 2 -NH-(CH 2 ) 6 -, -(CH 2 ) 3 -NH-(CH 2 ) 5 -, -(CH 2 ) 4 -NH-(CH 2 ) 4 -, -(CH 2 ) 5 -NH-(CH 2 ) 3 -, -(CH 2 ) 6 -NH-(CH 2 ) 2 -, -(CH 2 ) 7 -NH-CH 2 -, -CH(NH 2 )-CH 2 -, -CH 2 -CH(NH 2 )-, -CH(COOH)-CH 2 -, -CH 2 -CH(COOH) and -CH(R 3 )-, In particular, f is -(CH 2 ) 2 -(O-CH 2 -CH 2 ) 2 -, -CH 2 -(O-CH 2 -CH 2 ) 2 -CH 2 - and -(CH 2 ) 3 -O-CH 2 -CH 2 -O-CH 2 - is a residue selected from the group consisting of: Each e'' is a chemical bond, -NH-C(O)-, -C(O)-NH-, -C(O)-O- and -O-C(O)-, -NH-C(O)-NH-, -NH-C(S)-NH-, -C(O)-N(CH 3 )-, -N(CH 3 )-C(O)-, -NH-C(S)-, -C(S)-NH-, 【Chemistry 5】 wherein one wavy line indicates a conjugation site for f and the other wavy line indicates a conjugation site for at least one dye moiety (C), in particular e″ is —NH—C(O)—; m represents an integer from 0 to 8, preferably from 0 to 4, more preferably from 0 to 2, even more preferably 0 or 1, especially 1.

3. The compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, having the formula:

4. Spacer x 2 but has the following structure: -(CH 2 ) t -C(O)-NH-(CH 2 ) u -(O-CH 2 -CH 2 ) v -(CH 2 ) w -e''-, or -(CH 2 ) t -C(O)-NH-(CH 2 -CH 2 -O) v -CH 2 -e''- (In the formula, t is 1 or 2, in particular 2; u is an integer from 1 to 10, preferably from 1 to 3, in particular 2; v is an integer from 0 to 3, in particular 2; w is an integer from 0 to 2, in particular 0; e'' is as defined in claim 3) The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, having one of the following formulas:

5. Spacer x 2 but has the following structure: -(CH 2 ) 2 -C(O)-NH-(CH 2 ) 2 -(O-CH 2 -CH 2 ) 2 -e''- -(CH 2 ) 2 -C(O)-NH-(CH 2 ) 2 -(O-CH 2 -CH 2 ) 2 -NH-C(O)-CH 2 -(O-CH 2 -CH 2 ) n' -O-CH 2 -e''-、 -(CH 2 ) 2 -C(O)-NH-(CH 2 ) 2 -(O-CH 2 -CH 2 ) 2 -NH-[C(O)-CH((CH 2 ) 2 COOH)-NH] n'' -C(O)-CH((CH 2 ) 2 COOH)-e''-、 -(CH 2 ) 2 -C(O)-NH-(CH 2 ) 2 -(O-CH 2 -CH 2 ) 2 -NH-[C(O)-CH((CH 2 ) 4 NH 2 )-NH] n'' -C(O)-CH((CH 2 ) 4 NH 2 )-e''- or -(CH 2 ) 2 -C(O)-NH-(CH 2 ) 2 -(O-CH 2 -CH 2 ) 2 -NH-[C(O)-CH((CH 2 ) 4 N + (CH 3 ) 3 )-NH] n'' -C(O)-CH((CH 2 ) 4 N + (CH 3 ) 3 )-e''- + X - (In the formula, n' is an integer from 1 to 3; n'' is an integer from 0 to 2; X - is a pharmaceutically acceptable negatively charged counterion; Each e'' is a chemical bond, -NH-C(O)-, -C(O)-NH-, -C(O)-O- and -O-C(O)-, -NH-C(O)-NH-, -NH-C(S)-NH-, -C(O)-N(CH 3 )-, -N(CH 3 )-C(O)-, -NH-C(S)-, -C(S)-NH-, 【Chemistry 6】 wherein one wavy line indicates a conjugation site for f and the other wavy line indicates a conjugation site for at least one dye moiety (C), and in particular e″ is —NH—C(O)—. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, having one of the following formulas:

6. The radiometal chelator moiety (B) is: 【Chemistry 7】 【change】 wherein in each of the structures, one of the wavy lines represents a spacer x 1 The other wavy line indicates a conjugation site for at least one motif (A) that specifically binds to the cell membrane of a neoplastic cell via a spacer x 2 (c) shows a conjugation site for at least one dye moiety (C) via is selected from the group consisting of In particular, the chelating agent moiety is: 【Chemistry 8】 6. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein:

7. (A)-x 1 -(B)-x 2 -has the following structure: 【Chemistry 9】 The compound according to any one of claims 1 to 6, represented by: or a pharmaceutically acceptable salt thereof.

8. In formula (C), X 1 and X 4 are identical, and preferably C(R 6 8. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R is H, ...

9. In formula (C), X 2 and X 3 are identical, and preferably C(R 6 R 7 ), more preferably R 6 and R 7 are identical, and even more preferably LZ (L=C 1-10 Alkylene, even more preferably CH 2 and Z=H), or a pharmaceutically acceptable salt thereof.

10. In formula (C), Y is (L-) c Z 0 and preferably Y does not contain: 【Chemistry 10】 (In the formula, g is 1, 2, 3, or 4 (preferably 3 or 4, more preferably 3), and each R 9a is (L-) c Z or H; two R 9a can also form a carbocyclic ring having 5, 6, or 7 carbon atoms or a 4- to 7-membered heterocyclic ring; each c is independently 0 or 1; Each L is independently T 1 , -OT 1 -, -ST 1 -, -C(O)T 1 -, -C(O)OT 1 -, -OC(O)T 1 -, -C(O)NHT 1 -,-NHC(O)T 1 , or optionally -O-, -S-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NH-, -NHC(O)O- and T 1 C that is interrupted and / or terminated by one or more of 1-10 an alkylene group; T 1 is phenyl, naphthyl, indenyl, indanyl, tetralinyl, decalinyl, adamantyl, C 3-7 cycloalkyl, 4- to 7-membered heterocyclyl, or 7- to 11-membered heterobicyclyl, where T 1 is sometimes halogen, CN, C(O)R 8 , COOR 8 , OR 8 , C(O)N(R 8 R 8a ), S(O) 2 N(R 8 R 8a ), S(O)N(R 8 R 8a ), S(O) 2 R 8 , N(R 8 )S(O) 2 N(R 8a R 8b ), SR 8 , N(R 8 R 8a ), NO 2 ,OC(O)R 8 , N(R 8 )C(O)R 8a , N(R 8 )S(O) 2 R 8a , N(R 8 )S(O)R 8a , N(R 8 )C(O)N(R 8a R 8b ), N(R 8 )C(O)OR 8a , OC(O)N(R 8 R 8a ), oxo(=O) (wherein the ring is at least partially saturated), or C 1-6 Alkyl (C 1-6 alkyl is optionally substituted with one or more of the same or different halogens; R 8 , R 8a , R 8b is H or C 1-6 Alkyl (C 1-6 alkyl is optionally substituted with one or more of the same or different halogens; Each Z is independently H, halogen, CN, C(O)R 8 、C(O)OR 8 、C(O)O - 、OR 8 、C(O)N(R 8 R 8a )、S(O) 2 OR 8 、S(O) 2 O - 、S(O) 2 N(R 8 R 8a )、S(O)N(R 8 R 8a )、S(O) 2 R 8 、S(O)R 8 、N(R 8 )S(O) 2 N(R 8a R 8b )、SR 8 、N(R 8 R 8a )、NO 2 、P(O)(OR 8 ) 2 、P(O)(OR 8 )O - 、OC(O)R 8 、N(R 8 )C(O)R 8a 、N(R 8 )S(O) 2 R 8a 、N(R 8 )S(O)R 8a 、N(R 8 )C(O)N(R 8a R 8b )、N(R 8 )C(O)OR 8a 、or OC(O)N(R 8 R 8a )) and preferably Y is: 【Chemistry 11】 or 【Chemistry 12】 (g = 2, and each R 9a = H) The compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

11. In formula (C), a and b are the same and preferably 1, and more preferably R 1 and R 2 = SO 3 - The compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, wherein:

12. In formula (C), a and b are the same and 2, and preferably two adjacent R 1 and two adjacent R 2 The compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, wherein

13. In formula (C), R 3 and R 4 One of them is (L-) c Z Z0 and the other is (L-) c Z(L- = C 1-10 is alkylene, Z = H or SO 3 - and preferably c = 1), or a pharmaceutically acceptable salt thereof.

14. (L-) c Z Z0 But (C) x 2 or x 2 If is a chemical single bond, C binds to (B) 1-10 The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, which is alkylene.

15. The dye portion (C) has the following structure: 【Chemistry 13】 (In the formula, X - is a pharmaceutically acceptable negatively charged counterion; Y + is a pharmaceutically acceptable positively charged counterion; The wavy line indicates the site of conjugation to the rest of the compound.) The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

16. 16. The compound according to any one of claims 1 to 15, wherein the dye moiety (C) is a fluorescent dye moiety having an emission maximum in the range of 400 nm to 1000 nm.

17. The compound has the following chemical structure: 【Chemistry 14】 17. The compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, having the formula:

18. below: (a) a compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof; (b) a radioactive metal, preferably 89 Zr, 44 Sc, 111 In, 90 Y, 67 Ga, 68 Ga, 177 Lu, 99m Tc, 82 Rb, 64 Cu, 67 Cu, 153 Gd, 155 Gd, 157 Gd, 213 Bi, 225 Ac and 59 Fe, in particular 68 Ga; and optionally (c) one or more pharmaceutically acceptable carriers A composition comprising:

19. 19. The composition of claim 18 for use as a diagnostic agent.

20. 1. A method for assisting in the detection of neoplastic cells in a sample in vitro, comprising the steps of: (i) providing cells which are neoplastic or at risk of being neoplastic, in particular cells which are cancerous or at risk of being cancerous; (ii) administering to the cells a compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof, or a composition according to claim 18; (iii) detecting a fluorescent signal and / or a radioactive signal from the cells. The method comprising:

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