imaging agent

Abstract

The present invention relates to compounds of formula I or la: Y is EuK, -EuAF, -EuPG, -L-EuE; Z is a chelating moiety; other substituents are as defined herein. Also provided are formulations comprising such compounds, as well as imaging methods or cancer treatment methods comprising the use of such compounds or formulations.

Description

Technical Field

[0001] This invention relates to compounds that can be used as tracers, particularly tracers that can be used for targeting cancers such as prostate cancer. The tracers represent hybrid tracers comprising a probe, a dye portion, and a portion that may contain a radiolabeled component. Background Technology

[0002] Treatment of primary tumors and their metastases requires interventional molecular imaging techniques. Cancers such as prostate cancer have a relatively high incidence rate, and their treatment is technically challenging. Image-guided resection of primary tumors and lymph node metastases is considered to improve surgical outcomes.

[0003] One approach to providing image-guided resection involves providing molecules that can act as marker tracers. Prostate-specific membrane antigen (PSMA), a transmembrane protein expressed in prostate tissue, is a useful diagnostic and potential therapeutic target for prostate cancer. Furthermore, according to Fragomoni et al., J. Nuc. Med., June 1, 2018, 59(6), 871-877, PSMA expression has been observed in neovascularization systems of various non-prostate malignancies, increasing the potential for PSMA-based diagnostic and therapeutic applications outside of prostate cancer.

[0004] Glutamic acid-urea-lysine (EuK) carriers are known to target prostate-specific membrane antigen (PSMA), allowing tracers containing appropriately functionalized EuK carriers to be used for the detection of prostate cancer. Other EuX-based carriers are used in a similar manner. Exemplary molecules based on EuX carriers are disclosed, for example, in WO2010 / 108125 and WO 2013 / 082338. However, these molecules do have several drawbacks. For example, none of these molecules have been shown to be highly effective for hybrid imaging, where a single tracer facilitates both general preoperative detection and intraoperative fluorescence imaging.

[0005] Given the prospect of better oncological outcomes following radical surgical resection of tumors, there is a general need to develop more tracers for surgical guidance. Therefore, an object of the present invention is to provide compounds that can be used as tracers for the detection of cancers, such as prostate cancer and / or other cancers detectable with PSMA probes. In particular, one object of the present invention is to provide hybrid compounds that, through the combined use of fluorescent dyes and radiolabeling, allow for non-invasive imaging and / or imaging during surgery. Another object of the present invention is to provide compounds with predictive and tunable pharmacokinetic properties that facilitate the efficient separation of cancers (e.g., prostate cancer and / or other cancers detectable with PSMA probes) while preventing unwanted uptake from background organs. Summary of the Invention

[0006] This invention provides compounds for use as tracers. Specifically, the compounds of this invention bind a probe (e.g., a PSMA probe) to a fluorophore and a portion that can be radiolabeled (e.g., by chelating a radioactive nucleotide). When these compounds contain a radiolabeled nucleotide, they can be considered as hybrid tracers because the probe portion is coupled to two complementary labels: a fluorophore and a radiolabel.

[0007] According to a first aspect, the present invention provides compounds of formula I or Ia, or pharmaceutically acceptable salts thereof:

[0008]

[0009] R1 is selected from sulfonates, carboxyl groups, phosphonates, amines, azides, and H, and R2 is selected from sulfonates, carboxyl groups, phosphonates, amines, azides, and H; or R1 and R2 together form an aryl group optionally substituted with 1 to 4 groups (e.g., one or two groups), each group independently selected from sulfonates, carboxyl groups, phosphonates, amines, and azides. R3 is selected from sulfonates, carboxyl groups, phosphonates, amines, azides, and H, and R4 is selected from sulfonates, carboxyl groups, phosphonates, amines, azides, and H; or R3 and R4 together form an aryl group optionally substituted with 1 to 4 groups (e.g., one or two groups), each group independently selected from sulfonates, carboxyl groups, phosphonates, amines, and azides. R5, R6, R7, R8, R9, and R 10 Each group is independently selected from sulfonates, carboxylates, phosphonates, amines, azides, CH3, CH2CH3, and H. V is -CH2- or -CH2CH2O-. W is -CH2- or -CH2CH2O-. Y1 is -EuK, -EuFA, -EuPG, -L1-EuK, -L1-EuFA, -L1-EuPG, or -L3-EuE. Y2 is -L4-EuK, -L4-EuFA, -L4-EuPG, -EuE, or -L2-EuE. Z is the chelate moiety; for example, Z can be -MAS3, -MAG3, -DOTA-GA, -DOTA, -DTPA, -L2-MAS3, -L2-MAG3, -L2-DOTA-GA, -L2-DOTA, or -L2-DTPA. L1 is of the formula -NH-R. 12 -C(O)- connector, where R 12 It is a bonded, substituted, or unsubstituted alkyl group; for example, L1 can be a lysine residue, ornithine residue, aspartic acid residue, glutamic acid residue, or -NH-(C0-C7 alkyl)-C(O)-. L2 is of the formula -C(O)-R. 13 -NH- connector, where R 13It is a bond, or a substituted or unsubstituted alkyl group; for example, L2 can be a bond, a lysine residue, an ornithine residue, an aspartic acid residue, a glutamic acid residue, or -NH-(C0-C7 alkyl)-C(O)-. L3 is of the formula -NH-R 14 -NH- connector, where R 14 It can be a substituted or unsubstituted alkyl group; for example, L3 can be a lysine residue, an ornithine residue, or -NH-(C4-C7 alkyl)-NH-. L4 is of the formula -C(O)-R 15 -C(O)- connector, where R 15 It is a substituted or unsubstituted alkyl group; for example, L4 can be an aspartic acid residue, a glutamic acid residue, or -C(O)-(C4-C7 alkyl)-C(O)-. n is 2 or 3. m is 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21. p is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or 21.

[0010] The first aspect of the compound can be used as a mixed tracer, wherein "Y1" or "Y2" contains a probe, "Z" may contain a radiolabel (e.g., a chelated radioactive nucleotide), and the portion between them contains a fluorophore. The dye linker molecule has an amino acid-type structure, wherein Formula I has a "Y1" attached to the carbon terminus of the fluorophore and a "Z" attached to the nitrogen terminus of the fluorophore; while Formula 1a has a "Z" attached to the carbon terminus of the fluorophore and a "Y2" attached to the nitrogen terminus of the fluorophore. Using dye structures as linker molecules offers many advantages. For example, in such molecular designs, the fluorophore is oriented in a predictable manner, which is determined by the substituents R1 to R2. 10 This is highly advantageous in cases leading to asymmetric fluorophores. The latter feature allows the fluorophore to complement the EuX carrier and become part of the tracer pharmacophore. In particular, the linker dye can be used to modulate the interaction with the auxiliary hydrophobic pocket within the PSMA.

[0011] A second aspect of the invention provides compounds of formula II or IIa, or pharmaceutically acceptable salts thereof:

[0012]

[0013]

[0014] Among them, R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 V, W, Z, L3, m, n, and p are as defined in the first aspect. The compounds in the second aspect represent synthetic intermediates of the compounds in the first aspect.

[0015] A third aspect of the invention provides a compound of formula III or IIIa, or a pharmaceutically acceptable salt thereof:

[0016]

[0017] Among them, R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 V, W, Y1, Y2, m, n, and p are as defined in the first aspect. The compounds in the second aspect represent synthetic intermediates of the compounds in the first aspect.

[0018] The fourth aspect provides formulations comprising the compounds of the present invention and optionally a pharmaceutically acceptable carrier. In one embodiment, the compound of the present invention is the compound of the first aspect.

[0019] The fifth aspect provides a method for imaging a tumor, comprising administering a compound of the first aspect or an agent of the second aspect to a subject and imaging the tumor after a predetermined time.

[0020] The sixth aspect provides a method of treating cancer, comprising administering a compound of the first aspect or an agent of the second aspect. In one embodiment, the compound comprises a chelated radiolabeled substance.

[0021] The seventh aspect provides the use of the compounds of the first aspect or the formulations of the second aspect in imaging.

[0022] The eighth aspect provides a compound for use as a first aspect of a drug or a formulation for use as a second aspect. In one embodiment, the compound comprises a chelated radiolabeled substance.

[0023] The ninth aspect provides a compound of the first aspect or an agent of the second aspect for treating cancer. In one embodiment, the compound comprises a chelated radiolabeled substance.

[0024] The invention will now be further described with reference to the following embodiments and accompanying drawings. These are not intended to limit the invention, but are merely examples of the invention. Attached Figure Description

[0025] The embodiments of the present invention will be further described below with reference to the accompanying drawings, wherein:

[0026] Figure 1 The general reaction process for synthesizing the compounds of this invention is described in section 1.

[0027] Figure 2Results of plasma protein interaction assays for exemplary mixed tracer compounds are presented. A) Percentage of plasma protein binding relative to the indicated mixed tracer binding to serum albumin. All mixed tracers showed significant differences (p < 0.05) unless otherwise specified (ns). B) and C) demonstrate the stability of the mixed tracers in serum albumin as a function of time, based on absorbance in B) and fluorescence in C).

[0028] Figure 3 The image shows PSMA-related staining in LNCaP cells cultured with the mixed tracer EuK(SO3)-Cy5-MAS3. A) Bright-field image of the cultured cells. B) Overlay of confocal images of the cultured cells, where red represents Cy5-related signals of the PSMA-targeting tracer, blue represents the cell nucleus, and green represents lysosomes in the cytoplasm. C) Overlay of confocal and bright-field images.

[0029] Figure 4 provides a comparison of nuclear imaging and quantitative biodistribution of exemplary mixed tracers, demonstrating tumor binding of the exemplary mixed tracers. A) In vivo SPECT imaging. Coronal SPECT images of prostate tumor-bearing mice injected with any of the PSMA mixed tracer analogs. Organs are represented as Lu (lung), Li (liver), G (gallbladder), S (spleen), K (kidney), B (bladder), and T (tumor). B) Biodistribution of the mixed tracer matrix compared to the clinically approved tracer PSMAI&S. C) Biodistribution of the mixed tracer EUK(SO3)-CY5-MAS3 is provided. Biodistribution patterns 2 hours after injection of the different mixed tracers from the matrix compared to the reference tracer PSMAI&S are described as percentages of the injected dose per gram of tissue (%ID / g).

[0030] Figure 5 Biodistribution parameters and tumor uptake of exemplary mixed tracers are described. Overviews include: A) the percentage of injection activity found in all resected organs; the percentage of injection activity retained 2 hours after tracer injection (all ns); B) the amount of radioactivity still present in the blood pool; and C) no significant difference in hepatic clearance among the compounds. To highlight tumor visibility against background tissue, it is well known that background tissue blocks the signal in the prostate region, providing D) the tumor-to-fat ratio and E) the tumor-to-blood ratio for individual mixed tracers. Significance is described as: none (not significant) * (p ≤ 0.05) ** (p ≤ 0.01) *** (p ≤ 0.001).

[0031] Figure 6Ex vivo fluorescence imaging of tumors in prostate tissue from mice injected with an exemplary mixed tracer is shown. A) Photographs of tumor locations (within dashed lines) in relation to healthy prostate tissue (*), bladder (**), seminal vesicles (***), and abdominal adipose tissue (#). Fluorescence images of EuK-Cy5-MAS3, C) EuK(SO3)-Cy5-MAS3, D) EuK-Cy5-(SO3)MAS3, E) EuK(Ar)-Cy5-MAS3, F) EuK-Cy5-(Ar)MAS3, and G) EuK(Ar)-Cy5-(Ar)MAS3 obtained using clinical laparoscopy modified for Cy5 imaging. Detailed Implementation

[0032] Throughout this specification and claims, the words “comprising” and “including,” and variations thereof, mean “containing but not limited to,” and are not intended to exclude other parts, additives, components, wholes, or steps. Throughout this specification and claims, the singular encompasses the plural unless the context requires otherwise. In particular, where the indefinite article is used, the specification should be understood to consider both the plural and singular unless the context requires otherwise.

[0033] Features, integrals, characteristics, compounds, chemical parts, or groups described in connection with a particular aspect, embodiment, or example of the invention should be understood to be applicable to any other aspect, embodiment, or example described herein, unless incompatible therewith. All features disclosed in this specification (including any appended claims, abstract, and drawings), and / or all steps of any method or process so disclosed, may be combined in any combination, except for at least some such features and / or steps that are mutually exclusive. The invention is not limited to the details of any of the foregoing embodiments. The invention extends to any novel one or any novel combination of features disclosed in this specification (including any appended claims, abstract, and drawings), or to any novel one or any novel combination of steps of any method or process so disclosed.

[0034] The reader’s attention is drawn to all papers and documents related to this application that were submitted concurrently with or prior to this specification, which are made publicly available together with this specification, and the contents of all such papers and documents are incorporated herein by reference.

[0035] All publications, patent applications, patents, and other references mentioned herein are incorporated herein by reference in their entirety. In case of any conflict, this specification (including definitions) shall prevail.

[0036] definition

[0037] The following explanations of terms and methods are provided to better describe this disclosure and to guide those skilled in the art in practicing this disclosure.

[0038] This invention relates particularly to imaging. The term "imaging" includes providing a visual representation of a sample by detecting radiation. The radiation may be a product of the radioactive decay of a radiolabeled substance. The radiation may be a product of fluorescence. The visual representation can be provided by electronically processing the detected radiation, for example by performing positron emission tomography (PET), single-photon emission computed tomography (SPECT), scintillation, (optionally intraoperative) gamma-ray tracing / imaging, or (optionally intraoperative) beta-ray tracing. The visual representation can also be provided by visual detection, for example by observing a sample exposed to high-frequency electromagnetic radiation that fluoresces at visible wavelengths (e.g., from about 390 nm to 700 nm). The visual representation can also be provided by fluorescence spectroscopy.

[0039] This invention relates particularly to the treatment of diseases. The term "treatment" and the therapies included in this invention include the following and combinations thereof: (1) preventing, e.g., delaying the onset and / or progression of an event, state, symptom, or condition, for example, in the case of maintenance therapy or secondary prevention, or in the case of at least one clinical or subclinical symptom; (2) preventing or delaying the occurrence of clinical symptoms of an event, state, symptom, or condition in animals (e.g., humans) that may have or are susceptible to the state, symptom, or condition but have not yet experienced or exhibited clinical or subclinical symptoms of the state, symptom, or condition; and / or (3) alleviating and / or curing an event, state, symptom, or condition (e.g., causing the resolution of at least one of the event, state, symptom, or condition or its clinical or subclinical symptoms, curing a patient, or alleviating a patient's symptoms). The benefit to the patient to be treated may be statistically significant, or at least perceptible to the patient or physician. It should be understood that a drug may not produce a clinical effect in every patient receiving drug treatment; therefore, treatment may fail or be only partially successful in any individual patient or even in a specific patient population, and the meanings of the terms "treatment," "prevention," and "inhibitor," as well as related terms, should be understood accordingly. The compositions and methods described herein may be used to treat and / or prevent the aforementioned conditions.

[0040] The term "prevention" includes therapeutic approaches aimed at maintaining health or inhibiting or delaying the onset and / or progression of an event, state, symptom, or condition, such as reducing the likelihood of its occurrence. The result of prevention can be, for example, maintaining health or delaying the onset and / or progression of an event, state, symptom, or condition. It should be remembered that treatment may fail in any individual patient, or even in a specific patient population, and this paragraph should be understood accordingly.

[0041] The terms "probe" or "targeting moiety" refer to a portion or carrier that targets PSMA through affinity-type interactions. Exemplary probes include a moiety with the molecular formula EuX, which is a glutamate salt of another amino acid or analogue, such as EuK, EuFA, EuPG, or EuE, linked to another urea via a bridging urea.

[0042] The term "tracer" refers to a molecule that comprises a targeting portion and an imaging label. A tracer may include two imaging labels, for example, as a hybrid tracer having two different types of imaging labels. For instance, in a hybrid tracer, the probe portion may be coupled to two labels that can be used for complementary purposes: a fluorophore label and a radiolabel.

[0043] The terms "inhibition" (and "suppression") encompass the delay, cessation, or reduction of the incidence, risk, and / or severity of an event, state, symptom, or condition. Therefore, inhibiting an event, state, symptom, or condition may include delaying or stopping the onset and / or progression of such an event, state, symptom, or condition, and reducing the risk of its occurrence. The probe EuX can be considered an inhibitor of PSMA.

[0044] The term "chelating moiety" includes residues of a chelating agent, such as bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (CBTE2a), cyclohexyl-1,2-diaminotetraacetic acid (CDTA), 4-(1,4,8,11-tetraazabicyclotetradecane-1-yl)-methylbenzoic acid (CPTA), N′-[5-[acetyl(hydroxy)amino]pentyl]-N-[5-[[4-[5-aminopentyl-(hydroxy)amino]-4-oxobutyryl]pentyl]-N-hydroxybutyramide (DFO), 4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (DO2A), 1 4,7,10-Tetraazacyclododecane-N,N',N”,N”'-Tetraacetic acid (DOTA), 2-[1,4,7,10-Tetraazacyclododecane-4,7,10-triacetic acid]-glutaric acid (DOTAGA), N,N'-dipyridoxyethylenediamine-N,N'-diacetic acid-5,5'-bis(phosphoric acid) (DPDP), diethylenetriaminepentaacetic acid (DTPA), ethylenediamine-N,N'-tetraacetic acid (EDTA), ethylene glycol-O,O-bis(2-aminoethyl)-N,N,N',N'-tetraacetic acid (EGTA), N,N-bis(hydroxybenzyl)-ethylenediamine-N,N'-diacetic acid (HBED), hydroxyethyldiaminetriacetic acid (HEDTA), 1- (p-Nitrobenzyl)-1,4,7,10-tetraazacyclodecane-4,7,10-triacetate (HP-DOA3), 6-hydrazyl-N-methylpyridine-3-carboxamide (HYNIC), 1,4,7-triazacyclononane-1-succinic acid-4,7-diacetic acid (NODASA), 1-(1-carboxy-3-carboxypropyl)-4,7-(carboxyl)-1,4,7-triazacyclononane (NODAGA), 1,4,7-triazacyclononanetriacetic acid (NOTA), 4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (TE2A), 1,4,8,11-tetraazacyclododecane-1,4,8,11-tetraacetic acid Acids include TETA, terpyridine-bis(methyleneaminotetraacetic acid (TMT), 1,4,7,10-tetraazacyclo-tetrazane-N,N',N”,N”'-tetraacetic acid (TRITA), triethylenetetraminehexaacetic acid (TTHA), N,N′-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18-crown-6 (H2macropa), and 4-amino-4-{2-[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydro-pyridin-2-ylmethyl)-carbamoyl]-ethyl}pimelic acid bis-[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydro-pyridin-2-ylmethyl)-amide] (THP).This residue is obtained by covalently binding the carboxyl group contained in the chelating agent to the remainder of the compound via an ester bond or an amide bond, preferably an amide bond.

[0045] As used herein, the term "alkyl" includes straight-chain or branched alkyl moieties having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. This term includes, for example, methyl, ethyl, propyl (n-propyl or isopropyl), butyl (n-butyl, sec-butyl, or tert-butyl), pentyl, hexyl, etc. Specifically, alkyl can be "C1-C4 alkyl," i.e., alkyl with 1, 2, 3, or 4 carbon atoms; or "C1-C6 alkyl," i.e., alkyl with 1, 2, 3, 4, 5, or 6 carbon atoms; or "C1-C3 alkyl," i.e., alkyl with 1, 2, or 3 carbon atoms. The term "lower alkyl" includes alkyl with 1, 2, 3, or 4 carbon atoms.

[0046] As used herein, the term "cycloalkyl" includes an alicyclic moiety having 3, 4, 5, or 6 carbon atoms. This group can be a bridged or polycyclic ring system. More commonly, cycloalkyl groups are monocyclic. The term includes groups such as cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0047] As used herein, the term "heterocyclic alkyl" includes a saturated heterocyclic moiety having 3, 4, 5, 6, or 7 cyclic carbon atoms and 1, 2, 3, 4, or 5 cyclic heteroatoms selected from nitrogen, oxygen, phosphorus, and sulfur. For example, a heterocyclic alkyl group may contain 3, 4, or 5 cyclic carbon atoms and 1 or 2 cyclic heteroatoms selected from nitrogen and oxygen. The group can be a polycyclic system, but is more commonly monocyclic. The term also includes groups such as azirrobutyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, ethylene oxide, pyrazolyl, imidazolyl, indoleazinyl, piperazinyl, thiazolyl, morpholinyl, thiomorpholinyl, and quinolinyl.

[0048] Unless otherwise stated, the term "aryl" refers to a polyunsaturated aromatic substituent, which may be a monocyclic or polycyclic ring (preferably 1-3 rings) fused together or covalently linked. The term "heteroaryl" refers to an aryl group (or ring) containing 1-4 heteroatoms selected from N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom is optionally quaternized. Heteroaryl groups can be attached to the rest of the molecule via carbon or heteroatoms. Non-limiting examples of aryl and heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrole, 2-pyrrole, 3-pyrrole, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isooxazolyl, 4-isooxazolyl, 5-isooxazolyl, 2-thiazolyl, 4- -Thiazolyl, 5-thiazolyl, 2-furanyl, 3-furanyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-benzothiazolyl, purinel, 2-benzimidazolyl, 5-indolyl, 1-isoquinolinyl, 5-isoquinolinyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolinyl, and 6-quinolinyl. The substituents in each of the above aryl and heteroaryl ring systems are selected from the following acceptable substituent groups. "Arylidene" and "heteroaryl" refer to divalent residues derived from aryl and heteroaryl groups, respectively.

[0049] As used herein, the term "alkoxy" includes -O-alkyl, wherein the alkyl group is straight-chain or branched and contains 1, 2, 3, 4, 5, or 6 carbon atoms. In one type of embodiment, the alkoxy group has 1, 2, 3, or 4 carbon atoms, for example, 1, 2, or 3 carbon atoms. The term includes, for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, pentoxy, hexoxy, etc. The term "lower alkoxy" includes alkoxy groups having 1, 2, 3, or 4 carbon atoms. Alkoxy groups, particularly lower alkoxy groups (e.g., alkoxy groups having 2 carbon atoms), can be provided as polyalkoxy groups, i.e., as straight-chain or branched (e.g., straight-chain) repeating alkoxy units.

[0050] As used herein, the term "substituted" means that one or more, particularly up to five, and more particularly one, two, or three hydrogen atoms in the moiety are independently substituted by the corresponding number of said substituents. Unless otherwise stated, exemplary substituents include -OH, -CN, -NH2, =O, -halogen, -C1-C6 alkyl, -C2-C6 alkenyl, -C1-C6 haloalkyl, -C1-C6 haloalkoxy, and -C2-C6 haloalkenyl, -C1-C6 alkylcarboxylic acids (e.g., –CH3COOOH or -COOH). When said substituent is -C1-C6 alkyl or -C1-C6 haloalkyl, the C1-C6 chain is optionally interrupted by an ether bond (-O-) or an ester bond (-C(O)O-). Exemplary substituents for substituted alkyl groups may include -OH, -CN, -NH2, =O, -halogen, -CO2H, -C1-C6 haloalkyl, -C1-C6 haloalkoxy, and -C2-C6 haloalkyl, -C1-C6 alkylcarboxylic acids (e.g., –CH3COOOH or -COOH). For example, exemplary substituents for alkyl groups may include -OH, -CN, -NH2, =O, and -halogen.

[0051] Of course, it should be understood that substituents are only located at chemically possible positions, and those skilled in the art can determine, experimentally or theoretically, whether a particular substitution is possible without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable if bonded to a carbon atom having an unsaturated bond (e.g., an alkene bond). Furthermore, it should be understood that the substituents described herein can themselves be substituted with any substituent that meets the foregoing limitations on suitable substitution as recognized by those skilled in the art.

[0052] When steric mechanics determines the position of a substituent on a group, the isomer with the lowest conformational energy may be preferred. For example, the preferred state for carbon cyanides (e.g., Cy dyes) may be the all-trans conformation, as this represents the ground state (see, for example, AM Kolesnikov and EAMikhailenko, Russian Chemical Reviews, (1987), 56, 275–287; W. West et al., Journal of Physical Chemistry, (1967), 71, 1316–1326; and PJ Wheatley, Journal of the Chemical Society, (1959), 4096–4100).

[0053] When a compound, part, method, or product is described as "optionally" having a certain feature, this disclosure includes both such a compound, part, method, or product having that feature and such a compound, part, method, or product not having that feature. Therefore, when a part is described as "optionally substituted," this disclosure includes both unsubstituted and substituted parts.

[0054] When two or more parts are described as being selected “independently” or “each independently” from a list of atoms or groups, it means that these parts can be the same or different. Therefore, the determination of each part is independent of the determination of one or more other parts.

[0055] As used herein, the term "pharmaceutically acceptable" includes compounds, materials, compositions, and / or dosage forms that, to a reasonable medical judgment, are suitable for use in contact with human or animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, and in proportion to a reasonable benefit / risk ratio. This term includes acceptability for both human and veterinary purposes.

[0056] The term "pharmaceutically acceptable salt" refers to a salt comprising a compound prepared with a relatively non-toxic acid or base, depending on the specific substituents present on the compound described herein. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such a compound in its neutral form with a sufficient amount of the desired base (whether pure or in a suitable inert solvent). Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salts, or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such a compound in its neutral form with a sufficient amount of the desired acid (whether pure or in a suitable inert solvent). Examples of pharmaceutically acceptable acid addition salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrocarbonic acid, phosphoric acid, monohydrophosphoric acid, dihydrophosphoric acid, sulfuric acid, monohydrosulfuric acid, hydroiodic acid, or phosphorous acid, as well as salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid. Salts of amino acids, such as arginine salts, and salts of organic acids, such as glucuronic acid or galacturonic acid, are also included (see, for example, Berge et al., “Pharmaceutical Salts”, Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain compounds of the present invention contain both basic and acidic functional groups, which allows the compound to be converted into a basic or acid addition salt.

[0057] Therefore, the compounds of the present invention can exist in the form of salts of pharmaceutically acceptable acids. The present invention includes such salts. Examples of such salts include hydrochlorides, hydrobromic acids, sulfates, methanesulfonates, nitrates, maleates, acetates, citrates, fumarates, tartrates (e.g., (+)-tartrates, (-)-tartrates, or mixtures thereof (including racemic mixtures), succinates, benzoates, and salts with amino acids (such as glutamic acid). These salts can be prepared by methods known to those skilled in the art.

[0058] Some compounds of this invention have asymmetric carbon atoms (optical centers) or double bonds; racemates, diastereomers, tautomers, geometric isomers, and monomeric isomers are all included within the scope of this invention. The compounds of this invention do not include those known in the art that are too unstable to be synthesized and / or isolated.

[0059] The compounds of the present invention may also contain atomic isotopes in non-natural proportions on one or more atoms constituting such compounds. For example, the compounds may have a stable isotope enriched at a level (e.g., at least 50% or at least 75%) on one or more atoms of the compound, such as deuterium ( 2 H) or carbon-13 ( 13 C). For example, the compound may include a radioactive isotope, such as tritium (C). 3 H), Iodine-125 ( 125 I) or carbon-14 ( 14 C). All isotopic variants of the compounds of this invention, whether or not they are radioactive, are included within the scope of this invention.

[0060] As used herein, the term "radiolabeling" refers to radioactive isotopes that readily form cations. Exemplary radiolabeling includes... 44 Sc、 47 Sc、 51 Cr 52m Mn, 58 Co、 52 Fe、 56 Ni、 57 Ni、 62 Cu、 64 Cu、 67 Cu、 66 Ga、 68 Ga、 67 Ga、 89 Zr、 90 Y、 89 Y、 94m Tc, 99m Tc, 97 Ru、 105 Rh、 109 Pd,111 Ag、 110m In、 111 In、 113m In、 114m In、 117m Sn、 121 Sn、 127 Te、 142 Pr、 143 Pr、 149 Pm, 151 Pm, 149 Tb, 153 Sm、 157 Gd, 161 Tb, 166 Ho、 165 Dy、 169 Er、 169 Yb、 175 Yb、 172 Tm、 177 Lu、 186 Re、 188 Re、 191 Pt, 197 Hg, 198 Au、 199 Au、 212 Pb, 203 Pb, 211 At、 212 Bi、 213 Bi、 223 Ra、 225 Ac and 227 Th, or containing 18 F-cation molecules, for example 18 F-[AlF] 2+ Exemplary radioactive markings also include 44 Sc、 47 Sc、 64 Cu、 67 Cu、 68 Ga、 90 Y、 111 In、 161 Tb, 166 Ho、 177 Lu、 188 Re、 212 Pb, 212 Bi、 213 Bi、 225 Ac and 227 Th, or containing 18 F-cation molecules. Preferred radiolabeling may include... 68 Ga、 90 Y、 177 Lu、212 Bihe 213 Bi. The radioactive label can be a gamma emitter, i.e., a radioactive label that emits gamma radiation when it decays; a beta emitter, i.e., a radioactive label that emits beta particles when it decays; or an alpha emitter, i.e., a radioactive label that emits alpha particles when it decays. Exemplary gamma emitters include 99m Tc and 111 In. An exemplary beta emitter includes 90 Y、 166 Ho and 68 Ga、 177 Lu. Exemplary alpha emitters include 225 Ac、 224 Ra and 213 Bi. "Chelated radiolabel" refers to a radiolabel (e.g., a radioactive cation) complexed with a polydentate ligand. The chelation portion described herein refers to a polydentate ligand used according to this disclosure. Exemplary polydentate ligands used according to this disclosure include -MAS3, -MAG3, -DOTA-GA, -DOTA, and -DTPA.

[0061] As used herein, the term "pharmaceutical formulation" includes a formulation comprising at least one active compound and optionally one or more other pharmaceutically acceptable ingredients (e.g., a pharmaceutically acceptable carrier). A pharmaceutical formulation is also a pharmaceutical composition when it comprises two or more active compounds, or at least one active compound and one or more other pharmaceutically acceptable ingredients. Unless the context otherwise requires, all references to "formulation" herein refer to a pharmaceutical formulation.

[0062] As used herein, the term "product" or "product of the invention" includes any product containing the compounds of the invention. In particular, the term "product" relates to compositions and formulations comprising the compounds of the invention, such as pharmaceutical compositions.

[0063] As used herein, the term "therapeuticly effective amount" refers to the amount of drug or agent that, within a reasonable pharmacological judgment, is calculated (or will) provide a desired therapeutic response in a mammal (animal or human). This therapeutic response may, for example, be used to cure, delay the progression of a disease, symptom, or condition, or to prevent the disease, symptom, or condition.

[0064] As used herein, the term "imaging effective amount" refers to the amount of a compound or formulation that, within reasonable clinical or experimental experience, is calculated (or will) provide a sufficient response for imaging (e.g., based on the detection of radioactive decay or fluorescence). Compounds or formulations used for imaging may be provided in an imaging effective amount.

[0065] compound

[0066] The compounds of the present invention can be used as tracers. In particular, the compounds of the present invention bind a probe (e.g., a PSMA probe) to a fluorophore and a radiolabelable portion (e.g., by chelating a radioisotope). When these compounds contain a radiolabel, they can be considered as hybrid tracers because the probe portion is coupled to two complementary labels: a fluorophore and a radiolabel.

[0067] These compounds can be considered small-molecule mixed PSMA tracers that benefit from a general (target carrier moiety)-(hydrophobic dye moiety)-(chelate moiety) design. As described herein, the efficacy of such tracers can be optimized by fine-tuning the individual moieties of the compound. Furthermore, these findings indicate that this design concept remains valid when, for example, the target carrier, dye, or chelate is altered. For example, the dye moiety may contain Cy5 or Cy7 analogs, and the chelate moiety may contain chelate moieties disclosed herein, such as MAS3 or DOTAGA. The choice of dye affects the wavelength used for surgical guidance (Cy5 is far-infrared, Cy7 is near-infrared). Appropriate selection of the chelate moiety adapts to the chelation of different radiolabeled materials. For example, chelating agents that can be used with alpha emitters (e.g., 225 Ac、 224 Ra、 213 Bi), which can be used with therapeutic isotopes; while chelating agents (e.g., gamma emitters) can be used with gamma emitters. 99m Tc, 111 In), it can be used with radiolabeled imaging isotopes. Most chelates are capable of coordinating different radiolabels. For example, DOTAGA can chelate... 111 In、 68 Ga、 177 Lu et al.; MAS3 can chelate 99 mTc, 188 Re, etc.

[0068] Targeting Carrier: To generate an optimized PSMA-targeting compound, the compound needs to be designed such that the targeting carrier moiety (e.g., -EuX (X is K, FA, PG, or E)) should be carefully positioned within the enzyme. The targeting carrier primarily utilizes aspartic acid (S1 position) and glutamate (S1' position) binding sites, while urea can bind to Zn. 2+ Coordination.

[0069] Hydrophobic dye unit: Appropriate hydrophobic (aromatic) substituents can bind to an auxiliary hydrophobic pocket located adjacent to the S1 position, thereby further enhancing acceptor affinity. Therefore, many PSMA-targeted radiotracers contain spacer molecules that facilitate secondary binding to the auxiliary hydrophobic pocket. Furthermore, it has been demonstrated that the appropriate placement of at least one anionic moiety (e.g., -SO3-) can further enhance acceptor affinity. This paper presents PSMA-targeting compounds using bifunctional dye moieties as linker molecule, representing a paradigm shift in the design of fluorescent PSMA tracers. This design concept has been demonstrated for bifunctional Cy5 and Cy7 analogs and therefore appears to be applicable to all bifunctional anthocyanin dye analogs.

[0070] Chelating Unit: In small molecule PSMA tracers, radioisotope-bound chelates are traditionally located outside the pharmacophore. Many different chelates have been reported to be used with various radioisotopes, ranging from diagnostics to therapy. Bifunctional dyes replace the traditional spacers in the compounds of this invention. The novel hybrid tracer design of the compounds of this invention also accommodates the integration of different chelates (e.g., MAS3 and DOTAGA). As will be understood by those skilled in the art, this teaching can be extended to accommodate other common chelates (e.g., MAG3, DOTA, NOA) and matching isotopes.

[0071] On one hand, the present invention provides compounds of formula I (or formula Ia) as described above, or pharmaceutically acceptable salts, stereoisomers, or prodrugs thereof. In embodiments, R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 One or more of V, W, Y1, Y2, Z, L1, L2, L3, L4, m, n, and p are as described in the following paragraphs:

[0072] At least one of the substituents R1, R2, R5, R7, and R8 in the first indole moiety may be different from the substituents R3, R4, R6, R9, and R8 in the second indole moiety. 10 At least one of the following, thereby providing an asymmetric dye moiety. For the purposes of this invention, the dye moiety represents a first indole moiety, a second indole moiety, a polymethyl linker, and substituents R1-R. 10 In the context of dye moieties, asymmetry means that for at least one pair of corresponding substituents in the first and second indole moieties, the substituents are different. That is, in asymmetric dye moieties, at least one of the following conditions applies: R1 is different from R3, R2 is different from R4, R5 is different from R6, or R7 and R8 are different from R9 and R... 10 For example, R1 may be different from R3. For example, R2 may be different from R4.

[0073] We have determined that compounds of formula I or Ia with an asymmetric dye moiety exhibit enhanced utility as tracers. For example, compounds with at least one of R1 and R2 (but not R3 and R4) carrying a charge (e.g., a negative charge, such as sulfonates) have shown surprisingly good clearance and tumor specificity. Without wishing to be bound by any theory, it is believed that these advantages may be due to alterations in binding activity, the effect of the dye as part of the compound's pharmacophore (as evidenced by, for example, a lower IC50), and alterations in renal clearance, which could potentially provide reduced toxicity.

[0074] R1 can be selected from sulfonate, carboxyl, phosphonate, amine, azide, and H. R1 can be selected from sulfonate, carboxyl, phosphonate, and H. R1 can be selected from sulfonate, carboxyl, and phosphonate. R1 can be selected from amine, azide, and H. R1 can be selected from amine and azide. R1 may be sulfonate. R1 may be carboxyl. R1 may be phosphonate. R1 may be amine. R1 may be azide. R1 may be H.

[0075] R2 can be selected from sulfonate, carboxyl, phosphonate, amine, azide, and H. R2 can be selected from sulfonate, carboxyl, phosphonate, and H. R2 can be selected from sulfonate, carboxyl, and phosphonate. R2 can be selected from amine, azide, and H. R2 can be selected from amine and azide. R2 may be sulfonate. R2 may be carboxyl. R2 may be phosphonate. R2 may be amine. R2 may be azide. R2 may be H.

[0076] R1 and R2 together can form an aryl group, which is substituted by one or two groups independently selected from sulfonates, carboxyl groups, phosphonates, amines, and azides. R1 and R2 together can also form an unsubstituted aryl group. The aryl group may be or contain a phenyl group. The aryl group may be or contain a naphthyl group.

[0077] R3 can be selected from sulfonate, carboxyl, phosphonate, amine, azide, and H. R3 can be selected from sulfonate, carboxyl, phosphonate, and H. R3 can be selected from sulfonate, carboxyl, and phosphonate. R3 can be selected from amine, azide, and H. R3 can be selected from amine and azide. R3 may be sulfonate. R3 may be carboxyl. R3 may be phosphonate. R3 may be amine. R3 may be azide. R3 may be H.

[0078] R4 can be selected from sulfonate, carboxyl, phosphonate, amine, azide, and H. R4 can be selected from sulfonate, carboxyl, phosphonate, and H. R4 can be selected from sulfonate, carboxyl, and phosphonate. R4 can be selected from amine, azide, and H. R4 can be selected from amine and azide. R4 may be a sulfonate. R4 may be a carboxyl group. R4 may be a phosphonate. R4 may be an amine. R4 may be an azide. R4 may be H.

[0079] R3 and R4 together can form an aryl group, wherein the aryl group is substituted by one or two groups independently selected from sulfonates, carboxyl groups, phosphonates, amines, and azides. R3 and R4 together can also form an unsubstituted aryl group. The aryl group may be or contain a phenyl group. The aryl group may be or contain a naphthyl group.

[0080] At least one of R1 and R2 may not be H. At least one of R3 and R4 may not be H. At least one of R1, R2, R3 and R4 may not be H.

[0081] Specifically, at least one of R1 and R2 (e.g., R1) may be selected from sulfonates, carboxyl groups, phosphonates, amines, and azides; and R3 and R4 may be H. For example, at least one of R1 and R2 (e.g., R1) may be selected from sulfonates, carboxyl groups, and phosphonates; and R3 and R4 may be H.

[0082] R5 can be selected from sulfonates, carboxyl groups, phosphonates, amines, azides, CH3, CH2CH3, and H. R5 can be selected from sulfonates, carboxyl groups, phosphonates, and H. R5 can be selected from sulfonates, carboxyl groups, and phosphonates. R5 can be selected from amines, azides, and H. R5 can be selected from amines and azides. R5 may be a sulfonate. R5 may be a carboxyl group. R5 may be a phosphonate. R5 may be an amine. R5 may be an azide. R5 may be H.

[0083] R6 can be selected from sulfonates, carboxyl groups, phosphonates, amines, azides, CH3, CH2CH3, and H. R6 can be selected from sulfonates, carboxyl groups, phosphonates, and H. R6 can be selected from sulfonates, carboxyl groups, and phosphonates. R6 can be selected from amines, azides, and H. R6 can be selected from amines and azides. R6 may be a sulfonate. R6 may be a carboxyl group. R6 may be a phosphonate. R6 may be an amine. R6 may be an azide. R6 may be H.

[0084] R7, R8, R9 and R 10 Each of these can be independently selected from sulfonates, carboxyl groups, phosphonates, amines, azides, CH3, CH2CH3, and H. R7, R8, R9, and R 10Each of these can be independently selected from CH3, CH2CH3, and H. R7, R8, R9, and R 10 Each of these can be independently selected from CH3 and CH2CH3. R7, R8, R9, and R... 10 Each of these can be CH3. R7, R8, R9, and R 10 Each of these can be CH2CH3. R7, R8, R9, and R... 10 Each of them can be H.

[0085] In one embodiment, no more than two of the substituents R1, R2, R5, R7, and R8 of the first indole moiety are sulfonates, carboxyl groups, or phosphonates; and / or the substituents R3, R4, R6, R9, and R8 of the second indole moiety are sulfonates, carboxyl groups, or phosphonates. 10 No more than two of them are sulfonates, carboxylates, or phosphonates. In one embodiment, at least three of R1, R2, R5, R7, and R8 are independently selected from CH3, CH2CH3, and H; and / or R3, R4, R6, R9, R 10 At least three of them are independently selected from CH3, CH2CH3 and H.

[0086] n can be 2. n can be 3.

[0087] V can be -CH2-. V can be -CH2CH2O-. m can be 4, 5, 6, 7, 8, 9, 10, 11, or 12. m can be 4, 5, 6, 7, or 8; for example, m can be 4, 5, or 6. For example, m can be 5.

[0088] W can be -CH2-. W can be -CH2CH2O-. p can be 4, 5, 6, 7, 8, 9, 10, 11, or 12. p can be 4, 5, 6, 7, or 8; p can be 4, 5, or 6. For example, p can be 5.

[0089] Y1 can be -EuK, -EuFA, -EuPG, -L1-EuK, -L1-EuFA, -L1-EuPG, or -L3-EuE; where EuK, EuFA, EuPG, and EuE are defined as in Table 1. Y1 can be -EuK, -EuFA, -EuPG, or -L3-EuE. Y1 can be -L1-EuK, -L1-EuFA, -L1-EuPG, or -L3-EuE. Y1 can be -EuK. Y1 can be -EuFA. Y1 can be -EuPG. Y1 can be -L1-EuK. Y1 can be -L1-EuFA. Y1 can be -L1-EuPG. Y1 can be -L3-EuE.

[0090] Y2 can be -L4-EuK, -L4-EuFA, -L4-EuPG, -EuE, or -L2-EuE; where EuK, EuFA, EuPG, and EuE are defined as in Table 1. Y2 can be -L4-EuK, -L4-EuAF, or -L4-EuPG. Y2 can be -EuE or -L2-EuE. Y2 can be -L2-EuK. Y2 can be -L2-EuFA. Y2 can be -L2-EuPG. Y2 can be -EuE. Y2 can be -L2-EuE.

[0091] Z can be a chelating moiety, which is a residue of a chelating agent as defined herein. For example, Z can be or contain bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (CBTE2a), cyclohexyl-1,2-diaminotetraacetic acid (CDTA), 4-(1,4,8,11-tetraazabicyclotetradecane-1-yl)-methylbenzoic acid (CPTA), N′-[5-[acetyl(hydroxy)amino]pentyl]-N-[5-[[4-[5-aminopentyl-(hydroxy)amino]-4-oxobutyryl]amino]pentyl]-N-hydroxybutyramide (DFO), 4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (DO2A), 1,4,7,10-tetraazabicyclodecadecane, etc. Dialkyl-N,N',N”,N”'-tetraacetic acid (DOTA), 2-[1,4,7,10-tetraazacyclododecane-4,7,10-triacetic acid]-glutaric acid (DOTAGA), N,N'-dipyridinoxyethylethylenediamine-N,N'-diacetic acid-5,5'-bis(phosphoric acid) (DPDP), diethylenetriaminepentaacetic acid (DTPA), ethylenediamine-N,N'-tetraacetic acid (EDTA), ethylene glycol-O,O-bis(2-aminoethyl)-N,N,N',N'-tetraacetic acid (EGTA), N,N-bis(hydroxybenzyl)-ethylenediamine-N,N'-diacetic acid (HBED), hydroxyethyldiaminetriacetic acid (HEDTA), 1-(p-nitrobenzyl)-1 4,7,10-Tetraazacyclodecane-4,7,10-triacetic acid ester (HP-DOA3), 6-hydrazino-N-methylpyridine-3-carboxamide (HYNIC), 1,4,7-triazacyclononane-1-succinic acid-4,7-diacetic acid (NODASA), 1-(1-carboxy-3-carboxypropyl)-4,7-(carboxyl)-1,4,7-triazacyclononane (NODAGA), 1,4,7-triazacyclononanetriacetic acid (NOTA), 4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (TE2A), 1,4,8,11-tetraazacyclododecane-1,4,8,11-tetraacetic acid (TETA) The residues of terpyridine-bis(methyleneaminotetraacetic acid (TMT), 1,4,7,10-tetraazacyclo-tetrazane-N,N',N”,N”'-tetraacetic acid (TRITA), triethylenetetraminehexaacetic acid (TTHA), N,N′-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18-crown-6 (H2macropa) and 4-amino-4-{2-[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydro-pyridin-2-ylmethyl)-carbamoyl]-ethyl}pimelic acid bis-[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydro-pyridin-2-ylmethyl)-amide] (THP).Z can be -MAS3, -MAG3, -DOTA-GA, -DOTA, -DTPA, -L2-MAS3, -L2-MAG3, -L2-DOTA-GA, -L2-DOTA, -L2-DTPA; where MAS3, MAG3, DOTA-GA, DOTA, and DTPA are defined as shown in Table 1. Z can be -MAS3, -MAG3, -DOTA-GA, -DOTA, -DTPA. Z can be -L2-MAS3, -L2-MAG3, -L2-DOTA-GA, -L2-DOTA, -L2-DTPA, Z can be -MAS3 or -MAG3. Z can be -DOTA-GA or -DOTA. Z can be -MAS3. Z can be -MAG3. Z can be -DOTA-GA. Z can be -DOTA. Z can be -DTPA. Z can be -L2-MAS3. Z can be -L2-MAG3. Z can be -L2-DOTA-GA. Z can be -L2-DOTA. Z can also be -L2-DTPA.

[0092] Table 1: Exemplary substituents Y1, Y2, and Z, wherein Indicates the connection point with the rest of the compound.

[0093]

[0094]

[0095]

[0096] L1 can be of formula -NH-R 12 -C(O)- connector, where R 12 It is a bond, or a substituted or unsubstituted alkyl group. For example, R 12 It can be a key. For example, R 12 It can be a substituted or unsubstituted straight-chain or branched alkyl moiety having 2, 3, 4, 5, 6, 7, or 8 carbon atoms. L1 can be a lysine residue, ornithine residue, aspartic acid residue, glutamic acid residue, or -NH-(C4-C7 alkyl)-C(O)- residue. L1 can be a lysine residue. L1 can be an ornithine residue. L1 can be an aspartic acid residue. L1 can be a glutamic acid residue. L1 can be -NH-(C4-C7 alkyl)-NH-, for example, L1 can be -NH-(C5 alkyl)-NH- or L1 can be -NH-(C6 alkyl)-NH-.

[0097] L2 can be expressed as -C(O)-R 13 -NH- connector, where R 13 It is a bond, or a substituted or unsubstituted alkyl group. For example, R 13It can be a key. For example, R 13 It can be a substituted or unsubstituted straight-chain or branched alkyl moiety having 2, 3, 4, 5, 6, 7, or 8 carbon atoms. L2 can be a lysine residue, ornithine residue, aspartic acid residue, glutamic acid residue, or a -C(O)-(C4-C7 alkyl)-NH- residue. L2 can be a lysine residue. L2 can be an ornithine residue. L2 can be an aspartic acid residue. L2 can be a glutamic acid residue. L2 can be -NH-(C4-C7 alkyl)-NH-, for example, L2 can be -NH-(C5 alkyl)-NH- or L2 can be -NH-(C6 alkyl)-NH-.

[0098] L3 can be of the formula -NH-R 14 -NH- connector, where R 14 It is a substituted or unsubstituted alkyl group. For example, R 14 It can be a substituted or unsubstituted straight-chain or branched alkyl moiety having 4, 5, 6, 7, or 8 carbon atoms. L3 can be a lysine residue, an ornithine residue, or a -NH-(C4-C7 alkyl)-NH- residue. L3 can be a lysine residue. L3 can be an ornithine residue. L3 can be -NH-(C4-C7 alkyl)-NH-, for example, L3 can be -NH-(C5 alkyl)-NH- or L3 can be -NH-(C6 alkyl)-NH-.

[0099] L4 can be represented by the formula -C(O)-R 15 -C(O)- connector, where R 13 It is a substituted or unsubstituted alkyl group. For example, R 13 It can be a substituted or unsubstituted straight-chain or branched alkyl moiety having 4, 5, 6, 7, or 8 carbon atoms. L4 can be an aspartic acid residue, a glutamic acid residue, or a -C(O)-(C4-C7 alkyl)-C(O)- residue. L4 can be an aspartic acid residue. L4 can be a glutamic acid residue. L4 can be -C(O)-(C4-C7 alkyl)-(CO)-, for example, L4 can be -C(O)-(C5 alkyl)-(CO)-, or L4 can be -C(O)-(C6 alkyl)-C(O)-.

[0100] The compound may be selected from the following:

[0101]

[0102]

[0103]

[0104]

[0105]

[0106]

[0107]

[0108] The compound may contain a chelated radiolabeled element. The chelated radiolabeled element may be selected from gamma emitters (e.g., gamma emitters). 99m Tc, 111 In), β-emitters (e.g.) 90 Y、 166 Ho、 68 Ga、 177 Lu) and α-emitters (e.g. 225 Ac、 224 Ra、 213 Bi), or combinations thereof. The chelated radioactive label may be a gamma emitter; for example... 99m Tc, 111 In, or combinations thereof. The chelated radioactive label may be a β-emitter; for example... 90 Y、 166 Ho、 68 Ga、 177 Lu, or a combination thereof. The chelated radioactive label may be an α-emitter; for example... 225 Ac、 224 Ra、 213 Bi, or combinations thereof.

[0109] On the other hand, the present invention provides compounds of formula II (or formula IIa) as described above, or pharmaceutically acceptable salts, stereoisomers, or prodrugs thereof. In embodiments, R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 One or more of V, W, Z, n, and p are as defined herein. Compounds of this aspect as compounds of formula II represent intermediates of compounds of formula I. Compounds of this aspect as compounds of formula IIa represent intermediates of compounds of formula Ia. Furthermore, such compounds can bind to a probe (e.g., to form -V...). m -C(O)- probes, rather than binding to -EuK, -EuFA, or -EuPG. Therefore, compounds of Formula II can represent useful intermediates for other mixed probes. Compounds of Formula II may contain chelated radiolabels as defined herein.

[0110] On the other hand, the present invention provides compounds of formula III (or formula IIIa) as described above, or pharmaceutically acceptable salts, stereoisomers, or prodrugs thereof. In embodiments, R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10One or more of V, W, Y, n, and p are as defined herein. These compounds can be linked to the Z portion using chemical methods known to those skilled in the art, thereby providing, for example, compounds of formula I (or formula Ia). Thus, compounds of formula III represent intermediates of compounds of formula I, and compounds of formula IIIa represent intermediates of compounds of formula Ia.

[0111] Preparation and application

[0112] The compounds of the present invention can be administered parenterally, for example, intravenously or intraprostatically. The compounds of the present invention can be administered intravenously. The compounds of the present invention can be administered intraprostatically. The compounds can be administered in the form of pharmaceutical preparations comprising the compounds in free form, or, for example, in a pharmaceutically acceptable dosage form of a pharmaceutically acceptable non-toxic organic or inorganic acid or base addition salt. Depending on the condition and imaging requirements, the compositions can be administered at different dosages.

[0113] Therefore, according to another aspect of the invention, a pharmaceutical formulation or composition is provided, comprising the compounds of the invention, optionally mixed with a pharmaceutically acceptable adjuvant, diluent, or carrier. The formulation or composition may also contain compounds that affect the in vivo pharmacokinetics of the compounds of the invention. The formulation may also contain at least one compound that blocks or reduces the uptake of the compounds of the invention in an organ (or multiple organs). For example, the formulation may also contain a compound that blocks or reduces the uptake of the compounds in the kidneys and / or salivary glands. Examples of such compounds include mannitol, 2-(phosphonomethyl)glutarate, monosodium glutamate, succinyl gelatin, and albumin fragments.

[0114] The injectable pharmaceutical formulations or compositions of the present invention may comprise pharmaceutically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions, as well as sterile powders for reconstitution into sterile injectable solutions or dispersions prior to use. Examples of suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils (e.g., olive oil), and injectable organic esters, such as ethyl oleate.

[0115] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifiers, and dispersants. The inhibitory effect on microorganisms can be ensured by adding various antibacterial and antifungal agents, such as parabens, chlorobutanol, or phenolic sorbic acid. It may also be desirable to include isotonic agents, such as sugars or sodium chloride.

[0116] Formulations based on this subject may also contain inactive ingredients. Suitable inactive ingredients are well known in the art and described in standard textbooks, such as Goodman and Gillman's *The Pharmacological Bases of Therapeutics*, 8... th Ed., Gilman et al, Eds. Pergamon Press (1990), and Remington's Pharmaceutical Sciences, 17 th Ed., Mack Publishing Co., Easton, Pa. (1990), the entire contents of which are incorporated herein by reference.

[0117] The formulations may be used in combination with other pharmaceutical dosage forms to enhance their efficacy in treating any of the conditions described herein. In this regard, the formulations of the present invention may be administered as part of a regimen that further includes any other drugs and / or pharmaceutical dosage forms known in the art as effective in treating any of these diseases. The additional pharmaceutical dosage forms may contain at least one compound that blocks or reduces the uptake of the compounds of the present invention in an organ (or multiple organs). For example, the additional pharmaceutical dosage forms may contain compounds that block or reduce the uptake of said compounds in the kidneys and / or salivary glands. Examples of such compounds include mannitol, 2-(phosphonomethyl)glutarate, monosodium glutamate, succinyl gelatin, and albumin fragments. These compounds will also be discussed in the following paragraphs.

[0118] Mannitol. In the kidneys, PSMA is expressed only in the proximal tubules (Urology (2007) 70:385–90), where osmotic diuretics (such as mannitol) exert their pharmacological effects. Because mannitol is not reabsorbed by the renal tubules, it is used to promote diuresis, thereby increasing osmotic pressure, promoting water excretion, and inhibiting the reabsorption of sodium, chloride, and other solutes by the renal tubules. Mannitol has therefore been used to reduce the dose of radiolabeled PSMA delivered to the kidneys (EJNMMI (2017) 44:2189–2194).

[0119] 2-(phosphonomethyl)glutaric acid (2-PMPA). 2-PMPA is a PSMA inhibitor that has been used by Kratochwil et al. to reduce renal accumulation due to subsequent administration. They concluded that PMPA blocks the PSMA receptor because they hypothesized that "subsequent administration of high doses of a PSMA competitor may no longer block tumor uptake, but may still displace 'un- or 'not yet-internalized' PSMA ligands from renal tubular cells, thereby increasing the expected tumor-to-renal dose ratio" (J Nucl Med (2015) 56:293–298). Later studies found that 2-PMPA is useful for co-injection with PSMAI&T, but it may reduce the effective dose because tumor uptake is also significantly reduced (Theranostics (2016) 12:849–861).

[0120] Monosodium glutamate (MSG). MSG is a well-studied food additive that stimulates salivation. Rousseau et al. used it to block the uptake of 68GaPSMA-11 by the salivary glands (J Nucl Med August 2018). They also observed a significant reduction in the renal uptake of this tracer in MSG-treated mice compared to the control group. Notably, tumor uptake was not significantly reduced (unlike co-injection of PMPA).

[0121] Succinyl gelatin. Studies have shown that the co-administration of basic compounds, particularly amino acids such as lysine and arginine, can significantly reduce the concentration of radioactivity in the kidneys, in some cases by up to 60% (J Nucl Med (1997) 38:1929–1933; J Nucl Med (2002) 46:181–194; Scand J Clin Lab Invest (1977) 37:477–486; Eur J Nucl Med (1998) 25:201–212). Utilizing this mechanism, researchers have successfully reduced renal accumulation by co-administering the peptide-based succinyl gelatin (GELO) plasma bulking agent gelofusine (J Nucl Med (2006) 47:528–533; J Nucl Med (2006) 47:432–436).

[0122] Albumin fragments. Macroproteins are well known to be involved in the binding and uptake of hydrophilic (radiolabeled) peptides (EJNMMI(2011)38:623-632). Albumin is a natural ligand of macroproteins, but only a small fraction of circulating albumin is filtered in the glomerulus. This small fraction is reabsorbed by processes such as macroprotein-mediated endocytosis. Albumin-derived peptide fragments are present in higher concentrations in the proximal tubules than intact albumin and are potent inhibitors of renal reabsorption of various radiolabeled peptides (J Nucl Med(2008)49:1506-1511; EJNMMI(2010)37:226-234).

[0123] Imaging and other applications

[0124] The compounds of the present invention are tracers for imaging. In particular, the compounds of the present invention (e.g., compounds of formula I or Ia) can be used to image tumors expressing PMSA (e.g., prostate cancer tumors). These tumors include prostate cancer tumors, kidney tumors, breast cancer tumors, gliomas, colorectal adenocarcinomas, transitional cell carcinomas, pancreatic ductal adenocarcinomas, and gastric adenocarcinomas.

[0125] Radiolabeled tracers containing PSMA probes have been used for PSMA-targeted diagnostic imaging in prostate cancer. For example, Eder et al, Bioconjug. Chem., 2012, 23(4), 688-697, describe […]. 68 Ga]PSMA-11PSMA PET, which utilizes a urea-based PSMA inhibitor... 68 Ga complexes are used to provide PET imaging for prostate cancer. In another example, 99mTc-PSMA I&S has been successfully used in image-guided surgery, but this compound does not allow for high-resolution visualization of the tumor or its edges. While these types of methods can be used to identify metastatic tumors, PET and other gamma-ray-based imaging techniques lack sufficient resolution to display the fine edges of tumors in vivo.

[0126] The compounds of the present invention, particularly those of formula I (or formula Ia), comprise the structure YFZ, wherein Y comprises a targeting carrier, Z comprises a radiolabeled substance (such as a chelated radionucleotide), and F comprises a fluorophore (such as an anthocyanin dye (Cy5 or Cy7) analog), each "-" representing a bond or linker. The probe can target PSMA, particularly PSMA expressed in tumors. The radiolabeled substance can be detected by γ / β-tracing / imaging and is useful for imaging in preoperative planning, intraoperative, and postoperative evaluation. The fluorophore can be detected by fluorescence imaging / tracing, a method that provides higher resolution and can be used to more clearly define tumor margins. This can be advantageous, for example, during tumor resection surgery, as it can help guide the removal of all diseased tissue while preserving healthy tissue.

[0127] The specific structure of the compounds may also be advantageous for imaging and other applications, particularly for compounds targeting PSMA. Exemplary PMSA-targeting compounds of this disclosure may advantageously exhibit relatively low uptake in background organs compared to existing PMSA-targeting compounds. For example, compounds of formula I (or formula Ia) having an asymmetric dye moiety (e.g., an asymmetric fluorophore F) may exhibit enhanced tracer efficacy. Without wishing to be bound by any theory, these advantages are believed to be due to altered binding affinity (e.g., a lower IC-50) and changes in renal clearance and salivary gland uptake due to the structure of the compounds, potentially providing reduced background and toxicity.

[0128] The probe (Y) typically targets the PSMA. For example, EuK and related probes provide a vector that targets the aspartic acid (S1 position) and glutamate (S1' position) binding sites of the PSMA. The PSMA also includes an associated hydrophobic pocket (Barinka et al., J. Med. Chem., 2008, 51, 7737-43; the contents of which are incorporated herein by reference in their entirety), and it is believed that the hydrophobic substituents of the dye moiety (fluorophore F) can bind to the auxiliary hydrophobic binding pocket. Therefore, altering the hydrophobic substituents of the dye moiety can be used to modulate receptor affinity. Charged substituents of the dye moiety (e.g., sulfonates) can also interact with the auxiliary pocket. Therefore, altering the charged substituents of the dye moiety can also be used to modulate receptor affinity.

[0129] One aspect provides a method for imaging a tumor, comprising administering a compound or formulation of the present invention to a subject and imaging the tumor after a predetermined time. Related aspects provide the use of the compound or formulation of the present invention in imaging.

[0130] The compounds of the present invention may be compounds of Formula I and / or compounds of Formula Ia. The predetermined time may be a predetermined time prior to intraoperative imaging. In some cases, imaging may be dynamic, in which case the predetermined time may be as low as 0 hours. The predetermined time, for example, prior to intraoperative imaging, may be at least 0 hours, at least 0.25 hours, at least 0.5 hours, or at least 1 hour. The predetermined time may not exceed 48 hours. For example, the predetermined time may not exceed 24 hours. The predetermined time is, for example, at least about 0.5 hours and no more than about 48 hours prior to intraoperative imaging. For example, the predetermined time may be at least about 1 hour (or about 2 hours) and no more than about 36 hours; for example, the predetermined time may be at least about 3 hours and no more than about 24 hours.

[0131] The compound (e.g., a compound of formula I or formula Ia) may contain a chelated radiolabel, and imaging may include imaging of radioactive decay. Imaging of radioactive decay may include at least one of positron emission tomography (PET), single-photon emission computed tomography (SPECT), scintillation gamma-ray tracing / imaging, beta-ray tracing, intraoperative gamma-ray tracing / imaging, or intraoperative beta-ray tracing. Imaging may include positron emission tomography (PET), single-photon emission computed tomography (SPECT), scintillation, (intraoperative) gamma-ray tracing / imaging, or (intraoperative) beta-ray tracing. For example, imaging may include PET or SPECT. Imaging may include PET. Imaging may include SPECT.

[0132] Imaging can include fluorescence imaging. Imaging can include fluorescence spectroscopy.

[0133] Imaging can include radioactive decay imaging and fluorescence imaging. Imaging can include imaging of radioactive decay prior to fluorescence imaging. For example, radioactive decay imaging can be performed before surgery, while fluorescence imaging can be performed during surgery.

[0134] The imaging method may further include administering to the subject at least one compound that blocks or reduces the uptake of the compound of the present invention in an organ (or multiple organs). This can reduce the background signal of the compound of the present invention in the relevant organ. For example, at least one compound can block or reduce the uptake of the compound of the present invention in the kidneys and / or salivary glands. Examples of such compounds include mannitol, 2-(phosphonomethyl)glutarate, monosodium glutamate, succinyl gelatin, and albumin fragments. The at least one blocking compound may be administered co-administered with the compound of the present invention (e.g., as a single dosage form), or the at least one blocking compound may be administered separately from the compound of the present invention.

[0135] The compounds and formulations of the present invention can also be used to treat diseases, particularly cancers (e.g., prostate cancer and / or other cancers expressing PSMA; such as kidney cancer, breast cancer, glioma, colorectal adenocarcinoma, transitional cell carcinoma, pancreatic ductal adenocarcinoma, or gastric adenocarcinoma). Such compounds (or formulations containing such compounds) may contain radiolabels disclosed herein. For example, the radiolabels may be selected from β-emitters (e.g.,...). 90 Y、 166 Ho、 68 Ga、 177 Lu) and α-emitters (e.g. 225 Ac、 224 Ra、 213 Bi). The compound provides a mixed tracer that can target PMSA, and when the compound contains a radiolabeled element (as a β-emitter and / or α-emitter), the compound can be used for radiotherapy. In such therapeutic applications, the primary role of the dye moiety can be to provide favorable pharmacokinetics (e.g., by prolonging the compound's circulating half-life, thus extending the time window for binding to PSMA).

[0136] One aspect provides the compounds or preparations of the present invention for use as medicines. A related aspect is the compounds or preparations of the present invention for treating cancer. Cancer can be a cancer comprising cells expressing PSMA. For example, cancer can be prostate cancer, kidney cancer, breast cancer, glioma, colorectal adenocarcinoma, transitional cell carcinoma, pancreatic ductal adenocarcinoma, or gastric adenocarcinoma; for example, cancer may be prostate cancer.

[0137] One aspect provides a method of treating cancer, comprising administering a compound or preparation of the invention to a patient in need of treatment. The cancer may be a cancer comprising cells expressing PSMA. For example, the cancer may be prostate cancer, kidney cancer, breast cancer, glioma, colorectal adenocarcinoma, transitional cell carcinoma, pancreatic ductal adenocarcinoma, or gastric adenocarcinoma; for example, the cancer may be prostate cancer.

[0138] The treatment methods disclosed herein may further include administering at least one compound that blocks or reduces the uptake of the compounds of the present invention in an organ (or multiple organs). This can reduce the background signal of the compounds of the present invention in the relevant organ. For example, at least one compound can block or reduce the uptake of the compounds of the present invention in the kidneys and / or salivary glands. Examples of such compounds include mannitol, 2-(phosphonomethyl)glutarate, monosodium glutamate, succinyl gelatin, and albumin fragments. The at least one blocking compound may be administered co-administered with the compounds of the present invention (e.g., as a single dosage form), or the at least one blocking compound may be administered separately from the compounds of the present invention.

[0139] Additional Implementation Plan

[0140] This invention and disclosure also include the subject matter of the following numbered items:

[0141] 1. Compounds of formula I or Ia:

[0142]

[0143]

[0144] in:

[0145] R1 is selected from sulfonates, carboxyl groups, phosphonates, amines, azides, and H; R2 is selected from sulfonates, carboxyl groups, phosphonates, amines, azides, and H; or R1 and R2 together form an aryl group, which is optionally substituted by 1 to 4 groups (e.g., one or two groups), each group being independently selected from sulfonates, carboxyl groups, phosphonates, amines, and azides.

[0146] R3 is selected from sulfonates, carboxyl groups, phosphonates, amines, azides, and H; R4 is selected from sulfonates, carboxyl groups, phosphonates, amines, azides, and H; or R3 and R4 together form an aryl group, which is optionally substituted by 1 to 4 groups (e.g., one or two groups), each of which is independently selected from sulfonates, carboxyl groups, phosphonates, amines, and azides.

[0147] R5, R6, R7, R8, R9 and R 10 Each group is independently selected from sulfonates, carboxyl groups, phosphonates, amines, azides, CH3, CH2CH3, and H;

[0148] V is -CH2- or -CH2CH2O-;

[0149] W is -CH2- or -CH2CH2O-;

[0150] Y1 is -EuK, -EuFA, -EuPG, -L1-EuK, -L1-EuFA, -L1-EuPG or -L3-EuE;

[0151] Y2 is -L4-EuK, -L4-EuFA, -L4-EuPG, -EuE, or -L2-EuE;

[0152] Z represents the chelate portion;

[0153] L1 is the formula -NH-R 12 -C(O)- connector, where R 12 It is a bond, or a substituted or unsubstituted alkyl group;

[0154] L2 is the formula -C(O)-R 13 -NH- connector, where R 13It is a bond, or a substituted or unsubstituted alkyl group;

[0155] L3 is the formula -NH-R 14 -NH- connector, where R 14 It is a substituted or unsubstituted alkyl group;

[0156] L4 is the formula -C(O)-R 15 -C(O)- connector, where R 15 It is a substituted or unsubstituted alkyl group;

[0157] n is 2 or 3;

[0158] m is 4-21; and

[0159] p is 3-21.

[0160] Or its pharmaceutically acceptable salt.

[0161] 2. The compound according to claim 1, wherein at least one of the substituents R1, R2, R5, R7, R8 of the first indole moiety is different from the substituents R3, R4, R6, R9, R8 of the second indole moiety. 10 At least one of them, thereby providing an asymmetric dye portion.

[0162] 3. The compound according to claim 1 or claim 2, wherein R1 is selected from sulfonates and H, and R2 is selected from sulfonates and H.

[0163] 4. The compound according to item 1 or item 2, wherein R1 is a sulfonate and R2 is H.

[0164] 5. The compound according to claim 1 or claim 2, wherein R1 and R2 together form an optionally substituted aryl group.

[0165] 6. The compound according to any one of the preceding claims, wherein R3 is selected from sulfonates and H, and R4 is selected from sulfonates and H.

[0166] 7. The compound according to any one of items 1-4, wherein R3 and R4 together form an optionally substituted aryl group.

[0167] 8. The compound according to any one of the preceding claims, wherein R5 is H and / or R6 is H.

[0168] 9. The compound according to any one of the preceding claims, wherein R7, R8, R9 and R 10 Each is either -CH3 or -H; optionally, R7, R8, R9, and R 10 Each is -CH3.

[0169] 10. The compound according to any one of the preceding claims, wherein V is -CH2- and / or W is -CH2-.

[0170] 11. The compound according to any one of the preceding claims, wherein m is 4-12; optionally, wherein m is 4, 5 or 6; further optionally, wherein m is 5.

[0171] 12. The compound according to any one of the preceding claims, wherein p is 3-12; optionally, wherein p is 3, 4 or 5; further optionally, wherein p is 4.

[0172] 13. The compound according to any one of the preceding claims, wherein n is 2.

[0173] 14. The compound according to any one of the preceding claims, wherein n is 3.

[0174] 15. The compound according to any one of the preceding claims, wherein Y1 is -EuK.

[0175] 16. The compound according to any one of the preceding claims, wherein Z is bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (CBTE2a), cyclohexyl-1,2-diaminotetraacetic acid (CDTA), 4-(1,4,8,11-tetraazabicyclotetradecane-1-yl)-methylbenzoic acid (CPTA), N′-[5-[acetyl(hydroxy)amino]pentyl]-N-[5-[[4-[5-aminopentyl-(hydroxy)amino]-4-oxobutyryl]amino]pentyl]-N-hydroxybutyramide (DFO), 4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (DO2A), 1,4,7 10-Tetraazacyclododecane-N,N',N”,N”'-tetraacetic acid (DOTA), 2-[1,4,7,10-tetraazacyclododecane-4,7,10-triacetic acid]-glutaric acid (DOTAGA), N,N'-dipyridinoxyethylethylenediamine-N,N'-diacetic acid-5,5'-bis(phosphoric acid) (DPDP), diethylenetriaminepentaacetic acid (DTPA), ethylenediamine-N,N'-tetraacetic acid (EDTA), ethylene glycol-O,O-bis(2-aminoethyl)-N,N,N',N'-tetraacetic acid (EGTA), N,N-bis(hydroxybenzyl)-ethylenediamine-N,N'-diacetic acid (HBED), hydroxyethyldiaminetriacetic acid (HEDTA), 1-(para-nitroglycerin) (Hydroxybenzyl)-1,4,7,10-tetraazacyclodecane-4,7,10-triacetate (HP-DOA3), 6-hydrazyl-N-methylpyridine-3-carboxamide (HYNIC), 1,4,7-triazacyclononane-1-succinic acid-4,7-diacetic acid (NODASA), 1-(1-carboxy-3-carboxypropyl)-4,7-(carboxyl)-1,4,7-triazacyclononane (NODAGA), 1,4,7-triazacyclononanetriacetic acid (NOTA), 4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (TE2A), 1,4,8,11-tetraazacyclododecane-1,4,8,11-tetraacetic acid (T ETA), terpyridine-bis(methyleneaminotetraacetic acid (TMT), 1,4,7,10-tetraazacyclo-tetrazane-N,N',N”,N”'-tetraacetic acid (TRITA), triethylenetetraminehexaacetic acid (TTHA), N,N′-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18-crown-6 (H2macropa) or 4-amino-4-{2-[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydro-pyridin-2-ylmethyl)-carbamoyl]-ethyl}pimelic acid bis-[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydro-pyridin-2-ylmethyl)-amide] (THP) residues.

[0176] 17. The compound according to any one of the preceding claims, wherein Z is -MAS3, -MAG3, -DOTA-GA, -DOTA or -DTPA; optionally Z is -MAS3.

[0177] 18. The compound according to any one of items 1-17, wherein the compound is selected from:

[0178]

[0179]

[0180]

[0181]

[0182]

[0183] 19. The compound according to any one of the preceding claims, further comprising a chelated radiolabeled marker, said radiolabeled marker optionally selected from... 44 Sc、 47 Sc、 51 Cr 52m Mn, 58 Co、 52 Fe、 56 Ni、 57 Ni、 62 Cu、 64 Cu、 67 Cu、 66 Ga、 68 Ga、 67 Ga、 89 Zr、 90 Y、 89 Y、 94m Tc, 99m Tc, 97 Ru、 105 Rh、 109 Pd, 111 Ag、 110m In、 111 In、 113m In、 114m In、 117m Sn、 121 Sn、 127 Te、 142 Pr、 143 Pr、 149 Pm, 151 Pm, 149 Tb, 153 Sm、 157 Gd, 161 Tb,166 Ho、 165 Dy、 169 Er、 169 Yb、 175 Yb、 172 Tm、 177 Lu、 186 Re、 188 Re、 191 Pt, 197 Hg, 198 Au、 199 Au、 212 Pb, 203 Pb, 211 At、 212 Bi、 213 Bi、 223 Ra、 225 Ac and 227 Th, or containing 18 F-cation molecules, for example 18 F-[AlF] 2+ Exemplary radioactive markings also include 44 Sc、 47 Sc、 64 Cu、 67 Cu、 68 Ga、 90 Y、 111 In、 161 Tb, 166 Ho、 177 Lu、 188 Re、 212 Pb, 212 Bi、 213 Bi、 225 Ac and 227 Th, or containing 18 F is a cationic molecule.

[0184] 20. The compound according to claim 19, wherein the chelated radiolabel is selected from γ-emitters, β-emitters, and α-emitters, or combinations thereof.

[0185] 21. The compound according to claim 19 or 20, wherein the chelated radiolabeled material is selected from... 68 Ga、 177 Lu and 99m Tc, or combinations thereof.

[0186] 22. A compound of formula II or IIa, or a pharmaceutically acceptable salt thereof:

[0187]

[0188] in:

[0189] R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 V, W, Z, L3, m, n and p are as defined in any one of claims 1 to 14, 16 or 17.

[0190] 23. The compound according to item 22, further comprising a chelated radiolabel as defined in any one of items 19 to 21.

[0191] 24. A compound of formula III or IIIa, or a pharmaceutically acceptable salt thereof:

[0192]

[0193] in:

[0194] R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 V, W, Y1, Y2, m, n and p are as defined in any one of claims 1 to 15.

[0195] 25. A formulation comprising any one of claims 1 to 21 and optionally a pharmaceutically acceptable carrier.

[0196] 26. The formulation according to item 25, further comprising at least one other anticancer compound.

[0197] 27. The formulation according to item 25 or the formulation according to item 25, further comprising a compound that affects the in vivo kinetics of the compound according to any one of items 1-21, optionally wherein said compound reduces renal retention of the compound according to any one of items 1-21.

[0198] 28. A method for imaging a tumor, comprising administering to a subject a compound of any one of items 1 to 21 or an agent of any one of items 25 to 27, and imaging the tumor after a predetermined time.

[0199] 29. The method according to item 28, wherein the predetermined time is at least about 1 hour and no more than about 48 hours.

[0200] 30. The method according to claim 28 or 29, wherein the tumor is a prostate cancer tumor, a kidney tumor, a breast cancer tumor, a glioma, a colorectal adenocarcinoma, a transitional cell carcinoma, a pancreatic ductal adenocarcinoma, or a gastric adenocarcinoma; optionally, wherein the tumor is a prostate cancer tumor.

[0201] 31. The method according to any one of claims 28 to 30, wherein the compound comprises a chelated radiolabeled substance, and the imaging comprises positron emission tomography (PET), single-photon emission computed tomography (SPECT), scintillation, intraoperative gamma tracing / imaging, or intraoperative beta tracing.

[0202] 32. The method according to item 31, wherein the imaging includes PET or SPECT.

[0203] 33. The method according to any one of items 28 to 32, wherein the imaging includes fluorescence imaging, and optionally wherein the imaging includes fluorescence spectroscopy.

[0204] 34. A method of treating cancer, comprising administering a compound as described in any one of items 1 to 21 or an agent as described in any one of items 25 to 27.

[0205] 35. The method according to claim 34, wherein the cancer is prostate cancer, kidney cancer, breast cancer, glioma, colorectal adenocarcinoma, transitional cell carcinoma, pancreatic ductal adenocarcinoma, or gastric adenocarcinoma, optionally wherein the cancer is prostate cancer.

[0206] 36. Use of any compound of any one of items 1 to 21 or any formulation of any one of items 25 to 27 in imaging.

[0207] 37. The compound of any one of items 1 to 21 or the preparation of any one of items 25 to 27, used as a medicine.

[0208] 38. The compound of any one of items 1 to 21 or the preparation of any one of items 25 to 27, for the treatment of cancer.

[0209] 39. The compound or preparation according to claim 38, wherein the cancer is prostate cancer, kidney cancer, breast cancer, glioma, colorectal adenocarcinoma, transitional cell carcinoma, pancreatic ductal adenocarcinoma, or gastric adenocarcinoma, optionally wherein the cancer is prostate cancer.

[0210] Example

[0211] Example 1: Synthesis of an exemplary compound

[0212] according to Figure 1The reaction scheme shown synthesizes the compound. In this reaction scheme, an anthocyanin fluorophore containing phthalimide generates 2) an amine-containing anthocyanin fluorophore in 1) the release of the amine. Binding to EuK results in 3) a fluorescent-only targeting precursor. The subsequent addition of the MAS3-NHS chelate generates 4) a mixed tracer precursor containing both fluorescence and a chelate for radiolabeling. 5) Deprotection of the EuK moiety leads to activation of the targeting EuK. a–g summarize the different indole substituents of the dyes used in the matrix. Although this synthesis has been demonstrated for the probe EuK and the chelate MAS3, those skilled in the art will understand that this synthesis can be readily applied to other probes (e.g., EuAF or EuPG) and other chelates (e.g., MAG3, DOTA-GA, DOTA, DTPA).

[0213] Using a standard solid-phase synthesis strategy, asymmetric Cy5 dyes with slightly modified structures (1a–1g; hereinafter named Phth-Cy5-COOH(1a); Phth(SO3)-Cy5-COOH(1b); Phth-Cy5-(SO3)COOH(1c); Phth(SO3)-Cy5-(SO3)COOH(1d); Phth(Ar)-Cy5-COOH(1e); Phth-Cy5-(Ar)COOH(1f); Phth(Ar)-Cy5-(Ar)COOH(1g)) were synthesized to investigate the properties of each cyanofluorophore. These dyes form mixed tracer analogs. To do this, several synthetic steps were performed: the phthalimide moiety was converted to a free amine, and a slightly modified Gabriel synthesis was used on these dyes to give 2a–2g. Therefore, the MAS3 chelate (including a six-carbon spacer with an activated carboxylic acid) forms a simple amide bond via NHS activation coupling, producing 3a-3g. Using PyBOP, the KuE(tBu)3 targeting moiety is continuously added to the carboxylic acid group, producing compound 4a-4g. To definitively determine the mixed tracers, three tert-butyl esters on the KuE moiety were removed by stirring in TFA:H2O 95:5, yielding tracers 5a-5g and 5a-5g (hereinafter named EuK-Cy5-MAS3(5a); EuK-(SO3)Cy5-MAS3(5b); EuK-Cy5-(SO3)MAS3(5c); EuK(SO3)-Cy5-(SO3)MAS3(5d); EuK(Ar)-Cy5-MAS3(5e); EuK-Cy5-(Ar)MAS3(5f); EuK(Ar)-Cy5-(Ar)MAS3)(5g) (Scheme 1). The above results yielded a matrix of mixed tracers with slight structural modifications to modulate chemical, photophysical, in vitro, and in vivo characterization.

[0214] Example 2: Other Synthetic Methods

[0215] EuK(Z)-(OtBu)3

[0216] Inspired by Khan et al. J Medl Chem 42(6), 951-956, the contents of which are incorporated herein by reference in their entirety. H-Glu(OtBu)-OtBu·HCl (1.0 g, 3.38 mmol), 4-nitrophenylchloroformate (682 mg, 3.38 mmol), and triethylamine (943 μL, 6.76 mmol) were dissolved in dry DCM (20 mL). The mixture was refluxed under N2 atmosphere for 25 min, then stirred at room temperature for 60 min. A white precipitate formed, which disappeared upon addition of H-Lys(Z)-OtBu·HCl (1387 mg, 3.72 mmol) and triethylamine (943 μL, 6.76 mmol). The solution turned yellow, was refluxed for 10 min, and then stirred at room temperature for 90 min. TLC showed complete conversion of the starting material, and the mixture was concentrated to a small volume under vacuum. Add ethyl acetate (80 ml), and stir the suspension at room temperature for 16 hours, then filter through a glass filter (P3). Wash the white precipitate with ethyl acetate, combine the filtrate and supernatant, and concentrate under vacuum to give a yellow oil. Perform column chromatography using a gradient of ethyl acetate / hexane from 1:5 to 1:2 for 6 column volumes, followed by 1 column volume of 100% ethyl acetate. Combine the correct fractions and lyophilize to give the title compound as a pale yellow oil. MALDI-TOF m / z [M+Na] + Calculated value: 644.8, measured value: 644.6.

[0217] EuK(NH2)-(OtBu)3

[0218] Inspired by Makowski et al., Liebigs Ann. Chem., 1985, 1451-1464, the contents of which are incorporated herein by reference in their entirety. EuK(Z)-(OtBu)3 (1.9 g, 3.06 mmol), ammonium formate (385 mg, 6.11 mmol), and Pd / C (19 mg) were refluxed in anhydrous ethanol (40 mL) at 100 °C for 120 min under N2 atmosphere. The reaction mixture was allowed to cool to room temperature, and the suspension was filtered through diatomaceous earth. The diatomaceous earth was then washed with anhydrous ethanol (50 mL). The solvent was removed under vacuum to give a yellow oil. The title compound was purified by HPLC in quantitative yield as a pale yellow oil. MALDI-TOF m / z [M+H] + Calculated value: 487.6; Measured value: 488.1 [M+Na] + Calculated value: 510.6, measured value: 510.2.

[0219] DOTAGA-Ahx-COOH

[0220] DOTA-GA(OtBu)4 (50 mg, 71.34 μmol) was dissolved in DMSO (2 mL), followed by the addition of HSPyU (26.4 mg, 64.2-0.54 μmol) and DiPEA (62 μL, 356.38 μmol), and the solution was stirred at room temperature for 15 min. Subsequently, a suspension of 6-aminocaproic acid (9.4 mg, 71.34 μmol) and additional DiPEA (12.4 μL, 71.34 μmol) was added, and the solution was stirred for 105 min. Acetonitrile / H2O 9:11 was added, and the mixture was purified by preparative HPLC. After mixing the correct fractions, the mixture was freeze-dried to give a white solid. MALDI-TOF m / z [M+H] + Calculated value: 815.1, measured value: 814.3.

[0221] MAS3-Ahx-COOH

[0222] Synthesized according to standard SPPS, starting with Fmoc-Ahx-WANG resin (1.0 g). Following subsequent deprotection and coupling steps, the product was cleaved from the resin by gently shaking it in 100% TFA at room temperature for 180 min. The filtrate was collected, and the solvent was evaporated. The crude compound was purified by HPLC and lyophilized. MALDI-TOF[M+H] + Calculated value: 508.5; Measured value: 508.9.

[0223] MAG3-Ahx-COOH

[0224] Synthesized according to standard SPPS, starting with Fmoc-Ahx-WANG resin (1.48 g). Following subsequent deprotection and coupling steps, the product was cleaved from the resin by gently shaking it in 100% TFA at room temperature for 120 minutes. The filtrate was collected, and the solvent was evaporated. The crude compound was used without further purification. MALDI-TOF[M+H] + Calculated value: 418.5, measured value: 418.5.

[0225] Merrifield Resin

[0226] As performed by Lopalco et al., Org Biomol Chem., 2009, 7(5), 856-9, chloromethyl polystyrene resin (8.3 g, 15.0 mmol), N-Boc-aminophenol (9.4 g, 45.0 mmol), tetrabutylammonium iodide (1.7 g, 4.5 mmol), and CsCO3 (14.7 g, 45.0 mmol) were dissolved in acetone and refluxed overnight at 70 °C under N2 atmosphere. The resin was then thoroughly washed with DMF (100 mL), H2O (100 mL), DCM (100 mL), and Et2O (100 mL) and dried under vacuum. A 4-(4-nitrobenzyl)pyridine test was used to determine the completion of the reaction.

[0227] Indole-COOH

[0228] 2,3,3-Trimethyl-3H-indole (5.3 g, 33 mmol) and 6-bromohexanoic acid (5.9 g, 30 mmol) were heated in 75 mL of 1,2-dichlorobenzene at 95–120 °C for 72 hours. Et₂O (75 mL) was added, and the mixture was filtered. The residue was washed with Et₂O and dried under vacuum to give the title compound. The crude product was used directly in the next reaction without any further purification.

[0229] sulfonyl indole-COOH

[0230] Potassium 2,3,3-trimethyl-3H-indole-5-sulfonate (6.9 g, 25 mmol) and 6-bromohexanoic acid (7.3 g, 37.5 mmol) were stirred in 1,2-dichlorobenzene (40 mL) at 95 °C for 72 hours. The resulting solid compound was collected and pulverized. After washing with Et₂O, the crude product was obtained and used directly in the next reaction.

[0231] Indole-C4Phth

[0232] 2,3,3-Trimethyl-3H-indole (1.9 mL, 11.8 mmol) and 1-(4-bromobutyl)pyrrolidine-2,5-dione (10.0 g, 35.4 mmol) were heated in sulfolane (20.0 mL) at 90 °C for 48 hours under N2 atmosphere. The reaction mixture was precipitated in EtOAc, and the suspension was filtered. The residue was washed twice with Et2O and EtOAc.

[0233] Vacuum drying yielded the title compound as a pale yellow solid. This product was used in the next reaction without further purification.

[0234] sulfonyl indole-Phth

[0235] Sulfoindole (BB3) (6.925 g, 25.0 mmol) and 1-(4-bromobutyl)pyrrolidine-2,5-dione (17.6 g, 75.0 mmol) were heated in sulfolane (40.0 mL) at 90 °C under a nitrogen atmosphere for 24 hours. The reaction mixture was precipitated by adding methanol (5 mL), followed by filtration of the suspension. The residue was washed twice with Et2O and EtOAc. The title compound was dried under vacuum as a pink solid, ready for use without further purification.

[0236] Phth(SO3)-Cy7-COOH

[0237] Indole-COOH (1771 mg, 5.00 mmol) and glutaraldehyde dinitrile HCl (1566 mg, 5.50 mmol) were dissolved in a mixture of Ac₂O / AcOH (1:1; 50 mL) and stirred overnight at 60 °C. The next morning, the mixture was heated to 120 °C for 1 hour. The mixture was then allowed to cool to room temperature. After cooling, the resulting hemicyanins were precipitated in ice-cold MTBE / hexane (1:1; 1000 mL). The precipitate was washed twice with MTBE / hexane. Meanwhile, Merrifield resin (1590 mg, 2.65 mmol) was prepared by adding a TFA / DCM mixture (20:80; 40 mL) and bubbling with N₂ for 1 hour. Subsequently, the resin was washed three times with DCM (40 mL) before adding a DiPEA / DCM mixture (25:75; 40 mL). Before washing three times with DCM (40 mL), N2 was bubbled through for 20 minutes. The precipitated hemicyanine was then dissolved in DMF / DCM (1:1; 40 mL) and added to the resin. The resin was bubbled through with N2 for 1 hour before washing with various DMF / DCM compositions. Sulfonoindole-Phth (550 mg, 1.25 mmol) was dissolved in pyridine / Ac2O (3:1; 40 mL) and added to the resin, and the mixture was shaken overnight. A liquid was obtained, and the resin was washed with various DMF / DCM mixtures. The resulting dye-containing mixture was concentrated under vacuum before purification with DCVC (EtOAc / MeOH 0-60%). The relevant fractions were combined, concentrated, and further purified by HPLC. This purification yielded the title compound as a green solid. MALDI-TOF m / z [M+H] + Calculated value: 777.0; Measured value: 776.7.

[0238] Phth(SO3)-Cy7-EuK(OtBu)3

[0239] Phth(SO3)-Cy7-COOH (10.0 mg, 12.89 μmol) was dissolved in DMF (500 μL), followed by the addition of DiPEA (38.31 mg, 296.41 μmol) and PyBOP (60.36 mg, 115.99 μmol). After stirring for 2 minutes, EuK(OtBu)3 (56.56 mg, 115.99 μmol) dissolved in 500 μL of DMF was added. The mixture was stirred at room temperature for 60 minutes under a nitrogen atmosphere. Subsequently, H2O was added, and the crude product was purified by HPLC. The relevant fractions were combined, and the resulting mixture was concentrated under vacuum to give the title compound as a green solid. MALDI-TOF m / z [M+H] + Calculated value: 1246.6; Measured value: 1246.5.

[0240] Amine(SO3)-Cy7-EuK(OtBu)3

[0241] Phth(SO3)-Cy7-EuK(OtBu)3 (7.0 mg, 5.62 μmol) was dissolved in methylamine (33 wt% in EtOH; 10 mL) and stirred at room temperature for 1.5 hours. Subsequently, residual methylamine and EtOH were removed under vacuum. A mixture of H2O / MeCN was added before purifying the crude product by HPLC. The relevant fractions were combined and concentrated under vacuum to give the title compound as a green solid. MALDI-TOF m / z [M+H] + Calculated value: 1116.6; Measured value: 1116.3.

[0242] MAS3(SO3)-Cy7-EuK(OtBu3)

[0243] MAS3-Ahx-COOH (3.14 mg, 6.18 μmol) was dissolved in DMSO (500 μL), followed by the addition of DiPEA (7.26 mg, 56.20 μmol) and PyBOP (14.6 mg, 28.10 μmol). After stirring for 5 minutes, amine (SO3)-Cy7-EuK(OtBu)3 (6.27 mg, 5.62 μmol) dissolved in DMSO (500 μL) was added to the mixture. The resulting reaction mixture was stirred at room temperature for 1 h, followed by the addition of H2O and purification by HPLC. The relevant fractions were combined and concentrated under vacuum to give the title compound as a green solid. MALDI-TOF m / z [M+H] + Calculated value: 1607.0; Measured value: 1606.8.

[0244] MAS3(SO3)-Cy7-EuK

[0245] MAS3(SO3)-Cy7-EuK(OtBu)3 (2.0 mg, 1.24 μmol) was dissolved in TFA / H2O (95:5; 3.0 mL) and stirred at room temperature for 1 hour. Subsequently, residual TFA and H2O were removed under vacuum, and the crude product was purified by HPLC. This resulted in the acquisition of the title compound in a green solid form. MALDI-TOF m / z [M+H] + Calculated value: 1438.7; Measured value: 1438.6.

[0246] Phth-Cy7-(SO3)COOH

[0247] Sulfoindole-COOH (2166.69 mg, 5.00 mmol) and glutaraldehyde dionitrile HCl (1566 mg, 5.50 mmol) were dissolved in a mixture of Ac₂O / AcOH (1:1; 50 mL) and stirred overnight at 60 °C. The next morning, the mixture was heated to 120 °C for 1 hour. The mixture was then allowed to cool to room temperature. After cooling, the resulting hemicyanin precipitated in Et₂O (1000 mL). The precipitate was washed twice with Et₂O (300 mL). Meanwhile, Merrifield resin (1590 mg, 2.65 mmol) was prepared by adding a TFA / DCM mixture (20:80; 40 mL) and bubbling with N₂ for 1 hour. Subsequently, the resin was washed three times with DCM (40 mL) before adding a DiPEA / DCM mixture (25:75; 40 mL). Before washing three times with DCM (40 mL), N2 was bubbled through for 20 minutes. The precipitated hemicyanine was then dissolved in DMF / DCM (1:1; 40 mL) and added to the resin. The resin was bubbled through with N2 for 1 hour before washing with various DMF / DCM compositions. Indole-Phth (451.83 mg, 1.25 mmol) was dissolved in pyridine / Ac2O (3:1; 40 mL) and added to the resin, and the mixture was shaken overnight. The liquid was obtained and the resin was washed with various DMF / DCM mixtures. The resulting dye-containing mixture was concentrated under vacuum, precipitated with Et2O (400 mL), and washed twice with Et2O (200 mL). The solvent was then removed under vacuum and purified by HPLC. This purification yielded the title compound as a green solid. MALDI-TOF m / z [M+H]+ calculated 777.0, found 776.6.

[0248] Phth-Cy7-(SO3)EuK(OtBu)3

[0249] Phth-Cy7-(SO3)COOH (10.0 mg, 12.89 μmol) was dissolved in DMF (500 μL), followed by the addition of DiPEA (38.31 mg, 296.41 μmol) and PyBOP (60.36 mg, 115.99 μmol). After stirring for 2 minutes, EuK(OtBu)3 (56.56 mg, 115.99 μmol) dissolved in 500 μL of DMF was added. The mixture was stirred at room temperature for 60 minutes under a N2 atmosphere. Subsequently, H2O was added, and the crude product was purified by HPLC. The relevant fractions were combined, and the resulting mixture was concentrated under vacuum to give the title compound as a green solid. MALDI-TOF m / z [M+H] + Calculated value: 1246.6; Measured value: 1246.5.

[0250] Amine-Cy7-(SO3)EuK(OtBu)3

[0251] Phth-Cy7-(SO3)EuK(OtBu)3 (15.8 mg, 12.68 μmol) was dissolved in methylamine (33 wt% in EtOH; 10 mL) and stirred at room temperature for 2 hours. Subsequently, residual methylamine and EtOH were removed under vacuum. A mixture of H2O / MeCN was added before purifying the crude product by HPLC. The relevant fractions were combined and concentrated under vacuum to give the title compound as a green solid. MALDI-TOF m / z [M+H] + Calculated value: 1116.5; Measured value: 1116.3.

[0252] MAS3-Cy7-(SO3)EuK(OtBu)3

[0253] MAS3-Ahx-COOH (7.77 mg, 15.28 μmol) was dissolved in DMSO (100 μL), followed by the addition of DiPEA (17.96 mg, 138.95 μmol) and PyBOP (36.17 mg, 69.48 μmol). After stirring for 5 minutes, amine-Cy7-(SO3)EuK(OtBu)3 (15.5 mg, 13.90 μmol) dissolved in DMSO (900 μL) was added to the mixture. The resulting reaction mixture was stirred at room temperature for 75 minutes, followed by the addition of H2O and purification by HPLC. The relevant fractions were combined and concentrated under vacuum to give the title compound as a green solid. MALDI-TOF m / z [M+H] + Calculated value: 1607.0; Measured value: 1606.9.

[0254] MAS3-Cy7-(SO3)EuK

[0255] MAS3-Cy7-(SO3)EuK(OtBu)3 (22.3 mg, 13.90 μmol) was dissolved in TFA / H2O (95:5; 3.0 mL) and stirred at room temperature for 1 hour. Subsequently, residual TFA and H2O were removed under vacuum, and the crude product was purified by HPLC. This resulted in the acquisition of the title compound in the form of a green solid. MALDI-TOF m / z [M+H] + Calculated value: 1438.7; Measured value: 1438.6.

[0256] Phth(SO3)-Cy5-COOH

[0257] In short, indole-COOH (1.1 g, 4.0 mmol) and malondialdehyde diphenylamine hydrochloride (1.2 g, 4.4 mmol) were dissolved in AcOH / Ac2OH (1:1, 30.0 mL) and stirred at 60–120 °C for 12 hours under a N2 atmosphere. The crude product was dissolved in DMF:DCM (1:1, 70.0 mL). Merrifield resin (1.2 g, 2.0 mmol) was swollen in DCM for 5 minutes, then bubbled in 20% TFA in DCM (50.0 mL) for 1 hour to deprotect the amine in the resin, and bubbled in 20% DiPEA in DCM (50.0 mL) for 15 minutes to remove excess TFA. Sulfoindole-Phth (440.0 mg, 1.0 mmol) was dissolved in pyridine / Ac2O (3:1, 40.0 mL). The dye was precipitated from the mixture in diethyl ether and purified by column chromatography (DCM: MeOH, 20-100% MeOH, 8 CVs). After combining the correct fractions, the solvent was removed under vacuum, and the remaining solid was further purified by preparative reversed-phase HPLC. The title compound was a blue solid. m / z [M+H] + Calculated value: 750.93, measured value: 750.60.

[0258] Phth(SO3)-Cy5-EuK(OtBu3)

[0259] Phth(SO3)-Cy5-COOH (3.0 mg, 4.00 μmol), EuK(NH2)-(OtBu)3 (23.4 mg in 249.2 μL of MeOH, 48 μmol), PyBOP (47.8 mg, 92.00 μmol), and DiPEA (16.0 μL, 122.67 μmol) were stirred in DMF (800 μL) at room temperature for 90 minutes. The mixture was then kept at -21°C overnight. The mixture was purified by HPLC. MALDI-TOF m / z [M+H] + Calculated value: 1220.6; Measured value: 1219.6.

[0260] Amine(SO3)-Cy5-EuK(OtBu)3

[0261] Phth(SO3)-Cy5-CONH.EuK(OtBu)3 (10 mg) was dissolved in methylamine (33 wt% in EtOH; 10 mL) and stirred at room temperature for 2 hours. Afterward, residual methylamine and EtOH were removed under vacuum. A mixture of H2O / MeCN was added before purifying the crude product by HPLC. The relevant fractions were combined and concentrated under vacuum to give the title compound as a blue solid. MALDI-TOF m / z [M+H] + Calculated value: 1090.5; Measured value: 1090.2.

[0262] Phth-Cy5-(SO3)COOH

[0263] Sulfoindole-COOH (1.1 g, 2.4 mmol) and malondialdehyde diphenylamine hydrochloride (678.0 mg, 2.6 mmol) were dissolved in AcOH / Ac2OH (1:1, 15.0 mL) and stirred at 60–120 °C for 12 hours under N2 atmosphere. The crude product was dissolved in DMF (70.0 mL). Merrifield resin (720.0 mg, 1.2 mmol) was swollen in DCM for 5 minutes, then bubbled in DCM (20% TFA; 25.0 mL) for 1 hour, and then bubbled in DCM (20% DiPEA; 25.0 mL) for 15 minutes. Indole-Phth (264.0 mg, 0.6 mmol) was dissolved in pyridine / Ac2O (3:1, 20.0 mL). Sulfoindole-Phth (264.0 mg, 0.6 mmol) was dissolved in pyridine / Ac₂O (3:1, 20.0 mL). The dye was precipitated from the mixture in diethyl ether and purified by DCVC (EtOAc:MeOH, 0-100% MeOH, 25 fractions, 100.0 mL each). After combining the correct fractions, the solvent was removed under vacuum, and the remaining solid was further purified by preparative reversed-phase HPLC, yielding the title compound as a blue solid. m / z [M+H] + Calculated value: 750.93, measured value: 750.78. Phth-Cy5-(SO3)EuK(OtBu)3

[0264] Phth-Cy5-(SO3)COOH (4.0 mg, 5.33 μmol), EuK(NH2)-(OtBu)3 (23.4 mg in 249.2 μL of MeOH, 48 μmol), PyBOP (25.0 mg, 48 μmol), and DiPEA (21.3 μL, 122.67 μmol) were stirred in DMF (800 μL) at room temperature for 90 minutes. The mixture was then kept at -21°C overnight. The mixture was purified by HPLC. MALDI-TOF m / z [M+H] + Calculated value: 1220.6, measured value: 1219.6.

[0265] Amine-Cy5-(SO3)EuK(OtBu)3

[0266] Phth-Cy5-(SO3)CONH.EuK(OtBu)3 (6 mg, 4.92 μmol) was dissolved in methylamine (33 wt% in EtOH; 10 mL) and stirred at room temperature for 2 hours. Subsequently, residual methylamine and EtOH were removed under vacuum. A mixture of H2O / MeCN was added before purifying the crude product by HPLC. The relevant fractions were combined and concentrated under vacuum to give the title compound as a blue solid. MALDI-TOF m / z [M+H] + Calculated value: 1089.4; Measured value: 1089.6.

[0267] DOTAGA(OtBu)4(SO3)-Cy5-EuK(OtBu)3

[0268] Amine (SO3)-Cy5-EuK(OtBu)3 (1.7 mg, 1.56 μmol) was dissolved in DMSO (500 μL). PyBOP (1.6 mg, 3.12 μmol, 100 μg / μL in DMSO) was added to DOTAGA(OtBu)4-Ahx-COOH (1.8 mg, 2.19 μmol), and DMSO was added to a final volume of 100 μL. This was then added to the amine (SO3)-Cy5-EuK(OtBu)3 solution. DiPEA (2.7 μL, 15.61 μmol) was then added, and the mixture was stirred at room temperature for 30 minutes. The mixture was diluted with CH3CN and H2O (0.1% TFA v / v) and purified by HPLC. MALDI-TOF m / z [M+H] + Calculated value: 1885.5, measured value: 1885.2. DOTAGA(SO3)-Cy5-EuK

[0269] DOTAGA(OtBu)4(SO3)-Cy5-EuK(OtBu)3 was dissolved in TFA / TIPS / H2O 95 / 2.5 / 2.5 (5 mL). After stirring for 2 hours, the solvent was evaporated, and the residue was purified by HPLC. MALDI-TOF m / z [M+H] + Calculated value: 1492.8; Measured value: 1492.7.

[0270] DOTAGA(OtBu)4-Cy5-(SO3)EuK(OtBu)3

[0271] Amine-Cy5-(SO3)-EuK(OtBu)3 (2.4 mg, 2.20 μmol) was dissolved in DMSO (500 μL). PyBOP (2.3 mg, 4.41 μmol, stock solution in 100 μg / μL DMSO) was added to DOTAGA(OtBu)4-Ahx-COOH (2.5 mg, 3.09 μmol), and DMSO was added to a final volume of 100 μL. This was then added to the amine-Cy5-(SO3)-EuK(OtBu)3 solution. DiPEA (3.9 μL, 22.04 μmol) was then added, and the mixture was stirred at room temperature for 30 minutes. The mixture was diluted with CH3CN and H2O (0.1% TFA v / v) and purified by HPLC. MALDI-TOF m / z [M+H] + Calculated value: 1885.5, measured value: 1885.2. DOTAGA-Cy5-(SO3)EuK

[0272] DOTAGA(OtBu)4(SO3)-Cy5-EuK(OtBu)3 was dissolved in TFA / TIPS / H2O 95 / 2.5 / 2.5 (5 mL). After stirring for 2 hours, the solvent was evaporated, and the residue was purified by HPLC. MALDI-TOF m / z [M+H] + Calculated value: 1492.8; Measured value: 1492.7.

[0273] MAG3(SO3)-Cy5-EuK(OtBu3)

[0274] Amine (SO3)-Cy5-EuK(OtBu)3 (10 mg, 9.18 μmol), MAG3-Ahx-COOH (15 mg, 36.73 μmol), PyBOP (19 mg, 36.73 μmol), and DiPEA (16 μL, 91.83) were dissolved in DMF (1 mL). The mixture was stirred at room temperature for 30 minutes, then kept overnight at -21 °C. The crude product was purified by HPLC, and the correct fractions were then combined and lyophilized to give a blue solid. MALDI-TOF m / z [M+H] + Calculated value: 1489.9, measured value: 1489.8.

[0275] MAG3(SO3)-Cy5-EuK

[0276] MAG3(SO3)-Cy5-EuK(OtBu)3 was dissolved in TFA / H2O (95:5; 2.0 mL) and stirred at room temperature for 1 hour. Subsequently, residual TFA and H2O were removed, and the crude product was purified by HPLC. This resulted in the acquisition of the title compound as a blue solid. MALDI-TOF m / z [M+H] + Calculated value: 1321.6, measured value: 1321.6.

[0277] MAG3-Cy5-(SO3)EuK(OtBu)3

[0278] Amine-Cy5-(SO3)EuK(OtBu)3 (1.4 mg, 1.24 μmol), MAG3-Ahx-COOH (2.1 mg, 4.96 μmol), PyBOP (2.6 mg, 4.96 μmol), and DiPEA (2.2 μL, 12.40 μmol) were dissolved in DMF (250 mL), and the mixture was stirred at room temperature. The product was observed after 60 minutes, and the reaction mixture was stored at -21 °C. MALDI-TOF m / z [M+H] + Calculated value: 1489.9, measured value: 1489.8.

[0279] MAG3-Cy5-(SO3)EuK was dissolved in TFA / H2O. After stirring for 1 hour, the title compound was formed. MALDI-TOF m / z [M+H] + Calculated value: 1322.6; Measured value: 1322.4.

[0280] Phth(SO3)-Cy5-(SO3)EuK(OtBu3)

[0281] Phth(SO3)-Cy5-(SO3)COOH (10.0 mg, 12.06 μmol) and EuK(OtBu)3 (64 mg, 120.63 μmol) were dissolved in DMSO. PyBOP (63 mg, 120.63 μmol), DiPEA (53 μL, 301.57 μmol), and DMSO (300 μL) were added, and the mixture was stirred at room temperature for 1 hour. After dilution with CH3CN and H2O (0.1% TFA v / v), the mixture was purified by HPLC, and the correct fractions were then combined and lyophilized. MALDI-TOF m / z [M+2H] + Calculated value: 1298.6; Measured value: 1300.3.

[0282] Amine(SO3)-Cy5-(SO3)EuK(OtBu)3

[0283] Phth(SO3)-Cy5-(SO3)EuK(OtBu)3 was dissolved in methylamine (33 wt% in EtOH; 20 mL) and stirred for 3.5 hours. Afterward, the remaining methylamine and EtOH were removed under vacuum. A mixture of H2O / MeCN was added before purifying the crude product by HPLC. The relevant fractions were combined and concentrated under vacuum to give the title compound as a blue solid. MALDI-TOF m / z [M+2H] + The calculated value is 1170.5, and the measured value is 1169.9.

[0284] MAS3(SO3)-Cy5-(SO3)EuK(OtBu3)

[0285] Amine (SO3)-Cy5-(SO3)EuK(OtBu)3 (4.0 mg, 3.42 μmol), MAS3-Ahx-COOH (21 mg, 41.07 μmol), PyBOP (21.3 mg, 41.07 μmol), and DIPEA (11.8 μL, 68.44 μmol) were dissolved in DMSO (1.5 mL) and stirred for 2.5 h. After dilution with CH3CN and H2O (0.1% TFA v / v), the mixture was purified by HPLC, and the correct fractions were then combined and lyophilized. MALDI-TOF m / z [M+H] + Calculated value: 1660.0, measured value: 1660.6.

[0286] MAS3(SO3)-Cy5-(SO3)EuK3

[0287] MAS3(SO3)-Cy5-(SO3)EuK(OtBu)3 was stirred in TFA / H2O 95:5 (2 mL) for 1 hour. The solvent was evaporated, and the crude product was purified by HPLC and lyophilized. MALDI-TOF m / z [M+2H] + Calculated value: 1492.7, measured value: 1492.4.

[0288] Example 3: Photophysical properties of exemplary compounds

[0289] The photophysical properties of fluorophores are important for fluorophores used in mixing tracers (such as the compounds of this invention). These photophysical properties are influenced by the compound structure.

[0290] The brightness of a dye—one of the most important photophysical characteristics in a clinical setting—is the product of its molar extinction coefficient (ε) and quantum yield (ΦF). Therefore, we began by establishing a linear concentration range (7.5–0.25 M) in PBS and measuring the corresponding absorbance to determine the ε for each fluorophore (Table 1). Adding a sulfonate to the carboxylic acid-containing indoline moiety reduced the ε, and using an additional aryl moiety on the anthocyanin backbone significantly reduced the dye solubility, thus also greatly decreasing their ε. This is not surprising, as it has been previously established that benzo[e]indole-containing dyes are insoluble in H₂O or PBS due to their relatively flat and hydrophobic core, leading to dye-dye stacking via van der Waals forces (through the formation of J-aggregates).

[0291] The ΦF of a fluorophore is the relationship between the number of emitted photons and the number of photons absorbed by a single molecule. Therefore, it is also known as emission efficiency. These data were measured in PBS, and the values ​​are significantly lower than those in DMSO, but are more representative in the in vivo environment. From this data (Table 2), it can be concluded that the addition of an electron-withdrawing sulfonic acid groups to an indole moiety negatively impacts the emission efficiency of the fluorophore. Perhaps an asymmetric anthocyanin nucleus, due to its significant electron density imbalance, is an inefficient fluorophore. Consistent with our expectations—the ΦF of benzo[e]indole-containing fluorophores is reduced due to the formation of J-aggregates caused by benzo[e]indole. Typically, Phth(Ar)-Cy5-(Ar)COOH (containing two benzo[e]indole moieties) does not exhibit the lowest ΦF among fluorophores containing three benzo[e]indole moieties. The only trend observed from the above data is that free fluorophores benefit from symmetrical substituents on the anthocyanin structure (when emission efficiency is optimized). However, this trend was not observed in the mixed tracers. As shown in Table 2, EuK(SO3)-Cy5-MAS3 has the highest ΦF of the matrix.

[0292] Table 2. Selected photophysical properties of exemplary Cy5 dyes and mixed tracer analogs

[0293]

[0294]

[0295] * Due to severe buildup, λ could not be measured. ex / λ em

[0296] Brightness is determined by using the ε of the corresponding fluorophore of the mixed tracer.

[0297] Since the brightness of a fluorophore (or mixed tracer) is one of its most important photophysical properties, we calculated the brightness of all fluorophores. To gain a deeper understanding of the optical properties of the mixed tracers, brightness was determined using the ε of their respective fluorophores in PBS. These approximations of tracer performance clearly show that, based on ΦF measurements, the highest observed brightness is characterized by tracers EuK(SO3)-Cy5-MAS3 and EuK-Cy5-MAS3, respectively. Therefore, among these six exemplary mixed tracers, EuK(SO3)-Cy5-MAS3 contains the most favorable photophysical characteristics.

[0298] Example 4: Lipophilicity and serum interaction of exemplary compounds

[0299] The lipophilicity of a compound can be represented by its partition coefficient (logP), which affects pharmacodynamics and pharmacokinetics because it is a major determinant of the compound's absorption, distribution, metabolism, and excretion (ADME properties). Therefore, this criterion is widely used as an early indicator for preclinical evaluation. We calculated the logD values ​​(clogD) at pH 7.4 as follows: EuK-Cy5-MAS3-7.15, EuK(SO3)-Cy5-MAS3-10.31, EuK-Cy5-(SO3)MAS3-10.31, EuK(Ar)-Cy5-MAS3-6.16, EuK-Cy5-(Ar)MAS3-6.16, and EuK(Ar)-Cy5-(Ar)MAS3-5.17.

[0300] Serum binding, more precisely plasma protein binding (PPB), is a useful pharmacokinetic characteristic for preliminary evaluation of tracer candidates before in vivo use, as PPB is directly related to, for example, blood retention and clearance. PPB values ​​for all mixed tracers ranged from 52% to 89%, with an average of 76 ± 12%. Figure 2A). Mixed tracers containing benzo[e]indole (EuK(Ar)-Cy5-MAS3, EuK-Cy5-(Ar)MAS3, and EuK(Ar)-Cy5-(Ar)MAS3) tended to have the highest PPB. Interestingly, despite the reduction in lipophilicity by adding a sulfonate group, the PPBs of EuK(SO3)-Cy5-MAS3 and EuK-Cy5-(SO3)MAS3 were significantly higher than that of EuK-Cy5-MAS3. This can be explained by the Sudlow site (a binding pocket in subdomain IIA of albumin), which binds a large number of heterocyclic anions. There was no significant correlation between clogD and serum binding. Although PPB is related to lipophilicity, we do not expect that PPB can be predicted based on clogD because PPB cannot be explained by a single physicochemical property.

[0301] Not only is serum binding significant, but stability in serum is also valuable. This property is another component of the preclinical evaluation system for mixed tracers, as it indicates the in vivo stability of the compounds. Clearly, any compound that degrades in the in vivo environment is unsuitable for clinical use. All mixed tracers were incubated in serum at 37°C for 24 hours, and their absorbance and fluorescence were measured.

[0302] Time-process data indicates ( Figure 2 For both B and 2C), the absorbance signal of most mixed tracers decreased slightly (residual absorbance after 24 hours was 74.6 ± 0.4–84.6 ± 2.7%). The fluorescence signal weakened to 67.1 ± 7.3–88.7 ± 5.6% after 24 hours. Significantly reduced serum stability was observed only in EuK(Ar)-Cy5-(Ar)MAS3 (p < 0.05). All other exemplary tracers showed little difference.

[0303] Table 3 summarizes the results of lipophilicity, plasma protein binding, and serum stability for five exemplary compounds.

[0304] Table 3. Characteristics of Exemplary Hybrid Tracers

[0305]

[0306] **use 99m Measurement of Tc-labeled mixed tracers

[0307] Example 5: Serum confocal microscopy and receptor affinity of exemplary compounds

[0308] Fluorescence confocal microscopy is used as a tool for receptor localization. For example... Figure 3As shown, typical PSMA-targeting tracers brightly stain the extracellular receptor. IC50 was determined in a competitive binding assay using human LNCaP cells and EuK[ 125 I]I-BA is used as a competitive radioligand. EuK(Ar)-Cy5-(Ar)MAS3 has the lowest affinity (345±31), while four of the other five tracers are in the same nanomolar range (113–175 nM). The only IC 50 The improved tracer is EuK(SO3)-Cy5-MAS3, with an affinity of 19±6.

[0309] This indicates that the introduction of the benzene (Ar) moiety does not increase affinity compared to EuK-Cy5-MAS, while the introduction of the anionic sulfonate moiety (SO3) at the carbon end of the anthocyanin backbone increases PSMA affinity. Furthermore, the affinity of EuK-(SO3)Cy5-MAS3 is more than 9 times higher than that of EuK-Cy5(SO3)MAS3. Without being bound by any theory, this suggests that in the interaction with the amphiphilic inlet funnel of PSMA, the anthocyanin backbone of the exemplary mixed tracer allows hydrogen bonds to supplement hydrophobic interactions by carefully placing the anionic moiety (e.g., the sulfonate moiety).

[0310] Table 4: Use of human LNCaP cells and EuK [ 125 I]I-BA as an exemplary hybrid tracer for competing radioligands, PSMA affinity; n≥3

[0311] Mixed tracers <![CDATA[IC 50 (μM)]]> EuK-Cy5-MAS3 175.3±61.6 <![CDATA[EuK(SO3)-Cy5-MAS3]]> 19.2±5.8 <![CDATA[EuK-Cy5-(SO3)MAS3]]> 118.8±117.4 <![CDATA[EuK(SO3)-Cy5-(SO3)MAS3]]> 18.3±8.0 <![CDATA[EuK(Ar)-Cy5-MAS3]]> 113.1±35.9 <![CDATA[EuK-Cy5-(Ar)MAS3]]> 164.4±76.9 <![CDATA[EuK(Ar)-Cy5-(Ar)MAS3]]> 344.7±30.9

[0312] Example 6: Exemplary Compounds in Vivo

[0313] In vivo characteristics of orthotopically transplanted PC346C cells were investigated in male BALB / c nude mice. Multiple analyses were performed on these mice, including in vivo nuclear imaging (SPECT) of tumor-bearing mice, in vivo optical imaging (fluorescence imaging) of tumor-bearing mice, ex vivo biodistribution (percentage per gram of injected dose (%ID / g)) of healthy and tumor-bearing mice, and ex vivo fluorescence imaging.

[0314] In tumor-bearing BALB / c nude mice, SPECT imaging with a mixed tracer matrix produced clear nuclear images of tumors containing EuK(Ar)-Cy5-(Ar)MAS3, EuK(SO3)-Cy5-MAS3, and EuK-Cy5-(SO3)MAS3. Figure 4AFollowing analysis of the in vitro biodistribution, it immediately became apparent that the tumor uptake of the mixed tracers synthesized herein was similar to or superior to that of the clinically approved PSMA I&S. Perhaps more importantly, all the mixed PSMA analogs exhibited significantly reduced renal and splenic accumulations compared to PSMA I&S and PSMA-11 analogs (renal accumulation: range 10.8 ± 9.79–21.8 ± 14.2 ID / g, compared to 186.0 ± 23.0 for PSMA I&S and 124.8 ± 32.3–221.0 ± 24.4 for PSMA-11 analogs). The prolonged renal tracer clearance is likely a direct result of the renal tracer portion, and this significant reduction observed in the exemplary tracers of the present invention suggests the potential of these compounds for surgical guidance applications. Furthermore, nonspecific accumulation appears to be quite low for all the exemplary tracers of the present invention, which is another advantageous characteristic.

[0315] The tracer with the highest PPB concentration had the highest blood retention (6.91 ± 1.08 ID / g; EuK(Ar)-Cy5-MAS3), while the two tracers with clogD below -10 were almost entirely excreted from the blood. Figure 5 B). Interestingly, the hepatic clearance rates were lowest for the most lipophilic tracer (0.87 ± 0.66; EuK(Ar)-Cy5-(Ar)MAS3) and one of the penultimate lipophilic tracers (0.94 ± 0.13; EuK(Ar)-Cy5-MAS3). Figure 5 C). Substituents on EuK-containing indoles significantly reduced liver clearance (19.93 ± 13.44 EuK-Cy5-MAS3; 2.90 ± 2.06 EuK(SO3-)-Cy5-MAS3; 1.10 ± 0.14 EuK(Ar)-Cy5-MAS3; 0.87 ± 0.66 EuK(Ar)-Cy5-(Ar)MAS3). Further analysis of biodistribution data revealed that the correlation between PPB and clearance appeared to be contrary to previous assumptions, as the percentage of injection activity across all resected organs decreased with increasing PPB. Figure 5 A).

[0316] Because the prostate is surrounded by adipose tissue, an important consideration when designing mixed tracers for prostate-specific surgery is the tumor-to-fat ratio (T / F). Furthermore, the tumor-to-blood ratio (T / B) provides a general interpretation of the signal-to-background ratio. Clearly, for these exemplary mixed tracers, increasing lipophilicity does not improve the T / F and T / B ratios. Moreover, EuK-Cy5-MAS3 (2542±287) and EuK-Cy5-(SO3)MAS3 (2658±659) showed the best T / F ratios. Figure 5D), but they are not significantly different from each other. Except for EuK(SO3)-Cy5-MAS3(1713), the T / B ratios of all compounds are ( Figure 5 E) were all very low. Furthermore, ex vivo fluorescence imaging of tumor-bearing prostate tissue showed that, except for EuK-Cy5-(Ar)MAS3, the prostate tissue outlines of all mixed tracers were clearly defined. Figure 6 ).

[0317] The results indicate that the presence of different functional groups on the dye moiety and the location of these functional groups directly affect the pharmacokinetics of the tracer. Serum binding can be achieved by introducing exemplary anions (e.g., SO3). - Reinforced with ar groups or Ar groups. SO3 - Different numbers and positions of groups do not produce different serum binding, but this effect can be clearly observed for Ar groups. For example, single Ar functionalization on the EuK side of the dye moiety produces the highest serum binding, even compared to tracers with Ar units on each side. This trend appears consistent with the increased blood pool values ​​observed on biomaps and the increased uptake by blood, liver, and salivary glands. Although SO3 - High serum binding does not appear to reflect pool and in vivo strength (which is typically much lower), but in vivo data again indicate that single SO3 on the EuK side of the dye... - Functionalization yields the highest tissue uptake. Therefore, asymmetric dye-part junctions appear to be a key characteristic of in vivo tracer performance. Interestingly, these trends were not clearly observed in terms of renal retention and tumor uptake, implying that tracer pharmacokinetics may be the most critical component in tracer selection. Therefore, tracers providing reduced background signal are beneficial. In some cases, the lack of background signal appears to be a decisive factor.

Claims

1. Compounds of Formula I: I in: R1 is selected from sulfonic acid group and H, and R2 is H; or R1 and R2 together form a phenyl group; R3 is selected from sulfonic acid group and H, and R4 is H; or R3 and R4 together form a phenyl group; R5 and R6 are both H; R7, R8, R9 and R 10 Each is CH3; V is -CH2-; W is -CH2-; Y1 is -EuK; Z is a residue of -MAS3 or -MAS3; n is 2; m is 5; and p is 4. Where -EuK is ;and Among them, -MAS3 is ; in" "Indicates the connection point with the rest of the compound; Or its pharmaceutically acceptable salt.

2. The compound according to claim 1, wherein at least one of the substituents R1, R2, R5, R7, R8 of the first indole moiety is different from the substituents R3, R4, R6, R9, R8 of the second indole moiety. 10 At least one of them, thereby providing an asymmetric dye portion.

3. The compound according to claim 1: Where R1 is a sulfonic acid group and R2 is H; or R1 and R2 together form a phenyl group; or Where R3 is a sulfonic acid group and R4 is H; or R3 and R4 together form a phenyl group.

4. The compound according to claim 1, wherein Z is -MAS3.

5. The compound according to claim 1, wherein the compound is selected from: ; ; ; ; ; ; and .

6. The compound according to any one of claims 1 to 5, further comprising a chelated radiolabeled substance.

7. The compound of claim 6, wherein the chelated radiolabeled material is selected from... 44 Sc、 47 Sc、 51 Cr 52m Mn, 58 Co、 52 Fe、 56 Ni、 57 Ni、 62 Cu、 64 Cu、 67 Cu、 66 Ga、 68 Ga、 67 Ga、 89 Zr、 90 Y、 89 Y、 94m Tc, 99m Tc, 97 Ru、 105 Rh、 109 Pd, 111 Ag、 110m In、 111 In、 113m In、 114m In、 117m Sn、 121 Sn、 127 Te、 142 Pr、 143 Pr、 149 Pm, 151 Pm, 149 Tb, 153 Sm、 157 Gd, 161 Tb, 166 Ho、 165 Dy、 169 Er、 169 Yb、 175 Yb、 172 Tm、 177 Lu、 186 Re、 188 Re、 191 Pt, 197 Hg, 198 Au、 199 Au、 212 Pb, 203 Pb, 211 At、 212 Bi、 213 Bi、 223 Ra、 225 Ac and 227 Th, or containing 18 F is a cationic molecule.

8. Compounds of formula II or pharmaceutically acceptable salts thereof: II in: R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 V, W, Z, m, n and p are as defined in any one of claims 1 to 4.

9. Compounds of formula III or their pharmaceutically acceptable salts: III in: R1, R2, R3, R4, R5, R6, R7, R8, R9, R 10 V, W, Y1, m, n and p are as defined in any one of claims 1 to 4.

10. A formulation comprising the compound of any one of claims 1 to 7 and optionally a pharmaceutically acceptable carrier.

11. Use of the compound of any one of claims 1 to 7 or the formulation of claim 10 in the preparation of a pharmaceutical agent for tumor imaging.

12. The use according to claim 11, wherein the tumor is a prostate cancer tumor, a kidney tumor, a breast cancer tumor, a glioma, a colorectal adenocarcinoma, a transitional cell carcinoma, a pancreatic ductal adenocarcinoma, or a gastric adenocarcinoma; and / or The compound comprises a chelated radiolabeled substance, and the imaging comprises positron emission tomography, single-photon emission computed tomography, scintillation, gamma ray tracing / imaging, or beta tracing; and / or The imaging described therein includes fluorescence imaging.

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