Pharmaceutical compositions for diagnosis and / or therapy

Abstract

The present disclosure relates to pharmaceutical compositions for diagnosis and / or treatment comprising a compound having the structure of Formula (If) or a pharmaceutically acceptable salt thereof.

Description

[0001] This application is a divisional application of Chinese Patent Application 201880056196.2 filed on December 11, 2018 with the title “PSMA Ligands for Imaging and Endoradiotherapy”. TECHNICAL FIELD

[0002] The present disclosure relates to imaging and endoradiotherapy of prostate-specific membrane antigen (PSMA)-related diseases. Compounds are provided that bind to or inhibit PSMA and are further carrying at least one moiety suitable for radiolabeling. Also provided are medical uses of such compounds.

[0003] Several documents, including patent applications and manufacturer’s manuals, are cited in the specification of this application. The disclosures of these documents in their entireties are hereby incorporated by reference into the present disclosure. In particular, all references are incorporated by reference as if each individual document was specifically and individually indicated to be incorporated by reference herein. BACKGROUND

[0004] Over the past decades, prostate cancer (PCa) remains the most common male malignancy with low survival rates and high incidence. Due to its overexpression in prostate cancer (Silver, D. A., et al. Prostate-specific membrane antigen expression in normal and malignant human tissues. Clinical Cancer Research, 1997. 3(1): p. 81-85), prostate-specific membrane antigen (PSMA) or glutamate carboxypeptidase II (GCP II) has proven to be an excellent target for the development of highly sensitive radiolabeled agents for PCa endoradiotherapy and imaging (Afshar-Oromieh, A. et al. The diagnostic value of PET / CT imaging with the 68Ga-labelled PSMA ligand HBED-CC in the diagnosis of recurrent prostate cancer. European journal of nuclear medicine and molecular imaging, 2015. 42(2): p. 197-209; M. et al. Preclinical Evaluation of a Tailor-Made DOTA-Conjugated PSMA Inhibitor with Optimized Linker Moiety for Imaging and Endoradiotherapy of Prostate Cancer. Journal of Nuclear Medicine, 2015. 56(6):914-920; Robu, S. et al. Preclinical evaluation and first patient application of 99mTc-PSMA-I&S for SPECT imaging and radioguided surgery in prostate cancer. Journal of Nuclear Medicine, 2016: jnumed.116.178939; Weineisen, M. et al. Development and first in human evaluation of PSMA I&T-A ligand for diagnostic imaging and endoradiotherapy of prostate cancer. Journal of Nuclear Medicine, 2014. 55(Suppl 1):1083-1083; Rowe, S. et al. PET imaging of prostate-specific membrane antigen in prostate cancer: current state of the art and future challenges. Prostate cancer and prostatic diseases, 2016; Maurer, T. et al. Current use of PSMA-PET in prostate cancer management. Nature Reviews Urology, 2016). Prostate-specific membrane antigen is an extracellular hydrolase whose catalytic center includes two zinc(II) ions with bridging hydroxo ligands. It is highly upregulated in metastatic and hormone-refractory prostate cancer, but physiological expression has also been reported in kidney, salivary glands, small intestine, brain, and to a lesser extent in healthy prostate tissue.In the intestine, PSMA facilitates folate uptake by converting pteroyl poly-gamma-glutamate to pteroyl glutamate (folate). In the brain, it hydrolyzes N-acetyl-L-aspartyl-L- glutamate (NAAG) to N-acetyl-L-aspartate and glutamate. In normal and diseased prostate, the enzymatic function of PSMA has not been clarified.

[0005] PSMA targeting molecules usually comprise a binding unit comprising a zinc binding group (e.g. urea (Zhou, J. et al. NAAG peptidase inhibitors and their potential for diagnosis and therapy. Nature Reviews Drug Discovery, 2005, 4(12): p. 1015-1026), phosphinate or phosphoramidate) attached to the P1 glutamate moiety, which guarantees high affinity and specificity for PSMA and is usually also associated with effector functionality (Machulkin, A.E. et al. Small-molecule PSMA ligands. Current state, SAR and perspectives. Journal of drug targeting, 2016: p. 1-15). The effector moiety is more flexible and can tolerate structural modifications to some extent. The entrance channel of PSMA has two other prominent structural features that are very important for ligand binding. The first is the arginine patch, which is a positively charged region on the entrance funnel wall and is a structural explanation for the preference of PSMA for negatively charged functionality at the P1 position. After binding, the coordinated reset of the arginine side chains can lead to the opening of the S1 hydrophobic pocket, which is the second important structure, which has been shown to accommodate the iodosobenzyl group of several urea-based inhibitors, thus rendering them with high affinity for PSMA (Barinka, C. et al. Interactions between Human Glutamate Carboxypeptidase II and Urea-Based Inhibitors: Structural Characterization of the Complexes. Journal of medicinal chemistry, 2008, 51(24): p. 7737-7743).

[0006] ​Zhang et al. found a distal binding site of PSMA, which can be used in a bicoordinate binding mode (Zhang, A. X. et al. A remote arene-binding site on prostate specific membrane antigen revealed by antibody-recruiting small molecules. Journal of the American Chemical Society, 2010, 132(36): 12711-12716). The so-called arene binding site is a simple structural motif formed by the side chains of Arg463, Arg511 and Trp541 and is part of the PSMA entrance lid. The partial binding of the arene binding site by a distal inhibitor can lead to a great increase in the inhibitor affinity to PSMA due to avidity effects. For the interaction with PSMA in this way PSMA I&T was developed (see Figure 1 ), although there was no crystal structure analysis of the binding mode available. According to Zhang et al. the essential feature is a linker unit (in the case of PSMA I&T octanedioic acid), which promotes the open conformation of the PSMA entrance lid, so that the arene binding site is accessible. It has also been shown that the structural composition of the linker has a major influence on tumor targeting and biological activity as well as on imaging contrast and pharmacokinetics (Liu, T. et al. Spacer length effects on in vitro imaging and surface accessibility of fluorescent inhibitors of prostate specific membrane antigen, Bioorganic & medicinal chemistry letters, 2011, 21(23): 7013-7016), which are essential properties for high imaging quality and effective targeted intracavitary radiotherapy.

[0007] Currently, two classes of PSMA-targeting inhibitors are used in clinical settings. One class consists of tracers having chelating units for the complexation of PSMA I&T or related compounds with radionuclides (Kiess, AP et al., Prostate-specific membrane antigen as a target for cancer imaging and therapy, Quarterly Journal of Nuclear Medicine and Molecular Imaging: Official publication of the Italian Society of Nuclear Medicine [and] the International Association of Radiopharmaceuticals (IAR) [and] its departments, 2015, 59(3): 241). The other class consists of small molecules comprising a targeting unit and an effector molecule. Depending on the radionuclide / halogen used, radiolabeled PSMA inhibitors can be used for imaging or intracavitary radiotherapy. Among small molecule inhibitors with chelating agents used for imaging, the most commonly used formulations for selective PSMA imaging are PSMAHBED-CC (Eder, M. et al., 68Ga-complex lipophilicity and the targeting property of a urea-based PSMA inhibitor for PET imaging, Bioconjugate chemistry, 2012, 23(4): 688-697), PSMA-617 ( M. et al., Preclinical Evaluation of a Tailor-Made DOTA-Conjugated PSMA Inhibitor with Optimized Linker Moiety for Imaging and Endoradiotherapy of Prostate Cancer, Journal of Nuclear Medicine, 2015, 56(6): 914-920) and PSMA I&T (Weineisen, M. et al., Development and first in human evaluation of PSMA I&T-A ligand for diagnostic imaging and endoradiotherapy of prostate cancer, Journal of Nuclear Medicine, 2014, 55(Supplement 1): 1083-1083). PSMA HBED-CC or PSMA-11 is one of the earliest PSMA inhibitors, and because the chelator HBED-CC is not feasible for therapeutic applications, it is currently used for imaging. However, due to18 F possesses unique physical properties and advantages for PET imaging, such as a longer half-life, low positron energy, leading to higher image resolution and the possibility of large-scale generation in cyclotrons. Therefore, some research groups are focusing on its use for PCa imaging. 18 Development of F-labeled urea inhibitors. 18 F-labeled urea-based PSMA inhibitors [ 18 F]DCFPyl has shown promising results in detecting primary and metastatic PCa (Rowe, SP et al., PSMA-Based [ 18 [F]DCFPyL PET / CT Is Superior to Conventional Imaging for Lesion Detection in Patients with Metastatic Prostate Cancer, Molecular Imaging and Biology, 2016: 1-9), and is superior to [in comparative studies]. 68 Ga]PSMA-HBED-CC (Dietlein, M. et al., Comparison of [ 18 F]DCFPyL and [68Ga]Ga-PSMA-HBED-CC for PSMA-PET imaging in patients with relapsed prostate cancer, Molecular Imaging and Biology, 2015, 17(4): pages 575-584).

[0008] PSMA DKFZ 617 ( M. et al., Preclinical Evaluation of a Tailor-Made DOTA-Conjugated PSMA Inhibitor with Optimized Linker Moiety for Imaging and Endoradiotherapy of Prostate Cancer, Journal of Nuclear Medicine, 2015, 56(6):914-920, Becker, A. et al., Nephro-and hepatotoxicity after radioligand therapy of metastatic castrate-resistant prostate cancer with 177Lu-PSMA-617, Journal of Nuclear Medicine, 2016, 57(Suppl 2):1430-1430; Rahbar, K. et al Response and tolerability of a single dose of 177Lu-PSMA-617 in patients with metastatic castration-resistant prostate cancer: a multicenter retrospective analysis, Journal of Nuclear Medicine, 2016:p.jnumed.116.173757) and PSMA I&T (Weineisen, M. et al., Development and first in human evaluation of PSMA I&T - A ligand for diagnostic imaging and endoradiotherapy of prostate cancer, Journal of Nuclear Medicine, 2014, 55(Suppl 1):1083-1083, Eiber, M. et al., Systemic radioligand therapy with 177Lu-PSMA I&T in patients with metastatic castration-resistant prostate cancer, Journal of Nuclear Medicine, 2016, 57(Suppl 2):61-61; Schottelius, M.et al., [111In]PSMA-I&T: expanding the spectrum of PSMA-I&T applications towards SPECT and radioguided surgery, EJNMMI research, 2015, 5(1):1) used in a clinical setting for palliative care of prostate cancer patients. Chelating units DOTA and associated DOTAGA not only allow for imaging but also for therapeutic applications, as the potential range of radiometal chelates includes... 111 In、 177 Lu、 90 Y and 213 Bi et al. [ 111 PSMA I&T has been used clinically in radioguided surgery to assist surgeons in the removal of malignant tissue (Schottelius, M. et al.,

[111] PSMA-I&T: expanding the spectrum of PSMA-I&T applications towards SPECT and radioguided surgery, EJNMMI research, 2015, 5(1):1). Similarly, the recently developed and clinically tested PSMA inhibitor PSMA I&S (imaging and surgery) has shown encouraging results (Robu, S. et al., Preclinical evaluation and first patient application of 99mTc-PSMA-I&S for SPECT imaging and radioguided surgery in prostate cancer, Journal of Nuclear Medicine, 2016: p.jnumed.116.178939).

[0009] use[ 177 Intracavitary radiotherapy performed by PSMA I&T has demonstrated high efficacy, tolerability, and safety in patients receiving up to 7.4 GBq over four cycles. Dosimetry data of the organs obtained indicate that the kidneys and salivary glands, in particular, received the highest doses after tumor injury. For PSMA DKFZ 617 and [ 18F]DCFPyL showed similar radiation values (Rowe, S.P. et al., PSMA-Based [18F]DCFPyL PET / CT Is Superior to Conventional Imaging for Lesion Detection in Patients with Metastatic Prostate Cancer. Molecular Imaging and Biology, 2016: pp. 1-9; Delker, A. et al., Dosimetry for 177Lu-DKFZ-PSMA-617: a new radiopharmaceutical for the treatment of metastatic prostate cancer, European journal of nuclear medicine and molecular imaging, 2016, 43(1): pp. 42-51; Kabasakal, L. et al., Pre-therapeutic dosimetry of normal organs and tissues of 177Lu-PSMA-617 prostate-specific membrane antigen (PSMA) inhibitor in patients with castration-resistant prostate cancer, European journal of nuclear medicine and molecular imaging, 2015, 42(13): pp. 1976-1983; Yadav, M.P. et al., 177Lu-DKFZ-PSMA-617 therapy in metastatic castration resistant prostate cancer: safety, efficacy, and quality of life assessment, European journal of nuclear medicine and molecular imaging, 2016: pp. 1-11).The physiological expression of PSMA (Silver, DA, et al., Prostate-specific membrane antigen expression in normal and malignant human tissues, Clinical Cancer Research, 1997, 3(1): 81-85) and renal excretion of radiolabeled compounds explain these increases. Incidental renal and hematological toxicities following administration are usually reversible; however, there are reasonable concerns about chronic toxicities, particularly in patients with overall long-term survival, such as those with PCa. Therefore, appropriate concepts are needed to reduce unnecessary radiation exposure while simultaneously increasing tumor uptake.

[0010] In view of the above, the main technical problem of the present invention can be considered to be providing means and methods to reduce radiation-induced side effects of PSMA-targeted radiolabeled diagnostics and treatments. Another technical problem can be considered to be providing means and methods to increase tumor uptake of such diagnostic and therapeutic agents. More generally, the technical problem can be considered to be providing improved PSMA binders.

[0011] The technical problem is solved by the subject matter outlined in the appended claims and further explained in detail below. Summary of the Invention

[0012] Specifically, in a first aspect, the present invention provides compounds of formula (I) or pharmaceutically acceptable salts thereof.

[0013]

[0014] in:

[0015] m is an integer from 2 to 6, preferably from 2 to 4, and more preferably 2;

[0016] n is an integer from 2 to 6, preferably from 2 to 4, and more preferably 2 or 4;

[0017] R 1L It is CH2, NH or O, preferably NH;

[0018] R 2L It is C or P(OH), preferably C;

[0019] R 3L It is CH2, NH or O, preferably NH;

[0020] X 1 The bonds are selected from amide bonds, ether bonds, thioether bonds, ester bonds, thioester bonds, urea bridges, and amine bonds, and are preferably amide bonds;

[0021] L1 It is a divalent linking group having a structure selected from oligoamides, oligoethers, oligosulfides, oligopolyesters, oligosulfides, oligoureas, oligo(ether-amide), oligo(sulfide-amide), oligo(ester-amide), oligo(sulfide-amide), oligo(urea-amide), oligo(ether-sulfide), oligo(ether-ester), oligo(ether-sulfide), oligo(ether-urea), oligo(sulfide-ester), oligo(sulfide-sulfide), oligo(sulfide-urea), oligo(sulfide-urea), oligo(ester-sulfide), oligo(ester-urea), and oligo(sulfide-urea), preferably a divalent linking group having a structure selected from oligoamides and oligo(ester-amide), and the linking group may carry the EDS group;

[0022] X 2 The bonds are selected from amide bonds, ether bonds, thioether bonds, ester bonds, thioester bonds, urea bridges, and amine bonds, and are preferably amide bonds;

[0023] R 2 It is an optionally substituted aryl or optionally substituted aralkyl group, which may be substituted on its aromatic ring by one or more substituents selected from halogens (preferably I) and -OH;

[0024] R 3 It is an optionally substituted aryl or optionally substituted aralkyl group, which may be substituted on its aromatic ring by one or more substituents selected from halogens (preferably I) and -OH;

[0025] r is 0 or 1, preferably 1;

[0026] p is 0 or 1;

[0027] q is 0 or 1;

[0028] And preferably, p + q = 1;

[0029] R 4 Selected from aryl and group EDS;

[0030] X 3 Selected from amide bonds, ether bonds, thioether bonds, ester bonds, thioester bonds, urea bridges, amine bonds, and groups of the following formula.

[0031] The labeled bond on the carbonyl group will X 3 Connect to R M Another marker key will be X 3 Connected to the remainder of the compound of formula (I);

[0032] And preferably, it is an amide bond;

[0033] R MIt is a marker group, the marker group comprising a chelating group, the chelating group optionally comprising a chelated non-radioactive or radioactive cation;

[0034] Furthermore, the group EDS is contained at least once in the compound of formula (I) and has a structure selected from (E-1A), (E-1B), (E-2A), and (E-2B):

[0035]

[0036] in

[0037] The bond that links the group EDS to the remainder of the compound of formula (I) is marked;

[0038] s is 1, 2 or 3, preferably 1 or 2, more preferably 1;

[0039] t is 1, 2 or 3, preferably 1 or 2, more preferably 2;

[0040] For s > 1, R 5A Each occurrence is independently an electron-withdrawing substituent, preferably selected from -NO2 and -COOH, and more preferably -COOH, wherein R 5A The bond between the benzene ring and the benzene ring represents s groups R. 5A Replace the s hydrogen atoms at any position on the benzene ring;

[0041] For s>1, R 5B Each occurrence is independently a substituent with a lone pair of electrons on an atom of the benzene ring shown in the directly linked formula (E-1B), said substituent preferably selected from -OH and -NH2, and more preferably –NH2, and wherein R 5B The bonds between the benzene ring and the benzene ring represent s groups R. 5B Replace the s hydrogen atoms at any position on the benzene ring;

[0042] For t > 1, R 6A Each occurrence is independently an electron-withdrawing substituent, preferably selected from -NO2 and -COOH, and more preferably -COOH, wherein R 6A The bond between the benzene ring and the benzene ring represents t groups R. 6A Replace t hydrogen atoms at any position on the benzene ring; and

[0043] For t>1, R 6BEach occurrence is independently a substituent with a lone pair of electrons on an atom of the benzene ring shown in the directly linked formula (E-1B), said substituent preferably selected from -OH and -NH2, and more preferably –OH, and wherein R 6B The bond between the benzene ring and the benzene ring represents t groups R. 6B Replace t hydrogen atoms at any position on the benzene ring.

[0044] The introduction of EDS substituents as described above (where the aromatic ring contains one or more substituents with high electron density selected from electron-withdrawing substituents and substituents with lone pairs of electrons) produces several unexpected advantages. These advantages include increased affinity, improved internalization, enhanced tumor cell retention, less nonspecific binding, reduced renal accumulation, and increased tumor uptake.

[0045] In particular, the reduction in nonspecific uptake in organs other than the prostate leads to less unnecessary radiation and fewer radiation-induced side effects.

[0046] To illustrate these advantages, we will refer to the properties of the compounds referred to herein as PSMA-71 and PSMA-66, which will be discussed in detail below.

[0047] Specifically, nanomolar affinity (5.3 ± 2.0 nM vs. 7.9 ± 2.4 nM) and significantly improved internalization (206.8 ± 1.7% vs. 75.5 ± 1.6%) demonstrate [ 177 Lu]PSMA-71 is superior to [ 177 Lu]PSMA I&T. Biodistribution data indicate that, compared with [ 177 Compared to Lu]PSMA I&T, [ 177 Lu]PSMA-71 showed significantly higher tumor uptake (14.29±0.89 vs 4.06±1.12% ID / g, respectively), while renal accumulation was similar (32.36±2.49 vs 34.66±17.20% ID / g, respectively).

[0048] Similarly, lower nanomolar affinity (3.8 ± 0.3 nM vs. 7.9 ± 2.4 nM), significantly improved internalization (297.8 ± 2.0% vs. 75.5 ± 1.6%), increased in vitro tumor cell retention (90.1 ± 3.5% vs. 62.8 ± 0.4%, 60 min culture), lower in vivo nonspecific binding, reduced renal accumulation (117.5 ± 6.9% ID / g vs. 128.9 ± 10.7% ID / g), and more than two-fold increased tumor uptake (10.0 ± 0.4% vs. 4.7 ± 1.0% ID / g) demonstrate [ 177 Lu]PSMA-66 and [177 Lu]PSMA I&T is superior in comparison.

[0049] As described above, salts of the compounds of the present invention, including those of formula (I) (and preferred embodiments thereof), are also applicable to the present invention. It should be understood that these salts are generally pharmaceutically acceptable salt forms of these compounds, which can be formed, for example, by protonating an atom with a readily protonable lone pair of electrons (e.g., an amino group) with an inorganic or organic acid, or by forming a salt of a carboxylic acid group with a physiologically acceptable cation known in the art. Exemplary base addition salts include, for example, alkali metal salts, such as sodium or potassium salts; alkaline earth metal salts, such as calcium or magnesium salts; ammonium salts; aliphatic amine salts, such as trimethylamine, triethylamine, dicyclohexylamine, ethanolamine, diethanolamine, triethanolamine, procaine, meglumine, diethanolamine, or ethylenediamine; aralkylamine salts, such as N,N-dibenzylethylenediamine or benetamine; heterocyclic aromatic amine salts, such as pyridine, methylpyridine, quinoline, or isoquinoline; quaternary ammonium salts, such as tetramethylammonium, tetraethylammonium, benzyltrimethylammonium, benzyltriethylammonium, benzyltributylammonium, methyltrioctylammonium, or tetrabutylammonium; and basic amino acid salts, such as arginine or lysine. Exemplary acid addition salts include, for example, inorganic acid salts such as hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, phosphate (e.g., phosphate, hydrogen phosphate, or dihydrogen phosphate), carbonate, bicarbonate, or perchlorate; organic acid salts such as acetate, propionate, butyrate, valerate, hexanoate, heptate, octanoate, cyclopentanepropionate, undecanoate, lactate, maleate, oxalate, fumarate, tartrate, malate, citrate, nicotinate, benzoate, salicylate, or ascorbate; sulfonates such as methanesulfonate, ethanesulfonate, 2-hydroxyethanesulfonate, benzenesulfonate, p-toluenesulfonate (toluenesulfonate), 2-naphthalenesulfonate, 3-phenylsulfonate, or camphorsulfonate; and acidic amino acid salts such as aspartate or glutamate.

[0050] Other pharmaceutically acceptable examples of salts include, but are not limited to, acetates, alginates, alginates, ascorbic acid salts, aspartate salts, benzenesulfonates, benzoates, bicarbonates, bisulfates, tartrates, borates, bromides, butyrates, calcium edetate, camphorates, camphor sulfonates, camsylate, carbonates, chlorides, citrates, clavulanates, cyclopentanepropionate, diglucuronide, dihydrochloride, dodecyl sulfate, edetate, ethanediosulfate, and propionate dodecyl sulfate. Alkyl sulfates, esylates, esylates, formates, fumarates, gluceptates, gluconates, gluconates, glutamates, glycerol phosphates, glycolylarsanilates, hemisulfates, heptanates, hexanoates, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroiodide, 2- Hydroxyethanesulfonate, hydroxynaphthate, iodide, isothionate, lactate, lacturonate, laurate, lauryl sulfate, malate, maleate, malonate, mandelate, mesylate, methanesulfonate, methyl sulfate, mucilage, 2-naphthalenesulfonate, naphthalenesulfonate, nicotinate, nitrate, N-methylglucosamine ammonium salt, oleate, oxalate, bis(hydroxynaphthalene) salt (embosate), palmitate, pantothenate, pectate, persulfate, 3-phenylpropionate, phosphate / hydrophosphate, picrate, neopentanoate, polygalacturonate, propionate, salicylate, stearate, sulfate, hypoacetate, succinate, tannate, tartrate, teoclate, tosylate, triethyliodide, undecanoate, valerate (see, e.g., SMBerge et al., "Pharmaceutical") Salts, J. Pharm. Sci., 66, pp. 1-19 (1977)).

[0051] It should be understood that throughout this specification, unless otherwise stated, the term "compound" includes solvates, polymorphs, prodrugs, codrugs, eutectics, tautomers, racemates, enantiomers or diastereomers, or mixtures thereof.

[0052] When the compounds of the present invention are provided in crystalline form, the structure may include solvent molecules. The solvent is typically a pharmaceutically acceptable solvent and includes water (hydrates) or organic solvents, etc. Examples of possible solvates include ethanolates and isopropanolates.

[0053] The term "complex drug" refers to two or more therapeutic compounds linked by covalent chemical bonds. For a detailed definition, see, for example, N. Das et al., European Journal of Pharmaceutical Sciences 41, 2010, 571-588.

[0054] The term "eutectic" refers to a multicomponent crystal in which all components are solid under ambient conditions in their pure form. These components coexist in stoichiometric or non-stoichiometric form of target molecules or ions (i.e., the compounds of this invention) and one or more neutral molecular eutectic forming agents. For a detailed discussion, see, for example, Ning Shan et al., Drug Discovery Today, 13(9 / 10), 2008, 440446 and DJ Good et al., Cryst. Growth Des., 9(5), 2009, 2252–2264.

[0055] The compounds of the present invention may also be provided in the form of a prodrug, i.e., a compound that is metabolized in vivo into an active metabolite. Suitable prodrugs are, for example, esters. Specific examples of suitable groups are given in paragraphs

[0082] to

[0118] of US 2007 / 0072831 under the headings of prodrugs and protecting groups.

[0056] Regarding the pH-dependent charged state exhibited by the compounds of the present invention, it should be understood that all possible charged states are included. In this regard, the preferred pH range is 0 to 14.

[0057] With regard to the net charge of the compounds of the present invention, it should be understood that the compounds are provided in an electrically neutral form. This is achieved by one or more counterions, preferably counterions as defined above for the term "salt".

[0058] In formula (I), m is an integer from 2 to 6. Preferably, m is from 2 to 4, and more preferably 2. R 1L It is CH2, NH, or O, preferably NH. R 2L It is C or P(OH), preferably C. 3L It is CH2, NH, or O, preferably NH. Therefore, where m is 2, R 1L It is NH, R 2L It is C and R 3L Compounds of formula (I) of NH or their salts are also preferred.

[0059] n is an integer from 2 to 6, preferably from 2 to 4, more preferably 2 or 4, and most preferably 2.

[0060] Therefore, where m is 2, n is 2 or 4, and R 1LIt is NH, R 2L It is C and R 3L Compounds of formula (I) of NH or salts thereof are particularly preferred. More preferred are those in which m is 2, n is 2, and R is 2. 1L It is NH, R 2L It is C and R 3L It is a compound of formula (I) of NH or a salt thereof.

[0061] X in equation (I) 1 Selected from amide bonds (i.e., -C(O)-NH-), ether bonds (i.e., -O-), thioether bonds (i.e., -S-), ester bonds (i.e., -C(O)-O-), thioester bonds (i.e., -C(S)-O- or -C(O)-S-), urea bridges (i.e., –NH-C(O)-NH-), and amine bonds (i.e., –NH-). X 1 Preferably, it is an amide bond.

[0062] Furthermore, in equation (I), it is even more preferable that n is 2 and X 1 It is an amide bond, wherein the carbon atom of the amide bond –C(O)-NH- is attached to the group -(CH2). n -; or n is 4 and X 1 It is an amide bond, in which the carbon atom of the amide bond –C(O)-NH- is attached to the group -(CH2). n -. More preferably, n is 2 and X 1 It is an amide bond, in which the carbon atom of the amide bond –C(O)-NH- is attached to the group -(CH2). n -

[0063] Therefore, compounds of formula (I) and their salts are particularly preferred, wherein in formula (I), m is 2, n is 2, and R is 2. 1L It is NH, R 2L It is C, R 3L It is NH, and X 1 It is an amide bond, in which the carbon atom of the amide bond –C(O)-NH- is attached to the group -(CH2). n -

[0064] L in equation (I) 1It is a divalent linking group having a structure selected from oligoamides, oligoethers, oligosulfides, oligopolyesters, oligosulfides, oligoureas, oligo(ether-amide), oligo(sulfide-amide), oligo(ester-amide), oligo(sulfide-amide), oligo(urea-amide), oligo(ether-sulfide), oligo(ether-ester), oligo(ether-sulfide), oligo(ether-urea), oligo(sulfide-ester), oligo(sulfide-sulfide), oligo(sulfide-urea), oligo(sulfide-urea), oligo(ester-sulfide), oligo(ester-urea), and oligo(sulfide-urea). Preferably, it is a divalent linking group having a structure selected from oligoamides and oligo(ester-amide), more preferably, it is a divalent linking group having an oligoamide structure. The linking group may contain the EDS group.

[0065] In L 1 The term "oligomeric" used in the definitions of oligomeric amides, oligomeric ethers, oligomeric sulfides, oligopolyesters, oligomeric sulfides, oligoureas, oligomeric (ether-amide), oligomeric (sulfide-amide), oligomeric (ester-amide), oligomeric (sulfide-amide), oligomeric (urea-amide), oligomeric (ether-sulfide), oligomeric (ether-ester), oligomeric (ether-sulfide), oligomeric (ether-urea), oligomeric (sulfide-ester), oligomeric (sulfide-sulfide), oligomeric (sulfide-urea), oligomeric (sulfide-urea), oligomeric (ester-sulfide), oligomeric (ester-urea), and oligomeric (sulfide-urea) should preferably be understood to refer to a group having 2 to 20 subunits, more preferably 2 to 10 subunits, connected by bonds of the same type specified in the term. Those skilled in the art will understand that when two different types of bonds are indicated in parentheses, both types of bonds are included in the group involved (e.g., in "oligomeric (ester-amide)", both ester and amide bonds are included).

[0066] More preferably, L 1 It has a structure selected from oligoamides and oligo(ester-amides), wherein the oligoamide contains a total of 1 to 5, more preferably 1 to 3, and most preferably 1 or 2 amide bonds in its main chain, and the oligo(ester-amide) contains a total of 2 to 5, more preferably 2 to 3, and most preferably 2 amide and ester bonds in its main chain. In a particularly preferred embodiment, L 1 This represents a divalent linking group having an oligoamide structure, wherein the oligoamide structure contains one or two amide bonds in its main chain.

[0067] In addition, L 1 It may contain the group EDS as defined herein (i.e., a group with a high electron density substituent or "electron-dense substituent"), that is, with L 1 The covalently linked group EDS. Preferably, the optionally present group EDS is attached as a substituent to the divalent linking group L. 1 The main chain, L 1Having a structure selected from oligomers, oligoethers, oligosulfides, oligopolyesters, oligosulfides, oligoureas, oligo(ether-amides), oligo(sulfide-amides), oligo(ester-amides), oligo(sulfide-amides), oligo(urea-amides), oligo(ether-sulfide), oligo(ether-ester), oligo(ether-sulfide), oligo(ether-urea), oligo(sulfide-ester), oligo(sulfide-sulfide), oligo(sulfide-urea), oligo(sulfide-urea), oligo(ester-sulfide), oligo(ester-urea), and oligo(sulfide-urea), preferably having a structure selected from oligomers and oligo(ester-amides), and most preferably having an oligomer structure, wherein the main chain extends into the X in the compound of formula (I). 1 With X 2 Between. Furthermore, in this respect, L 1 The further preferred definition also applies, that is, more preferably, L 1 The product has a structure selected from oligoamides and oligo(ester-amides), wherein the oligoamide contains a total of 1 to 5, more preferably 1 to 3, and most preferably 1 or 2 amide bonds in its main chain, and the oligo(ester-amide) contains a total of 2 to 5, more preferably 2 to 3, and most preferably 2 amide and ester bonds in its main chain. In a particularly preferred embodiment, L 1 This represents a divalent linking group having an oligoamide structure, wherein the oligoamide structure contains one or two amide bonds in its main chain.

[0068] Based on the above, L 1 It can contain one or more, for example, two or three EDS groups. However, it is preferred that L... 1 Without the EDS or L group 1 It contains a group EDS, and more preferably L 1 It contains a group called EDS.

[0069] If L 1 If the component contains the group EDS (more preferably, the EDS contains a single group EDS), then preferably, the group EDS has a structure selected from (E-1A), (E-2A), and (E-2B). More preferably, the group EDS has a structure selected from (E-2A) and (E-2B), and most preferably, it has the structure (E-2A).

[0070] Those skilled in the art will understand that L 1 The presence of the EDS group can serve as information about the possible position of this group in the compound according to the invention. 1 The fact that L can contain the EDS group does not apply to L 1The presence of other groups on the main chain, such as substituted or additional substituents, imposes a restriction. For example, it is preferred that the linking group L... 1 It comprises one or more, for example two, groups attached to its main chain as substituents, said groups being independently selected from -OH, -OCH3, -COOH, -COOCH3, -NH2, and -NHC(NH)NH2. More preferably, the linking group L 1 It contains one or more, for example two, COOH groups attached to its main chain as substituents.

[0071] In equation (I), X 2 The bonds are selected from amide bonds, ether bonds, thioether bonds, ester bonds, thioester bonds, urea bridges, and amine bonds, and are preferably amide bonds. More preferably, the nitrogen atom of the amide bond –C(O)-NH- is attached to L. 1 .

[0072] Therefore, X is also preferred. 1 and X 2 All are amide bonds, especially amide bonds arranged in the preferred orientation as further defined above.

[0073] Based on the above, the preferred option is –X in equation (I). 2 -L 1 -X 1 - Some have a structure selected from the following:

[0074] *-C(O)-NH-R 7 -NH-C(O)-R 8 -C(O)-NH-(L-1),

[0075] *-C(O)-NH-R 9A -NH-C(O)-R 10A -C(O)-NH-R 11A -NH-C(O)- (L-2A), and

[0076] *-C(O)-NH-R 9B -C(O)-NH-R 10B -C(O)-NH-R 11B -NH-C(O)- (L-2B);

[0077] The amide bond marked with * is connected to the R in formula (I). 2 On the carbon atom, and in which

[0078] R 7 R 8 R 9A R 9B R 11A and R11B Independently selected from optionally substituted C2 to C10 alkanediyl groups, preferably optionally substituted straight-chain C2 to C10 alkanediyl groups, each of which may be substituted by one or more substituents independently selected from -OH, -OCH3, -COOH, -COOCH3, -NH2, -NHC(NH)NH2 and the EDS group, and

[0079] R 10A and R 10B The substituent is selected from optional substituted C2 to C10 alkyldiyl groups, preferably optional substituted straight-chain C2 to C10 alkyldiyl groups, and optional substituted C6 to C10 arenediyl groups, preferably phenylene groups, wherein the alkyldiyl and arenediyl groups may each be substituted by one or more substituents independently selected from -OH, -OCH3, -COOH, -COOCH3, -NH2, -NHC(NH)NH2 and the EDS group. 10A Preferably, it is an optional substituted C2 to C10 alkyldiyl as defined above, more preferably an optional substituted straight-chain C2 to C10 alkyldiyl. R 10B Preferably, it is an optional substituted C6 to C10 aryl dimethyl group as defined above, more preferably a phenylene group, such as p-phenylene.

[0080] Among the groups of formulas (L-1), (L-2A), and (L-2B), R is preferred. 7 The optional substituent is -COOH, R 8 The optional substituents present are the groups EDS and R. 9A and R 9B The optional substituent is -COOH, R 10A The optional substituent is the group EDS, and R 11A and R 11B The substituents that may be present are -COOH.

[0081] It is also preferred that each group of formulas (L-1) and (L-2A) carries at least one substituent as described above, preferably as R. 8 and R 10A The substituent group is EDS. Also in this context, it is preferred that the EDS group has a structure selected from (E-1A), (E-2A), and (E-2B). More preferably, the EDS group has a structure selected from (E-2A) and (E-2B), and most preferably, it has the structure (E-2A).

[0082] Furthermore, it is preferred that R in formula (L1) does not include carbon atoms contained in optional substituents. 7 and R 8The total number of carbon atoms in it is 6 to 20, more preferably 6 to 16; excluding carbon atoms contained in optionally present substituents, R of formula (L2A) 9A R 10A and R 11A The total number of carbon atoms in it is 6 to 20, more preferably 6 to 16; and in the absence of carbon atoms contained in optional substituents, R of formula (L2B) 9B R 10B and R 11B The total number of carbon atoms in it is 6 to 20, more preferably 6 to 16.

[0083] From the information provided above regarding n and X 1 From the information regarding the preferred meaning, it can be understood that a further preferred option is: if n is 4, then -X in equation (I) 2 -L 1 -X 1 -Having a structure (L1), and if n is 2, then –X in equation (I) 2 -L 1 -X 1 - It has a structure (L-2A) or (L-2B).

[0084] Based on the above definition, it is more preferable that –X in equation (I) is... 2 -L 1 -X 1 - Some have a structure selected from the following:

[0085] *-C(O)-NH-CH(COOH)-R 12 -NH-C(O)-R 13 -C(O)-NH-(L-3),

[0086] *-C(O)-NH-CH(COOH)-R 14 -NH-C(O)-R 15 -C(O)-NH-R 16 -CH(COOH)-NH-C(O)-

[0087] (L-4), and

[0088] *-C(O)-NH-CH(COOH)-R 17 -C(O)-NH-R 18 -C(O)-NH-R 19 -CH(COOH)-NH-C(O)-

[0089] (L-5);

[0090] The keys marked with * are connected to equation (I) containing R.2 carbon atoms,

[0091] R 12 and R 14 Independently selected from straight-chain C2 to C6 alkyldiyl groups, preferably selected from straight-chain C3 to C6 alkyldiyl groups.

[0092] R 13 It is a straight-chain C2 to C10 alkyldiyl, preferably a straight-chain C4 to C8 alkyldiyl.

[0093] R 15 and R 16 Independently selected from straight-chain C2 to C6 alkyldiyl groups, preferably selected from straight-chain C2 to C4 alkyldiyl groups.

[0094] And R 13 and R 15 Each may have an EDS group as a substituent, and preferably, R 13 and R 15 Each contains an EDS group as a substituent.

[0095] R 17 It is a straight-chain C2 to C6 alkyldiyl, preferably a straight-chain C2 to C4 alkyldiyl.

[0096] R 18 It is a phenylene oxide, such as p-phenylene oxide, and

[0097] R 19 It is a straight-chain C2 to C6 alkyldiyl, preferably a straight-chain C2 to C4 alkyldiyl.

[0098] In this context, it is preferable that it can be connected to R. 13 and R 15 The group EDS has a structure selected from (E-1A), (E-2A), and (E-2B). More preferably, the group EDS has a structure selected from (E-2A) and (E-2B), and most preferably, it has the structure (E-2A).

[0099] Furthermore, it is preferred that R in formula (L-3) does not include the carbon atom contained in the EDS group as a substituent. 12 and R 13 The total number of carbon atoms in formula (L-4) is 6 to 16, more preferably 6 to 14; and in the absence of carbon atoms contained in the substituent group EDS, R in formula (L-4) 14 R 15 and R 16 The total number of carbon atoms in it is 6 to 16, more preferably 6 to 14.

[0100] From the information provided above regarding n and X1 From the information regarding the preferred meaning, it can be understood that, particularly preferred, is: if n is 4, then -X in equation (I) 2 -L 1 -X 1 -Has a structure (L-3); and if n is 2, then –X in equation (I) 2 -L 1 -X 1 - It has a structure (L-4) or (L-5).

[0101] Particularly preferred is that in equation (I), n is 2, and –X 2 -L 1 -X 1 - Some have one of the following structures:

[0102] *-C(O)-NH-CH(COOH)-(CH2)4-NH-C(O)-CH(EDS)-CH2-C(O)-NH-(CH2)3-CH(CO OH)-NH-C(O)-(L-6)

[0103] *-C(O)-NH-CH(COOH)-(CH2)2-C(O)-NH-Ph-C(O)-NH-(CH2)3-CH(COOH)-NH-C(O)-(L-7)

[0104] The keys marked with * are connected to equation (I) containing R. 2 The carbon atom, EDS is the group EDS as defined herein, including its preferred embodiments, and Ph is a phenylene group.

[0105] In the same context, it is preferred that the group EDS has a structure selected from (E-1A), (E-2A), and (E-2B). More preferably, the group EDS has a structure selected from (E-2A) and (E-2B), and most preferably, it has the structure (E-2A).

[0106] In equation (I), R 2 It is an optionally substituted aryl group or an optionally substituted aralkyl group, preferably an optionally substituted aralkyl group. It should be understood that the term "aralkyl" as used herein refers to an alkyl group in which the hydrogen atom is replaced by an aryl group as a substituent. Preferably, an aralkyl group is a group in which one aryl group is bonded to an alkyl diene. R 2 The aryl or aralkyl group represented may be substituted on its aromatic ring with one or more substituents selected from halogens (preferably I) and -OH. The aryl moiety of the aryl group and the aralkyl group is preferably selected from phenyl and naphthyl, such as 2-naphthyl. The alkyl dienyllium moiety of the aralkyl group is preferably C1-C4 alkyl dienyllium, more preferably a -CH2- group. Therefore, R2 More preferably selected from optionally substituted -CH2-phenyl and optionally substituted -CH2-naphthyl, particularly optionally substituted –CH2-(2-naphthyl). The optionally substituted -CH2-(2-naphthyl) is R 2 A particularly preferred choice.

[0107] The optionally substituted aryl group and the aryl moiety of the optionally substituted aralkyl group, including their preferred embodiments, can be substituted by one or more substituents selected from halogens (preferably I) and -OH. Therefore, one or more, for example, two or three substituents selected from halogens (preferably I) and -OH can be present. However, it is preferred that R... 2 It is not replaced.

[0108] Based on the above, it should be understood that R 2 Most preferably, it is unsubstituted -CH2-(naphthyl), and the naphthyl is most preferably 2-naphthyl, thereby providing R as –CH2-(2-naphthyl). 2 .

[0109] In equation (I), R 3 It is an optionally substituted aryl group or an optionally substituted aralkyl group, preferably an optionally substituted aralkyl group. The aryl group or aralkyl group may be substituted on its aromatic ring with one or more substituents selected from halogens (preferably I) and -OH. The aryl moiety of the aryl group and aralkyl group is preferably selected from phenyl and naphthyl, for example, 2-naphthyl. More preferably, the aryl moiety of the aryl group and the aralkyl group is phenyl. The alkyl dienyllium moiety of the aralkyl group is preferably C1-C4 alkyl dienyllium, more preferably a -CH2- group. Therefore, R 3 More preferably, it is an optionally substituted -CH2-phenyl.

[0110] The optionally substituted aryl group and the aryl moiety of the optionally substituted aralkyl group, including their preferred embodiments, can be substituted by one or more substituents selected from halogens (preferably I) and -OH. Therefore, one or more, for example, two or three substituents selected from halogens (preferably I) and -OH can be present. Preferably, R 3 It can be substituted by a substituent -OH, or by a combination of a substituent -OH and a substituent -I.

[0111] Therefore, R is particularly preferred. 3 It is a -CH2-phenyl group that is substituted on the benzene ring by a substituent -OH, or by a combination of a substituent -OH and a substituent -I, and most preferably, the substituent -OH is located at the para position of the benzene ring relative to the -CH2- group.

[0112] Based on the above, preferably, in equation (I), R2 It is a group of the following formula

[0113] or

[0114] And R 3 It is a group of the following formula

[0115] or

[0116] in The markers will respectively mark R 2 and R 3 The bonds that connect to the rest of the compound of formula (I).

[0117] Even better is R 2 and R 3 The combination of R, where R 2 It is a group of the following formula

[0118]

[0119] And R 3 It is a group of the following formula

[0120]

[0121] in The markers will respectively mark R 2 and R 3 Bonds that connect to the rest of the compound.

[0122] In equation (I), r can be 0 or 1, preferably r is 1.

[0123] Furthermore, as mentioned above, p is 0 or 1, and q is 0 or 1, and preferably, p + q = 1. More preferably, p is 0 and q is 1.

[0124] R in equation (I) 4 Selected from aryl and group EDS. The aryl group is preferably selected from phenyl and naphthyl, such as 2-naphthyl. Therefore, R 4 More preferably, it is selected from phenyl, naphthyl (e.g., 2-naphthyl) and the EDS group. Most preferably, it is the EDS group.

[0125] If R 4 If the group is EDS, then preferably, the group EDS has a structure selected from (E-1A), (E-2A) and (E-1B).

[0126] X 3 Selected from amide bonds, ether bonds, thioether bonds, ester bonds, thioester bonds, urea bridges, amine bonds, and groups of the following formula.

[0127] The labeled bond on the carbonyl group will X 3 Connect to R M Another marker key will be X 3 It connects to the rest of the molecule.

[0128] Preferably, X 3 Selected from amide bonds and groups of the following formula

[0129] The labeled bond on the carbonyl group will X 3 Connect to R M Another marker key will be X 3 It connects to the rest of the molecule.

[0130] In a more preferred embodiment, X 3 It is an amide bond -C(O)-NH-, in which the carbon atom is attached to R. M .

[0131] R M It is a marker group, which includes a chelating group, which optionally includes a chelated non-radioactive or radioactive cation.

[0132] Those skilled in the art will understand that the above definition (according to the definition, R) M (Contains chelating groups) covering R M In the case of a chelating group; in this case, the chelating group is usually directly related to X. 3 Combine;

[0133] And covering R M In cases where the chelating group includes, for example, another linker portion; in this case, the chelating group can be indirectly connected to X through this other linker portion. 3 Combine.

[0134] R M The provided chelating groups are suitable for forming chelates with radioactive or non-radioactive cations. Suitable chelating groups for various cations are well known in the art and can be used in the context of this invention.

[0135] The chelating group optionally comprising a chelating non-radioactive or radioactive cation is preferably selected from chelating groups comprising at least one of the following:

[0136] (i) a macrocyclic structure having 8 to 20 ring atoms, wherein two or more, preferably three or more, of the ring atoms are selected from oxygen, sulfur, and nitrogen atoms; and

[0137] (ii) A non-cyclic open-chain structure having 8 to 20 main chain atoms, wherein 2 or more, preferably 3 or more, of the main chain atoms are heteroatoms selected from oxygen, sulfur and nitrogen atoms.

[0138] Exemplary chelating groups, and therefore also exemplary groups R. M , is a residue selected from the following chelating agents: bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (CBTE2a), cyclohexyl-1,2-diaminetetraacetic acid (CDTA), 4-(1,4,8,11-tetraazabicyclotetradecyl-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 (CBTE2a), cyclohexyl-1,2-diaminetetraacetic acid (CDTA), 4-(1,4,8,11-tetraazabicyclotetradecyl-1-yl)-methylbenzoic acid (CPTA), N'-[5-[acetyl(hydroxy)amino]pentyl]-N-hydroxybutyramide (DFO), 4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclotetradecane (CDTA), cyclohexyl-1,2-diaminetetraacetic acid ... Cyclo[6.6.2]hexadecane (DO2A), 1,4,7,10-tetraazacyclododecyl-N,N',N”,N”'-tetraacetic acid (DOTA), 2-[1,4,7,10-tetraazacyclododecyl-4,7,10-triacetic acid]-glutaric acid (DOTAGA), N,N'-dipyridoxyethylenediamine-N,N'-diacetic acid-5,5'-bis(phosphate) (DPDP), diethylenetriaminepentaacetic acid (DTPA), ethylenediamine-N,N'-tetraacetic acid (EDTA), ethylene glycol-O,O-bis(2-aminoethyl)-N,N,N',N'-tetraacetic acid (E GTA), 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 (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-(carbonyloxy)-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), tripyridylbis(methyleneaminotetraacetic acid) (TMT), 1,4,7,10-tetraazacyclotridecane-N,N',N”,N”'-tetraacetic acid (TRITA), triethylenetetraminehexaacetic acid (TTHA), N,N′-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18-crown ether-6(H 2macropa) and 4-amino-4-{2-[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydropyridin-2-ylmethyl)carbamoyl]ethyl} pimelic acid bis[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydropyridin-2-ylmethyl)-amide](THP);

[0139] The residues are provided by covalently bonding the carboxyl group contained in the chelating agent to the remainder of the compound via an ester or amide bond, preferably an amide bond. Those skilled in the art will understand that, in formula (I), the ester or amide bond can be X in this case. 3 Includes or may preferably be composed of X 3 express.

[0140] Among these chelating agents, DOTA and DOTAGA are preferred.

[0141] Therefore, it is also preferred that R in equation (I) M -X 3 - is a group of the following formula

[0142]

[0143] Among them, the use of The marked bond is attached to the remainder of the compound of formula (I), and the chelating group may comprise a chelated non-radioactive or radioactive cation.

[0144] The exemplary radioactive cations optionally chelated by the chelating group are 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 cations, or those containing 18 F-cation molecules, for example 18 F-[AlF] 2+ .

[0145] The preferred chelated cations are selected from 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 227Th cations, or containing 18 F is a cationic molecule.

[0146] In formula (I), the EDS group is included at least once, such that it can include, for example, one, two, or three EDS groups. Preferably, the compound or salt according to the invention includes one or two EDS groups. As described above, the one or more EDS groups can be derived from L 1 Carry, and / or may be carried by R 4 represent.

[0147] The most preferred compound of formula (I) or a salt thereof is one comprising a linking group L 1 Compounds or salts thereof carrying one EDS group, including the preferred embodiments as described above, and compounds or salts thereof containing two EDS groups, one of which is composed of R 4 The other is represented by L (i.e., r is 1). 1 Carry, including its best implementation as described above.

[0148] As described above, the group EDS has a structure selected from (E-1A), (E-1B), (E-2A), and (E-2B):

[0149]

[0150] in

[0151] The bond that links the group EDS to the remainder of the compound of formula (I) is marked;

[0152] s is 1, 2 or 3, preferably 1 or 2, more preferably 1;

[0153] t is 1, 2 or 3, preferably 1 or 2, more preferably 2;

[0154] For s>1, R 5A Each occurrence is independently an electron-withdrawing substituent, preferably selected from -NO2 and -COOH, and more preferably -COOH, wherein R 5A The bond between the benzene ring and the benzene ring represents s groups R. 5A Replace the s hydrogen atoms at any position on the benzene ring;

[0155] For s>1, R 5B Each occurrence is independently a substituent with a lone pair of electrons on an atom of the benzene ring shown in the directly linked formula (E-1B), said substituent preferably selected from -OH and -NH2, and more preferably –NH2, and wherein R 5B The bonds between the benzene ring and the benzene ring represent s groups R. 5B Replace s hydrogen atoms at any position on the benzene ring;

[0156] For t>1, R6A Each occurrence is independently an electron-withdrawing substituent, preferably selected from -NO2 and -COOH, and more preferably -COOH, wherein R 6A The bond between the benzene ring and the benzene ring represents t groups R. 6A t hydrogen atoms replacing any position on the benzene ring; and

[0157] For s>1, R 6B Each occurrence is independently a substituent with a lone pair of electrons on an atom of the benzene ring shown in the directly linked formula (E-1B), said substituent preferably selected from -OH and -NH2, and more preferably -OH, wherein R 6B The bonds between the benzene ring and the benzene ring represent t groups R. 6B t hydrogen atoms replacing any position on the benzene ring.

[0158] Typically, it is preferred that, in the EDS group (E-1A), for s > 1, the substituent R 5A It is the same and selected from -NO2 and -COOH, and more preferably -COOH; and in the EDS group (E-2A), for t>1, the substituent R 6A They are the same and selected from -NO2 and -COOH, and more preferably -COOH.

[0159] Similarly, it is generally preferred that, in the EDS group (E-1B), for s > 1, the substituent R 5B They are identical and selected from –OH and –NH2, and more preferably –NH2; and in the EDS group (E-2B), for t>1, the substituent R 6B They are the same and selected from -OH and –NH2, and more preferably –OH.

[0160] Therefore, it is further preferred that the compound of formula (I) contains the group EDS having formula (E-2A):

[0161]

[0162] in The bond that links the group EDS to the remainder of the compound of formula (I) is marked; and

[0163] t is 1 or 2, and R 6A It is -NO2 or -COOH.

[0164] In the context of this invention, the following groups are most preferably used as the EDS group.

[0165]

[0166] Based on the above, the preferred compound of formula (I) is represented by the following formula (Ia).

[0167]

[0168] Where n, X 1 L 1 X 2 R 2 R 3 R 4 q, p, X 3 and R M As defined above, including preferred embodiments thereof, wherein the group EDS is contained at least once and has the structure as defined above, including the preferred embodiments thereof.

[0169] Formula (Ib) illustrates a more preferred compound of formula (I).

[0170]

[0171] Where n, X 1 L 1 X 2 R 2 R 3 R 4 X 3 and R M As defined above, including preferred embodiments thereof, wherein the group EDS is contained at least once and has the structure as defined above, including the preferred embodiments thereof.

[0172] Formula (Ic) illustrates a more preferred compound of formula (I).

[0173]

[0174] Where n, X 1 L 1 X 2 R 4 X 3 and R M As defined above, including preferred embodiments thereof, wherein the group EDS is contained at least once and has the structure as defined above, including the preferred embodiments thereof.

[0175] The following formulas (Id) and (Ie) illustrate the more preferred compound of formula (I):

[0176]

[0177] Where R 9A R 10A R 11AR 4 X 3 and R M As defined above, including their preferred embodiments, and wherein (i)R 4 It is a group EDS having the structure defined above, including its preferred embodiment, or (ii)R 10A EDS having a group having the structure defined above, including its preferred embodiment, or both (i) and (ii) are applicable;

[0178]

[0179] Where R 9B R 10B R 11B R 4 X 3 and R M As defined above, including their preferred embodiments, and wherein R 4 It is a group EDS having the structure as defined above, including its preferred embodiments.

[0180] The following formulas (If) and (Ig) illustrate the more preferred compound of formula (I).

[0181]

[0182] Where R 9A R 10A R 11A R 4 X 3 and R M As defined above, including their preferred embodiments, and wherein (i)R 4 It is a group EDS having the structure defined above, including its preferred embodiment, or (ii)R 10A EDS having a group having the structure defined above, including its preferred embodiment, or both (i) and (ii) are applicable;

[0183]

[0184] Where R 9B R 10B R 11B R 4 X 3 and R M As defined above, including their preferred embodiments, and wherein R 4 It is a group EDS having the structure as defined above, including its preferred embodiments.

[0185] In a preferred embodiment, the chelating group has a bonded radionuclide bond, and the radionuclide emits alpha radiation. Radionuclides emitting alpha radiation include... 212 Bi、 213 Bihe 225 Acc.

[0186] As described above, the introduction of electron-deficient substituents significantly increases internalization capacity. This characteristic, as confirmed by in vitro experiments, leads to higher tumor uptake, particularly longer retention in tumor tissue (see Examples). Because the complex of the chelating agent and the alpha-particle-emitting radionuclide is readily disintegrated by physical recoil, the prolonged intracellular retention characteristic reduces the likelihood of the radionuclide freely circulating in vivo, thereby improving safety and reducing unnecessary radiation exposure.

[0187] The particularly preferred compounds of the present invention are as follows:

[0188] DOTAGA–y(3-I)fk(L-Asu[KuE]-2,4-DNBA)(PSMA-36):

[0189]

[0190] DOTAGA-F(4-NH2)y-2-nal-k(Suc-N 5 -orn-C 4 -EuE)(PSMA-49):

[0191]

[0192] DOTAGA-F(4-NO2)-y-2-nal-k(Suc-N 5 -orn-C 4 -EuE)(PSMA-52):

[0193]

[0194] 2,4-DNBA-Dap(DOTAGA)-y-2-nal-k(Suc-N 5 -orn-C 4 -EuE)(PSMA-53):

[0195]

[0196] DOTAGA-F(4-NH2)y-2-nal-e(Abz-N 5 -orn-C 4 -EuE)(PSMA-60):

[0197]

[0198] DOTAGA-F(4-NH2)y-2-nal-k(d[N 5 -orn-C 4 -EuE]-2,4-DNBA)(PSMA-61):

[0199]

[0200] DOTAGA-F(4-NH2)y-2-nal-k(d[N 5 -orn-C 4 -EuE]-TMA)(PSMA-62):

[0201]

[0202] 2,4-DNBA-Dap(DOTAGA)y-2-nal-e(Abz-N 5 -orn-C 4 -EuE)(PSMA-65):

[0203]

[0204] DOTAGA-Dap(TMA)y-2-nal-k(d[N 5 -orn-C 4 -EuE]-TMA)(PSMA-66):

[0205]

[0206] DOTAGA-2-Nal-y-2-nal-k(d[N 5 -orn-C 4 -EuE]-TMA)(PSMA-71):

[0207]

[0208] DOTAGA-F(4-NH2)y-2-nal-k(d[N 5 -orn-C 4 -EuE]-3,5-DHBA)(PSMA-78):

[0209]

[0210] The advantageous properties of these compounds of the present invention can be seen from the data in Tables 1 and 2 below, which show that... Figure 1 It is shown graphically in the middle.

[0211] Table 1. Overview of all in vitro study parameters for EuK-based PSMA inhibitors. E represents Glu, u represents urea, and K represents Lys. Lowercase single letters (e.g., "y") indicate the D form of the corresponding amino acid. The half-maximal inhibitory concentration (IC50) of the PSMA inhibitor is also listed. 50 Using LNCaP cells (1.5*10) 5 Cells / well, 1 hour, 4°C, HBSS + 1% BSA) and as radioligands ([ 125 I]I-BA)KuE, determined in competitive binding assays. Internalized radioactivity (activity) is expressed as [%] against ([ 125 Relative cellular uptake of I]I-BA)KuE (1.25*10) 5 Cells / well, PLL coated plate, for ([ 125 I]I-BA)KuE, c=0.2nm, and for 177 Lu-labeled PSMA inhibitor, c = 1.0 nm. (DMEM / F-12 + 5% BSA, 37 °C, 60 min). Data corrected for nonspecific binding (10 μm 2-PMPA). IC 50 Internalization data are expressed as mean ± SD (n = 3). Lipophilicity is expressed as logP (distribution coefficient in n-octanol / PBS) of the radiolabeled PSMA inhibitor. LogP data are expressed as mean ± SD (n = 6). After logarithmic plotting and correction, albumin binding (HSA) is expressed as [%] (n = 1). Configurations describe the simplified N-terminal to C-terminal structural composition of the peptide spacer and linker in the absence of chelating agents. nd = not determined. "-||-" indicates simplified conjugation.

[0212]

[0213] Table 2. Overview of all in vitro study parameters for EuE-based PSMA inhibitors. Half-maximal inhibitory concentration (IC50) of PSMA inhibitors. 50 Using LNCaP cells (1.5*10) 5 Cells / well, 1 hour, 4°C, HBSS + 1% BSA) and as radioligands ([ 125 I]I-BA)KuE, determined in a competitive binding assay. Internalized radioactivity is expressed as [%] relative to ([ 125 Relative cellular uptake of I]I-BA)KuE (1.25*10) 5 Cells / well, PLL coated plate, for ([ 125 I]I-BA)KuE, c=0.2nm, and for 177Lu-labeled PSMA inhibitor, c = 1.0 nm. (DMEM / F-12 + 5% BSA, 37 °C, 60 min). Data corrected for nonspecific binding (10 μm²-PMPA). IC 50 Internalization data are expressed as mean ± SD (n = 3). Lipophilicity is expressed as logP (distribution coefficient in n-octanol / PBS) of the radiolabeled PSMA inhibitor. LogP data are expressed as mean ± SD (n = 6). After logarithmic plotting and correction, albumin binding (HSA) is expressed as [%] (n = 1). Configurations describe the simplified N-terminal to C-terminal structural composition of the peptide spacer and linker in the absence of chelating agents. nd = not determined. "-||-" indicates simplified conjugation.

[0214]

[0215]

[0216] The preferred labeling schemes for these most preferred compounds are defined above.

[0217] In another aspect, the present invention provides pharmaceutical compositions comprising, or consisting of, one or more compounds or salts of the present invention disclosed above.

[0218] In another aspect, the present invention provides diagnostic compositions comprising, or consisting of, one or more compounds or salts of the present invention disclosed above.

[0219] In another aspect, the present invention provides therapeutic compositions comprising, or consisting of, one or more compounds or salts of the present invention disclosed above.

[0220] The pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, excipient, and / or diluent. Examples of suitable pharmaceutical carriers, excipients, and / or diluents are well known in the art and include phosphate-buffered saline solutions, water, emulsions such as oil / water emulsions, various types of wetting agents, sterile solutions, etc. Compositions containing such carriers can be formulated using known conventional methods. These pharmaceutical compositions can be administered to a subject at an appropriate dose. Administration of appropriate compositions can be accomplished in various ways, such as intravenous, intraperitoneal, subcutaneous, intramuscular, local, intradermal, intranasal, or intrabronchial administration. Particularly preferred is administration by, for example, injection and / or delivery to a site in the pancreas, delivery into a cerebral artery, or direct entry into brain tissue. The composition can also be administered directly to a target site, for example, via a biological ballistic delivery to an external or internal target site, such as the pancreas or brain. Dosing regimens are determined by the attending physician and clinical factors. As is well known in the medical field, the dosage for any patient depends on many factors, including the patient's body size, body surface area, age, the specific compound to be administered, sex, time and route of administration, overall health condition, and other medications administered concurrently. Pharmaceutically active substances can be present in doses ranging from 0.1 ng to 10 mg / kg body weight; however, doses below or above this exemplary range can be contemplated, particularly taking into account the factors described above.

[0221] With respect to the aforementioned disclosed pharmaceutical compositions, diagnostic compositions, and therapeutic compositions comprising one or more compounds of the present invention, it is preferred that no other pharmaceutically active compounds, diagnostically active compounds, or therapeutically active compounds are present. Alternatively, other compounds with therapeutic, diagnostic, or pharmaceutical activity, such as anticancer agents, may be present.

[0222] The combination of therapeutic treatments with the compounds of this invention may have synergistic or cumulative therapeutic effects, similar to those achieved with […]. 177 LuDOTATATE radiotherapy combined with chemotherapy or immunotherapy is used to treat neuroendocrine tumors. (Comparison) 177 Lu PRRT and oral chemotherapy agent capecitabine (Xeloda; Genentech) in combination and alone [ 177The first phase 3 study of Lu-octreotate began in 2017 at Erasmus MC in Rotterdam (van Essen M, Krenning EP, Kam BL, de Herder WW, van Aken MO, Kwekkeboom DJ, Report on short-term side effects of treatments with 177Lu-octreotate in combination with capecitabine in seven patients with gastroenteropancreatic neuroendocrine tumors. Eur J Nucl Med Mol Imaging. 2008; 35:743–748).

[0223] Further research on a combination therapy, termed peptide receptor chemoradionuclide therapy (PRCRT), has recently been published (Kong G, Callahan J, Hofman MS, et al. High clinical and morphologic response using 90Y-DOTA-octreotate sequenced with 177Lu-DOTA-octreotate induction peptide receptor chemoradionuclide therapy (PRCRT) for bulky neuroendocrine tumors. Eur J Nucl Med Mol Imaging. 2017; 44:476–489). Similar "combination therapy" approaches will be developed in the near future to improve the efficiency of PSMA-targeted radioligand therapy.

[0224] In another aspect, the present invention provides one or more compounds or salts of the present invention disclosed above for medical use.

[0225] Preferred uses in medicine include nuclear medicine, such as nuclear diagnostic imaging, also known as nuclear molecular imaging, and / or targeted radiotherapy for diseases associated with overexpression, preferably overexpression of PSMA in diseased tissues.

[0226] In another aspect, the present invention provides compounds or salts of the present invention as defined above, which are used in methods for the diagnosis and / or staging of cancer, preferably prostate cancer.

[0227] Preferred indications include the detection or staging of cancers (e.g., but not limited to high-grade gliomas, lung cancer, especially prostate cancer and metastatic prostate cancer), the detection of metastatic disease in patients with intermediate to high-risk primary prostate cancer, and the detection of metastatic sites in patients with biochemically recurrent prostate cancer, even in cases of low serum PSA levels. Another preferred indication is the imaging and visualization of angiogenesis.

[0228] Cancer is the preferred indication for medical treatment, especially radiation therapy. Prostate cancer is a particularly preferred indication.

[0229] In another aspect, the present invention provides compounds or salts of the present invention as defined above, which are used in methods for the diagnosis and / or staging of cancer, preferably prostate cancer.

[0230] With regard to the embodiments characterized in this application, particularly in the claims, it is intended that each embodiment mentioned in the dependent claims be combined with each embodiment of each claim (independent or dependent claim) referenced by the dependent claims. For example, where independent claim 1 describes three alternatives A, B, and C, dependent claim 2 describes three alternatives D, E, and F, and dependent claim 3, which references claims 1 and 2, describes three alternatives G, H, and I, it should be understood that, unless otherwise specified, this application expressly discloses embodiments corresponding to the following combinations: A, D, G; A, D, H; A, D, I; A, E, G; A, E, H; A, E, I; A, F, G; A, F, H; A, F, I; B, D, G; B, D, H; B, D, I; B, E, G; B, E, H; B, E, I; B, F, G; B, F, H; B, F, I; C, D, G; C, D, H; C, D, I; C, E, G; C, E, H; C, E, I; C, F, G; C, F, H; C, F, I.

[0231] Similarly, and where no alternatives are listed in the independent and / or dependent claims, it should be understood that if a dependent claim refers back to multiple prior claims, then any combination of the subject matter thereby covered is considered explicitly disclosed. For example, if independent claim 1, dependent claim 2 refers back to claim 1, and dependent claim 3 refers back to both claims 2 and 1, then the combination of the subject matter of claims 3 and 1 is explicitly and clearly disclosed, as is the combination of the subject matter of claims 3, 2, and 1. In the presence of another dependent claim 4 that refers to any one of claims 1 to 3, then the combination of the subject matter of claims 4 and 1, claims 4, 2 and 1, claims 4, 3 and 1, and claims 4, 3, 2, and 1 is explicitly and clearly disclosed.

[0232] Specifically, the present invention provides the subject matter outlined in the following items.

[0233] 1. A compound of formula (I) or a pharmaceutically acceptable salt thereof,

[0234]

[0235] in:

[0236] m is an integer from 2 to 6, preferably from 2 to 4, and more preferably 2;

[0237] n is an integer from 2 to 6, preferably from 2 to 4, and more preferably 2 or 4;

[0238] R 1L It is CH2, NH or O, preferably NH;

[0239] R 2L It is C or P(OH), preferably C;

[0240] R 3L It is CH2, NH or O, preferably NH;

[0241] X 1 The bonds are selected from amide bonds, ether bonds, thioether bonds, ester bonds, thioester bonds, urea bridges, and amine bonds, and are preferably amide bonds;

[0242] L 1 It is a divalent linking group having a structure selected from oligoamides, oligoethers, oligosulfides, oligopolyesters, oligosulfides, oligoureas, oligo(ether-amide), oligo(sulfide-amide), oligo(ester-amide), oligo(sulfide-amide), oligo(urea-amide), oligo(ether-sulfide), oligo(ether-ester), oligo(ether-sulfide), oligo(ether-urea), oligo(sulfide-ester), oligo(sulfide-sulfide), oligo(sulfide-urea), oligo(sulfide-urea), oligo(ester-sulfide), oligo(ester-urea), and oligo(sulfide-urea), preferably a divalent linking group having a structure selected from oligoamides and oligo(ester-amide), and the linking group may carry the EDS group;

[0243] X 2 The bonds are selected from amide bonds, ether bonds, thioether bonds, ester bonds, thioester bonds, urea bridges, and amine bonds, and are preferably amide bonds;

[0244] R 2 It is an optionally substituted aryl or optionally substituted aralkyl group, which may be substituted on its aromatic ring by one or more substituents selected from halogens (preferably I) and -OH;

[0245] R 3It is an optionally substituted aryl or optionally substituted aralkyl group, which may be substituted on its aromatic ring by one or more substituents selected from halogens (preferably I) and -OH;

[0246] r is 0 or 1, preferably 1;

[0247] p is 0 or 1;

[0248] q is 0 or 1;

[0249] And preferably, p + q = 1;

[0250] R 4 The aryl group and group EDS are selected from the optional substituted aryl group, wherein the aryl group may be substituted on its aromatic ring by one or more substituents selected from halogen (preferably I), -OH and –NH2;

[0251] X 3 Selected from amide bonds, ether bonds, thioether bonds, ester bonds, thioester bonds, urea bridges, amine bonds, and groups of the following formula.

[0252] The labeled bond on the carbonyl group will X 3 Connect to R M Another marker key will be X 3 Connected to the remainder of the compound of formula (I);

[0253] And preferably, it is an amide bond;

[0254] R M It is a marker group, the marker group comprising a chelating group, the chelating group optionally comprising a chelated non-radioactive or radioactive cation;

[0255] Furthermore, the group EDS is contained at least once in the compound of formula (I) and has a structure selected from (E-1A), (E-1B), (E-2A), and (E-2B):

[0256]

[0257]

[0258] in

[0259] The bond that links the group EDS to the remainder of the compound of formula (I) is marked;

[0260] s is 1, 2 or 3, preferably 1 or 2, more preferably 1;

[0261] t is 1, 2 or 3, preferably 1 or 2, more preferably 2;

[0262] For s > 1, R 5A Each occurrence is independently an electron-withdrawing substituent, preferably selected from -NO2 and -COOH, and more preferably -COOH, wherein R 5A The bond between the benzene ring and the benzene ring represents s groups R. 5A Replace the s hydrogen atoms at any position on the benzene ring;

[0263] For s>1, R 5B Each occurrence is independently a substituent with a lone pair of electrons on an atom of the benzene ring shown in the directly linked formula (E-1B), said substituent preferably selected from -OH and -NH2, and more preferably –NH2, and wherein R 5B The bonds between the benzene ring and the benzene ring represent s groups R. 5B Replace the s hydrogen atoms at any position on the benzene ring;

[0264] For t > 1, R 6A Each occurrence is independently an electron-withdrawing substituent, preferably selected from -NO2 and -COOH, and more preferably -COOH, wherein R 6A The bond between the benzene ring and the benzene ring represents t groups R. 6A Replace t hydrogen atoms at any position on the benzene ring; and

[0265] For t>1, R 6B Each occurrence is independently a substituent with a lone pair of electrons on an atom of the benzene ring shown in the directly linked formula (E-1B), said substituent preferably selected from -OH and -NH2, and more preferably –OH, and wherein R 6B The bond between the benzene ring and the benzene ring represents t groups R. 6B Replace t hydrogen atoms at any position on the benzene ring.

[0266] 2. The compound or salt according to item 1, wherein m is 2, n is 2 or 4, and R 1L It is NH, R 2L It is C, and R 3L It is NH.

[0267] 3. The compound or salt according to item 1 or 2, wherein n is 2.

[0268] 4. The compound or salt according to any one of items 1 to 3, wherein X 1 It is an amide bond.

[0269] 5. The compound or salt according to item 4, wherein n is 2 and X 1 It is an amide bond, wherein the carbon atom of the amide bond –C(O)-NH- is attached to the group (CH2).n .

[0270] 6. The compound or salt according to any one of items 1 to 5, wherein L 1 It is a divalent linking group having a structure selected from oligoamides and oligo(ester-amides), wherein the oligoamide contains a total of 1 to 5, more preferably 1 to 3, and most preferably 1 or 2 amide bonds in its main chain, and the oligo(ester-amide) contains a total of 2 to 5, more preferably 2 to 3, and most preferably 2 amide and ester bonds in its main chain, and the linking group may carry the EDS group.

[0271] 7. The compound or salt according to item 6, wherein L 1 This represents a divalent linking group having an oligoamide structure containing one or two amide bonds in its main chain, and the linking group may have the EDS group.

[0272] 8. The compound or salt according to any one of claims 1 to 7, wherein the linking group L 1 It contains a group called EDS.

[0273] 9. The compound or salt according to any one of items 1 to 8, wherein X 2 It is an amide bond.

[0274] 10. The compound or salt according to item 9, wherein X 2 It is an amide bond, wherein the nitrogen atom of the amide bond –C(O)-NH- is attached to L 1 .

[0275] 11. A compound or salt according to any one of items 1 to 10, wherein -X in formula (I) 2 -L 1 -X 1 - Some have structures selected from the following:

[0276] *-C(O)-NH-R 7- NH-C(O)-R 8 -C(O)-NH-(L-1),

[0277] *-C(O)-NH-R 9A -NH-C(O)-R 10A -C(O)-NH-R 11A -NH-C(O)-(L-2A), and

[0278] *-C(O)-NH-R 9B -C(O)-NH-R 10B -C(O)-NH-R 11B-NH-C(O)-(L-2B);

[0279] The amide bond marked with * is connected to the R in formula (I). 2 carbon atoms, and in which

[0280] R 7 R 8 R 9A R 9B R 11A and R 11B Independently selected from optionally substituted C2 to C10 alkyldiyl groups, preferably optionally substituted straight-chain C2 to C10 alkyldiyl groups, each of which may be substituted by one or more substituents independently selected from -OH, -OCH3, -COOH, -COOCH3, -NH2, -NHC(NH)NH2 and the EDS group, and

[0281] R 10A and R 10B The substituent is selected from optionally substituted C2 to C10 alkyldiyl groups, preferably optionally substituted straight-chain C2 to C10 alkyldiyl groups, and optionally substituted C6 to C10 aryldiyl groups, preferably phenylene groups, wherein the alkyldiyl and aryldiyl groups may each be substituted by one or more substituents independently selected from -OH, -OCH3, -COOH, -COOCH3, -NH2, -NHC(NH)NH2 and the EDS group. 10A Preferably, it is an optional substituted C2 to C10 alkyldiyl as defined above, more preferably an optional substituted straight-chain C2 to C10 alkyldiyl. R 10B Preferably, it is an optional substituted C6 to C10 aryl dimethyl group as defined above, more preferably a phenylene group, such as p-phenylene.

[0282] 12. The compound or salt according to item 11, wherein, without including the carbon atom contained in the optionally present substituent, R of formula (L1) 7 and R 8 The total number of carbon atoms in it is 6 to 20, more preferably 6 to 16; excluding carbon atoms contained in optionally present substituents, R of formula (L2A) 9A R 10A and R 11A The total number of carbon atoms in it is 6 to 20, more preferably 6 to 16; and in the absence of carbon atoms contained in optional substituents, R of formula (L2B) 9B R 10B and R 11B The total number of carbon atoms in it is 6 to 20, more preferably 6 to 16.

[0283] 13. The compound or salt according to item 11 or 12, wherein

[0284] The part –X 2 -L 1 -X 1 -Having a structure (L-1), and R 8 EDS containing at least one substituent, or

[0285] The part –X 2 -L 1 -X 1 -Having a structure (L-2A), and R 10A EDS containing at least one substituent.

[0286] 14. The compound or salt according to claim 7, wherein the fraction –X 2 -L 1 -X 1 -Has a structure selected from the following:

[0287] *-C(O)-NH-CH(COOH)-R 12- NH-C(O)-R 13- C(O)-NH-(L-3),

[0288] *-C(O)-NH-CH(COOH)-R 14 -NH-C(O)-R 15 -C(O)-NH-R 16 -CH(COOH)-NH-C(O)-

[0289] (L-4), and

[0290] *-C(O)-NH-CH(COOH)-R 17 -C(O)-NH-R 18 -C(O)-NH-R 19 -CH(COOH)-NH-C(O)-

[0291] (L-5);

[0292] The keys marked with * are connected to equation (I) containing R. 2 carbon atoms,

[0293] R 12 and R 14 Independently selected from straight-chain C2 to C6 alkyldiyl groups, preferably selected from straight-chain C3 to C6 alkyldiyl groups.

[0294] R 13 It is a straight-chain C2 to C10 alkyldiyl, preferably a straight-chain C4 to C8 alkyldiyl.

[0295] R15 and R 16 Independently selected from straight-chain C2 to C6 alkyldiyl groups, preferably selected from straight-chain C2 to C4 alkyldiyl groups.

[0296] And R 13 and R 15 Each may have an EDS group as a substituent, and more preferably, R 13 and R 15 Each contains an EDS group as a substituent.

[0297] R 17 It is a straight-chain C2 to C6 alkyldiyl, preferably a straight-chain C2 to C4 alkyldiyl.

[0298] R 18 It is a phenylene oxide, such as p-phenylene oxide, and

[0299] R 19 It is a straight-chain C2 to C6 alkyldiyl, preferably a straight-chain C2 to C4 alkyldiyl.

[0300] 15. The compound or salt according to item 14, wherein, without including the carbon atom contained in the EDS group as a substituent, R in formula (L-3) 12 and R 13 The total number of carbon atoms in formula (L-4) is 6 to 16, more preferably 6 to 14; and in the absence of carbon atoms contained in the substituent group EDS, R in formula (L-4) 14 R 15 and R 16 The total number of carbon atoms in it is 6 to 16, more preferably 6 to 14.

[0301] 16. The compound or salt according to any one of items 1 to 15, wherein R 2 It is an optionally substituted aralkyl group, wherein the optionally substituted aralkyl group is selected from optionally substituted -CH2-phenyl and optionally substituted -CH2-naphthyl, more preferably optionally substituted –CH2-(2-naphthyl), wherein the phenyl and the naphthyl are optionally substituted with substituents selected from halogens (preferably I) and -OH.

[0302] 17. The compound or salt according to claim 16, wherein R 2 It is an aralkyl group of the formula -CH2-naphthyl, more preferably -CH2-(2-naphthyl).

[0303] 18. The compound or salt according to any one of items 1 to 17, wherein R 3It is an optionally substituted aralkyl group, wherein the optionally substituted aralkyl group is selected from optionally substituted -CH2-phenyl and optionally substituted –CH2-naphthyl, more preferably optionally substituted –CH2-phenyl, wherein the phenyl and the naphthyl are optionally substituted with substituents selected from halogens (preferably I) and -OH.

[0304] 19. The compound or salt according to item 18, wherein R 3 It is an aralkyl group of the formula -CH2-phenyl, wherein the benzene ring is substituted by a substituent -OH, or by a combination of a substituent OH and a substituent –I.

[0305] 20. The compound or salt according to any one of items 1 to 15,

[0306] Where R 2 It is a group of the following formula

[0307] or

[0308] And R 3 It is a group of the following formula

[0309] or

[0310] in The markers will respectively mark R 2 and R 3 The bonds that connect to the rest of the compound of formula (I).

[0311] 21. The compound or salt according to item 20,

[0312] Where R 2 It is a group of the following formula

[0313]

[0314] And R 3 It is a group of the following formula

[0315]

[0316] in The markers will respectively mark R 2 and R 3 Bonds that connect to the rest of the molecule.

[0317] 22. The compound or salt according to any one of items 1 to 21, wherein r is 1.

[0318] 23. A compound or salt according to any one of items 1 to 22, wherein p is 0 and q is 1.

[0319] 24. The compound or salt according to any one of items 1 to 23, wherein R 4 Selected from phenyl, optionally naphthyl, and EDS groups.

[0320] 25. The compound or salt according to any one of claims 24, wherein R 4 It is selected from naphthyl (more preferably 2-naphthyl) and the EDS group.

[0321] 26. The compound or salt according to any one of items 1 to 25, wherein X 3 It is an amide bond or a group of the following formula

[0322] The labeled bond on the carbonyl group will X 3 Connect to R M And another marker key will X 3 Connected to the rest of the molecule.

[0323] 27. The compound or salt according to item 26, wherein X 3 It is an amide bond -C(O)-NH-, in which the carbon atom is attached to R. M .

[0324] 28. The compound or salt according to any one of items 1 to 27, wherein R M It may optionally include a chelating group that chelates a non-radioactive or radioactive cation.

[0325] 29. The compound or salt according to any one of claims 1 to 28, wherein the chelating group comprises at least one of the following:

[0326] (i) a macrocyclic structure having 8 to 20 ring atoms, wherein two or more, preferably three or more, of the ring atoms are selected from oxygen, sulfur, and nitrogen atoms; and

[0327] (ii) A non-cyclic open-chain structure having 8 to 20 main chain atoms, wherein 2 or more, preferably 3 or more, of the main chain atoms are heteroatoms selected from oxygen, sulfur and nitrogen atoms.

[0328] 30. The compound or salt according to any one of claims 1 to 29, wherein the chelating group is a residue of a chelating agent selected from: bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (CBTE2a), cyclohexyl-1,2-diaminetetraacetic acid (CDTA), 4-(1,4,8,11-tetraazacyclotetradecyl-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-tetraazacyclododecyl-N,N',N”,N”'-tetraacetic acid (DOTA), 2-[1,4,7,10-tetraazacyclododecyl-4,7,10-triacetic acid]-glutaric acid (DOTAGA), N,N'-dipyridoxyethylenediamine-N,N'-diacetic acid-5,5'-bis(phosphate) (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 (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-(carbonyloxy)-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), tripyridylbis(methyleneaminotetraacetic acid) (TMT), 1,4,7,10-tetraazacyclotridecane-N,N',N”,N”'-tetraacetic acid (TRITA), triethylenetetraminehexaacetic acid (TTHA), N,N′-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18-crown ether-6(H 2macropa ) and 4-amino-4-{2-[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydropyridin-2-ylmethyl)carbamoyl]ethyl} pimelic acid bis[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydropyridin-2-ylmethyl)-amide](THP);

[0329] The residues are provided by covalently bonding the carboxyl group contained in the chelating agent to the remainder of the compound via an ester bond or an amide bond, more preferably an amide bond.

[0330] 31. The compound or salt according to claim 30, wherein the chelating agent is selected from DOTA and DOTAGA.

[0331] 32. The compound or salt according to item 30 or 31, wherein X 3 It is an amide bond that connects the chelating group to the rest of the molecule.

[0332] 33. The compound or salt according to claim 32, wherein R M -X 3 - is a group of the following formula

[0333]

[0334] Among them The tagged bond is connected to the remainder of the compound of formula (I), and the chelating group may comprise a chelated non-radioactive or radioactive cation.

[0335] 34. The compound or salt according to any one of claims 1 to 33, wherein the chelating group comprises a chelated cation, preferably a chelated radioactive cation, said chelated radioactive cation being 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 cations, or those containing 18 F-cation molecules, for example 18 F-[AlF] 2+ .

[0336] 35. The compound or salt according to claim 34, wherein the chelating group comprises a chelated cation selected from... 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 cations, or those containing 18 F is a cationic molecule.

[0337] 36. A compound or salt according to any one of items 1 to 35, wherein the compound of formula (I) comprises one or both groups EDS.

[0338] 37. The compound or salt according to claim 36, wherein the compound of formula (I) comprises the linking group L 1 It carries one EDS group, or contains two EDS groups, one of which is composed of R. 4 Represents and another by L 1 carry.

[0339] 38. The compound or salt according to any one of items 1 to 37, wherein in the EDS group (E-1A), for s > 1, the substituent R 5A They are identical and selected from -NO2 and -COOH; and wherein, in the EDS group (E-2A), for t > 1, the substituent R 6A They are the same and are selected from -NO2 and -COOH.

[0340] 39. The compound or salt according to any one of claims 1 to 38, wherein in the EDS group (E-1B), for s > 1, the substituent R 5B They are identical and selected from -OH and -NH2; in the EDS group (E-2B), for t>1, the substituent R 6B They are the same and are selected from -OH and –NH2.

[0341] 40. A compound or salt according to any one of claims 1 to 39, said compound or salt comprising a group EDS having the formula (E-2A):

[0342]

[0343] in The bond that links the group EDS to the remainder of the compound of formula (I) is marked; and

[0344] t is 1 or 2, and R 6A Selected from -NO2 and -COOH.

[0345] 41. A compound according to any one of items 1 to 38, wherein said group EDS has the formula (E-3).

[0346]

[0347] in The group EDS is marked to be attached to the rest of the compound of formula (I).

[0348] 42. The compound according to any one of items 1 to 41, wherein the compound has the following formula (Ia), or a pharmaceutically acceptable salt thereof:

[0349]

[0350] Where n, X 1 L 1 X 2 R 2 R 3 R 4 q, p, X 3 and R M As defined in the foregoing, and wherein the group EDS is contained at least once and has the structure defined in the foregoing.

[0351] 43. The compound according to claim 42, wherein the compound has the following formula (Ib), or a pharmaceutically acceptable salt thereof:

[0352]

[0353] Where n, X 1 L 1 X 2 R 2 R 3 R 4 X 3 and R M As defined in the foregoing, and wherein the group EDS is contained at least once and has the structure defined in the foregoing.

[0354] 44. The compound according to any one of claims 43, wherein the compound has the following formula (Ic), or a pharmaceutically acceptable salt thereof:

[0355]

[0356] Where n, X 1 L 1 X 2 R 4 X 3 and R M As defined in the foregoing, and wherein the group EDS is contained at least once and has the structure defined in the foregoing.

[0357] 45. The compound according to claim 44, wherein the compound has the following formula (Id) or (Ie), or a pharmaceutically acceptable salt thereof:

[0358]

[0359] Where R 9A R 10A R 11A R 4 X 3 and R M As defined in the foregoing, and where (i)R 4It is a group EDS having the structure defined in the foregoing items, or (ii)R 10A EDS having a group having the structure defined in the foregoing, or both (i) and (ii) apply;

[0360]

[0361] Where R 9B R 10B R 11B R 4 X 3 and R M As defined in the foregoing, and where R 4 It is a group EDS having the structure defined in the foregoing items.

[0362] 46. ​​The compound according to claim 45, wherein the compound has the following formula (If) or (Ig), or a pharmaceutically acceptable salt thereof:

[0363]

[0364] Where R 9A R 10A R 11A R 4 X 3 and R M As defined in the foregoing, and where (i)R 4 It is a group EDS having the structure defined in the foregoing items, or (ii)R 10A EDS having a group having the structure defined in the foregoing, or both (i) and (ii) apply;

[0365]

[0366] Where R 9B R 10B R 11B R 4 X 3 and R M As defined in the foregoing, and where R 4 It is a group EDS having the structure defined in the foregoing items.

[0367] 47. The compound or salt thereof according to any one of claims 1, wherein said compound or salt has one of the following formulas:

[0368]

[0369]

[0370]

[0371]

[0372] 48. A pharmaceutical composition or diagnostic composition comprising, or consisting of, one or more compounds or salts according to any one of items 1 to 47.

[0373] 49. A compound or salt according to any one of items 1 to 47, used for the diagnosis and / or treatment of (a) cancer, including prostate cancer; or (b) angiogenesis / angiogenesis.

[0374] The attached diagram illustrates:

[0375] Figure 1 :[ nat / 177 Lu]PSMA I&T and [ nat / 177 Lu]PSMA-62 and [ nat / 177 Feature network diagram of Lu]PSMA-66.

[0376] Figure 2 The selected 177 The kinetics of externalization of Lu-labeled PSMA inhibitors from LNCaP cells. (1.25*10) 5 Cells / well were incubated with the corresponding radioligand (c = 1.0 nm) in DMEM solution (5% BSA) at 37 °C for 1 h. Afterward, the supernatant was removed and the cells were washed once with DMEM solution (5% BSA, 37 °C). Replacement was then performed with either A) DMEM solution only (5% BSA) or B) blocking DMEM solution (5% BSA, 10 μm 2-PMPA). Total internalized radioactivity at t = 0 min was corrected for nonspecific binding (10 μm 2-PMPA) and normalized to 100%. All data are expressed as mean ± SD (n = 3).

[0377] Figure 3 2.5 to 3.0 MBq (0.15 to 0.25 nmol) of [ 177 Lu]PSMA-66 and [ 177 Biodistribution of Lu]PSMA I&T in CB-17SCID mice carrying LNCaP tumors (%ID / g) (n = 4 respectively).

[0378] Figure 4 CB-17SCID mice carrying LNCaP tumors were injected with approximately 10.3 MBq (0.19 nmol tracer) of […]. 68Following Ga]PSMA-36, maximum intensity projection (MIP) of μPET scans (dynamic scan, total frames 1 to 1.5 hpi) (top left). Dynamic PET data (90-minute acquisition time, OSEM 3D reconstruction) of blood pool (heart), kidney, tumor, muscle, lacrimal gland, and salivary gland from CB-17SCID mice carrying LNCaP tumors. 68 The TAC (logarithmic plot) of Ga]PSMA-36 is in %ID / mL.

[0379] Figure 5 CB-17SCID mice carrying LNCaP tumors were injected with approximately 11 and 13 MBq (0.15 to 0.25 nmol tracer), respectively. 68 Following Ga-labeled PSMA inhibitors PSMA-62 and PSMA-66, the maximum intensity projection (MIP) of μPET scans (dynamic scans, total frames 1 to 1.5 hpi) (top left). Derived from two... 68 The Ga-labeled tracer was used in dynamic PET data (90-minute acquisition time, OSEM 3D reconstruction) of the blood pool (heart), kidney, tumor, and muscle of CB-17SCID mice carrying LNCaP tumors. 68 TAC (logarithmic plot) of Ga-labeled PSMA inhibitors, in %ID / mL.

[0380] Figure 6 : 2.5 to 6.0 MBq (0.15 to 0.25 nmol) of [ 177 Lu]PSMA-62、[ 177 Lu]PSMA-66, [ 177 Lu]PSMA-71 and [ 177 Biodistribution of Lu]PSMA I&T in CB-17SCID mice carrying LNCaP tumors (%ID / g) (n = 4 respectively). Detailed Implementation

[0381] The following examples illustrate the present invention.

[0382] Example 1: Materials and Methods

[0383] 1. General information

[0384] Fmoc-(9-fluorenylmethoxycarbonyl-) and all other protected amino acid analogs were purchased from Bachem (Bubendorf, Switzerland) or Iris Biotech (Marktredwitz, Germany). 2-Chlorotriphenylmethylchloro(2-CTC) resin was obtained from PepChem (Tübingen, Germany). The chelating agent DOTAGA anhydride was supplied by Chematech (Dijon, France). PSMA-DKFZ-617 was purchased from ABX Advanced Compounds (Radeberg, Germany). All necessary solvents and other organic reagents were purchased from Alfa Aesar (Karlsruhe, Germany), Sigma-Aldrich (Munich, Germany), or VWR (Darmstadt, Germany). Solid-phase synthesis of the peptides was performed manually using an Intelli-Mixer syringe shaker (Neolab, Heidelberg, Germany). Analytical reversed-phase high-performance liquid chromatography (RP-HPLC) was performed using a Shimadzu gradient RP-HPLC system (Shimadzu Deutschland GmbH, Neufahrn, Germany) on a Nucleosil 100C18 column (5 μm, 125 × 4.0 mm, CS GmbH, Langerwehe, Germany). Peptide analysis was performed at a constant flow rate of 1 mL / min using different gradients of 0.1% (v / v) trifluoroacetic acid (TFA) aqueous solution (solvent A) and 0.1% TFA (v / v) acetonitrile (MeCN) solution (solvent B) (specific gradients are cited in the text). A Shimadzu SPD 20A Prominence UV / VIS detector (Shimadzu Deutschland GmbH) was used at λ = 220 nm and 254 nm. HSA binding was determined using a Chiralpak HSA (5μm, 50x 3mm) analytical column (Daicel Chemical Industries) attached to a Chiralpak HSA (5μm, 10x 3mm) guard column (Daicel Chemical Industries), purchased from Chiral Technologies Europe (Illkirch, France). Nonlinear regression analysis of HSA binding was performed using OriginPro2016G (Northampron, USA). Retention time t was cited in this paper. RAnd the capacity factor K'. Preparative RP-HPLC of peptides was obtained using a Multospher 100RP 18-5 column (250×20mm, CS GmbH) on a Shimadzu RP-HPLC system at a constant flow rate of 5 mL / min. Analytical and preparative radioactive RP-HPLC of radioiodinated reference ligands were performed using a Nucleosil 100C18 column (5 μm, 125×4.0 mm). Radioactivity was detected by connecting the outlet of a UV spectrophotometer to a NaI(Tl) trap scintillation counter from EG&G Ortec (Munich, Germany). As previously reported in [1,2] analysis. 68 Ga-and 177 Lu-labeled compounds. In expression L Electrospray ionization mass spectrometry (ESI-MS) spectra were acquired using a CMS mass spectrometer (Advion Ltd., Harlow, UK) and a Varian 500-MS IT mass spectrometer (Agilent Technologies, Santa Clara, USA). For the Bradford Assay, the S9 fraction was separated using a JASCO Germany GmbH (Gross-Umstadt, Germany) V-630 UV-Vis spectrophotometer and centrifuged in an Avanti JXN-26 centrifuge (Beckman Coulter GmbH (Krefeld, Germany)). The determination of radioactive S9 metabolites was performed using a Heraeus PICO 17 centrifuge (Thermo Fisher Scientific Messtechnik GmbH, Munich, Germany). NMR data were obtained using an AV 300 (300 MHz) or AV 400 (400 MHz) receiver from Bruker (Billerica, USA) with a 300 kHz applied. The NMR data were obtained using a Biometra UNOThermoblock (Biometra, The S9 portion was incubated in Deutschland for in vitro metabolite analysis.

[0385] 2. Synthesis scheme (SP)

[0386] SP-1: 2-CTC Resin Loading: 2-CTC resin (1.6 mmol / g) was loaded for 2 hours at room temperature (RT) with Fmoc-AA-OH (1.5 equivalences) in anhydrous dichloromethane (DCM) containing N,N-diisopropylethylamine (DIPEA) (4.5 equivalences). The remaining triphenylmethyl chloride was capped for 15 minutes by adding 2 mL / g methanol (MeOH). Afterward, the resin was filtered, thoroughly washed with DCM (2×), dimethylformamide (DMF) (2×), and MeOH (2×), and stored under vacuum overnight. Loading was determined using weight variation.

[0387]

[0388] Formula 1. Determination of resin loading: m 总 : Mass of loaded resin (Fmoc-AA-OH and HCl); M As : Molar mass of amino acids; m 净重量 The quality of the resin used; M HCl Molar mass of hydrochloric acid

[0389] SP-2: Peptide synthesis via TBTU / HOBt coupling: A solution (8 ml / g resin) of Fmoc-AA-OH (2.0 equivalents), N,N,N′,N′-tetramethyl-O-(benzotriazol-1-yl)urea tetrafluoroborate (TBTU) (2.0 equivalents), N-hydroxybenzotriazole (HOBt) (2.0 equivalents), and DIPEA (4.5 equivalents) in DMF was added to the resin-bound free amine peptide at room temperature and shaken for 2 hours, followed by washing with DMF (6×). Coupling with secondary or aromatic amines was performed using different protocols. Fmoc-AA-OH (3.0 equivalents) was dissolved together with 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridine 3-oxohexafluorophosphate (HATU) (3.0 equivalents), 1-hydroxy-7-azabenzotriazole (HOAt) (3.0 equivalents), and DIPEA (6.0 equivalents) in DMF (8 mL / g resin), and stirred for 15 minutes. The pre-activated solution was added to the peptide bound to the resin, and the mixture was shaken at room temperature for 2 hours. After the reaction was complete, the resin was washed with DMF (6×). Generally, the synthesis of all peptide scaffolds is carried out according to the previously described (Weineisen, M.; Schhottelius, M.; Simecek, J.; Eiber, M.; Schwaiger, M.; Wester, H., Development and first in human evaluation of PSMA I&T-A ligand for diagnostic imaging and endoradiotherapy of prostate cancer, Journal of Nuclear Medicine, 2014, 55, 1083-1083; Weineisen, M.; Simecek, J.; Schhottelius, M.; Schwaiger, M.; Wester, H.-J, Synthesis and preclinical evaluation of DOTAGA-conjugated PSMA ligands for functional imaging and endoradiotherapy of prostate cancer, EJNMMI research, 2014, 4, 1).

[0390] SP-3: Deprotection of Fmoc on resin: Fmoc-protected peptides bound to the resin were treated with 20% piperidine DMF solution (v / v) for 5 minutes, followed by a second treatment for 15 minutes. The resin was then thoroughly washed with DMF (8×).

[0391] SP-4: Deprotection of Dde on resin: An N-(1-(4,4-dimethyl-2,6-dioxocyclohexyl)ethyl)(Dde)-protected peptide (1.0 equivalent) was dissolved in a 2.0% solution of hydrazine monohydrate (N2H4·H2O) DMF (v / v). After 15 minutes, the deprotected peptide, if bound to the resin, was washed with DMF (6×) or precipitated in diethyl ether (Et2O) to give the crude product. If both Fmoc and Dde protecting groups were present and only Dde deprotection was necessary, the resin-loaded peptide was treated for 3 hours at room temperature with a solution containing NH2OH·HCl (630 mg), imidazole (460 mg), DCM (0.5 mL), DMF (0.5 mL), and N-methyl-2-pyrrolidone (NMP) (2.5 mL). The resin-loaded peptide was then washed with DMF (6×).

[0392] SP-5: Alloc / allyl deprotection on resin: Alloc / allyl protecting groups were removed from the peptides bound to the resin using a DCM solution (6.0 mL) containing triisopropylsilane (TIPS) (50.0 equivalents) and (triphenyl)palladium (0)(Pd(PPh3)4) (0.3 equivalents). The resin was treated with this solution for 1.5 hours at room temperature. Finally, the resin was washed with DCM (3×) to remove Pd(PPh3)4.

[0393] SP-6: tBu / Boc Deprotection: The tert-butyl (tBu) / tert-butyloxycarbonyl (Boc) protecting groups were removed by dissolving the crude product in TFA (approximately 500 μL) and stirring at room temperature for 40 minutes. Subsequently, TFA was almost completely removed using a nitrogen stream. After precipitation in Et₂O, the crude product was centrifuged and the supernatant was removed. The dried pellets were further used in the following synthetic steps.

[0394] SP-7.1: A) Cleavage of peptides from resin while retaining side-chain protecting groups: Dissolve the fully protected, resin-bound peptide in DCM / trifluoroethanol (TFE) / acetic acid (AcOH) (6 / 3 / 1; v / v / v) and shake for 30 minutes. Filter the solution and redissolve the resin in another cleavage solution for 30 minutes. Combine the fractions and concentrate the solvent under reduced pressure. Redissolve the filtrate in toluene and concentrate under reduced pressure to remove AcOH. Precipitate in water or Et₂O to produce crude side-chain protected peptides.

[0395] SP-7.2: B) Cleavage of the peptide from the resin while simultaneously deprotecting all acid-sensitive protecting groups: Dissolve the fully protected resin-bound peptide in a mixture of TFA / TIPS / water (95 / 2.5 / 2.5; v / v / v) and shake for 30 minutes. Filter the solution and treat the resin again in the same manner for 30 minutes. Then, combine the fractions and concentrate the solvent under a constant nitrogen flow. Precipitate the crude peptide in Et2O and allow it to dry overnight.

[0396] SP-8: Deacetylation of the carbohydrate moiety: Deacetylation was performed at room temperature by dissolving the PSMA inhibitor in MeOH containing KCN (0.5 equivalents) (Herzig, J.; Nudelman, A.; Gottlieb, HE; ​​Fischer, B., Studies in sugar chemistry. 2. A simple method for O-deacylation of polyacylated sugars, The Journal of Organic Chemistry, 1986, 51, 727-730), with stirring overnight. The final product was purified by RP-HPLC.

[0397] SP-9: Preparation of non-radioactive metal-complexed PSMA inhibitors:

[0398] SP-9.1: nat Ga compounds: for the preparation of nat Ga III The chelate was formed by mixing 50 μL of an aqueous (aq.) solution of a 2.0 mmol PSMA inhibitor and 50 μL of an aqueous solution of a 2.0 mmol Ga(NO3)3 and heating at 40 °C for 30 min. The formation of the chelate was assessed using RP-HPLC and ESI-MS. The resulting 1.0 mmol solution was diluted and used for in vitro IC50. 50 Measurement and HSA binding.

[0399] SP-9.2: nat Lu compounds: The corresponding PSMA inhibitors were prepared from a 2.0 mmol / L aqueous solution containing 2.5 mol excess LuCl3 (20 mmol / L aqueous solution). nat Lu III The complex was prepared and heated to 95°C for 30 minutes. After cooling, the mixture was confirmed by RP-HPLC and ESI-MS. nat Lu III - Formation of chelates. The resulting natLu complex was then diluted in a 1.0 mm aqueous solution and used in vitro IC without further treatment. 50 Research.

[0400] 3. Building blocks for PSMA-36 and EuE-based PSMA inhibitors

[0401] Di-tert-butyl(((s)-6-amino-1-(tert-butoxy)-1-oxohex-2-yl)carbamoyl)-l-glutamate

[0402]

[0403] Molecular formula: C 24 H 45 N3O7

[0404] Molecular weight: 487.64

[0405] ((OtBu)KuE(OtBu)2)(1): As previously described, a tert-butyl-protected Lys-urea-Glu-binding motif (EuK) was synthesized via solution fusion [3]. Briefly, a solution of DCM containing L-di-tert-butylglutamate·HCl (2.0 g, 7.71 mmol, 1.0 equivalence) was cooled on ice for 30 min and then treated with trimethylamine (TEA) (2.69 mL, 19.28 mmol, 2.5 equivalence) and 4-(dimethylamino)pyridine (DMAP) (3.3 mg, 0.3 mmol, 0.04 equivalence). After an additional 5.0 min of stirring, 1,1'-carbonyldiimidazole (CDI) (1.38 g, 8.84 mmol, 1.1 equivalence) was dissolved in DCM and added slowly over 30 min. The reaction mixture was further stirred overnight and allowed to warm to room temperature. The reaction was terminated with 8 mL of saturated (sat.) NaHCO3 solution, followed by washing with water (2×) and brine (2×), and drying over saturated Na2SO4 solution. Residual solvent was removed under vacuum, and the crude product (S)-di-tert-butyl-2-(1H-imidazolium-1-carboxamido)glutarate was usable without further purification. RP-HPLC (10 to 90% B within 15 minutes): t R =12.2 minutes; K'=5.8. Theoretical monoisotopic mass (C 17 H 27 N3O5): 353.4; Measured value: m / z = 376.1 [M+Na] +The crude product (S)-di-tert-butyl-2-(1H-imidazolium-1-carboxamido)glutarate (2.72 g, 7.71 mmol, 1.0 equivalent) was dissolved in 1,2-dichloroethane (DCE) and cooled on ice for 30 min. TEA (2.15 mL, 15.42 mmol, 2.0 equivalent) and H-Lys(Cbz)-OtBu·HCl (2.87 g, 7.71 mmol, 1.0 equivalent) were added to the solution, and the solution was stirred overnight at 40 °C. The remaining solvent was evaporated, and the crude product was purified by rapid silica gel chromatography using an eluent mixture containing ethyl acetate (EtOAc) / hexane / TEA (500 / 500 / 0.8; v / v / v). After solvent removal, (9R,13S)-tritert-butyl-3,11-dioxo-1-phenyl-2-oxa-4,10,12-triazapentadecan-9,13,15-tricarboxylic acid ester was obtained in colorless oil form. RP-HPLC (40 to 100% B within 15 minutes): t R =14.5 minutes; K'=6.25. Theoretical monoisotopic mass (C 32 H 51 N3O9): 621.8; Measured value: m / z = 622.3 [M+H] + To synthesize (OtBu)KuE(OtBu)2(1), (9R,13S)-tritert-butyl-3,11-dioxo-1-phenyl-2-oxa-4,10,12-triazapentadecan-9,13,15-tricarboxylic acid ester (3.4 g, 5.47 mmol, 1.0 equivalent) was dissolved in ethanol (EtOH) (75 mL), and palladium (0.34 g, 0.57 mmol, 0.1 equivalent) (10%) supported on activated carbon was added to this solution. The flask containing the reaction mixture was first purged with a stream of hydrogen, and the solution was stirred overnight at room temperature under light hydrogen pressure (gas bag). The crude product was purified by diatomaceous earth, and the solvent was evaporated under vacuum. The expected product 1 (1.9 g, 3.89 mmol, yield 71.6%) was obtained in the form of a waxy solid. RP-HPLC (10 to 90% B within 15 minutes): t R =12.6 minutes; K'=6.4. Theoretical monoisotopic mass (C 24 H 45 N3O7) = 487.6; Measured value: m / z = 488.3 [M+H] + 510.3 [M+Na] + .

[0406] (s)-5-(tert-butoxy)-4-(3-((s)-1,5-ditert-butoxy-1,5-dioxopentan-2-yl)ureo)-5-oxopentanoic acid ((OtBu)EuE(OtBu)2)(2):

[0407]

[0408] Molecular formula: C 23 H 40 N2O9

[0409] Molecular weight: 488.58

[0410] Following the method described in 1[3], tert-butyl-protected Glu-urea-Glu-binding motifs (EuE) were synthesized by replacing HL-Lys(Cbz)-OtBu·HCl with HL-Glu(OBzl)-OtBu·HCl. The desired product (4.10 g, 8.39 mmol, 84% yield) was obtained in the form of a waxy and highly hygroscopic solid. RP-HPLC (10 to 90% B within 15 min): t R =11.3 minutes; K′=7.69. Theoretical monoisotopic mass (C 23 H 49 N₂O₉) = 488.3; Measured value: m / z = 489.4 [M+H] + 516.4 [M+Na] + .

[0411] (s)-NHFmoc-Asu(OtBu)-OBzl(5): HOAt (21.8 mg, 0.16 mmol, 1.5 equivalence), HATU (61.0 mg, 161.0 μmol, 1.5 equivalence), and DIPEA (73.2 μL, 0.48 mmol, 4.5 equivalence) were added to a DMF solution of (s)-Fmoc-Asu(OtBu)-OH (50 mg, 107.0 μmol, 1.0 equivalence). After stirring at room temperature for 15 minutes, benzyl alcohol (22.2 μL, 0.32 mmol, 3.0 equivalence) was further added, and the solution was stirred overnight. Finally, the solvent was removed under vacuum. The reaction was analyzed by RP-HPLC (10 to 90% B over 15 minutes): t R =17.1 minutes; K' = 7.55.5 theoretical monoisotopic mass (C 34 H 39 NO6) = 557.28; Measured value: m / z = 580.7 [M+Na] + .

[0412] (s)-NHFmoc-Asu-OBzl(6): Crude product 5 was deprotected at room temperature with a stirred mixture (v / v) of TFA (95%) and DCM (5%) for 45 min using tBu. After solvent evaporation, crude product 6 was purified using preparative RP-HPLC (60 to 80% B within 15 min): t R=9.3 minutes; K' = 8.9.6. Theoretical monoisotopic mass (C 30 H 31 NO6) = 501.22; Measured value: m / z = 524.5 [M+Na] + .

[0413] OBzl-(s)-Fmoc-Asu[(OtBu)KuE(OtBu)2](7): HOBt (20.9 mg, 0.15 mmol, 1.5 equivalence), TBTU (36.3 mg, 15.5 μmol, 1.5 equivalence), and DIPEA (79.4 μL, 59.7 mg, 0.46 mmol, 4.5 equivalence) were added to a DMF solution of 6 (51.8 mg, 10.3 μmol, 1.0 equivalence). After stirring for 15 minutes, 1 (75.6 mg, 15.5 μmol, 1.5 equivalence) was added, and the mixture was stirred for another 20 hours at room temperature. The crude product 7 was purified by preparative RP-HPLC (70 to 80% B within 15 minutes). R =8.9 minutes; K' = 1.97.7. Theoretical monoisotopic mass (C 54 H 74 N4O 12 = 970.53; Measured value: m / z = 971.8 [M+H] + .

[0414] (s)-Fmoc-Asu[(OtBu)KuE(OtBu)2](8): For the deprotection of benzyl alcohol (Bzl), 7 (57.2 mg, 65.0 μmol, 1.0 equivalent) was dissolved in EtOH (2.0 mL) and palladium (10%) (5.72 mg, 9.0 μmol, 0.1 equivalent) supported on activated carbon was added. The flask was purged with a stream of hydrogen beforehand and the solution was stirred under light hydrogen pressure (gas bag). After stirring for 70 minutes, the crude product was filtered through diatomaceous earth, EtOH was vacuum evaporated, and the product was purified by preparative RP-HPLC (70 to 70.5% B within 15 minutes): t R =6.5 minutes; theoretical monoisotopic mass (C) of K' = 0.54.5. 47 H 68 N4O 12 = 880.48; Measured value: m / z = 881.8 [M+H] + .

[0415] OPfp-(s)-Fmoc-Asu[(OtBu)KuE(OtBu)2](9):

[0416]

[0417] Molecular formula: C 53 H 67 F5N4O 12

[0418] Molecular weight: 1047.13

[0419] To a dry DMF solution (13.6 mg, 15.4 μmol, 1.0 equivalent), DIC (4.77 μL, 1.94 mg, 30.8 μmol, 2.0 equivalent) and PfpOH (5.67 mg, 30.8 μmol, 2.0 equivalent) were added. After stirring for 5 minutes, pyridine (2.49 μL, 31.0 μmol, 2.0 equivalent) was added, and the solution was stirred overnight at room temperature. The reaction at step 9 was analyzed by RP-HPLC (10 to 90% B over 15 minutes). R =17.2 minutes; K'=7.6.9 theoretical monoisotopic mass (C 53 H 67 F5N4O 12 =1046.47; Measured value: m / z = 1069.8 [M+Na] + .

[0420] NHS-2,4-dinitrobenzoate (NHS-DNBA) (27):

[0421]

[0422] Molecular formula: C 11 H7N3O8

[0423] Molecular weight: 309.19

[0424] To a dry THF solution of 2,4-dinitrobenzoic acid (DNBA) (10.0 mg, 47.1 μmol, 1.0 equivalence), N,N'-dicyclohexylcarbodiimide (DCC) (9.7 mg, 47.1 μmol, 1.0 equivalence) and N-hydroxysuccinimide (NHS) (10.8 mg, 94.3 μmol, 2.0 equivalence) were added, and the reaction mixture was stirred overnight. The crude product was purified by RP-HPLC. RP-HPLC (10 to 90% B within 15 min): t R =10.21 minutes; K'=4.1. Theoretical monoisotopic mass (C 11 H7N3O8) = 309.02; Measured value: Not detected in ESI-MS.

[0425] DOTAGA-3-iodo-D-Tyr-D-Phe-D-Lys-OH(DOTAGA-y(3-I)fk)(30):

[0426]

[0427] Molecular formula: C 43 H 61 IN8O 14

[0428] Molecular weight: 1040.91

[0429] The synthesis of 30 was accomplished using a solid-phase strategy, as previously described [2,3]. In brief: the initial starting point was 2-CTC resin loading according to SP-1 of Fmoc-D-Fmoc-D-Lys(Boc)-OH. Following lysine conjugation, Fmoc was deprotected according to SP-3, and Fmoc-D-phenylalanine was coupled using SP-2. Fmoc-D-Tyr(3-I)-OH was coupled using the same procedure. After the reaction was complete, the Fmoc protecting group was cleaved according to SP-3, and the resin-bound peptide was condensed with a chelating agent using DOTAGA-anhydride (2.0 equivalences) and DIPEA (2.0 equivalences) in DMF. The reaction was stirred at room temperature for 48 hours. Finally, the crude product was cleaved from the resin according to SP-7.2, precipitated in Et2O, and centrifuged. The supernatant was removed, and 30 was purified using RP-HPLC. RP-HPLC (10% to 90% B in 15 min): t R = 6.2 minutes; K' = 2.1. Theoretical monoisotopic mass (C 43 H 61 IN8O 14 =1040.34; Measured value: m / z = 1040.5 [M+H] + m / z = 521.3[M+2H] 2+ m / z = 1063.4 = [M + Na] + .

[0430] DOTAGA–y(3-I)fk(L-Asu[KuE])(PSMA-8):

[0431]

[0432] Molecular formula: C 63 H 93 IN 12 O 23

[0433] Molecular weight: 1513.40

[0434] Add 9 (7.5 mg, 7.2 μmol, 1.5 equivalent) and DIPEA (3.3 μL, 21.6 μmol, 4.0 equivalent) to a DMF solution containing 30 (5.0 mg, 4.8 μmol, 1.0 equivalent). Stir the reaction solution overnight at room temperature. After the reaction is complete, remove the solvent under vacuum, and treat the crude product with a mixture of piperidine and DMF (20 / 80; v / v) for 15 min to achieve Fmoc deprotection. Reduce the solvent to approximately 300 μL by vacuum evaporation, precipitate in Et2O, and centrifuge. Treat the resulting granules according to SP-6 to remove tBu. Purify the final product (10% to 90% B within 15 min) by RP-HPLC: t R = 6.09 minutes; K' = 2.05. Theoretical monoisotopic mass (C 63 H 93 IN 12 O 23 =1512.55; Measured value: m / z = 1513.9 [M+H] + 757.8 [M+2H] 2+ .

[0435] DOTAGA–y(3-I)fk(L-Asu[KuE]-2,4-DNBA)(PSMA-36):

[0436]

[0437] Molecular formula: C 70 H 95 IN 14 O 28

[0438] Molecular weight: 1707.50

[0439] PSMA-36 was synthesized by dissolving PSMA-8 (3.0 mg, 3.3 μmol, 1.0 equivalent) in DMF and adding 27 (4.1 mg, 13.2 μmol, 4.0 equivalent) and DIPEA (2.3 μL, 13.2 μmol, 4.0 equivalent). The solution was stirred at room temperature for 10 hours, and the final product was purified by RP-HPLC (10 to 50% B within 15 minutes). R =12.12 minutes; K′=5.06. Theoretical monoisotopic mass (C 70 H 95 IN 14 O 28 =1706.55; Measured value: m / z = 1707.8 [M+H] + 854.7 [M+2H] 2+ .

[0440] [ nat Lu]DOTAGA-y(3-I)fk(L-Asu[KuE]-2,4-DNBA)([ nat Lu]PSMA-36): RP-HPLC (10% to 60% B within 15 minutes): t R = 9.81 minutes; K' = 3.91. Theoretical monoisotopic mass (C 70 H 92 IN 14 O 28 Lu) = 1878.47; Measured value: m / z = 1879.9 [M+H] + .

[0441]

[0442] Schematic diagram of PSMA-36 synthesis. (a) HOAt, HATU, DIPEA, benzyl alcohol, [DMF]; (b) 95% TFA, 5% DCM; (c) 1, HOBt, TBTU, DIPEA, [DMF]; (d) Pd / C (10%), H2, [EtOH]; (e) DIC, PFP, pyridine, [DMF]; (f) 30, DIPEA, [DMF]; (g) 20% pyridine-DMF solution, [DMF]; (h) TFA; (i) 27, DIPEA [DMF]

[0443] 4. Synthesis of EuE-based PSMA inhibitors PSMA-52 and PSMA-53

[0444] DOTAGA-F(4-NO2)-y-2-nal-k(Suc-N 5 -orn-C 4 -EuE)(PSMA-52):

[0445]

[0446] Molecular formula: C 76 H 100 N 14 O 29

[0447] Molecular weight: 1673.71

[0448] As described in SP-1, initial resin loading was performed using Fmoc-D-Orn(NHDde)-OH. After Fmoc deprotection according to SP-3, 2 (1.5 equivalents) was coupled to d-Orn(NHDde) according to SP-2. In the next step, the Dde protecting group was cleaved according to SP-4, and the free amino group was treated with succinic anhydride (4.0 equivalents) and DIPEA (1.5 equivalents) dissolved in DMF. The reaction mixture was reacted overnight at room temperature. Next, Fmoc-D-Lys-OtBu·HCl (1.5 equivalents) was coupled according to SP-2, and Fmoc deprotection was performed as described in SP-3. The subsequent conjugation with the Fmoc-protected amino acids Fmoc-D-2-Nal-OH, Fmoc-D-Tyr(OtBu)-OH, and Fmoc-L-Phe(4-NO2)-OH was carried out as described in SP-2. In the final step, the N-terminal Fmoc-deprotected amino acid was conjugated with a chelating agent using DOTAGA anhydride (2.0 equivalences) and DIPEA (2.0 equivalences). The reaction was stirred at room temperature for 48 hours. After the reaction with DOTAGA anhydride was complete, the peptide was cleaved from the resin according to SP-7.2, the crude product was precipitated in Et2O, centrifuged, and the supernatant was removed. The final product was purified by RP-HPLC. RP-HPLC (10% to 60% B in 15 min): t R = 9.71 minutes; K' = 3.86. Theoretical monoisotopic mass (C 76 H 100 N 14 O 29 =1672.68; Measured value: m / z = 1673.0 [M+H] + .

[0449] [ nat Lu]DOTAGA-F(4-NO2)-y-2-nal-k(Suc-N 5 -orn-C 4 -EuE)([ nat Lu]PSMA-52): RP-HPLC (10% to 60% B within 15 minutes): t R = 9.4 minutes; K' = 3.7. Theoretical monoisotopic mass (C 76 H 97 N 14 O 29 Lu) = 1844.6; Measured value: m / z = 1846.0 [M+H] + .

[0450] 2,4-DNBA-Dap(DOTAGA)-y-2-nal-k(Suc-N 5 -orn-C4 -EuE)(PSMA-53):

[0451]

[0452] Molecular formula: C 77 H 100 N 16 O 32

[0453] Molecular weight: 1761.73

[0454] As described in SP-1, initial resin loading was performed using Fmoc-D-Orn(NHDde)-OH. Following Fmoc deprotection according to SP-3, 2 (1.5 equivalents) was coupled to D-Orn(NHDde) according to SP-2. In the next step, the Dde protecting group was cleaved according to SP-4, and the free amino group was treated with succinic anhydride (4.0 equivalents) and DIPEA (1.5 equivalents) dissolved in DMF. The reaction mixture was allowed to react overnight at room temperature. Next, Fmoc-D-Lys-OtBu·HCl (1.5 equivalents) was coupled according to SP-2, and Fmoc deprotection was performed as described in SP-3. As described in SP-2, subsequent conjugation with the Fmoc-protected amino acids Fmoc-D-2-Nal-OH, Fmoc-D-Tyr(OtBu)-OH, and Fmoc-L-Dap(NHDde)-OH was carried out. Following the coupling of Fmoc-L-Dap(NHDde)-OH, Fmoc deprotection was achieved as described in SP-3. Next, the free amino group was conjugated with 2,4-dinitrobenzoic acid (2,4-DNBA) using 2,4-DNBA (2.0 equivalence), HOBt (2.0 equivalence), TBTU (2.0 equivalence), and DIPEA (4.0 equivalence) in DMF. After the reaction was complete, Dde deprotection was achieved using SP-5. In the final step, the N-terminal free amino acid L-Dap was conjugated with a chelating agent using DOTAGA anhydride (2.0 equivalence) and DIPEA (2.0 equivalence). The reaction was stirred at room temperature for 48 hours. After the reaction with DOTAGA anhydride was complete, the peptide was cleaved from the resin according to SP-7.2, the crude product was precipitated in Et2O, centrifuged, and the supernatant was removed. The final product was purified by RP-HPLC.

[0455] RP-HPLC (10% to 60% B within 15 minutes): t R =11.71 minutes; K'=4.86. Theoretical monoisotopic mass (C 77 H 100 N 16 O 32 =1760.67; Measured value: m / z = 1762.1 [M+H]+ .

[0456] [ nat Lu]2,4-DNBA-Dap(DOTAGA)-y-2-nal-k(Suc-N 5 -orn-C 4 -EuE)([ nat Lu]PSMA-53):RP-HPLC (10% to 60% B within 15 minutes):t R =8.3 minutes; K' = 3.15. Theoretical monoisotopic mass (C 77 H 97 N 16 O 32 Lu) = 1932.59; Measured value: m / z = 1933.7 [M+H] + .

[0457]

[0458] A schematic diagram of the general synthetic procedure for EuE-based PSMA inhibitors PSMA-52 and PSMA-53, taking PSMA-52 as an example. (a) 20% piperidine DMF solution, 2, HOBt, TBTU, DIPEA [DMF]; (b) succinic anhydride, DIPEA [DMF]; (c) Fmoc-D / L-Lys-OAll·HCl, HOBt, TBTU, DIPEA [DMF]; (d) 20% piperidine DMF solution, Fmoc-D-2-Nal-OH, HOBt, TBTU, DIPEA [DMF]; (e) 20% piperidine DMF solution, Fmoc-D-Tyr(OtBu)-OH, HOBt, TBTU, DIPEA [DMF]; (f) 20% piperidine DMF solution, Fmoc-D-Phe(4-NH2)-OH, HOBt, TBTU, DIPEA [DMF]; (g) Dota galvanic anhydride, DIPEA [DMF]; (h) TFA;

[0459] 5. Synthesis of PSMA-61 and PSMA-62

[0460] DOTAGA-F(4-NH2)y-2-nal-k(d[N 5 -orn-C 4 -EuE]-2,4-DNBA)(PSMA-61):

[0461]

[0462] Molecular formula: C 83 H 105 N 17 O32

[0463] Molecular weight: 1852.84

[0464] As described in SP-1, the initial resin loading was performed using Fmoc-D-Orn(NHDde)-OH. Following Fmoc deprotection according to SP-3, 2 (1.5 equivalents) was coupled to D-Orn(NHDde) according to SP-2. In the next step, the Dde protecting group was cleaved according to SP-4, and the free amino group was treated with Fmoc-D-Asp-OAll·HCl (1.5 equivalents) according to SP-2. Fmoc-D-Asp-OAll·HCl was deprotected according to SP-3, and 2,4-DNBA (1.5 equivalents), HOBt (2.0 equivalents), TBTU (2.0 equivalents), and DIPEA (4.0 equivalents) in DMF were conjugated with 2,4-DNBA. After the reaction was complete, allyl deprotection was performed according to SP-5. The next steps involved repeated conjugation of SP-2 with Fmoc-D-Lys-OtBu·HCl (1.5 equivalents), Fmoc-D-2-Nal-OH, Fmoc-D-Tyr(OtBu)-OH, and Fmoc-L-Phe(4-NHBoc)-OH. In the final step, the N-terminal Fmoc deprotected amino acid L-Phe(4-NHBoc)-OH was conjugated with a chelating agent using DOTAGA anhydride (2.0 equivalents) and DIPEA (2.0 equivalents). The reaction was stirred at room temperature for 48 hours. After the reaction with DOTAGA anhydride was complete, the peptide was cleaved from the resin according to SP-7.2, the crude product was precipitated in Et2O, centrifuged, and the supernatant was removed. The final product was purified by RP-HPLC.

[0465] RP-HPLC (10% to 90% B within 15 minutes): t R = 6.40 minutes; K' = 2.2. Theoretical monoisotopic mass (C 83 H 105 N 17 O 32 =1851.71; Measured value: m / z = 1852.5 [M+H] + 926.7 [M+2H] 2+ .

[0466] [ nat Lu]DOTAGA-F(4-NH2)y-2-nal-k(d[N 5 -orn-C 4 -EuE]-2,4-DNBA)

[0467] ([ natLu]PSMA-61): RP-HPLC (10% to 90% B within 15 minutes): t R = 8.22 minutes; K' = 3.11. Theoretical monoisotopic mass (C 83 H 102 N 17 O 32 Lu) = 2023.63; Measured value: m / z = 1013.1 [M+2H] 2+ . DOTAGA-F(4-NH2)y-2-nal-k(d[N 5 -orn-C 4 -EuE]-TMA)(PSMA-62):

[0468]

[0469] Molecular formula: C 85 H 107 N 15 O 32

[0470] Molecular weight: 1850.86

[0471] As described in SP-1, the initial resin loading was performed using Fmoc-D-Orn(NHDde)-OH. Following Fmoc deprotection according to SP-3, 2 (1.5 equivalents) was coupled to D-Orn(NHDde) according to SP-2. In the next step, the Dde protecting group was cleaved according to SP-4, and the free amino group was treated with Fmoc-D-Asp-OAll·HCl (1.5 equivalents) according to SP-2. The amino group of Fmoc-D-Asp-OAll·HCl was deprotected by Fmoc according to SP-3 and protected with Dde-OH (2.0 equivalents) and DIPEA (4.0 equivalents) in DMF. The reaction was stirred overnight. Subsequently, allyl deprotection of D-Asp was completed using SP-5. The next steps involved repeated conjugation of Fmoc-D-Lys-OtBu·HCl (1.5 equivalents), Fmoc-D-2-Nal-OH, Fmoc-D-Tyr(OtBu)-OH, and Fmoc-L-Phe(4-NHBoc)-OH with SP-2. To conjugate TMA to D-Asp, selective Dde deprotection was performed using SP-4 to yield the free amino group. TMA was coupled using TMA (2.0 equivalents), HOBt (1.5 equivalents), TBTU (1.5 equivalents), and DIPEA (10 equivalents) in DMF. The reaction was stirred at room temperature for 8 hours. Following TMA conjugation, Fmoc deprotection of Fmoc-L-Phe(4-NHBoc)-OH was performed using SP-3. In the final step, the N-terminal Fmoc-deprotected amino acid L-Phe(4-NHBoc)-OH was conjugated with a chelating agent using DOTAGA anhydride (2.0 equivalences) and DIPEA (2.0 equivalences). The reaction was stirred at room temperature for 48 hours. After the reaction with DOTAGA anhydride was complete, the peptide was cleaved from the resin according to SP-7.2, the crude product was precipitated in Et2O, centrifuged, and the supernatant was removed. The final product was purified by RP-HPLC. RP-HPLC (10 to 70% B in 15 min): t R = 7.48 minutes; K' = 2.74. Theoretical monoisotopic mass (C 85 H 107 N 15 O 32 =1849.72; Measured value: m / z = 1850.5 [M+H] + 925.7 [M+2H] 2+ .

[0472] [ nat Lu]DOTAGA-F(4-NH2)y-2-nal-k(d[N 5 -orn-C 4 -EuE]-TMA)([ natLu]PSMA-62): RP-HPLC (10 to 70% B within 15 minutes): t R =7.27 minutes; K' = 2.64. Theoretical monoisotopic mass (C 85 H 104 N 15 O 32 Lu) = 2021.64; Measured value: m / z = 1012.3 [M+2H] 2+ .

[0473] 6. Synthesis of PSMA-65, PSMA-66 and PSMA-71

[0474] 2,4-DNBA-Dap(DOTAGA)y-2-nal-e(Abz-N 5 -orn-C 4 -EuE)(PSMA-65):

[0475]

[0476] Molecular formula: C 79 H 96 N 16 O 32

[0477] Molecular weight: 1781.72

[0478] As described in SP-1, initial resin loading was performed using Fmoc-D-Orn(NHDde)-OH. Following Fmoc deprotection according to SP-3, 2 (1.5 equivalents) was coupled to D-Orn(NHDde) according to SP-2. In the next step, the Dde protecting group was cleaved according to SP-4, and the free amino group was treated with Fmoc-4-Abz-OH (1.5 equivalents), HOAt (1.5 equivalents), HATU (1.5 equivalents), and DIPEA (4.0 equivalents) in DMF. The reaction was stirred overnight at room temperature. In the next step, Fmoc deprotection was performed on the Abz residues according to SP-3. The next step involved repeated conjugation with Fmoc-D-Glu-OtBu, Fmoc-D-2-Nal-OH, Fmoc-D-Tyr(OtBu)-OH, and Fmoc-L-Dap(Dde)-OH according to SP-2. Following Fmoc deprotection of Fmoc-L-Dap(Dde)-OH according to SP-3, 2,4-DNBA was coupled using 2,4-DNBA (1.5 equivalences), HOBt (2.0 equivalences), TBTU (2.0 equivalences), and DIPEA (4.0 equivalences) in DMF. After the reaction was complete, Dde deprotection of the L-Dap(Dde)- residues was performed according to SP-4, and the residues were conjugated to a chelating agent using DOTAGA anhydride (2.0 equivalences) and DIPEA (2.0 equivalences). The reaction was stirred at room temperature for 48 hours. After the reaction with DOTAGA anhydride was complete, the peptide was cleaved from the resin according to SP-7.2, the crude product was precipitated in Et2O, centrifuged, and the supernatant was removed. The final product was purified by RP-HPLC. RP-HPLC (10 to 60% B in 15 min): t R =10.2 minutes; K'=4.1. Theoretical monoisotopic mass (C 79 H 96 N 16 O 32 =1780.64; Measured value: m / z = 1781.3 [M+H] + .

[0479] [ nat Lu]2,4-DNBA-Dap(DOTAGA)y-2-nal-e(Abz-N 5 -orn-C 4 -EuE)

[0480] ([ nat Lu]PSMA-65): RP-HPLC (10 to 60% B within 15 minutes): t R = 9.8 minutes; K' = 3.9. Theoretical monoisotopic mass (C 79 H 93 N16 O 32 Lu) = 1952.56; Measured value: m / z = 1954.0 [M+H] + .

[0481] DOTAGA-Dap(TMA)y-2-nal-k(d[N 5 -orn-C 4 -EuE]-TMA)(PSMA-66):

[0482]

[0483] Molecular formula: C 88 H 107 N 15 O 37

[0484] Molecular weight: 1966.89

[0485] As described in SP-1, the initial resin loading was performed using Fmoc-D-Orn(NHDde)-OH. After Fmoc deprotection according to SP-3, 2 (1.5 equivalents) was coupled to D-Orn(NHDde) according to SP-2. In the next step, the Dde protecting group was cleaved according to SP-4, and the free amino group was treated with Fmoc-D-Asp-OAll·HCl (1.5 equivalents) according to SP-2. The amino group of Fmoc-D-Asp-OAll·HCl was deprotected by Fmoc according to SP-3 and protected with 2.0 equivalents of Dde-OH and 4.0 equivalents of DIPEA in DMF. The reaction was stirred overnight. Subsequently, allyl deprotection of D-Asp was completed using SP-5. The next step involved repeated conjugations with Fmoc-D-Lys-OtBu·HCl (1.5 equivalents), Fmoc-D-2-Nal-OH, Fmoc-D-Tyr(OtBu)-OH, and Fmoc-L-Dap(Dde)-OH according to SP-2. To conjugate TMA to D-Asp and L-Dap, selective Dde deprotection was performed using SP-4 to yield free amino groups. TMA was coupled using TMA (4.0 equivalents), HOBt (3.0 equivalents), TBTU (3.0 equivalents), and DIPEA (20 equivalents) in DMF. The reaction was stirred at room temperature for 8 hours. Following TMA conjugation, Fmoc deprotection of Fmoc-L-Dap(TMA)-OH was performed using SP-3. In the final step, the N-terminal Fmoc-deprotected amino acid L-Dap was conjugated with a chelating agent using DOTAGA anhydride (2.0 equivalences) and DIPEA (2.0 equivalences). The reaction was stirred at room temperature for 48 hours. After the reaction with DOTAGA anhydride was complete, the peptide was cleaved from the resin according to SP-7.2, the crude product was precipitated in Et2O, centrifuged, and the supernatant was removed. The final product was purified by RP-HPLC. RP-HPLC (10 to 70% B within 15 minutes): t R = 7.48 minutes; K' = 2.74. Theoretical monoisotopic mass (C 88 H 107 N 15 O 37 ) = 1965.70; Measured value: m / z = 1966.4 [M+H] + 984.1[M+2H] 2+ .

[0486] [ nat Lu]DOTAGA-Dap(TMA)y-2-nal-k(d[N 5 -orn-C 4 -EuE]-TMA)(PSMA-66): RP-HPLC (10 to 70% B within 15 minutes): tR =7.46 minutes; K' = 2.73. Theoretical monoisotopic mass (C 88 H 108 N 15 O37Lu)=2137.62; Measured value: m / z=1070.4[M+2H] 2+ .

[0487] DOTAGA-2-Nal-y-2-nal-k(d[N 5 -orn-C 4 -EuE]-TMA)(PSMA-71):

[0488]

[0489] As described in SP-1, the initial resin loading was performed using Fmoc-D-Orn(NHDde)-OH. After Fmoc deprotection according to SP-3, 2 (1.5 equivalents) was coupled to D-Orn(NHDde) according to SP-2. In the next step, the Dde protecting group was cleaved according to SP-4, and the free amino group was treated with Fmoc-D-Asp-OAll·HCl (1.5 equivalents) according to SP-2. The amino group of Fmoc-D-Asp-OAll·HCl was deprotected by Fmoc according to SP-3 and protected with Dde-OH (2.0 equivalents) and DIPEA (4.0 equivalents) in DMF at room temperature. The reaction was stirred overnight. Subsequently, allyl deprotection of D-Asp was completed using SP-5. The next step involved repeated conjugations with Fmoc-D-Lys-OtBu·HCl (1.5 equivalents), Fmoc-D-2-Nal-OH, Fmoc-D-Tyr(OtBu)-OH, and Fmoc-L-2-Nal-OH according to SP-2. To conjugate TMA to D-Asp, selective Dde deprotection was performed using SP-4 to yield the free amino group. TMA was coupled using TMA (2.0 equivalents), HOBt (1.5 equivalents), TBTU (1.5 equivalents), and DIPEA (10 equivalents) in DMF. The reaction was stirred at room temperature for 8 hours. Following TMA conjugation, Fmoc deprotection of Fmoc-L-2-Nal-OH was completed using SP-3. In the final step, the N-terminal Fmoc-deprotected amino acid L-2-Nal-OH was conjugated with a chelating agent using DOTAGA anhydride (2.0 equivalences) and DIPEA (2.0 equivalences). The reaction was stirred at room temperature for 48 hours. After the reaction with DOTAGA anhydride was complete, the peptide was cleaved from the resin according to SP-7.2, the crude product was precipitated in Et2O, centrifuged, and the supernatant was removed. The final product was purified by RP-HPLC. RP-HPLC (10 to 80% B within 15 minutes): t R=7.57 minutes; K' = 2.79. Theoretical monoisotopic mass (C 89 H 108 N 14 O 32 =1884.73; Measured value: m / z = 1886.1 [M+H] + 943.5 [M+2H] 2+ .

[0490] [ nat Lu]DOTAGA-2-Nal-y-2-nal-k(d[N 5 -orn-C 4 -EuE]-TMA)([ nat Lu]PSMA-67): RP-HPLC (10 to 90% B within 15 minutes): t R =7.81 minutes; K' = 2.91. Theoretical monoisotopic mass (C 89 H 105 N 14 O 32 Lu) = 2056.64; Measured value: m / z = 1029.7 [M+2H] 2+ .

[0491] 7. Radiolabeling

[0492] 68 Ga-mark: will 68 Ge / 68 The Ga generator was eluted with an aqueous HCl solution (1.0 mL), thereby transferring a 1.25 mL fraction containing approximately 80% radioactivity (600 to 800 MBq) to a reaction flask (ALLTECH, 5 mL). This flask was pre-filled with the corresponding compound (5.0 nmol) and an aqueous solution of 2-(4-(2-hydroxyethyl)-1-piperazinyl)-ethanesulfonic acid (HEPES) (950 μL, 2.7 mL). The reaction flask was heated at 95 °C for 5 min, and then the radiolabeled compound was immobilized on a pretreated SPE column (C8light, SepPak). After pre-rinsing the column with water (10 mL), the radiolabeled PSMA inhibitor was eluted from the column with a mixture of EtOH and water (1 / 1; v / v), phosphate-buffered saline (PBS) (1.0 mL), and then with water (1.0 mL). At the end of the radiolabeling process, the EtOH was evaporated under vacuum, and the tracer was used without further purification. Radiochemical purity was controlled using Radio-TLC (1.0 m sodium citrate buffer and 0.06 m NH4OAc / MeOH buffer (1 / 1; v / v)).

[0493] 177Lu labeling: Following the previous description [5], with only minor modifications, the preparation... 177 Lu-labeled compounds are used without further purification. In short, the appropriate tracer (0.75 to 1.0 nmol, 7.5 to 10 μL) is added to NH4OAc buffer (10 μL, 1.0 mM, pH = 5.9). 177 LuCl3 (10 to 40 MBq; AS > 3000 GBq / mg, 740 MBq / mL, 0.04 mHCl, ITG, Garching, Germany) was added, and finally the mixture was filled with a trace amount of pure water (maximum 100 μL) (Merck, Darmstadt, Germany). The reaction mixture was heated at 95 °C for 40 minutes, and the radiochemical purity was determined by radioactive TLC.

[0494] 125 I-label: In short, the tin alkylation precursor (SnBu3-BA)(OtBu)KuE(OtBu)2(PSMA-45) (approximately 0.1 mg) was dissolved in peracetic acid (20 μL), [ 125 I]NaI (5.0 μL, approx. 21.0 MBq) (74 TBq / mmol, 3.1 GBq / mL, 40 mm NaOH, Hartmann Analytic, Braunschweig, Germany), MeCN (20 μL), and AcOH (10 μL). The reaction solution was incubated at room temperature for 10 min, loaded into a column (C18 Sep Pak Plus, pretreated with 10 mL MeOH and 10 mL water), and washed with water (10 mL). After elution with a 1 / 1 mixture (v / v) of EtOH and MeCN (2.0 mL), the solution was evaporated to dryness under a gentle nitrogen flow and treated with TFA (200 μL) for 30 min, followed by evaporation of the TFA. 125 The crude product of I]I-BA)KuE was purified by radio-RP-HPLC (20 to 40% B within 20 minutes): t R =13.0 minutes; K'=6.2.

[0495] 8. Determination of HSA binding

[0496] HSA binding experiments were performed as previously described [6]. The mobile phase consisted of a binary gradient system with a constant total flow rate of 0.5 mL / min. Mobile phase A was a 50 mm pH 6.9 NH4OAc solution, and mobile phase B was 2-propanol (RP-HPLC grade, VWR, Germany). The gradient of mobile phase A was 100% from 0 to 3 min, and mobile phase B was set to 20% from 3 min to the end of each run. On each experimental day, the column was calibrated with nine reference materials to confirm performance and establish nonlinear regression. The PSMA inhibitor was dissolved at a concentration of 0.5 mg / mL in a mixture of 2-propanol and NH4OAc buffer (50 mm pH 6.9) (1 / 1; v / v). At each run, 10 μL of the inhibitor-containing solution was injected into the RP-HPLC system, and the retention time was measured. References for HSA binding [%] were obtained from Valko et al. or Yamazaki et al. [6,7]. Nonlinear regression was established using OriginPro 2016G.

[0497] 9. Determination of lipophilicity

[0498] Lipophilicity: Radiolabeled PSMA inhibitor (0.5 to 1.0 MBq) dissolved in PBS (500 μL, pH = 7.4) was added to n-octanol (500 μL) in a reaction flask (1.5 mL), and the mixture was vortexed vigorously for 3 minutes (n = 6). To quantify phase separation, the mixture was centrifuged at 6000 g for 5 minutes (Biofuge 15, Heraus Sepatech, Osterode, Germany). The radioactivity of a sample (100 μL) from each phase was measured using a gamma counter to obtain logP. (o / w) value.

[0499] 10. Cell experiments

[0500] Cell Culture: PSMA-positive LNCAP cells (300265; Cell Lines Service GmbH) were cultured in Dulbecco Modified Eagle Medium / Nutrient Mixture F-12 (1 / 1) (DMEM-F12, Biochrom) supplemented with fetal bovine serum (FCS) (10%, Biochrom) and maintained at 37°C in a humid CO2 atmosphere (5%). The day before all experiments using LNCaP cells (24 hours ± 2 hours), the cultured cells were harvested and centrifuged using a mixture of trypsin / EDTA (0.05% / 0.02%) and PBS. After centrifugation, the supernatant was discarded, and the cell pellet was resuspended in medium. Cells were then counted using a hemocytometer (Neubauer) and seeded into 24-well plates. IC50 was determined by transferring 150,000 cells / mL to each well of a 24-well plate. 50 The internalization rate was obtained by transferring 125,000 cells / mL per well into a 24-well PLL coated plate.

[0501] 11. An affinity (IC 50 )

[0502] After removing the culture medium, cells were treated once with HBSS (500 μL, Hank's Balanced Salt Solution, Biochrom, Berlin, Germany, with 1% BSA added) and placed on ice for 15 minutes to equilibrate in HBSS (200 μL, 1% BSA). Next, cells were added with HBSS (1% BSA, control) or at progressively increasing concentrations of the corresponding ligand (10-1 ligands in HBSS (1% BSA)). -10 Up to 10 -4 A solution of m) (25 μL per well) was prepared, followed by the addition of ([ 125 Cells were incubated with HBSS (1% BSA) solution containing 25 μL of KuE (2.0 nm). All experiments were performed at least three times for each concentration. After incubation on ice for 60 minutes, the experiments were terminated by removing the culture medium and continuously rinsing with HBSS (200 μL). The culture medium from both steps was combined into one fraction representing the amount of free radioligands. Cells were then dissolved in NaOH (250 μL, 1.0 nm) and combined with HBSS (200 μL) from the subsequent washing step. Quantification of bound and free radioligands was performed using a gamma counter.

[0503] 12. Internalization

[0504] After removing the culture medium, the cells were washed once with DMEM-F12 solution (500 μL, 5% BSA) and equilibrated at 37°C in DMEM-F12 solution (200 μL, 5% BSA) for at least 15 minutes. Then, each well was blocked by treating with either DMEM-F12 solution (25 μL, 5% BSA) or 2-PMPA solution (25 μL, 100 μm). Next, the appropriate amounts of [unspecified ingredients] were added. 68 Ga or 177 Cells were incubated with Lu-labeled PSMA inhibitors (25 μL; 2.0 nm and 10 nm, respectively) at 37 °C for 5, 15, 30, and 60 min, respectively. Experiments were terminated by placing 24-well plates on ice for 3 min and continuously removing the culture medium. Each well was washed with HBSS (250 μL), and fractions from the first two steps were combined, representing the amount of free radioligands. Surface-bound radioactivity was removed by incubating cells with ice-cold 2-PMPA solution (250 μL, 10 μM in PBS) for 5 min, followed by a wash with ice-cold PBS (250 μL). Internalized radioactivity was determined by incubating cells in NaOH (250 μL, 1.0 M) and combining the fractions from the subsequent wash with NaOH (250 μL, 1.0 M). Each experiment (control and block) was repeated three times for each time point. Free, surface-bound, and internalized radioactivity were quantified using a gamma counter.

[0505] 13. Externalization

[0506] The externalization kinetics of a radiolabeled PSMA inhibitor were determined using LNCaP cells prepared in a manner similar to that described for the internalization assay. After an initial cell washing step with DMEM-F12 solution (5% BSA), cells were rehydrated at 37°C for at least 15 minutes. Subsequently, LNCaP cells were incubated with the corresponding radiolabeled peptide (25 μL, 10.0 nm) at 37°C for 60 minutes, with a total volume of 250 μL per well. After 60 minutes, the supernatant containing unbound free fractions was removed, and the total amount of radioactivity added was measured using a gamma counter to calculate the total amount of radioactivity added. Acid washing was avoided in subsequent externalization and recycling studies to ensure enzyme integrity. To determine the recycling rate, fresh DMEM-F12 solution (250 μL, 5% BSA) was added to the cells to allow for reinternalization. Conversely, reinternalization was inhibited by adding a DMEM-F12-solution containing 2-PMPA (225 μL DMEM-F12 (5% BSA) and 25 μL 100 μM 2-PMPA-solution (PBS)). Cells were then incubated at 37°C for 0, 20, 40, and 60 min. The supernatant was then removed, and cells were washed with ice-cold HBSS (250 μL). The combination of the supernatant and the accompanying wash with HBSS (200 μL) constituted the externalized radioligands at the time points studied. Furthermore, cells were then washed twice with ice-cold 2-PMPA HBSS solution (250 μL, 10 μM), and the results were combined to represent the fraction of membrane-bound radioligands. The internalized fraction was determined by dissolution in NaOH (250 μL, 1.0 M) as described for the internalization assay. The radioactivity of free, externalized, membrane-bound, and internalized radioligands was quantified using a gamma counter.

[0507] 14. Animal experiments

[0508] All animal experiments were conducted in accordance with the German General Animal Welfare Regulations (Deutsches Tierschutzgesetz, approval number 55.2-1-54-2532-71-13). For the tumor model, LNCaP cells (approximately 10 7 (10 cells) were suspended in serum-free DMEM-F12 medium and Matrigel (1 / 1; v / v) (BD Biosciences, Germany) and seeded onto the right shoulder of 6- to 8-week-old male CB-17SCID mice (Charles River Laboratories, Sulzfeld, Germany). Animals were used for experiments after the tumors reached a diameter of 4 to 8 mm.

[0509] 15. PET

[0510] Imaging experiments were performed using Siemens Inveon small animal PET, and the data were analyzed using the relevant Inveon ResearchWorkplace software. Mice were anesthetized with isoflurane and injected via the tail vein with approximately 4.0 to 17 MBq of [a specific drug / method / treatment]. 68 Ga-labeled compounds (approximately 150 to 300 μL) were injected into the bed 90 minutes later for dynamic imaging. Static blockade images were acquired within a 15-minute acquisition time after 1 h p. PSMA blockade was achieved by co-injection of 8 mg / kg of 2-PMPA solution (PBS). All images were reconstructed using the OSEM3D algorithm without scanner or attenuation correction.

[0511] 16. Biodistribution

[0512] Approximately 4.0 to 12.0 MBq (approximately 150 to 300 μL) of the corresponding amount 68 Ga or 177 Lu-labeled PSMA inhibitors were injected into the tail vein of male CB-17SCID mice carrying LNCaP tumors, and the mice were sacrificed after a specific time period (n=4, respectively). Selected organs were removed, weighed, and measured in a γ counter.

[0513] 17. References in Example 1

[0514] 1. J., et al., A Monoreactive Bifunctional TriazacyclononanePhosphinate Chelator with High Selectivity for Gallium-68. ChemMedChem, 2012.7(8):p.1375-1378.

[0515] 2.Weineisen,M.,et al.,Development and first in human evaluation ofPSMA I&T-Aligand for diagnostic imaging and endoradiotherapy of prostatecancer.Journal of Nuclear Medicine,2014.55(supplement 1):p.1083-1083.

[0516] 3.Weineisen,M.,et al.,Synthesis and preclinical evaluation of DOTAGA-conjugated PSMA ligands for functional imaging and endoradiotherapy ofprostate cancer.EJNMMI research,2014.4(1):p.1.

[0517] 4.Weineisen,M.,et al.,68Ga-and 177Lu-Labeled PSMA I&T:Optimization ofaPSMA-Targeted Theranostic Concept and First Proof-of-Concept HumanStudies.Journal of Nuclear Medicine,2015.56(8):p.1169-1176.

[0518] 5.Sosabowski,J.K.and S.J.Mather,Conjugation of DOTA-like chelatingagents to peptides and radiolabeling with trivalent metallicisotopes.Nat.Protocols,2006.1(2):p.972-976.

[0519] 6.Valko,K.,et al.,Fast gradient HPLC method to determine compoundsbinding to human serum albumin.Relationships with octanol / water andimmobilized artificial membrane lipophilicity.Journal of pharmaceuticalsciences,2003.92(11):p.2236-2248.

[0520] 7. Yamazaki, K. and M. Kanaoka, Computational prediction of the plasmaprotein-binding percent of diverse pharmaceutical compounds. Journal of pharmaceutical sciences, 2004.93(6): p.1480-1494.

[0521] Example 2: Results

[0522] 1. Influence of the introduction of 2,4-dinitrobenzoic acid into the PSMA I&T linker region

[0523] DOTAGA-y(3-I)fk(Sub-KuE)(PSMA I&T):

[0524]

[0525] DOTAGA–y(3-I)fk(L-Asu[KuE]-2,4-DNBA)(PSMA-36):

[0526]

[0527] Table 3

[0528]

[0529] →Affinity is slightly higher, and internalization is increased by 251%.

[0530] 2. Change of the binding motif from EuK to EuE and the peptide spacer from -y(3-I)fk- to -y-2-nal-k-

[0531] DOTAGA-y(3-I)fk(Sub-KuE)(PSMA I&T):

[0532]

[0533] DOTAGA-y-2-nal-k(Suc-N 5 -orn-C 4 -EuE)(PSMA-46):

[0534]

[0535] Table 4

[0536]

[0537] → Compared to the reference compound PSMA I&T, the improved reference compound PSMA-46 exhibits higher internalization and improved affinity. Therefore, based on the structure of PSMA-46, electron-deficient aromatic residues were introduced in further development steps.

[0538] 3. Introduction of 4-nitrophenylalanine and 2,4-DNBA into the peptide spacer of PSMA-46

[0539] DOTAGA-y-2-nal-k(Suc-N 5 -orn-C 4 -EuE)(PSMA-46):

[0540]

[0541] DOTAGA-F(4-NO2)-y-2-nal-k(Suc-N 5 -orn-C 4 -EuE)(PSMA-52):

[0542]

[0543] 2,4-DNBA-Dap(DOTAGA)-y-2-nal-k(Suc-N 5 -orn-C 4 -EuE)(PSMA-53):

[0544]

[0545]

[0546] →Although the affinity remains similar, the introduction of 4-nitrophenylalanine slightly increases internalization, but internalization can potentially be significantly increased by introducing another nitro group via 2,4-DNBA.

[0547] →To increase internalization, two electron-withdrawing groups are preferred.

[0548] 4. Introduction of 4-aminophenylalanine

[0549] DOTAGA-F(4-NH2)y-2-nal-k(Suc-N 5 -orn-C 4 -EuE)(PSMA-49):

[0550]

[0551] DOTAGA-F(4-NH2)y-2-nal-k(d[N 5 -orn-C 4-EuE]-2,4-DNBA)(PSMA-61):

[0552] DOTAGA-F(4-NH2)y-2-nal-k(d[N 5 -orn-C 4 -EuE]-TMA)(PSMA-62):

[0553]

[0554] Table 6

[0555]

[0556] → Both 2,4-DNBA and benzotriic acid modifications can further increase internalization.

[0557] 5. Introduction of electron-deficient groups in the peptide spacer

[0558] DOTAGA-F(4-NH2)y-2-nal-e(Abz-N 5 -orn-C 4 -EuE)(PSMA-60)

[0559]

[0560] 2,4-DNBA-Dap(DOTAGA)y-2-nal-e(Abz-N 5 -orn-C 4 -EuE)(PSMA-65):

[0561]

[0562] Table 7

[0563]

[0564] → Electron-deficient aromatic modification of 2,4-DNBA can increase internalization.

[0565] 6. Benzenetricarboxylic acid is introduced into the linker and the peptide spacer of PSMA inhibitors

[0566] DOTAGA-F(4-NH2)y-2-nal-k(d[N 5 -orn-C 4 -EuE]-TMA)(PSMA-62):

[0567]

[0568] DOTAGA-Dap(TMA)y-2-nal-k(d[N 5-orn-C 4 -EuE]-TMA)(PSMA-66):

[0569]

[0570] Table 8

[0571]

[0572] →4-Aminophenylalanine exhibits similar affinity to Dap(TMA) upon exchange, but its internalization ability is slightly reduced. Since both ligands appear very promising, both tracers were evaluated in further experiments.

[0573] →The conclusion drawn from these experiments is that electron-deficient aromatic residues are transferable and can increase internalization while maintaining high affinity.

[0574] 7. With [ 177 Lu]PSMA I&T and [ 177 Lu]PSMA-617, the compound [ 177 Lu]PSMA-62 and [ 177 Effect of internalization on cell retention in vitro for Lu]PSMA-66.​

[0575] [ 177 Lu]PSMA-66 exhibited the highest intracellular radioactivity in tumor cells after 1 hour, followed by [ 177 Lu]PSMA-62, however, it was found that [ 177 Lu]PSMA-62's internalization is higher than [ 177 Lu]PSMA-66 (343.9% vs. 297.8%, respectively). Notably, even when reinternalization was blocked with 100 μm 2-PMPA solution, intracellular clearance [ 177 Lu]PSMA-66 was also lower than all other compounds studied. If reinternalization was blocked, compared to the reference [ 177 The difference between Lu]PSMAI&T and others is more than double.

[0576] [ 177 Lu]PSMA-66 has nine free carboxyl groups, which equals nine negative intracellular charges (pH = 7.4). This high charge characteristic of the compound may explain its prolonged intracellular retention time due to the electrostatic repulsion of negatively charged cell membranes.

[0577] 8. In vivo experiments: Biodistribution

[0578] Table 9: CB-17SCID mice (n=4, , respectively) carrying LNCaP tumor xenografts at 1 h pi [ 177 Lu]PSMA-49、[ 177 Lu]PSMA-62 and [ 177Biodistribution data of LuPSMA-66 (%ID / g). Injections of 3.5 MBq to 5.5 MBq were performed respectively. 177 Lu-labeled radioligands (0.15 to 0.25 nmol tracer).

[0579]

[0580] After 1 hour pi (4.69 ± 0.95%) [ 177 Compared to tumor uptake by Lu]PSMA I&T, tumor radioactivity is significantly increased through improved internalization and affinity. For example, […]. 177 Lu]PSMA-16、[ 177 Lu]PSMA-40 and [ 177 As observed in Lu]PSMA-41, the extension of the peptide spacer with 4-amino-D-phenylalanine leads to high renal uptake, and this modification has been confirmed to increase renal accumulation.

[0581] Compared to the reference, the introduction of benzopyridine into [ 177 The linker of Lu]PSMA-62 resulted in decreased renal uptake (106.45 ± 17.18% vs. 162.96 ± 23.20%, respectively) and a slight decrease in tumor uptake. This was due to the direct association with [ 177 Compared to Lu]PSMA-49, [ 177 The higher internalization rate of LuPSMA-62 meant that the lower tumor uptake was unexpected. It remains unclear to what extent internalization contributes to tumor uptake and whether it is less important than affinity. 177 Lu]PSMA-49 and [ 177 A direct comparison with Lu]PSMA-62 shows that affinity is more important because [ 177 Lu]PSMA-49 has a higher affinity for PSMA (2.5±0.6 nm vs. 4.0±0.2 nm).

[0582] Table 10: After 24 hours (pi) [ 177 Lu]PSMA I&T、[ 177 Lu]PSMA-62 and [ 177 Lu]PSMA-66 and [ 177 Biodistribution data (%ID / g) of LuPSMA-71 in CB-17SCID mice (n=4, , respectively) carrying LNCaP tumor xenografts. Injection of 3.5 MBq to 5.5 MBq of the corresponding... 177 Lu-labeled radioligands (0.15 to 0.25 nmol tracer).

[0583]

[0584] The results in Table 10 show the tracers evaluated. 177 Lu]PSMA-62、[ 177 Lu]PSMA-66 and [ 177 Significant differences were observed between Lu]PSMA-71. Regarding renal clearance, a decrease in renal uptake was observed for all ligands compared to 1 hpi (Table 9). Although [ 177 Lu]PSMA I&T showed the highest renal uptake after 24 hours post-pi, but [ 177 The lowest level of LuPSMA-62 was observed, consistent with renal clearance observed in PET studies. [24h pi] 177 Tumor uptake of Lu]PSMA-61 remained almost constant over 23 hours (8.00 ± 0.75 vs. 7.70 ± 1.35% ID / g, 1 h p and 24 hp.i., respectively). Although in terms of internalization and affinity, [ 177 Lu]PSMA-62 and [ 177 Lu]PSMA-66 showed similar in vitro parameters, but from 1 h pi to 24 h pi, [ 177 Lu]PSMA-66 tumor uptake and [ 177 Lu]PSMA-62 showed a greater reduction (10.00±0.44 vs 5.73±1.39% ID / g, 1h pi and 24h pi, respectively). Stronger tumor preservation and more favorable tumor-to-liver and tumor-to-muscle ratios resulted in [ 177 Lu]PSMA-62 and [ 177 Compared to PSMA-66, Lu]PSMA-71 showed superior performance. PSMA-71 exhibited the highest tumor uptake and also the highest HSA binding value. Although 24-hour renal uptake was lower than [ 177 Similar to Lu]PSMA I&T, [ 177 The tumor uptake of LuPSMA-71 was more than three times higher (4.06 ± 1.12 versus 14.29 ± 0.89% ID / g, respectively). 177 Lu]PSMA I&T and [ 177 Lu]PSMA-71).

[0585] in this regard,[ 177 Lu]PSMA-71 can be considered a tracer of particular value for intracavitary radiotherapy and is a candidate for clinical application.

[0586] 9. In vivo experiments: PET imaging

[0587] The effect of 2,4-dinitrobenzoic acid linker substitution on EuK-based inhibitors

[0588] The EuK-based inhibitor PSMA-36 was evaluated in small animal PET scans to examine the effect of 2,4-dinitrobenzoic acid in the linker on in vivo distribution.

[0589] The TAC logarithmic plot shows [ 68 Specific renal and tumor uptake of GaPSMA-36. A linear decrease in blood pool radioactivity and muscle region suggests low nonspecific binding and rapid excretion. Accumulation in the tumor remained stable during the observation period. Despite […] 177 Compared to Lu]PSMA I&T, [ 177 Lu]PSMA-36 showed more than three times the internalization rate, but after 85 min pi, tumor uptake was only moderate, at 3.5% ID / mL. [ 68 The most significant difference compared to Ga]PSMA I&T was the high and stable uptake in the lacrimal and salivary glands, showing approximately 2% ID / mL in both regions. This is due to the difference compared to the reference [ 68 The only structural difference in Ga]PSMA I&T is the introduction of 2,4-dinitrobenzoic acid, so the linker modification must be the reason for this increased absorption. However, further research is needed to confirm this effect.

[0590] It is also worth noting that the clearance rate in these areas is slower compared to blood pools and muscle, suggesting the involvement of unique retention mechanisms. PSMA has been reported to be involved in angiogenesis during neovascularization in the mouse eye, thus potentially explaining […]. 68 Uptake of Ga]PSMA-36[1]. In clinical treatments of 177Lu-labeled PSMA inhibitors, the accumulation of tracers in the salivary glands is a common problem[2]. Drug uptake in the salivary glands depends on intracellular or extracellular pathways, most commonly on simple diffusion between the phospholipid bilayers of acinar cells. The concentration of the drug in saliva is primarily reflected by the free, non-ionized fraction in plasma associated with passive diffusion[3-5]. In this respect, passive diffusion seems highly unlikely to cause uptake in the salivary glands. Other mechanisms must be involved because EuK-based PSMA inhibitors are highly charged and therefore highly polar in vivo. Furthermore, passive diffusion in each region shows high background radioactivity during PET scans, which would not occur with most PSMA ligands, as rapid clearance removes the tracer from the blood pool.

[0591] Effects of benzopyridinic acid on EuE-based inhibitors

[0592] The replacement of PSMA ligands by electron-deficient aromatic systems leads to […].177 Lu]PSMA-62 and [ 177 The internalization rates of Lu]PSMA-66 were increased (343.9% and 297.8%, respectively). Therefore, the two ligands were evaluated and compared in a PET study.

[0593] Both tracers exhibited excellent tracer kinetics in terms of renal, muscle, and blood pool uptake. 68 Ga]PSMA-66 and [ 68 Ga]PSMA-62 showed slightly higher specific uptake in the kidney (45.3% ID / mL vs. 34.8% ID / mL, respectively). 68 Ga]PSMA-66 and [ 68 Ga]PSMA-62 showed higher renal accumulation compared to PET scans, which correlated well with biodistribution experiments. Radioactive TAC in muscle and blood pools showed linear uptake and continuous clearance from these compartments.

[0594] 10. References in Example 2

[0595] 1.Grant, CL, et al., Prostate specific membrane antigen (PSMA) regulates angiogenesis independently of VEGF during ocular neovascularization. PloS one, 2012.7(7):p.e41285.

[0596] 2.Kulkarni, HR, et al., PSMA-Based Radioligand Therapy for MetastaticCastration-Resistant Prostate Cancer: The Bad Berka Experience Since 2013. Journal of Nuclear Medicine, 2016.57 (Supplement 3): p.97S-104S.

[0597] 3.Haeckel, R., Factors influencing the saliva / plasma ratio of drugs. Annals of the New York Academy of Sciences, 1993.694(1):p.128-142.

[0598] 4.Jusko,W.J.and R.L.Milsap,Pharmacokinetic Principles of DrugDistribution in Salivaa.Annals of the New York Academy of Sciences,1993.694(1):p.36-47.

[0599] 5.Aps,J.K.and L.C.Martens,Review:the physiology of saliva andtransfer of drugs into saliva.Forensic science international,2005.150(2):p.119-131.

[0600] 6.Young,J.D.,et al.,68Ga-THP-PSMA:a PET imaging agent for prostatecancer offering rapid,room temperature,one-step kit-basedradiolabeling.Journal of Nuclear Medicine,2017:p.jnumed.117.191882.

[0601] 7.Wüstemann,T.,et al.,Design of Internalizing PSMA-specific Glu-ureido-based Radiotherapeuticals.Theranostics,2016.6(8):p.1085.

[0602] 8.Hao,G.,et al.,A multivalent approach of imaging probe design toovercome an endogenous anion binding competition for noninvasive assessmentof prostate specific membrane antigen.Molecular pharmaceutics,2013.10(8):p.2975-2985.

[0603] 9.Soret,M.,S.L.Bacharach,and I.Buvat,Partial-volume effect in PETtumor imaging.Journal of Nuclear Medicine,2007.48(6):p.932-945.

[0604] 10.Bao,Q.,et al.,Performance evaluation of the inveon dedicated PETpreclinical tomograph based on the NEMA NU-4 standards.Journal of NuclearMedicine,2009.50(3):p.401-408.

Claims

1. A pharmaceutical composition comprising a compound having the structure of formula (If) or a pharmaceutically acceptable salt thereof. (If) in: R M -X 3 It is a group of formula (M-2): (M-2)， Among them The marked bond is connected to the remainder of the compound of formula (If), and the group of formula (M-2) optionally contains a chelated non-radioactive or radioactive cation; R 4 It is a group having the following structure: ， Where R 5B It is -NH2 and s is 1; R 9A It is a straight-chain C2 to C10 alkyl diol substituted with -COOH. R 10A It is a straight-chain C2 alkyldiyl group, which is... replace, R 11A It is a straight-chain C2 to C10 alkyl diol substituted with -COOH.

2. The pharmaceutical composition according to claim 1, wherein the compound comprises a radioactive cation.

3. The pharmaceutical composition according to claim 1, wherein the compound comprises 177 Lu or its cations.

4. The pharmaceutical composition according to claim 1, wherein the compound comprises 225 Ac or its cations.

5. The pharmaceutical composition according to claim 1, wherein the compound comprises 161 Tb or its cations.

6. The pharmaceutical composition according to claim 1, wherein the compound comprises 212 Pb or its cations.

7. A pharmaceutical composition comprising a compound having the following structure or a pharmaceutically acceptable salt thereof: And the compound described therewith 225 Ac or its cation chelates; The pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

8. The pharmaceutical composition of claim 7, wherein the pharmaceutical composition comprises the compound or a pharmaceutically acceptable salt thereof, and further comprises one or more pharmaceutically acceptable carriers and / or diluents.

9. A pharmaceutical composition comprising a compound having the following structure or a pharmaceutically acceptable salt thereof: And the compound described therewith 161 Tb or its cation chelates The pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

10. The pharmaceutical composition of claim 9, wherein the pharmaceutical composition comprises the compound or a pharmaceutically acceptable salt thereof, and further comprises one or more pharmaceutically acceptable carriers and / or diluents.

11. A pharmaceutical composition comprising a compound having the following structure or a pharmaceutically acceptable salt thereof: And the compound described therewith 212 Pb or its cation chelates The pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

12. The pharmaceutical composition of claim 11, wherein the pharmaceutical composition comprises the compound or a pharmaceutically acceptable salt thereof, and further comprises one or more pharmaceutically acceptable carriers and / or diluents.

13. The pharmaceutical composition according to any one of claims 1-6, further comprising one or more pharmaceutically acceptable excipients.

14. The pharmaceutical composition according to any one of claims 1-6, further comprising one or more pharmaceutically acceptable carriers and / or diluents.

15. Use of the pharmaceutical composition according to any one of claims 1-14 in the preparation of a medicament for diagnosing and / or treating cancer.

16. The use according to claim 15, wherein the cancer is prostate cancer.

17. Use of the pharmaceutical composition according to any one of claims 1-14 in the preparation of a medicament for diagnosing and / or treating tumors.

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