CXCR4 targeted diagnostic and therapeutic agents with reduced species selectivity

Abstract

# imgabs0 #. The present disclosure relates to CXCR4 targeted diagnostic and therapeutic agents with reduced species selectivity for use in imaging and intraluminal radiotherapy of diseases involving chemokine receptor 4 (CXCR4). Compounds that bind to or inhibit hCXCR4 and mCXCR4 and also bear at least one moiety suitable for labeling are provided, preferably having the structure of formula (IV). Also provided are pharmaceutical uses of such compounds.

Description

[0001] This application is a divisional application of Chinese patent application 201980059915.0, filed on September 11, 2019, with the invention name “CXCR4 targeted diagnostic and therapeutic agents with reduced species selectivity”. Technical Field

[0002] The present disclosure relates to imaging and intracavitary radiation therapy of diseases involving chemokine receptor 4 (CXCR4). Compounds that bind to or inhibit CXCR4 and also carry at least one moiety suitable for labeling are provided. Medical uses of such compounds are also provided.

[0003] Several documents, including patent applications and manufacturer manuals, are cited in this specification. The disclosures of these documents, although not considered relevant to the patentability of the present invention, are incorporated herein by reference in their entirety. More specifically, all references are incorporated by reference to the same extent as if each individual document was specifically and individually indicated to be incorporated by reference. Background Art

[0004] The sole endogenous ligand, CXCL12 (formerly known as SDF-1α), binds to its cognate receptor, chemokine receptor 4 (CXCR4) [1], activating the downstream protein kinase B (AKT) / mitogen-activated protein kinase (MAPK) signaling pathway, leading to altered gene expression, actin polymerization, cytoskeletal rearrangements, and cell migration. The physiological functions of the CXCL12 / CXCR4 axis include embryogenesis (regulation of embryonic stem cell migration and localization), immune response (leukocyte trafficking to sites of inflammation), embryo implantation (localization of blastocysts in the maternal endometrium), hematopoiesis (homing and differentiation of hematopoietic stem / progenitor cells in the bone marrow), brain development, and neoangiogenesis [2-6]. As a result, genetic defects in mice lacking both CXCL12 and CXCR4 genes display the same lethal phenotype, with severe impairments in hematopoiesis and central nervous system (CNS) development [7].

[0005] The CXCR4 receptor has been implicated in a variety of diseases. For example, it was first identified as a co-receptor mediating HIV-1 entry into T cells [3]. Furthermore, in rheumatoid arthritis (RA), CXCR4-expressing CD4+ memory T cells accumulate in the inflamed synovium due to high local CXCL12 concentrations [8]. Chronic inflammation of the arterial wall characterized by chemokine-mediated leukocyte influx plays a central role in the pathogenesis of atherosclerosis [9]. The cytokine macrophage migration inhibitory factor (MIF) is a unique proinflammatory regulator of a variety of acute and chronic inflammatory diseases that contributes to lesion progression and plaque inflammation. These chemokine-like functions are mediated by the interaction of MIF with the chemokine receptors CXCR2 and CXCR4, suggesting an important role for CXCR4 in atherosclerotic plaque development, post-injury vascular remodeling, atherosclerotic plaque destabilization, and aneurysm formation

[10] .

[0006] In addition, CXCR4 is involved in B cell trafficking and tissue localization in patients with chronic leukemia

[11] and in the regulation of organ-specific metastasis in different breast cancer models

[12] . In addition, significant CXCR4 overexpression has been detected in more than 20 human tumor types, including hematopoietic malignancies, brain tumors, gastrointestinal cancers, and other cancer types [2,13-14]. Therefore, methods for early assessment of tumor metastatic potential and metastatic spread are valuable tools for treatment planning, monitoring, and control, as cancer metastasis is one of the key factors affecting patient life expectancy.

[0007] Given the undisputed relevance of CXCR4 as a diagnostic and therapeutic molecular target, a variety of CXCR4-targeting peptide and non-peptide antagonists have been developed over the past decade. Among them, the bicyclam AMD3100 (plerixafor / Mozobil) is the only compound approved by the FDA (2008) for stem cell mobilization and for the treatment of hematological and other cancers [7,15-17]. In preclinical studies using mouse models of various human hematological cancers and solid cancers, antitumor treatment with alternative small molecule CXCR4 antagonists such as AMD3465 [18-19] or MSX-122

[20] , peptide CXCL12 derivatives (CTCE-9908

[21] , BKT-140 [22-24], POL-5551 [25-27]) or anti-CXCR4-antibodies [28-29] has consistently led to prolonged overall survival, primarily by effectively preventing distant organ metastases

[30] . Another potent CXCR4 antagonist, LY2510924 (cyclo[Phe-Tyr-Lys(iPr)-D-Arg-2-Nal-Gly-D-Glu]-Lys(iPr)-NH2)[31-32], exhibits high antitumor activity in solid tumor and breast cancer metastasis models and is currently being evaluated in Phase II clinical studies (NCT01391130 and NCT1439568). Lactam cyclized heptapeptides are reported to be potent CXCR4 antagonists with high efficacy in the treatment of cancer, rheumatoid arthritis, pulmonary fibrosis, and HIV infection (WO 2008150689 A1). Recently, a disulfide-bridged cyclic heptapeptide antagonist (WO / 2011 / 092575A1) was shown to exhibit high in vivo stability [33-34], effectively inhibit lung metastasis in a melanoma model

[35] , and reduce the metastatic potential of HCC and osteosarcoma in mouse models

[36] . The modified analog (R29, Ac-Arg-Ala-[D-Cys-Arg-Phe-His-Pen]-COOH) effectively reversed the suppressive activity of T-regulatory cells in renal cancer

[37] .

[0008] Among these CXCR4-targeted antagonists, the T-140-based peptide is the first compound used for molecular imaging of CXCR4-expression in vivo; different analogs, whether used 18 F or 68 Ga radiolabeled or conjugated to fluorescent dyes, as well as the corresponding bimodal probes, have been used for non-invasive detection and visualization of CXCR4-expressing tissues using positron emission tomography (PET), optical or SPECT imaging [38-45].

[0009] In addition, cyclic pentapeptides based on the N-terminal sequence of CXCL12

[46] or, more importantly, the reduced size of the T-140 binding sequence ring (Gly-Nal-Arg-Arg-D-Tyr) (FC-131)

[47] have been extensively evaluated. Detailed structure-activity relationship (SAR) studies have highlighted the relevance of individual amino acid residues and their stereochemistry as well as amide bond methylation [48–53] for optimal CXCR4 affinity and antagonist activity. Based on these findings and our own SAR studies, our group developed the first CXCR4-targeted molecular imaging agent based on a pentapeptide, [ 68 Ga]pentixafor, replacing Arg with D-ornithine 2 (in FC131) and N-methylation of D-Orn leads to the CPCR4-scaffold (loop (D-Tyr 1 -D-[NMe]Orn 2 -Arg 3 -Nal 4 -Gly 5 )), which showed excellent binding affinity to CXCR4

[54] . Orn was further functionalized with a suitable linker (4-aminomethylbenzoic acid, AMBS) and DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) chelator. 2 side chain, to obtain pentixafor (cyclo (D-Tyr 1 -D-[NMe]Orn 2 (AMBS-DOTA)-Arg 3 -Nal 4 -Gly 5 ), also known as CPCR4.2) [54-56]; WO2007 / 096662, WO2009 / 027706, WO2011 / 131735). 68 Ga-labeled analogs, [ 68 Ga]pentixafor

[57] displays high affinity and selectivity for human CXCR4, rapid renal excretion, and minimal nonspecific background accumulation, thus allowing sensitive and high-contrast imaging of CXCR4-expressing tissues in vivo using PET. In addition to its successful use in patients with lymphoma

[58] , multiple myeloma[59–61], AML

[62] , SCLC

[63] , glioblastoma

[64] , or other solid tumors

[65] , 68 Ga]pentixafor-PET has recently been shown to be a valuable tool for monitoring inflammatory processes in vivo, for example after myocardial infarction[66–68] or stroke

[69] , in atherosclerosis[70–72] or other inflammatory diseases[73–74].

[0010] An alternative peptide backbone (cyclo(D-3-iodo-Tyr 1 -D-[NMe]Orn 2 -Arg 3 -Nal 4 -Gly 5 )) was used to synthesize the first CXCR4-targeted intracavitary radiotherapeutic agent, pentixather (cyclo(D-3-iodo-Tyr 1 -D-[NMe]Orn 2 (AMBS-DOTA)-Arg 3 -Nal 4 -Gly 5 )[75-76](WO2015 / 185162). First used in patients with multiple myeloma[ 177 Lu]- and [ 90 Peptide receptor targeted radionuclide therapy (PRRT) has shown very promising results with pentixather [77-78], so a clinical trial has recently been initiated to further evaluate the use of pentixather in the treatment of 68 Ga]pentixafor / [ 177 Lu / 90 Y]pentixather's CXCR4-targeted theranostic concept. In addition, ongoing preclinical studies are aimed at using [ 213 Bi / 225 Ac]pentixather determines the potential of alpha-therapy in the treatment of disseminated micrometastatic hematological cancers.

[0011] Although CXCR4-targeted theranostics have undisputed clinical potential, [ 68 Ga]pentixafor and [ 177 Lu / 90 [Y] Pentixafor / pentixather show remarkable selectivity for the human CXCR4 (hCXCR4) receptor, while showing little affinity for the murine receptor (mCXCR4). This has so far hampered their use as preclinical tools for studying CXCR4-related pathologies such as carcinogenesis or inflammatory diseases in mouse models. Recently, a pentapeptide scaffold with a modified N-alkylation pattern compared to pentixafor / pentixather has been introduced

[79] , and peptides based on this backbone have been shown to have hCXCR4 affinities in the low nanomolar to subnanomolar range. Recent studies have shown that, surprisingly, the analog with the highest hCXCR4 affinity (cyclo(D-Tyr 1 -D-[N-hexyl-6-guanidino]D-Ala 2 -Arg 3-Nal 4 -Gly 5 ); CPCR4.3) also binds to mCXCR4 with similarly high affinity. Based on these results, it has recently been shown that radioiodinated CPCR4.3 sensitively detects CXCR4-expressing immune cells in a mouse model of esophageal cancer

[80] .

[0012] Unfortunately, however, radioiodinated CPCR4.3 is not well suited for imaging purposes outside of autoradiographic studies; due to its lipophilicity, the peptide displays suboptimal pharmacokinetics characterized by significant hepatobiliary clearance, which has hampered its use in in vivo imaging studies. However, implementation of alternative labeling strategies, such as radiometallation via DOTA chelators (as in Pentixafor / Pentixather), requires identification of suitable attachment sites for further structural modifications within the peptide sequence. Summary of the invention

[0013] Based on this hypothesis, the present invention describes the design of novel CPCR4.3 analogs, which have 2 The present invention summarizes the methods for determining suitable linker moieties between the peptide core and the radiolabel / signaling unit and describes the design of the resulting novel CXCR4 targeting probes with high m / hCXCR4 affinity (reduced species selectivity) labeled with a wide range of radionuclides and / or fluorescent dyes for molecular imaging and therapeutic applications.

[0014] Therefore, in a first aspect, the present invention relates to a compound of the following formula (I) or a pharmaceutically acceptable salt thereof,

[0015]

[0016] in:

[0017] R 1 is an alkyl group, preferably a C1-C6 alkyl group, more preferably a methyl group;

[0018] R 2 is H or an alkyl group, which may be unsubstituted or substituted by at least one substituent selected from the group consisting of -NH2, -NH-C(=NH)-NH2, -C(O)NH2, -C(O)OH, -OH, -SH, -SCH3, and a 5- to 10-membered carbocyclic ring or a 5- to 10-membered heterocyclic ring containing oxygen, nitrogen or sulfur as a heteroatom, wherein the alkyl group is preferably a C1-C6 alkyl group;

[0019] R 3is H or an alkyl group, which may be unsubstituted or substituted by at least one substituent selected from the group consisting of -NH2, -NH-C(=NH)-NH2, -C(O)NH2, -C(O)OH, -OH, -SH, -SCH3, and a 5- to 10-membered carbocyclic ring or a 5- to 10-membered heterocyclic ring containing oxygen, nitrogen or sulfur as a heteroatom, wherein the alkyl group is preferably a C1-C6 alkyl group;

[0020] R 4 is H or an alkyl group, the alkyl group may be unsubstituted or substituted by at least one substituent selected from the group consisting of: -NH2, -NH-C(=NH)-NH2, -C(O)NH2, -C(O)OH, -OH, -SH, -SCH3, -SR A 、-SR 11 and a 5- to 10-membered carbocyclic ring or a 5- to 10-membered heterocyclic ring containing oxygen, nitrogen or sulfur as a heteroatom, wherein the alkyl group is preferably a C1-C6 alkyl group;

[0021] R 5 is H or I;

[0022] R 6 is at least one selected from -NH2, -NH-C(=NH)-NH2, -NH-R B 、-NH-C(=NH)-NH-R B 、-NH-C(=NH)-NH-R 14 and -NH-C(=NH)-NH-C(=O)-R 15 The alkyl group substituted by a substituent, wherein the alkyl group is preferably a C1-C8 alkyl group,

[0023] R 7 To R 10 Each independently is H or alkyl; preferably H or C1-C6 alkyl; and more preferably H;

[0024] R 11 Selected from -(CH2)-C(=O)-NH-R 12 -NH-C(=O)-CH3 and

[0025] -(CH2)-C(=O)-NH-(CH2) r -R 13 -(CH2) s -NH-C(=O)-CH3, where R 12 is a C2-C10 alkanediyl group, preferably a C3-C6 alkanediyl group, which may be substituted by a group -C(=O)-NH2, R 13 is phenylene, preferably 1,3-phenylene, and r and s are independently an integer selected from 0, 1 or 2, and are preferably both 1;

[0026] R 14 and R 15 Independently C1-C10 alkyl, preferably C2-C6 alkyl, which may be substituted by at least one substituent selected from -NH-C(=O)-CH3 and -C≡CH;

[0027] R A is a group comprising at least one selected from the following:

[0028] (i) a chelating moiety,

[0029] (ii) a chelate formed by the chelating moiety (i) and a chelated radioactive or non-radioactive cation,

[0030] (iii) a moiety carrying a covalently bound radioisotope or a precursor suitable for labelling with such a radioisotope,

[0031] (iv) a phosphonate moiety, and

[0032] (v) fluorescent labels, such as quantum dots or fluorescent dyes; and

[0033] R B is a group comprising at least one selected from the following:

[0034] (i) a chelating moiety,

[0035] (ii) a chelate formed by a chelating moiety and a chelated radioactive or non-radioactive cation,

[0036] (iii) a group carrying a covalently bound radioisotope or a precursor suitable for labelling with such a radioisotope,

[0037] (iv) a phosphonate moiety, and

[0038] (v) fluorescent labels, such as quantum dots or fluorescent dyes,

[0039] And R B It may also contain a group of formula (II)

[0040]

[0041] Its use The indicated bond is bound to the rest of the compound, where R 1 To R 5 and R 7 To R 10 As defined above, and wherein R 16 is an alkanediyl group, preferably a C1-C6 alkanediyl group, and R 17 is selected from -NH- and -NH-C(=NH)-NH-;

[0042] And among them

[0043] The compound of formula (I) comprises at least one selected from -SR A 、-NH-R B and -NH-C(=NH)-NH-R B substituent.

[0044] The compounds according to the first aspect of the invention differ from the CXCR4 binding agents known in the art in at least one characteristic. To further explain, one of these characteristics is that in the compounds of the invention R 6 More specifically, it is substituted with an alkyl group carrying a substituent selected from amino and guanidinium, which in turn may be further substituted. These further substituents include R B Part. R B Embodiments of the present invention include chelating moieties and other markers or groups suitable for marking. In other words, not only the carrying R 6 The nitrogen atom of the molecule is replaced, and by such replacement, an anchor point is introduced into the molecule, which provides further options for derivatization. Preferred derivatizations are further described below, including the introduction of moieties for diagnostic molecular imaging and intracavitary radiotherapy. The present invention demonstrates that such derivatization is possible without negatively interfering with target binding.

[0045] Another difference from the CXCR4 binding agents known in the art is that the group R 1 Instead of the side chain of ornithine, it is an alkyl group, preferably a methyl group.

[0046] Introducing the substituent R defined above 6 and R 1 The alkylation mode of the present invention is changed at R, also known as "transalkylation", which provides significant advantages, in addition to the advantages of 6 In addition to the selection of introducing the marker or a portion suitable for marking

[0047] A significant increase in affinity for the target human CXCR4 (hCXCR4), and

[0048] · Significant reduction in ligand binding species selectivity, resulting in 6 The modified peptides are effective for human (hCXCR4) and mouse (mCXCR4)

[0049] Almost the same high affinity for the receptor.

[0050] As described above, the present inventors aim to provide CXCR4 binding molecules for diagnostic molecular imaging and intracavitary radiotherapy. Surprisingly, it was found that in addition to R 6Another site outside of the R is suitable for functionalization with a signaling unit such as a radiolabel or a fluorescent dye. Such a second site is represented by R 4 Specifically, in R 4 Contains R A In those embodiments, using R 4 As anchor points for modifications, the modifications include R similar to those described above B , introducing a radioactive label, a moiety suitable for radioactive labeling, or other signaling unit suitable for molecular imaging.

[0051] According to the present invention, there is R A and R B At least one of them. Both of them can exist.

[0052] R in formula (I) 1 The alkyl group is preferably a straight chain alkyl group. The alkyl group is preferably a C1-C6 alkyl group, more preferably a C1 or C2 alkyl group, and most preferably a methyl group.

[0053] R in formula (I) 2 is H or an alkyl group, which may be unsubstituted or substituted by at least one substituent selected from the group consisting of -NH2, -NH-C(=NH)-NH2, -C(O)NH2, -C(O)OH, -OH, -SH, -SCH3, and a 5- to 10-membered carbon ring or a 5- to 10-membered heterocycle containing oxygen, nitrogen or sulfur as a heteroatom. The alkyl group is preferably a straight-chain alkyl group. With respect to the number of carbon atoms, it is preferred that the alkyl group is a C1-C6 alkyl group, more preferably a C2-C3 alkyl group, and most preferably a C3 alkyl group. As for the substituent group, it is preferred that R 2 is substituted by at least one group selected from -NH2 and guanidinium (-NH-C(=NH)-NH2), particularly preferably R 2 is a straight-chain C3 alkyl group, at the terminal C atom (i.e., R 2 The C atom farthest from the point of attachment) is substituted by one of -NH2 and guanidinium (-NH-C(=NH)-NH2). The most preferred is -CH2-CH2-CH2-NH-C(=NH)-NH2.

[0054] As will be appreciated by those skilled in the art, any 5-10 membered carbocycle mentioned herein may be formed by a monocycle having a number of ring members falling within a defined range, or by a condensed ring (e.g., two condensed rings) having a total number of ring members falling within a defined range. Similarly, any 5-10 membered heterocycle mentioned herein may be formed by a monocycle having a number of ring members falling within a defined range, or by a condensed ring (e.g., two condensed rings) having a total number of ring members falling within a defined range.

[0055] R in formula (I) 3is H or an alkyl group, which may be unsubstituted or substituted by at least one substituent selected from -NH2, -NH-C(=NH)-NH2, -C(O)NH2, -C(O)OH, -OH, -SH, -SCH3, and a 5- to 10-membered carbocyclic ring or a 5- to 10-membered heterocyclic ring containing oxygen, nitrogen or sulfur as a heteroatom. With respect to the number of carbon atoms, preferably the alkyl group is a C1-C6 alkyl group, more preferably a C1-C2 alkyl group, and most preferably a methyl group. With respect to the substituent group, preferably R 3 is substituted by a 5-10 membered carbon ring, more preferably substituted by a phenyl or naphthyl group. 3 The group is methyl substituted by a phenyl or a naphthyl group, and most preferred is the group -CH2-naphthyl, for example -CH2-(2-naphthyl),

[0056] R in formula (I) 4 is H or an alkyl group, the alkyl group may be unsubstituted or substituted by at least one substituent selected from the group consisting of: -NH2, -NH-C(=NH)-NH2, -C(O)NH2, -C(O)OH, -OH, -SH, -SCH3, -SR A 、-SR 11 and a 5- to 10-membered carbon ring or a 5- to 10-membered heterocyclic ring containing oxygen, nitrogen or sulfur as a heteroatom. The alkyl group is preferably a C1-C6 alkyl group, more preferably a C1-C2 alkyl group, and most preferably a methyl group. R is generally preferred 4 H and is selected from -SH, -SR A and-SR 11 Particularly preferred R 4 H, -CH2-SH, -CH2-SR A and -CH2-SR 11 .

[0057] R 11 Selected from -(CH2)-C(=O)-NH-R 12 -NH-C(=O)-CH3 and

[0058] -(CH2)-C(=O)-NH-(CH2) r -R 13 -(CH2) s -NH-C(=O)-CH3. 12 is a C2-C10 alkanediyl group, preferably a C3-C6 alkanediyl group, and more preferably a C4-C6 alkanediyl group. Preferably, the alkanediyl group is a straight chain group. The alkanediyl group may be substituted by a -C(=O)-NH2 group. 13 is phenylene, preferably 1,3-phenylene, and r and s are independently an integer selected from 0, 1 or 2, and are preferably both 1;

[0059] R in formula (I) 5 It is H or I, preferably H.

[0060] R in formula (I) 6 is at least one selected from -NH2, -NH-C(=NH)-NH2, -NH-R B 、-NH-C(=NH)-NH-R B 、-NH-C(=NH)-NH-R 14 and -NH-C(=NH)-NH-C(=O)-R 15 The alkyl group is preferably a straight chain alkyl group. Regarding the number of carbon atoms, the alkyl group is preferably a C1-C8 alkyl group, more preferably a C2-C6 alkyl group, and most preferably a C6 alkyl group. As for the substituent group, R 6 is selected from -NH-C(=NH)-NH2, -NH-C(=NH)-NH-R B 、-NH-C(=NH)-NH-R 14 and -NH-C(=NH)-NH-C(=O)-R 15 Therefore, it is particularly preferred that R 6 The group is a straight-chain C2-C6 alkyl group, and even more preferably a straight-chain C6 alkyl group, at the terminal C atom (i.e., R 6 The C atom farthest from the point of attachment) is replaced by a moiety selected from -NH-C(=NH)-NH2, -NH-C(=NH)-NH-R B 、-NH-C(=NH)-NH-R 14 and -NH-C(=NH)-NH-C(=O)-R 15 The group is substituted.

[0061] R 14 and R 15 is independently a C1-C10 alkyl group, preferably a C2-C6 alkyl group. The alkyl group is preferably a straight-chain alkyl group. The alkyl group, preferably a straight-chain alkyl group, may be substituted by at least one substituent selected from -NH-C(=O)-CH3 and -C≡CH. Preferably, the alkyl group is a straight-chain C1-C10 alkyl group, more preferably a straight-chain C2-C6 alkyl group, which is unsubstituted or has a C atom at its terminal (i.e., R in the above formula) 14 and R 15 The C atom farthest from the point of attachment of the molecule is substituted with a substituent selected from -NH-C(=O)-CH3 and -C≡CH.

[0062] R in formula (I) 7 To R 10 Each is independently H or alkyl; preferably H or C1-C6 alkyl; and more preferably H.

[0063] In line with the above, it will be appreciated that the compound of formula (I) is preferably represented by the following formula (III):

[0064]

[0065] Where R 1 , R 4 , R 5 and R 6 As defined in formula (I), including the preferred meanings thereof.

[0066] More preferably, the compound of formula (I) is represented by the following formula (IV):

[0067]

[0068] Where R 1 , R 4 , R 5 and R 6 As defined for formula (I), including the preferred meanings thereof.

[0069] As mentioned above, the compound of formula (I) or preferably the compound of formula (III) and (IV) comprises at least one selected from -SR A 、-NH-R B and -NH-C(=NH)-NH-R B Preferably, the compound of formula (I) or the preferred compounds of formula (III) and (IV) contain a substituent selected from -SR A 、-NH-R B and -NH-C(=NH)-NH-R B More preferably, the compound of formula (I) or the preferred compounds of formula (III) and (IV) contain a substituent selected from -SR A and -NH-C(=NH)-NH-R B Substituents (therefore no substituent -NH-R B ).

[0070] Therefore, in a preferred embodiment, the compound of formula (I) or preferably the compound of formula (III) and (IV) comprises the substituent -NH-R B and -NH-C(=NH)-NH-R B One of the following, more preferably -NH-C(=NH)-NH-R B , and there is no formula-SR A In another preferred embodiment, the compound of formula (I) or the preferred compounds of formula (III) and (IV) contain a substituent of formula -SR A Substituents, and no -NH-R Band -NH-C(=NH)-NH-R B Substituent.

[0071] In line with the above, a preferred subgroup of compounds according to the present invention has formula (Ia):

[0072]

[0073] Where R 1 To R 3 , R 5 and R 7 To R 10 As defined for formula (I), including the preferred meanings thereof.

[0074] R 6A For a selected from -NH-R B and -NH-C(=NH)-NH-R B The alkyl group substituted by a substituent is preferably substituted by -NH-C(=NH)-NH-R B Substitution, where R B As defined for formula (I), including its preferred meanings. The alkyl group is preferably a straight chain alkyl group. In terms of the number of carbon atoms, it is preferably a C1-C8 alkyl group, more preferably a C2-C6 alkyl group, and most preferably a C6 alkyl group. Therefore, particularly preferred R 6A The group is a straight-chain C2-C6 alkyl group, and even more preferably a straight-chain C6 alkyl group, at the terminal C atom (i.e., R 6 The C atom farthest from the attachment point) is replaced by a -NH-C(=NH)-NH-R B Most preferably, R 6A is -(straight chain C6 alkyl)-NH-C(=NH)-NH-R B .

[0075] R 4A is H or alkyl, which may be unsubstituted or substituted by a substituent selected from the group consisting of: -NH2, -NH-C(=NH)-NH2, -C(O)NH2, -C(O)OH, -OH, -SH, -SCH3, -SR 11 and a 5- to 10-membered carbocyclic ring or a 5- to 10-membered heterocyclic ring containing oxygen, nitrogen or sulfur as a heteroatom, wherein R 11 As defined for formula (I), including its preferred meanings. Alkyl is preferably C1-C6 alkyl, more preferably C1-C2 alkyl, and most preferably methyl. Generally preferred R 4A is H and is one selected from -SH and -SR 11 Particularly preferably, R 4A It is H, -CH2-SH and -CH2-SR 11 . R4A Most preferably, H. 11 As defined in formula (I), including the preferred meanings thereof.

[0076] Another preferred subgroup of compounds according to the invention has the formula (Ib):

[0077]

[0078] Where R 1 To R 3 , R 5 and R 7 To R 10 As defined for formula (I), including the preferred meanings thereof.

[0079] R 4B is replaced by a substituent -SR A Substituted alkyl, wherein R A As defined for formula (I), including its preferred meanings. Preferably, the substituted alkyl group is a C1-C6 alkyl group, more preferably a C1 or C2 alkyl group, and most preferably a methyl group. Therefore, it is particularly preferred that R 4B -CH2-SR A .

[0080] R 6B is a molecule selected from -NH2, -NH-C(=NH)-NH2, -NH-C(=NH)-NH-R 14 and -NH-C(=NH)-NH-C(=O)-R 15 The substituents are substituted alkyl, wherein R 14 and R 15 As defined for formula (I), including their preferred embodiments. Alkyl is preferably a straight chain alkyl. In terms of the number of carbon atoms, it is preferably a C1-C8 alkyl, more preferably a C2-C6 alkyl, and most preferably a C6 alkyl. Therefore, particularly preferred R 6B is a straight chain C2-C6 alkyl group, and even more preferably a straight chain C6 alkyl group, at the terminal C atom (i.e., R 6B The C atom furthest from the point of attachment of 6B It is -(straight-chain C6 alkyl)-NH-C(=NH)-NH2. 14 and R 15 As defined for formula (I), including the preferred meanings thereof.

[0081] In line with the above, preferably the compound of formula (Ia) is a compound of formula (IIIa):

[0082]

[0083] Where R 1 , R 4A , R 6A and R 5 As defined for formula (I) and formula (Ia), respectively, including their preferred meanings. Even further preferred compounds of formula (Ia) are compounds of formula (IVa):

[0084]

[0085] Where R 1 , R 4A , R 6A and R 5 as defined for formula (I) and formula (Ia), respectively, including their preferred meanings.

[0086] Similarly, preferred compounds of formula (Ib) are compounds of formula (IIIb):

[0087]

[0088] Where R 1 , R 4B , R 6B and R 5 As defined for formula (I) or formula (Ib), respectively, including their preferred meanings. Even further preferred compounds of formula (Ib) are compounds of formula (IVb):

[0089]

[0090] Where R 1 , R 4B , R 6B and R 5 as defined for formula (I) or formula (Ib), respectively, including their preferred meanings.

[0091] In formula (I) and preferred formulas (Ia), (Ib), (III), (IIIa), (IIIb), (IV), (IVa) and (IVb), R A is a group comprising at least one selected from the following:

[0092] (i) a chelating moiety,

[0093] (ii) a chelate formed by the chelating moiety (i) and a chelated radioactive or non-radioactive cation,

[0094] (iii) a moiety carrying a covalently bound radioisotope or a precursor suitable for labelling with such a radioisotope,

[0095] (iv) a phosphonate moiety, and

[0096] (v) Fluorescent labels, such as quantum dots or fluorescent dyes.

[0097] Usually, R A Including one of (i) to (v), i.e. not necessarily two or more of (i) to (v) are present in one compound according to the present invention. Among the parts (i) to (v), (i) and (ii) are preferred. Therefore, R is particularly preferred A Includes one of (i) and (ii).

[0098] Those skilled in the art will understand that, according to the above definition, R A Including at least one of the above (i) to (v) includes the following situations: R A Contains another part or multiple parts together with at least one of (i) to (v). According to a preferred embodiment, R A In addition to at least one of the above (i) to (v), a linking group that connects at least one of (i) to (v) to the rest of the compound may also be included. The linking group may be a divalent linking group that connects one of (i) to (v) to the rest of the compound, or a branched linking group that allows two or more of (i) to (v) to be connected to the rest of the compound. Preferably, R A Contains a divalent linking group.

[0099] Therefore, it is more preferred that R A Is the formula -L A1 -R A1 A group in which L A1 is a divalent linking group, and R A1 is a group selected from the following or comprises a group selected from the following: (i) a chelating moiety, (ii) a chelate formed by the chelating moiety (i) and a chelated radioactive or non-radioactive cation, (iii) a moiety carrying a covalently bound radioisotope or a precursor suitable for labeling with such a radioisotope, (iv) a phosphonate moiety, and (v) a fluorescent label, such as a quantum dot or a fluorescent dye. Among them, (i) and (ii) are more preferred.

[0100] Especially preferred -L A1 -R A1 A group selected from formula (Va) and formula (Vb):

[0101] -(CH2)-C(=O)-NH-R 24 -NH-R A1 (Va),

[0102] -(CH2)-C(=O)-NH-(CH2) y -R 25 -(CH2)z -NH-R A1 (Vb),

[0103] Where R 24 is a C2-C10 alkanediyl group, preferably a C3-C6 alkanediyl group, which is preferably linear and may be substituted by a group -C(=O)-NH2, R 25 is a phenylene group, preferably a 1,3-phenylene group, and y and z are independently an integer selected from 0, 1 or 2, and are preferably both 1. A1 As defined above, including its preferred meanings. Among them, -L is particularly preferred A1 -R A1 It is a group of formula (Vb).

[0104] The chelating part of (i) and (ii) is A and R A1 Suitable chelating moieties for various cations are well known in the art and can be used in the context of the present invention.

[0105] Preferably, the chelating moiety comprises at least one of the following

[0106] a macrocyclic structure having 8 to 20 ring atoms, 2 or more, preferably 3 or more of the ring atoms being selected from oxygen atoms, sulfur atoms and nitrogen atoms; and

[0107] A non-cyclic open-chain chelate 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 atoms, sulfur atoms and nitrogen atoms.

[0108] Preferred examples of chelating moieties are residues of chelating agents such as bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (CBTE2a), cyclohexyl-1,2-diaminetetraacetic acid (CDTA), 4-(1,4,8,11-tetraazacyclotetradec-1-yl)-methylbenzoic acid (CPTA), N'-[5-[acetyl(hydroxy)amino]pentyl]-N-[5-[[4-[5-aminopentyl(hydroxy)amino]-4-oxobutanoyl]amino]pentyl]-N-hydroxybutanediol. DFO, 4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (DO2A), 1,4,7,10-tetraazacyclododecane-N,N',N",N''-tetraacetic acid (DOTA), α-(2-carboxyethyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid or 2-[1,4,7,10-tetraazacyclododecane-4,7,10-triacetic acid]-glutaric acid (DOTAGA), N,N'-dipyridine Oxyethylenediamine-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-4,7,1 0-triacetate (HP-DOA3), 6-hydrazino-N-methylpyridine-3-carboxamide (HYNIC), 1,4,7-triazacyclononane-1-succinic acid-4,7-diacetic acid (NODASA), 1-(1-carboxy-3-carboxypropyl)-4,7-(carboxy (carbooxy))-1,4,7-triazacyclononane (NODAGA), 1,4,7-triazacyclononane triacetic 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), terpyridine bis(methyleneaminotetraacetic acid (TMT), 1,4,7,10-tetraazacyclotridecane-N,N',N",N"'-tetraacetic acid (TRITA), triethylenetetraaminehexaacetic acid (TTHA), N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18-crown-6 (H2macropa) and 4-amino-4-{2-[(3-hydroxy- The residue is obtained by covalently bonding the carboxyl group contained in the chelating agent to the rest of the compound through an ester bond or an amide bond (preferably an amide bond). For example, in formula (Va) and formula (Vb), the amide bond can be conveniently directly formed by the carboxyl group contained in the above exemplary chelating agent and R in these formulas. A or R A1 The nitrogen atoms are connected separately.

[0109] In a more preferred embodiment, in R A or R A1 In the context of the definition, the chelating moiety of (i) or (ii) is a residue of a chelating agent selected from DOTA and DOTAGA, and more preferably can be obtained by reacting the carboxyl groups contained in DOTA and DOTAGA and R in formula (Va) and (Vb). A1 The attached nitrogen atom forms an amide bond to obtain the residue.

[0110] In R A and R A1 In the context of the definition of, exemplary radioactive cations optionally chelated by the chelating moiety according to (ii) are selected from the group consisting of cations 44 Sc, 47 Sc, 51 Cr, 52m Mn, 58 Co. 52 Fe, 56 You, 57 You, 62 Cu, 64 Cu, 67 Cu, 66 Ga, 68 Ga, 67 Ga, 89 Zr, 90 Y. 89 Y. 94m Tc, 99m Tc,97 Such as 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 2. 169 Yb, 175 Yb, 172 Tm, 177 Lu, 186 Re, 188 Re, 191 Pt, 197 Hg, 198 Au, 199 Au, 212 Pb, 203 Pb, 211 At 212 Bi, 213 Bi, 223 Ra, 225 Ac and 227 Th, or contain 18 Cationic molecules of F, such as 18 F-[AlF] 2+ .

[0111] Preferred chelated cations are selected from the group consisting of 44 Sc, 47 Sc, 64 Cu, 67 Cu, 68 Ga, 90 Y. 111 In, 161 Tb, 166 Ho, 177 Lu, 188 Re, 212 Pb, 212 Bi, 213 Bi, 225 Ac and 227 Th, or contain18 F. Even further preferred are cationic molecules 68 Ga, 90 Y. 177 Lu, 212 Bihe 213 Bi.

[0112] As is known in the art, R A and R A1 The term "carrying a covalently bound radioactive isotope (preferably 18 F fluoride) or a part (iii) suitable for a precursor labeled with such a radioisotope. As an example of such a precursor, reference can be made to the formula -N + (CH3)2-CH2-BF3 - A group of formula -Ar-SiF(C(CH3)3)2, wherein Ar is a divalent aromatic group, preferably a phenylene group. As will be appreciated by a person skilled in the art, such a group of formula -Ar-SiF(C(CH3)3)2 can be conveniently bonded to the remainder of the compound according to the invention via a functional group which can be attached to Ar at the open valence shown in the formula and which is suitable for the covalent coupling of the group -Ar-SiF(C(CH3)3)2. For example, reference may be made to an amide bond or an ester bond, which may be formed, for example, in the case where the group -Ar-SiF(C(CH3)3)2 is provided as part of a benzoic acid derivative of formula -C(O)-C6H4-SiF(C(CH3)3)2.

[0113] As is known in the art, such as R A and R A1 The phosphonate moiety (iv) referred to in the definition of , ie a moiety comprising the structure -P(=O)(OH)2 or a salt thereof, may also be used.

[0114] As R A and R A1 As the fluorescent label (v) referred to in the definition, fluorescent dyes, such as Cy5 or Cy7, or quantum dots can be used.

[0115] In formula (I) and preferred formulas (Ia), (Ib), (III), (IIIa), (IIIb), (IV), (IVa) and (IVb), R B is a group comprising at least one selected from the following:

[0116] (i) a chelating moiety,

[0117] (ii) a chelate formed by a chelating moiety and a chelated radioactive or non-radioactive cation,

[0118] (iii) a group carrying a covalently bound radioisotope or a precursor suitable for labelling with such a radioisotope,

[0119] (iv) a phosphonate moiety, and

[0120] (v) fluorescent labels, such as quantum dots or fluorescent dyes,

[0121] And R B It may further contain a group of formula (II)

[0122]

[0123] Its use The indicated bond is bound to the rest of the compound, where R 1 To R 5 and R 7 To R 10 As defined for formula (I), including their preferred meanings, and wherein R 16 is an alkanediyl group, preferably a C1-C6 alkanediyl group, and R 17 is selected from -NH- and -NH-C(=NH)-NH-;

[0124] Usually, R B Including one of (i) to (v), it is not necessary that two or more of (i) to (v) are present in one compound according to the present invention.

[0125] Those skilled in the art will understand that, according to the above definition, R B Including at least one of the above (i) to (v) includes the following situations: R B Contains another part or multiple parts together with at least one of (i) to (v). According to a preferred embodiment, R B In addition to at least one of the above (i) to (v), a linking group that connects at least one of (i) to (v) to the rest of the compound may also be included. The linking group may be a divalent linking group that connects one of (i) to (v) to the rest of the compound, or a branched linking group that allows two or more of (i) to (v) to be connected to the rest of the compound. Preferably, R B Contains a divalent linking group.

[0126] Therefore, it is more preferred that R B Is the formula -L B1 -R B1 A group in which L B1 is a divalent linking group, and R B1is or comprises a group selected from (i) a chelating moiety, (ii) a chelate formed by the chelating moiety (i) and a chelated radioactive or non-radioactive cation, (iii) a moiety carrying a covalently bound radioisotope or a precursor suitable for labeling with such a radioisotope, (iv) a phosphonate moiety, and (v) a fluorescent label, such as a quantum dot or a fluorescent dye. In addition, R B1 It may further comprise a group of formula (II) as defined above.

[0127] L is particularly preferred B1 -R B1 A group selected from formula (VIa) to formula (VIf):

[0128] -R 18 -NH-R B1 (VIa),

[0129] -R 18 -NH-(C(=O)-R 19 -NH) u -R B1 (VIb),

[0130] -R 18 -NH-(C(=O)-R 20 -NH) v -R B1 (VIc),

[0131] -C(=O)-R 21 -NH-R B1 (VId),

[0132] -C(=O)-R 21 -NH-(C(=O)-R 22 -NH) w -R B1 (VIe),

[0133] -C(=O)-R 21 -NH-(C(=O)-R 23 -NH) x -R B1 (VIf),

[0134] in:

[0135] R 18 is a C1-C10 alkanediyl group, preferably a straight-chain alkanediyl group, more preferably a straight-chain C2-C6 alkanediyl group, and most preferably a straight-chain C6 alkanediyl group,

[0136] R 19is a C1-C10 alkanediyl group, preferably a straight-chain alkanediyl group, more preferably a straight-chain C2-C6 alkanediyl group, u is 1, 2 or 3, preferably 1 or 2,

[0137] R 20 is an oligoether moiety, preferably a linear oligoether moiety, more preferably a linear oligoether moiety having 5 to 10 carbon atoms and 2 or 3 oxygen atoms providing ether bonds, v is 1, 2 or 3, preferably 1 or 2,

[0138] R 21 is a C1-C10 alkanediyl group, preferably a straight-chain alkanediyl group, more preferably a straight-chain C2-C6 alkanediyl group, most preferably a straight-chain C6 alkanediyl group,

[0139] R 22 is a C1-C10 alkanediyl group, preferably a straight-chain alkanediyl group, more preferably a straight-chain C2-C6 alkanediyl group, w is 1, 2 or 3, preferably 1 or 2,

[0140] R 23 is an oligoether moiety, preferably a linear oligoether moiety, more preferably a linear oligoether moiety having 5 to 10 carbon atoms and 2 or 3 oxygen atoms providing ether bonds, x is 1, 2 or 3, preferably 1 or 2,

[0141] and R B1 As defined above.

[0142] The chelating part of (i) and (ii) is B and R B1 Suitable chelating moieties for various cations are well known in the art and can be used in the context of the present invention.

[0143] Preferably, the chelating moiety comprises at least one of the following

[0144] a macrocyclic structure having 8 to 20 ring atoms, 2 or more, preferably 3 or more of the ring atoms being selected from oxygen atoms, sulfur atoms and nitrogen atoms; and

[0145] A non-cyclic open-chain chelate 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 atoms, sulfur atoms and nitrogen atoms.

[0146] Preferred examples of chelating moieties are residues of chelating agents such as bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (CBTE2a), cyclohexyl-1,2-diaminetetraacetic acid (CDTA), 4-(1,4,8,11-tetraazacyclotetradec-1-yl)-methylbenzoic acid (CPTA), N'-[5-[acetyl(hydroxy)amino]pentyl]-N-[5-[[4-[5-aminopentyl(hydroxy)amino]-4-oxobutanoyl]amino]pentyl]-N-hydroxy succinamide (DFO), 4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (DO2A), 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid (DOTA), α-(2-carboxyethyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid or 2-[1,4,7,10-tetraazacyclododecane-4,7,10-triacetic acid]-glutaric acid (DOTAGA), N,N '-Dipyridyloxyethylenediamine-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-tetraazacyclo Decane-4,7,10-triacetate (HP-DOA3), 6-hydrazino-N-methylpyridine-3-carboxamide (HYNIC), 1,4,7-triazacyclononane-1-succinic acid-4,7-diacetic acid (NODASA), 1-(1-carboxy-3-carboxypropyl)-4,7-(carboxy)-1,4,7-triazacyclononane (NODAGA), 1,4,7-triazacyclononane triacetic 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), terpyridine bis(methyleneaminotetraacetic acid (TMT), 1,4,7,10-tetraazacyclotridecane-N,N',N",N"'-tetraacetic acid (TRITA), triethylenetetraaminehexaacetic acid (TTHA), N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18-crown-6 (H2macropa) and 4-amino-4-{2-[(3-hydroxy -1,6-dimethyl-4-oxo-1,4-dihydropyridin-2-ylmethyl)carbamoyl]ethyl}pimelate bis[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydropyridin-2-ylmethyl)-amide] (THP), the residue being obtained by covalently bonding the carboxyl group contained in the chelating agent to the rest of the compound via an ester bond or an amide bond (preferably an amide bond). For example, in formulas (VIa) to (VIf), the amide bond can be conveniently directly formed by the carboxyl group contained in the above exemplary chelating agents and R in these formulas. B or R B1 The nitrogen atoms are connected separately.

[0147] In a more preferred embodiment, in R B or R B1 In the context of the definition, the chelating moiety of (i) or (ii) is a residue of a chelating agent selected from DOTA and DOTAGA, and more preferably can be obtained by the carboxyl groups contained in DOTA and DOTAGA and R in formula (VIa) and formula (VIf). B1 The attached nitrogen atom forms an amide bond to obtain the residue.

[0148] In R B and R B1 In the context of the definition of, exemplary radioactive cations optionally chelated by the chelating moiety according to (ii) are selected from the group consisting of cations 44 Sc, 47 Sc, 51 Cr, 52m Mn, 58 Co. 52 Fe, 56 You, 57 You, 62 Cu, 64 Cu, 67 Cu, 66 Ga, 68 Ga, 67 Ga, 89 Zr, 90 Y. 89 Y. 94m Tc, 99m Tc,97 Such as 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 2. 169 Yb, 175 Yb, 172 Tm, 177 Lu, 186 Re, 188 Re, 191 Pt, 197 Hg, 198 Au, 199 Au, 212 Pb, 203 Pb, 211 At 212 Bi, 213 Bi, 223 Ra, 225 Ac and 227 Th, or contain 18 Cationic molecules of F, such as 18 F-[AlF] 2+ .

[0149] Preferred chelated cations are selected from the group consisting of 44 Sc, 47 Sc, 64 Cu, 67 Cu, 68 Ga, 90 Y. 111 In, 161 Tb, 166 Ho, 177 Lu, 188 Re, 212 Pb, 212 Bi, 213 Bi, 225 Ac and 227 Th, or contain18 F. Even further preferred are cationic molecules 68 Ga, 90 Y. 177 Lu, 212 Bihe 213 Bi.

[0150] As is known in the art, R B and R B1 The term "carrying a covalently bound radioactive isotope (preferably 18 F fluoride) or a moiety (iii) which is suitable for labelling with such a radioisotope.

[0151] As an example of such a precursor, reference may be made to the formula: + (CH3)2-CH2-BF3 - A group of formula -Ar-SiF(C(CH3)3)2, wherein Ar is a divalent aromatic group, preferably a phenylene group. As will be appreciated by a person skilled in the art, such a group of formula -Ar-SiF(C(CH3)3)2 can be conveniently bonded to the remainder of the compound according to the invention via a functional group which can be attached to Ar at the open valence shown in the formula and which is suitable for the covalent coupling of the group -Ar-SiF(C(CH3)3)2. For example, reference may be made to an amide bond or an ester bond, which can be formed, for example, when the group -Ar-SiF(C(CH3)3)2 is provided as part of a benzoic acid derivative of formula -C(O)-C6H4-SiF(C(CH3)3)2.

[0152] As is known in the art, such as R B and R B1 The phosphonate moiety (iv) referred to in the definition of , ie a moiety comprising the structure -P(=O)(OH)2 or a salt thereof, may also be used.

[0153] As R B and R B1 As the fluorescent label (v) referred to in the definition, fluorescent dyes, such as Cy5 or Cy7, or quantum dots can be used.

[0154] In addition, R B and R B1 may further contain a group of formula (II)

[0155]

[0156] Its use The indicated bond is bound to the rest of the compound, where R 1 To R 5 and R 7To R 10 As defined for formula (I), including their preferred meanings, and wherein R 16 is an alkanediyl group, preferably a C1-C6 alkanediyl group, and R 17 is selected from -NH- and -NH-C(=NH)-NH-.

[0157] In a second aspect, the present invention provides a pharmaceutical composition comprising or consisting of the compound or salt of the first aspect.

[0158] In a third aspect, the present invention provides a diagnostic composition comprising or consisting of a compound or salt of the first aspect.

[0159] The pharmaceutical composition may also include 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, water, emulsions, such as oil / water emulsions, various types of wetting agents, sterile solutions, etc. Compositions comprising such carriers can be formulated by known conventional methods. These pharmaceutical compositions can be administered to individuals at suitable doses. Administration of suitable compositions can be accomplished in different ways, such as by intravenous, intraperitoneal, subcutaneous, intramuscular, topical, intradermal, intranasal or intrabronchial administration. It is particularly preferred that the administration is carried out by, for example, injection and / or delivery to a site in the pancreas or in a cerebral artery or directly into brain tissue. The composition can also be administered directly to a target site, such as by gene gun delivery to an external or internal target site, such as the pancreas or the brain. The dosage regimen will be determined by the attending physician and clinical factors. As is well known in the medical arts, the dosage for any one patient depends on many factors, including the patient's size, body surface area, age, the specific compound to be administered, sex, time and route of administration, general health, and other drugs administered concurrently. The pharmaceutically active substance may be present in an amount of 0.1 ng to 10 mg / kg body weight per dose; however, doses below or above this exemplary range are contemplated, particularly in light of the factors noted above.

[0160] The above also applies mutatis mutandis to the diagnostic compositions.

[0161] Insofar as the pharmaceutical compositions, diagnostic compositions and therapeutic compositions disclosed above comprise one or more compounds of the invention, it is preferred that no other pharmaceutically active compounds, diagnostically active compounds or therapeutically active compounds are present. Alternatively, other therapeutically active, diagnostically active or pharmaceutically active compounds, such as anticancer agents, may be present.

[0162] In a fourth aspect, the present invention provides a compound or salt of the first aspect for use in medicine, preferably in nuclear medicine, for use in nuclear molecular imaging, optical imaging and intracavitary radiotherapy, preferably targeted intracavitary radiotherapy.

[0163] In a fifth aspect, the present invention provides a compound or salt of the first aspect for use in a method for treating or preventing cancer, cardiovascular disease, AIDS or inflammatory lesions. These medical uses are derived from the expression, tissue distribution and / or activity of CXCR4, as described in the introduction above.

[0164] In a sixth aspect, the present invention provides a compound or salt of the first aspect for use in a method for diagnosing and / or staging cancer.

[0165] With regard to the present description, in particular the embodiments characterized in the claims, it is intended that every embodiment mentioned in the dependent claims be combined with every embodiment of every claim to which the dependent claims are dependent (independent or 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 claim 3 dependent on claims 1 and 2 describes three alternatives G, H and I, it will be understood that, unless otherwise specifically stated, the specification explicitly 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.

[0166] Similarly, where independent and / or dependent claims do not recite alternatives, it will also be understood that if a dependent claim refers back to multiple prior claims, any combination of subject matter covered thereby is deemed to be explicitly disclosed. For example, where there is 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 clearly and explicitly disclosed, as is the combination of the subject matter of claims 3, 2, and 1. Where there is another dependent claim 4 that refers to any 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 clearly and explicitly disclosed. The same applies to the following items. DETAILED DESCRIPTION

[0167] Specifically, the present invention includes the following items:

[0168] 1. A compound of the following formula (I) or a pharmaceutically acceptable salt thereof,

[0169]

[0170] in:

[0171] R 1 is an alkyl group, preferably a C1-C6 alkyl group, more preferably a methyl group;

[0172] R 2 is H or an alkyl group, which may be unsubstituted or substituted by at least one substituent selected from the group consisting of -NH2, -NH-C(=NH)-NH2, -C(O)NH2, -C(O)OH, -OH, -SH, -SCH3, and a 5- to 10-membered carbocyclic ring or a 5- to 10-membered heterocyclic ring containing oxygen, nitrogen or sulfur as a heteroatom, wherein the alkyl group is preferably a C1-C6 alkyl group;

[0173] R 2 is H or an alkyl group, which may be unsubstituted or substituted by at least one substituent selected from the group consisting of -NH2, -NH-C(=NH)-NH2, -C(O)NH2, -C(O)OH, -OH, -SH, -SCH3, and a 5- to 10-membered carbocyclic ring or a 5- to 10-membered heterocyclic ring containing oxygen, nitrogen or sulfur as a heteroatom, wherein the alkyl group is preferably a C1-C6 alkyl group;

[0174] R 4 is H or an alkyl group, the alkyl group may be unsubstituted or substituted by at least one substituent selected from the group consisting of: -NH2, -NH-C(=NH)-NH2, -C(O)NH2, -C(O)OH, -OH, -SH, -SCH3, -SR A 、-SR 11 and a 5- to 10-membered carbocyclic ring or a 5- to 10-membered heterocyclic ring containing oxygen, nitrogen or sulfur as a heteroatom, wherein the alkyl group is preferably a C1-C6 alkyl group;

[0175] R 5 is H or I;

[0176] R 6 is at least one selected from -NH2, -NH-C(=NH)-NH2, -NH-R B 、-NH-C(=NH)-NH-R B 、-NH-C(=NH)-NH-R 14 and -NH-C(=NH)-NH-C(=O)-R 15 An alkyl group substituted with a substituent, wherein the alkyl group is preferably a C1-C8 alkyl group;

[0177] R 7 To R 10 Each independently is H or alkyl; preferably H or C1-C6 alkyl; and more preferably H;

[0178] R 11 Selected from -(CH2)-C(=O)-NH-R 12 -NH-C(=O)-CH3 and

[0179] -(CH2)-C(=O)-NH-(CH2) r -R 13 -(CH2) s -NH-C(=O)-CH3, where R 12 is a C2-C10 alkanediyl group, preferably a C3-C6 alkanediyl group, which may be substituted by a group -C(=O)-NH2, R 13 is phenylene, preferably 1,3-phenylene, and r and s are independently an integer selected from 0, 1 or 2, and are preferably both 1;

[0180] R 14 and R 15 Independently C1-C10 alkyl, preferably C2-C6 alkyl, which may be substituted by at least one substituent selected from -NH-C(=O)-CH3 and -C≡CH;

[0181] R A is a group comprising at least one selected from the following:

[0182] (i) a chelating moiety,

[0183] (ii) a chelate formed by the chelating moiety (i) and a chelated radioactive or non-radioactive cation,

[0184] (iii) a moiety carrying a covalently bound radioisotope or a precursor suitable for labelling with such a radioisotope,

[0185] (iv) a phosphonate moiety, and

[0186] (v) fluorescent labels, such as quantum dots or fluorescent dyes; and

[0187] R B is a group comprising at least one selected from the following:

[0188] (i) a chelating moiety,

[0189] (ii) a chelate formed by a chelating moiety and a chelated radioactive or non-radioactive cation,

[0190] (iii) a group carrying a covalently bound radioisotope or a precursor suitable for labelling with such a radioisotope,

[0191] (iv) a phosphonate moiety, and

[0192] (v) fluorescent labels, such as quantum dots or fluorescent dyes,

[0193] And R B It may also contain a group of formula (II)

[0194]

[0195] Its use The indicated bond is bound to the rest of the compound, where R 1 To R 5 and R 7 To R 10 As defined above, and wherein R 16 is an alkanediyl group, preferably a C1-C6 alkanediyl group, and R 17 is selected from -NH- and -NH-C(=NH)-NH-;

[0196] And among them

[0197] The compound of formula (I) comprises at least one selected from -SR A 、-NH-R B and -NH-C(=NH)-NH-R B substituent.

[0198] 2. The compound or salt described in item 1, wherein R 1 It is a C1-C6 alkyl group.

[0199] 3. The compound or salt described in item 2, wherein R 1 It's methyl.

[0200] 4. The compound or salt of any one of items 1 to 3, wherein R 2 It is a C1-C6 alkyl group substituted by a group selected from -NH2 and -NH-C(=NH)-NH2.

[0201] 5. The compound or salt described in item 4, wherein R 2 It is -CH2-CH2-CH2-NH-C(=NH)-NH2.

[0202] 6. The compound or salt of any one of items 1 to 5, wherein R 3 It is a methyl group substituted by a 5- to 10-membered carbon ring.

[0203] 7. The compound or salt according to item 6, wherein R 3 It is the group -(CH2)-naphthyl.

[0204] 8. The compound or salt of any one of items 1 to 7, wherein R 7 To R 10 For H.

[0205] 9. The compound or salt of any one of items 1 to 8, wherein R 5 For H.

[0206] 10. The compound according to any one of items 1 to 9, which is a compound of formula (III) or a pharmaceutically acceptable salt thereof:

[0207]

[0208] in

[0209] R 1 , R 4 , R 5 and R 6 As defined in any one of items 1 to 3 and 9.

[0210] 11. The compound described in item 10, which is a compound of formula (IV) or a pharmaceutically acceptable salt thereof:

[0211]

[0212] in

[0213] R 1 , R 4 , R 5 and R 6 As defined in any one of items 1 to 3 and 9.

[0214] 12. The compound or salt of any one of items 1 to 11, wherein R 4 is H or is selected from -SH, -SR A and-SR 11 C1-C6 alkyl substituted with a substituent, wherein R A and R 11 As defined in item 1.

[0215] 13. The compound or salt of item 12, wherein R 4 H, -CH2-SH, -CH2-SR A or -CH2-SR 11 , where R A and R 11 As defined in item 1.

[0216] 14. The compound or salt of any one of items 1 to 13, wherein R 6 is selected from -NH-C(=NH)-NH2, -NH-C(=NH)-NH-R B 、-NH-C(=NH)-NH-R 14 and -NH-C(=NH)-NH-C(=O)-R 15C1-C8 alkyl substituted by a group, wherein R B , R 14 and R 15 As defined in item 1.

[0217] 15. The compound or salt of item 14, wherein R 6 is a straight-chain C6 alkyl group, at the terminal C atom of which is substituted by a moiety selected from -NH-C(=NH)-NH2, -NH-C(=NH)-NH-R B 、-NH-C(=NH)-NH-R 14 and -NH-C(=NH)-NH-C(=O)-R 15 substituted by a group, wherein R B , R 14 and R 15 As defined in item 1.

[0218] 16. A compound or salt according to any one of items 1 to 15, comprising a -SR A and -NH-C(=NH)-NH-R B substituent.

[0219] 17. The compound according to any one of items 1 to 9, which is a compound of formula (Ia) or a pharmaceutically acceptable salt thereof:

[0220]

[0221] in

[0222] R 1 To R 3 , R 5 and R 7 To R 10 As defined in items 1 to 9,

[0223] R 4A is H or alkyl, which may be unsubstituted or substituted by a substituent selected from the group consisting of: -NH2, -NH-C(=NH)-NH2, -C(O)NH2, -C(O)OH, -OH, -SH, -SCH3, -SR 11 and a 5- to 10-membered carbocyclic ring or a 5- to 10-membered heterocyclic ring containing oxygen, nitrogen or sulfur as a heteroatom, wherein R 11 as defined in item 1, and

[0224] R 6A is selected from -NH-R B and -NH-C(=NH)-NH-R B The substituents are substituted alkyl, wherein R B As defined in item 1.

[0225] 18. The compound described in item 17, which is a compound of formula (IIIa) or a pharmaceutically acceptable salt thereof:

[0226]

[0227] Where R 1 , R 4A , R 6A and R 5 As defined in item 17.

[0228] 19. The compound according to item 18, which is a compound of formula (IVa) or a pharmaceutically acceptable salt thereof:

[0229]

[0230] Where R 1 , R 4A , R 6A and R 5 As defined in item 17.

[0231] 20. The compound or salt of any one of items 17 to 19, wherein R 4A Selected from H, -CH2-SH and -CH2-SR 11 , where R 11 As defined in item 1.

[0232] 21. The compound or salt of item 20, wherein R 4A For H.

[0233] 22. The compound or salt of any one of items 17 to 21, wherein R 6A It is a straight-chain C2-C6 alkyl group, at the terminal C atom of which is replaced by a -NH-C(=NH)-NH-R B Group substitution, where R B As defined in item 1.

[0234] 23. The compound or salt of item 22, wherein R 6A is -(straight chain C6 alkyl)-NH-C(=NH)-NH-R B , where R B As defined in item 1.

[0235] 24. The compound of any one of items 1 to 9, which is a compound of formula (Ib) or a pharmaceutically acceptable salt thereof:

[0236]

[0237] in

[0238] R 1 To R3 , R 5 and R 7 To R 10 As defined in items 1 to 9,

[0239] R 4B Is a -SR A Substituent-substituted alkyl, wherein R A as defined in item 1, and

[0240] R 6B is a molecule selected from -NH2, -NH-C(=NH)-NH2, -NH-C(=NH)-NH-R 14 and -NH-C(=NH)-NH-C(=O)-R 15 The substituents are substituted alkyl, wherein R 14 and R 15 As defined in item 1.

[0241] 25. The compound described in item 24, which is a compound of formula (IIIb) or a pharmaceutically acceptable salt thereof:

[0242]

[0243] Where R 1 , R 4B , R 6B and R 5 As defined in item 24.

[0244] 26. The compound of item 26, which is a compound of formula (IVb) or a pharmaceutically acceptable salt thereof:

[0245]

[0246] Where R 1 , R 4B , R 6B and R 5 As defined in item 24.

[0247] 27. The compound or salt of any one of items 24 to 26, wherein R 4B Is a -SR A The substituent is a C1-C6 alkyl group substituted with a substituent.

[0248] 28. The compound or salt of item 27, wherein R 4B Yes-CH2-SR A .

[0249] 29. The compound or salt of any one of items 24 to 28, wherein R 6Bis a molecule selected from -NH2, -NH-C(=NH)-NH2, -NH-C(=NH)-NH-R 14 and -NH-C(=NH)-NH-C(=O)-R 15 A straight chain C2-C6 alkyl group substituted with a substituent, wherein R 14 and R 15 As defined in item 1.

[0250] 30. The compound or salt of item 29, wherein R 6B It is -(straight chain C6 alkyl)-NH-C(=NH)-NH2.

[0251] 31. A compound or salt as described in any one of items 1 to 23, wherein R B Is the formula -L B1 -R B1 A group wherein L is a divalent linking group and R B1 is a group selected from the following or contains a group selected from the following

[0252] (i) a chelating moiety,

[0253] (ii) a chelate formed by the chelating moiety (i) and a chelated radioactive or non-radioactive cation,

[0254] (iii) a moiety carrying a covalently bound radioisotope or a precursor suitable for labelling with such a radioisotope,

[0255] (iv) a phosphonate moiety, and

[0256] (v) fluorescent labels, such as quantum dots or fluorescent dyes,

[0257] And R B1 A group of formula (II) as defined in item 1 may also be contained.

[0258] 32. The compound or salt of item 31, wherein -L B1 -R B1 A group selected from formula (VIa) to (VIf):

[0259] -R 18 -NH-R B1 (VIa),

[0260] -R 18 -NH-(C(=O)-R 19 -NH) u -R B1 (VIb),

[0261] -R 18 -NH-(C(=O)-R20 -NH) v -R B1 (VIc),

[0262] -(C=O)-R 21 -NH-R B1 (VId),

[0263] -(C=O)-R 21 -NH-(C(=O)-R 22 -NH) w -R B1 (VIe),

[0264] -(C=O)-R 21 -NH-(C(=O)-R 23 -NH) x -R B1 (VIf),

[0265] in:

[0266] R 18 is a C1-C10 alkanediyl group, preferably a straight-chain alkanediyl group, more preferably a straight-chain C2-C6 alkanediyl group, and most preferably a straight-chain C6 alkanediyl group,

[0267] R 19 is a C1-C10 alkanediyl group, preferably a straight-chain alkanediyl group, more preferably a straight-chain C2-C6 alkanediyl group, u is 1, 2 or 3, preferably 1 or 2,

[0268] R 20 is an oligoether moiety, preferably a linear oligoether moiety, more preferably a linear oligoether moiety having 5 to 10 carbon atoms and 2 or 3 oxygen atoms providing ether bonds, v is 1, 2 or 3, preferably 1 or 2,

[0269] R 21 is a C1-C10 alkanediyl group, preferably a straight-chain alkanediyl group, more preferably a straight-chain C2-C6 alkanediyl group, and most preferably a straight-chain C6 alkanediyl group,

[0270] R 22 is a C1-C10 alkanediyl group, preferably a straight-chain alkanediyl group, more preferably a straight-chain C2-C6 alkanediyl group, w is 1, 2 or 3, preferably 1 or 2,

[0271] R 23 is an oligoether moiety, preferably a linear oligoether moiety, more preferably a linear oligoether moiety having 5 to 10 carbon atoms and 2 or 3 oxygen atoms providing ether bonds, x is 1, 2 or 3, preferably 1 or 2,

[0272] And R B1 As defined in item 31.

[0273] 33. A compound or salt as described in any one of items 1 to 23, wherein R B It comprises (i) a chelating moiety or (ii) a chelate formed by the chelating moiety (i) and a chelated radioactive or non-radioactive cation.

[0274] 34. The compound or salt of item 31 or 32, wherein R B1 Comprising (i) a chelating moiety or (ii) a chelate formed by the chelating moiety (i) and a chelated radioactive or non-radioactive cation, or wherein R B1 It is (i) a chelating moiety or (ii) a chelate formed by the chelating moiety (i) and a chelated radioactive or non-radioactive cation.

[0275] 35. The compound or salt of item 33 or 34, wherein the chelating moiety comprises at least one of the following

[0276] a macrocyclic structure having 8 to 20 ring atoms, 2 or more, preferably 3 or more of the ring atoms being selected from oxygen atoms and nitrogen atoms; and

[0277] A non-cyclic open-chain chelate 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 atoms, sulfur atoms and nitrogen atoms.

[0278] 36. The compound or salt of any one of items 33 to 35, wherein the chelating moiety is a residue derived from a chelating agent selected from the group consisting of: bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (CBTE2a), cyclohexyl-1,2-diaminetetraacetic acid (CDTA), 4-(1,4,8,11-tetraazacyclotetradec-1-yl)-methylbenzoic acid (CPTA), N'-[5-[acetyl(hydroxy)amino]pentyl]-N-[5-[[4-[5-aminopentyl(hydroxy)amino]- [4-oxobutyryl]amino]pentyl]-N-hydroxysuccinamide (DFO), 4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (DO2A), 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid (DOTA), α-(2-carboxyethyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid or 2-[1,4,7,10-tetraazacyclododecane-4,7,10-triacetic acid]-glutaric acid (DOTAGA), N,N'-dipyridyloxyethylenediamine-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)-1,4,7,1 0-tetraazacyclodecane-4,7,10-triacetate (HP-DOA3), 6-hydrazino-N-methylpyridine-3-carboxamide (HYNIC), 1,4,7-triazacyclononane-1-succinic acid-4,7-diacetic acid (NODASA), 1-(1-carboxy-3-carboxypropyl)-4,7-(carboxy)-1,4,7-triazacyclononane (NODAGA), 1,4,7-triazacyclononane triacetic 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), terpyridine bis(methyleneaminotetraacetic acid (TMT), 1,4,7,10-tetraazacyclotridecane-N,N',N",N"'-tetraacetic acid (TRITA), triethylenetetraaminehexaacetic acid (TTHA), N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18-crown-ether- 6 (H2macropa) and 4-amino-4-{2-[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydropyridin-2-ylmethyl)carbamoyl]ethyl}heptanedioic acid bis[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydropyridin-2-ylmethyl)-amide] (THP), the residue being obtained by covalently bonding the carboxyl group contained in the chelating agent to the rest of the compound through an ester bond or an amide bond. .

[0279] 37. The compound or salt of claim 36, wherein the chelating agent is selected from DOTA and DOTAGA.

[0280] 38. The compound or salt of item 36, wherein R B Is the formula -L B1 -R B1 The group, -LR B1 As defined in item 32, and R B1 The carboxyl group of the chelating agent and R in any of the groups of formula (VIa) to (VIf) B1 The chelating moiety is derived from the chelating agent DOTA or DOTAGA by forming an amide bond between the attached nitrogen atoms.

[0281] 39. The compound or salt of any one of items 33 to 38, wherein R B comprising a chelate formed by a chelating moiety and a chelated radioactive cation, or wherein R B1 is or comprises a chelate formed by a chelating moiety and a chelated radioactive cation, and wherein the chelated radioactive cation is selected from the group consisting of 44 Sc, 47 Sc, 51 Cr, 52m Mn, 58 Co. 52 Fe, 56 You, 57 You, 62 Cu, 64 Cu, 67 Cu, 66 Ga, 68 Ga, 67 Ga,89 Zr, 90 Y. 89 Y. 94m Tc, 99m Tc, 97 Such as 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 2. 169 Yb, 175 Yb, 172 Tm, 177 Lu, 186 Re, 188 Re, 191 Pt, 197 Hg, 198 Au, 199 Au, 212 Pb, 203 Pb, 211 At 212 Bi, 213 Bi, 223 Ra, 225 Ac and 227 Th, or contain 18 Cationic molecules of F, such as 18 F-[AlF] 2+ .

[0282] 40. The compound or salt of item 39, wherein the radioactive cation is selected from the group consisting of 99m Tc, 186 Re, 188 Re, 44 Sc, 47 Sc, 64 Cu, 67 Cu, 68 Ga, 90 Y. 111 In,161 Tb, 166 Ho, 177 Lu, 188 Re, 212 Pb, 212 Bi, 213 Bi, 225 Ac and 227 Th, or contain 18 F cationic molecule.

[0283] 41. The compound or salt of any one of items 1-16 and 24-30, wherein R A Is the formula -L A1 -R A1 A group in which L A1 is a divalent linking group, and R A1 is a group selected from the following or contains a group selected from the following:

[0284] (i) a chelating moiety,

[0285] (ii) a chelate formed by the chelating moiety (i) and a chelated radioactive or non-radioactive cation,

[0286] (iii) a moiety carrying a covalently bound radioisotope or a precursor suitable for labelling with such a radioisotope,

[0287] (iv) a phosphonate moiety, and

[0288] (v) Fluorescent labels, such as quantum dots or fluorescent dyes.

[0289] 42. The compound or salt of item 41, wherein -L A1 -R A1 A group selected from formula (IVa) and formula (IVb):

[0290] -(CH2)-C(=O)-NH-R 24 -NH-R A1 (IVa),

[0291] -(CH2)-C(=O)-NH-(CH2) y -R 25 -(CH2) z -NH-R A1 (IVb),

[0292] Where R 24 is a C2-C10 alkanediyl group, preferably a C3-C6 alkanediyl group, which is preferably linear and may be substituted by a group -C(=O)-NH2,

[0293] R 25is phenylene, preferably 1,3-phenylene, and y and z are independently integers selected from 0, 1 or 2, and are preferably both 1, and R A1 As defined in item 41.

[0294] 43. The compound or salt of item 42, wherein -L A1 -R A1 It is a group of formula (IVb).

[0295] 44. The compound or salt of any one of items 1 to 16 and items 24 to 30, wherein R A It comprises (i) a chelating moiety or (ii) a chelate formed by the chelating moiety (i) and a chelated radioactive or non-radioactive cation.

[0296] 45. The compound or salt of any one of items 41 to 43, wherein R A1 Comprising (i) a chelating moiety or (ii) a chelate formed by the chelating moiety (i) and a chelated radioactive or non-radioactive cation, or wherein R A1 It is (i) a chelating moiety or (ii) a chelate formed by the chelating moiety (i) and a chelated radioactive or non-radioactive cation.

[0297] 46. ​​The compound or salt of any one of items 44 or 45, wherein the chelating moiety comprises at least one of the following

[0298] a macrocyclic structure having 8 to 20 ring atoms, 2 or more, preferably 3 or more of the ring atoms being selected from oxygen atoms and nitrogen atoms; and

[0299] A non-cyclic open-chain chelate 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 atoms and nitrogen atoms.

[0300] 47. The compound or salt of any one of items 44 to 46, wherein the chelating moiety is a chelating agent, for example: bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (CBTE2a), cyclohexyl-1,2-diaminetetraacetic acid (CDTA), 4-(1,4,8,11-tetraazacyclotetradec-1-yl)-methylbenzoic acid (CPTA), N'-[5-[acetyl(hydroxy)amino]pentyl]-N-[5-[[4-[5-aminopentyl(hydroxy)amino]-4-oxobutyryl [4-(2-hydroxyethyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraazacyclododecane-4,7,10-triacetic acid]-pentanedioic acid (DOT), [4-(2-hydroxyethyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid]-pentanedioic acid (DOT), [4-(2-hydroxyethyl) ... AGA), N,N'-dipyridyloxyethylenediamine-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-triacetate (HP-DOA3), 6-hydrazino-N-methylpyridine-3-carboxamide (HYNIC), 1,4,7-triazacyclononane-1-succinic acid-4,7-diacetic acid (NODASA), 1-(1-carboxy-3-carboxypropyl)-4,7-(carboxy)-1,4,7-triazacyclononane (NODAGA), 1,4,7-triazacyclononane triacetic 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), terpyridine bis(methyleneaminotetraacetic acid (TMT), 1,4,7,10-tetraazacyclotridecane-N,N',N",N"'-tetraacetic acid (TRITA), triethylenetetraaminehexaacetic acid (TTHA), N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18-crown-ether- 6 (H2macropa) and 4-amino-4-{2-[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydropyridin-2-ylmethyl)carbamoyl]ethyl}heptanedioic acid bis[(3-hydroxy-1,6-dimethyl-4-oxo-1,4-dihydropyridin-2-ylmethyl)-amide] (THP), the residue being obtained by covalently bonding the carboxyl group contained in the chelating agent to the rest of the compound through an ester bond or an amide bond. .

[0301] 48. The compound or salt of claim 47, wherein the chelating agent is selected from DOTA and DOTAGA.

[0302] 49. The compound or salt of item 47, wherein R A Is the formula -L A1 -R A1 The group, -L A1 -R A1 is a group of formula (IVb), and R A1 is by R A1 The chelating moiety is derived from the chelating agent DOTA or DOTAGA by forming an amide bond between the attached nitrogen atoms.

[0303] 50. The compound or salt of any one of items 44 to 49, wherein R A comprising a chelate formed by a chelating moiety and a chelated radioactive cation, or wherein R A1 is or comprises a chelate formed by a chelating moiety and a chelated radioactive cation, and wherein the chelated radioactive cation is selected from the group consisting of 44 Sc, 47 Sc, 51 Cr, 52m Mn, 58 Co. 52 Fe, 56 You, 57 You, 62 Cu, 64 Cu, 67 Cu, 66 Ga, 68 Ga, 67Ga, 89 Zr, 90 Y. 89 Y. 94m Tc, 99m Tc, 97 Such as 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 2. 169 Yb, 175 Yb, 172 Tm, 177 Lu, 186 Re, 188 Re, 191 Pt, 197 Hg, 198 Au, 199 Au, 212 Pb, 203 Pb, 211 At 212 Bi, 213 Bi, 223 Ra, 225 Ac and 227 Th, or contain 18 Cationic molecules of F, such as 18 F-[AlF] 2+ .

[0304] 51. The compound or salt of item 50, wherein the radioactive cation is selected from the group consisting of 99m Tc, 186 Re, 188 Re, 44 Sc, 47 Sc, 64 Cu, 67 Cu, 68 Ga, 90 Y. 111 In,161 Tb, 166 Ho, 177 Lu, 188 Re, 212 Pb, 212 Bi, 213 Bi, 225 Ac and 227 Th, or contain 18 F cationic molecule.

[0305] 52. A pharmaceutical composition comprising or consisting of a compound or salt according to any one of items 1 to 51.

[0306] 53. A diagnostic composition comprising or consisting of a compound or salt according to any one of items 1 to 51.

[0307] 54. A compound or salt according to any one of items 1 to 51 for use in medicine, preferably in nuclear medicine, in nuclear molecular imaging, or optical imaging, or in targeted intracavitary radiotherapy.

[0308] 55. A compound or salt according to any one of items 1 to 51 for use in a method for treating or preventing cancer, cardiovascular disease, AIDS or inflammatory disorders.

[0309] 56. A compound or salt according to any one of items 1 to 51 for use in a method for diagnosing and / or staging cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0310] The accompanying drawings show:

[0311] Figure 1 : Structure of a cyclic CXCR4-binding molecule.

[0312] Figure 2 : Fluorescence microscopy of P5h and P5f binding to Chem-1 cells expressing CXCR4. (A) P5h, 100 nM, 4°C. (B) P5h, 100 nM, 37°C. (C) P5f, 100 nM, 4°C. (D) P5f, 100 nM, 37°C. Cells were also stained with Hoechst [2 μg / ml] (nuclei, blue) and Lysotracker Green [2 μM] (lysosomes, green) for visual aid. Cy5 images were taken with an excitation time of 8 s. Original magnification ×20.

[0313] Figure 3: Fluorescence microscopy of P5g binding to Chem-1 cells expressing CXCR4. (A) P5g, 500nM, 4°C. (B) P5g, 500nM, at 4°C, in the presence of 100μM AMD3100 (C) P5g, 500nM, 37°C. (D) P5g, 500nM, at 37°C, in the presence of 100μM AMD3100. Cells were also stained with Hoechst [2μg / ml] (nuclei, blue) and Lysotracker Green [2μM] (lysosomes, green) for visual aid. Cy5 images were taken with an excitation time of 8s. Original magnification ×20.

[0314] Figure 4 : Fluorescence microscopy of P5h binding to Chem-1 cells expressing CXCR4. (A) P5h, 500 nM, 4°C. (B) P5h, 500 nM, at 4°C, in the presence of 100 μM AMD3100 (C) P5h, 500 nM, 37°C. (D) P5h, 500 nM, at 37°C, in the presence of 100 μM AMD3100. Cells were also stained with Hoechst [2 μg / ml] (nuclei, blue) and Lysotracker Green [2 μM] (lysosomes, green) for visual aid. Cy5 images were taken with an excitation time of 8 s. Original magnification ×20.

[0315] Figure 5 : Fluorescence microscopy of P5i binding to Chem-1 cells expressing CXCR4. (A) P5i, 500 nM, 4°C. (B) P5i, 500 nM, at 4°C, in the presence of 100 μM AMD3100 (C) P5i, 500 nM, at 37°C. (D) P5i, 500 nM, at 37°C, in the presence of 100 μM AMD3100. Cells were also stained with Hoechst [2 μg / ml] (nuclei, blue) and Lysotracker Green [2 μM] (lysosomes, green) for visual aid. Cy5 images were taken with an excitation time of 8 s. Original magnification ×20.

[0316] Figure 6: Fluorescence microscopy of P5f binding to Chem-1 cells expressing CXCR4. (A) P5f, 500 nM, 4°C. (B) P5f, 500 nM, at 4°C, in the presence of 100 μM AMD3100 (C) P5f, 500 nM, at 37°C. (D) P5f, 500 nM, at 37°C, in the presence of 100 μM AMD3100. Cells were also stained with Hoechst [2 μg / ml] (nuclei, blue) and Lysotracker Green [2 μM] (lysosomes, green) for visual aid. Cy5 images were taken with an excitation time of 8 s. Original magnification ×20.

[0317] Figure 7 : Fluorescence microscopy of Pc5-QD binding to different CXCR4-expressing cell lines. Left: human Daudi lymphoma cells, 100 nM Pc5-QD, 1 h, RT; middle: Eμ-Myc1080 mouse B-cell lymphoma cells, 25 nM Pc5-QD, 1 h, RT; right: Chem-1 cells expressing hCXCR4, 25 nM Pc5-QD, 1 h, RT. Original magnification ×20.

[0318] Figure 8 : Specificity of Pc5-QD binding to Chem-1 cells expressing hCXCR4. Upper side: uncoated QD, 25 nM, 1 h, RT (negative control) and corresponding optical microscopy images, lower side: Pc5-QD, 25 nM, 1 h, RT, and corresponding optical microscopy images. Original magnification ×20.

[0319] Fig. 9 :pi1 hour[ 125 I] Biodistribution of R1 in Black Six normal mice. White bars: tracer only (n=5), grey bars: co-injected with 50 μg AMD3100 (2 mg / kg, n=4). Data are expressed as % iD / g, mean ± SD.

[0320] Fig.10 :pi1 hour[ 125 I] Biodistribution of R1 in Black Six normal mice and CD-1 nude mice. White bars: Black Six mice (n=5), gray bars: CD-1 nude mice. Data are expressed as %iD / g, mean ± SD.

[0321] Fig.11 :pi1 hour[ 125 I]R1 and [ 125 I] Biodistribution of P5a in CD-1 nude mice. White bars: [ 125I]R1, tracer only (n=5); gray bars: [ 125 I] P5a, tracer only (n=5). Data are expressed as % iD / g, mean ± SD.

[0322] Fig.12 :pi1 hour[ 68 Biodistribution of Ga]P5c in CD-1 nude mice. White bars: tracer only (n=4), grey bars: co-injection of 50 μg AMD3100 (2 mg / kg, n=4). Data are expressed as %iD / g, mean±SD.

[0323] Fig.13 :pi1 hour[ 125 I]R1,[ 68 Ga]P5c and [ 68 Biodistribution of Ga]Pc5b in CD-1 nude mice. White bar: [ 125 I]R1(n=5), gray bars: [ 68 Ga]P5c(n=4), dark grey bars: [ 68 Ga]Pc5b (n=4). Data are expressed as %iD / g, mean ± SD.

[0324] Fig.14 : At 1 hour pi, littermates of CB-17SCID mice bearing Daudi lymphoma [ 68 Ga]P5c-PET (MIP). Animals received approximately 12 MBq of [ 68 Ga]P5c. Left: tracer only (red arrows indicate lymphoma xenografts); right: co-injection of 50 μg AMD3100.

[0325] Fig.15 : At 1 hour pi, healthy CB-17SCID mice littermates [ 68 Ga]Pc5b-PET (MIP). Animals received approximately 13 MBq of [ 68 Ga]Pc5b. A: tracer only, B: co-injection of 50 μg AMD3100.

[0326] The following examples illustrate the present invention:

[0327] Example 1

[0328] Chemicals and instruments

[0329] Solvents were purchased from Aldrich, Fluka, Merck and Prolabo, reagents were purchased from Aldrich, Fluka, Sigma, Merck, Acros and Lancaster, the quality was "synthetic", "according to analysis" or "HPLC grade", and were used without purification. Amino acids, derivatives for protecting groups and coupling reagents were purchased from Iris Biotech (Marktredwitz, Germany), Bachem (Bubendorf, Switzerland) or Carbolution (Saarbrücken, Germany). All reactions sensitive to oxygen or water were carried out under argon atmosphere.

[0330] Solid phase peptide synthesis was carried out in a plastic syringe (VWR) equipped with a filter. The reaction solution and wash solution were drawn into the syringe for mixing, and the syringe was shaken manually or using an Intelli-Mixer syringe shaker mixer (Neolab, Heidelberg, Germany). For the loading of the first amino acid, the required equivalent (eq.) was calculated based on the theoretical loading provided by the supplier.

[0331] Thin layer chromatography (TLC) was used for reaction control and R f The peaks were measured on aluminum foil coated with Silica 60, F254 (Merck). The peaks were detected under UV light (254 nm) or after staining with Mountain solution.

[0332] Preparative flash column chromatography was performed with a 50- to 100-fold mass excess of Silica 60 (particle size 0.040-0.063 mm, Merck) and an applied overpressure of 1-1.5 bar.

[0333] Analytical and semi-preparative reversed-phase high performance liquid chromatography (HPLC) were performed using the following apparatus:

[0334] a)Amersham Pharmacia Biotech: Basic, 10F, autosampler A 900, pump system P-900, detector UV-900, Software Unicorn Vers.3.00. Column: YMC-ODS-A C18 ( 5μm, 250mm×4.6mm), analytical type.

[0335] b)Amersham Pharmacia Biotech: Basic, 100F, pump system P-900, detector UV-900, Software Unicorn Vers.3.00. Column: YMC-ODS-A C18 ( 10μm, 250mm×20mm), semi-preparative.

[0336] c) Beckman: high pressure pump module 125, UV detector 166. Software System Gold; column: YMC-ODS-A C18 ( 5μm, 250mm×20mm), semi-preparative.

[0337] d) Sykam: Gradient HPLC system (Sykam GmbH, Eresing, Germany), 206PHD UV-visible detector (Linear TM Instruments Corporation, Reno, USA), Winnie 32 software. Columns: a) Nucleosil100C18 (5 μm, 125×4.0 mm) (CS Chromotographie Services GmbH, Langerwehe, Germany), analytical, and b) Multospher 100RP 18-5 (250×20 mm) (CS), semi-preparative. Radioactivity was detected by connecting the outlet of the UV photometer to a NaI(Tl) well-type scintillation counter of EG&G Ortec (München, Germany).

[0338] e) Shimadzu: Prominence Gradient HPLC system (Shimadzu, Duisburg, Germany).

[0339] As eluent, a mixture of H2O (solvent A) and acetonitrile (solvent B) containing 0.1 vol-% TFA was used. Different linear gradients were applied over 15-30 min (analytical) and 15-30 min (semi-preparative). The flow rates were 1 mL / min (analytical) and 8-9 mL / min (semi-preparative), respectively. UV detection was performed at 220 and 254 nm.

[0340] Electrospray mass spectrometry (ESI-MS) was performed using the following

[0341] a) Finnigan apparatus (Type LCQ in conjunction with HPLC-system Hewlett Packard HP 1100). Columns: a) YMC Hydrosphere C18 ( 3μm, 125mm×2.1mm), flow rate: 0.55ml / min; b) YMC-UltraHT-Hydrosphere C18 ( 2μm, 50mm×2.0mm), flow rate: 0.75mL / min; c) YMC Octyl C8 ( 5 μm, 250 mm×2.1 mm), flow rate: 0.35 mL / min. As eluent, a mixture of H2O and acetonitrile containing 0.1 vol-% formic acid was used for different linear gradients (7 min, 15 min, 40 min).

[0342] or b) Varian 500-MS IT mass spectrometer (Agilent Technologies, Santa Clara, USA).

[0343] Nuclear magnetic resonance (NMR) experiments were performed at 300 K using AV250, AV360 and DMX500 (Bruker), and chemical shifts (δ) were expressed in parts per million (ppm) as 1 H- and 13 C-shifted internal standard, using the residual signal of the solvent (for 1 H is CHCl3 (δ = 7.26 ppm) and for 13 C is CDCl3 (δ = 77.16 ppm); for 1 H is DMSO-d5 (δ = 2.50 ppm) and for 13 C is DMSO-d6 (δ=39.52 ppm). 1 H-Broadband Decoupling Recording 13 C-NMR spectrum. The data were processed using MestreC and MestreNova programs, respectively.

[0344] Fluorescence microscopy experiments were performed using a BioRevo BZ9000 fluorescence microscope (Keyence, Osaka, Japan).

[0345] Example 2

[0346] synthesis

[0347] 1. General Operation

[0348] GP1. Loading of trityl chloride polystyrene (TCP) resin

[0349] Peptide synthesis was performed using TCP-resin (0.9 mmol / g) following the standard Fmoc strategy. Fmoc-Xaa-OH (1.2 eq) was coupled to TCP resin with DIEA (2.5 eq) in anhydrous DCM (0.8 mL / g resin) for 1 h at room temperature. The remaining trityl chloride groups were capped by adding 1 mL / g (resin) of MeOH, DIEA (5:1; vzv) solution for 15 min. The resin was filtered, washed 5 times with DCM and 3 times with MeOH. After drying the resin under vacuum, the loading was determined gravimetrically and ranged from 0.4-0.9 mmol / g.

[0350] GP2. Fmoc deprotection

[0351] The resin-bound Fmoc peptide was treated with 20% piperidine in NMP (v / v) for 10 min and a second time for 5 min. The resin was washed 5 times with NMP.

[0352] GP3. N-methylation under Mitsunobu conditions

[0353] A solution of triphenylphosphine (5 eq.), DIAD (5 eq.) and MeOH (10 eq.) in anhydrous THF (1 mL / g resin) was added to the resin-bound Ns-protected peptide and shaken at room temperature for 10 minutes. The resin was filtered off and washed 3 times with anhydrous THF and 3 times with NMP.

[0354] GP4. HATU / HOAt coupling

[0355] A solution of Fmoc-Xaa-OH, HATU (2 eq.), HOAt (2 eq.), DIPEA (4 eq.) in NMP (1 mL / g resin) was added to the resin-bound peptide and shaken at room temperature for 3 hours and washed 5 times with NMP.

[0356] GP5. On-Resin Ns Deprotection

[0357] To deprotect Ns, the resin-bound Ns-peptide was stirred in a solution of inercaptoethanol (10 eq.) and DBU (5 eq.) in NMP (1 mL / g resin) for 5 min. The deprotection procedure was repeated once more and the resin was washed 5 times with NMP.

[0358] GP6. Cleavage of peptide from resin

[0359] For complete cleavage from the resin, the peptide was treated three times with a mixture of acetic acid / 2,2,2-trifluoroethanol / DCM (3 / 1 / 6, v / v / v) at room temperature for half an hour, and the solvent was evaporated under reduced pressure.

[0360] GP7. Peptide backbone cyclization

[0361] DPPA (3 eq.) was added to a solution of the peptide in DMF (1 mM peptide concentration) and NaHCO3 (5 eq.) at room temperature and stirred overnight or until no linear peptide was observed by ESI-MS. The solvent was evaporated under reduced pressure to a small volume, the peptide was precipitated in saturated NaCl solution, and washed twice in HPLC grade water.

[0362] GP8. Removal of Dde protecting group

[0363] Dde-protection was performed using 2% hydrazine in DMF at room temperature. After 30 min, the deprotected peptides were precipitated using water (Pbf / tBu / Boc-protected peptides) or ether (deprotected peptides) and dried in a desiccator before further functionalization.

[0364] GP9. Removal of acid-labile side chain protecting groups

[0365] The cyclized peptide was stirred in a solution of TFA, water and TIPS (95:2.5:2.5; v:v:v) at room temperature for one hour or until no more protected peptide was observed by ESI-MS, then precipitated in diethyl ether, washed twice with diethyl ether and dried.

[0366] GP10. N-Alkylation of Dde-6-aminohexanol

[0367] A solution of 2-nitrobenzenesulfonyl chloride (o-Ns-Cl, 4 equiv.) and sym-trimethylpyridine (10 equiv.) in NMP was added to the resin-bound peptide and stirred for 15 min. The resin was subsequently washed with NMP (3×) and THF abs. (3×). Then, a solution of PPh3 (5 equiv.) and Dde-6-aminohexanol in THF abs. was drawn into a syringe and a small amount of DIAD (5 equiv.) in THF abs. was slowly added. After the resin was incubated with the solution at room temperature for 30 min, the mixture was removed and the reaction steps were repeated again. The resin was washed with THF (3×) and NMP (5×). The o-Ns-protecting group was removed by adding a solution of mercaptoethanol (10 equiv.) and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU, 5 equiv.) in NMP for 5 min (2×). Finally, the resin was washed with NMP (5×).

[0368] GP11. Alkyl / Acylguanidinylation in Solution

[0369] The orthogonally deprotected peptide P0 was dissolved in DMF, followed by the addition of precursors (A6-A11, B4-B7) (1.5 eq.) and DIPEA (5 eq.). The solution was stirred at room temperature for 4 hours or until no starting material was detected (monitored by ESI-MS). The peptide was then precipitated in saturated aqueous sodium chloride solution (brine) and washed with water (2×).

[0370] GP12. Dde-Protection of Linear Amino Alcohols

[0371] DdeOH (1.05 eq.) was added to a DCM solution of the linear amino alcohol (1.0 eq.) and stirred overnight. The solution was washed with 1 M aqueous HCl (3×) and the combined organic layers were dried over Na2SO4. After evaporation of the solvent in vacuo, the Dde-protected amino alcohol was obtained as a yellow solid.

[0372] GP13. Alkylation of N,N′-di-Boc-1H-pyrazole-1-carboxamidine

[0373] N, N'-di-Boc-1H-pyrazole-1-carboximidamide (1.0 g, 3.2 mmol, 1.0 eq.), PPh3 (0.99 g, 3.8 mmol, 1.2 eq.) and the corresponding alcohol (1.2 eq.) were dissolved in 10 ml THF abs. DIAD (747 μl, 3.8 mmol, 1.2 eq.) was added dropwise to the vigorously stirred solution (room temperature) and then stirred at room temperature for 2 hours. After evaporating the solvent in vacuo, the crude product was purified by flash chromatography.

[0374] GP14. Conjugation of free amino functional groups with unprotected DOTA

[0375] DOTA (4 eq.), NHS (5 eq.) and EDCI (5 eq.) were dissolved in water and DIPEA (8 eq.) was added. After 15 minutes, each peptide (1 eq.) was added to an equal volume of water. The progress of the coupling reaction was monitored using RP-HPLC. After the reaction was complete, the solvent was evaporated in vacuo. The residue was resuspended in methanol, the suspension was centrifuged, and the product dissolved in the methanol supernatant was precipitated with ether, dried, and purified using preparative RP-HPLC.

[0376] GP15. nat Ga-, nat Cu- nat Lu-and nat Preparation of Y-DOTA reference compounds

[0377] To prepare nat Ga-and nat Cu-complexes were prepared by mixing equal volumes of 2 mM Ga(NO3)3 or Cu(NO3)2 in 1 M NaOAc buffer and 2 mM aqueous solutions of the respective peptides and heating to 95°C for 30 min. The corresponding metal chlorides were prepared by adding a 2.5 molar excess of the respective metal chloride dissolved in water to the peptides. nat Lu andnat After heating to 95°C for 30 minutes, the formation of each metal complex was confirmed by RP-HPLC and ESI-MS.

[0378] GP16. Conjugation of free amino functional groups with Cy5-dye

[0379] To a DMF solution of the amine-functionalized peptide (1.1 eq.) and each Cy5 dye analog (1 eq.), HATU (3 eq.) and DIPEA (5 eq.) were added successively. The progress of the coupling reaction was monitored using RP-HPLC. Upon completion of the reaction (usually within 30-60 min at room temperature), the crude product was immediately isolated from the reaction mixture using preparative RP-HPLC.

[0380] GP17. Solution-phase alkylation of Cys side chains

[0381] The thiol-functionalized peptide Pc0 (1 equivalent) and each 2-chloroacetyl-functionalized linker unit (C1-C4) or Boc-6-aminohexyl bromide (3 equivalents) were dissolved in acetonitrile / water (9 / 1; v / v), and DIPEA (4 equivalents) was added. The resulting suspension was stirred at room temperature for 12-18 hours. After the alkylation reaction was completed, the crude product was precipitated in water, washed and dried in vacuo.

[0382] 2. Synthesis of cyclic pentapeptide analogs R and R1

[0383] CPCR4.3(R)

[0384] As mentioned before 52 Preparation of CPCR4.3(c(y-(NHexGua)aR-Nal-G),R).

[0385]

[0386] Iodinated-CPCR4.3(R1)

[0387] Synthesis of the unlabeled reference compound iodo-CPCR4.3 was performed using N-iodosuccinimide (NIS) in acetonitrile / water 53 Briefly, CPCR4.3 was dissolved in a 1:1 (v / v) mixture of acetonitrile and water to obtain a 9 mM solution, and 0.6 equivalents of NIS were added.

[0388] After the reaction was completed (5-10 min at room temperature), iodo-CPCR4.3 was separated using semi-preparative RP-HPLC.

[0389]

[0390] 3. Synthesis of Precursors for Alkylation of the Hexylguanidine Side Chain in R

[0391] 2-(1-((2-Hydroxyethyl)amino)ethylidene)-5,5-dimethylcyclohexane-1,3-dione (A1)

[0392] A1 (1.3 g, 5.8 mmol, 83%) was prepared according to GP12 and obtained as a yellow solid.

[0393]

[0394] 1 H-NMR (360MHz, DMSO): δ = 13.26 (bs, 1H), 5.00 (bs, 1H), 3.58 (t, J = 5.2Hz, 2H), 3.51-3.46 (m, 2H), 2.48 (s, 3H), 2.26 (s, 4H), 0.94 (s, 6H). 13 C-NMR (90MHz, DMSO): δ = 196.3, 172.9, 106.9, 59.1, 52.4, 45.4, 29.8, 27.9, 17.6. HPLC(10-90%,15min):t R =4.40min,MS(ESI):m / z=226.1[M+H] + .

[0395] 2-(1-((2-Hydroxypropyl)amino)ethylidene)-5,5-dimethylcyclohexane-1,3-dione (A2)

[0396] A2 (1.5 g, 6.3 mmol, 90%) was prepared according to GP12 and obtained as a yellow solid.

[0397]

[0398] 1 H-NMR (360MHz, DMSO): δ = 13.25 (bs, 1H), 4.30 (bs, 1H), 3.51-3.45 (m, 4H), 2.48 (s, 3H), 2.26 (s, 4H), 1.71 (q, J = 5.2Hz, 2H), 0.94 (s, 6H). 13 C-NMR (90MHz, DMSO): δ=196.4,172.9,106.9,57.7,52.3,31.7,29.7,27.9,17.3. HPLC(10-90%,15min):t R =4.73min,MS(ESI):m / z=240.1[M+H] + .

[0399] 2-(1-((2-Hydroxybutyl)amino)ethylidene)-5,5-dimethylcyclohexane-1,3-dione (A3)

[0400] A3 (1.5 g, 5.9 mmol, 84%) was prepared according to GP12 and obtained as a yellow solid.

[0401]

[0402] 1 H-NMR ((360MHz, DMSO): δ = 13.27 (bs, 1H), 4.49 (t, J = 5.2Hz, 1H), 3.46-3.40 (m, 4H), 2.48 (s, 3H), 2.27 (s, 4H), 1.63-1.45 (m, 4H), 0.94 (s, 6H). 13 C-NMR (90MHz, DMSO): δ = 196.4, 172.8, 106.9, 52.3, 42.6, 29.8, 29.6, 27.9, 25.3, 17.4. HPLC(10-90%,15min):t R =5.01min,MS(ESI):m / z=254.1[M+H] + .

[0403] 2-(1-((2-Hydroxypentyl)amino)ethylidene)-5,5-dimethylcyclohexane-1,3-dione (A4)

[0404] A4 (1.7 g, 6.4 mmol, 91%) was prepared according to GP12 and obtained as a yellow solid.

[0405]

[0406] 1 H-NMR ((360MHz, DMSO): δ = 13.26 (bs, 1H), 3.44-3.37 (m, 4H), 2.48 (s, 3H), 2.27 (s, 4H), 1.58 (q, J = 5.2Hz, 2H), 1.47-1.33 (m, 4H), 0.94 (s, 6H). 13 C-NMR (90MHz, DMSO): δ = 196.4, 172.8, 106.9, 60.5, 52.3, 42.7, 32.0, 29.8, 28.4, 27.9, 22.9, 17.4. HPLC(10-90%,15min):t R =5.43min,MS(ESI):m / z=268.2[M+H] + .

[0407] 2-(1-((2-Hydroxyhexyl)amino)ethylidene)-5,5-dimethylcyclohexane-1,3-dione (A5)

[0408] A5 (2.5 g, 8.9 mmol, 85%) was prepared according to GP12 and obtained as a yellow solid.

[0409]

[0410] 1 H-NMR ((360MHz, DMSO): δ = 13.26 (bs, 1H), 4.36 (bs, 1H), 3.44-3.36 (m, 4H), 2.47 (s, 3H), 2 .26(s,4H),1.57(q,J=6.2Hz,2H),1.42(t,J=6.2Hz,2H),1.35-1.29(m,4H),0.94(s,6H). 13 C-NMR (90MHz, DMSO): δ = 196.4, 172.8, 106.9, 60.6, 52.3, 42.6, 32.4, 29.8, 28.5, 27.9, 26.2, 25.1, 17.4. HPLC(10-90%,15min):t R =6.47min,MS(ESI):m / z=282.2[M+H] + .

[0411] tert-Butyl-(((tert-butyloxycarbonyl)imino)(1H-pyrazol-1-yl)methyl)(2-((1-(4,4-dimethyl-2, 6-Dioxocyclohexylidene)ethyl)amino)ethyl)carbamate (A6)

[0412] N,N′-di-BOC-1H-pyrazole-1-carboximidamide was reacted with A1 according to GP13 and then purified by flash chromatography (DCM / EtOAc 2:1) to afford A6 (0.75 g, 1.45 mmol, 45%) as a yellow viscous oil.

[0413]

[0414] 1 H-NMR (360MHz, DMSO): δ = 13.25 (bs, 1H), 8.15 (bs, 1H), 7.87 (d, J = 1.5Hz, 1H), 6.59 (dd, J = 2.8Hz, 1.5Hz,1H),3.85-3.72(m,4H),2.49(s,3H),2.25(s,4H),1.42(s,9H),1.21(s,9H),0.93(s,6H). Rf=0.25 (DCM / EtOAc 2:1). HPLC(10-90%,7min):t R =4.27min,MS(ESI):m / z=541.1[M+Na] + ,518.0[M+H] + ,418.0[M-Boc+H] + ,318.1[M-2Boc+H] + .

[0415] tert-Butyl-(((tert-butyloxycarbonyl)imino)(1H-pyrazol-1-yl)methyl)(2-((1-(4,4-dimethyl-2, 6-Dioxocyclohexylidene)ethyl)amino)propyl)carbamate (A7)

[0416] Reaction of N,N′-di-BOC-1H-pyrazole-1-carboximidamide with A2 according to GP13, followed by purification by flash chromatography (DCM / EtOAc 2:1) afforded A7 (0.87 g, 1.64 mmol, 51%) as a yellow viscous oil.

[0417]

[0418] 1 H-NMR (360MHz, DMSO): δ = 13.29 (bs, 1H), 8.22 (bs, 1H), 7.87 (d, J = 1.5Hz, 1H), 6.60 (dd, J = 2.8Hz, 1.5Hz, 1H), 3.70-3 .64(m,2H),3.57-3.50(m,2H),2.46(s,3H),2.26(s,4H),1.98-1.89(m,2H),1.41(s,9H),1.22(s,9H),0.94(s,6H). 13 C-NMR (90MHz, DMSO): δ=196.2,173.8,155.9,151.8,143.6,130.5,109.5,106.8,82.2,81.7,52.0,42.4,32.9,29.7,27.8,27.8,27.3,17.2. Rf=0.42(DCM / EtOAc 2:1),HPLC(10-90%,7min):t R =4.41min,MS(ESI):m / z=554.1[M+Na] + ,532.1[M+H] + ,432.0[M-Boc+H] + ,332.0[M-2Boc+H] + .

[0419] tert-Butyl-(((tert-butyloxycarbonyl)imino)(1H-pyrazol-1-yl)methyl)(2-((1-(4,4-dimethyl-2, 6-Dioxocyclohexylidene)ethyl)amino)butyl)carbamate (A8)

[0420] Reaction of N,N′-di-BOC-1H-pyrazole-1-carboximidamide with A3 according to GP13, followed by purification by flash chromatography (DCM / EtOAc 2:1) afforded A8 (1.29 g, 2.37 mmol, 74%) as a yellow viscous oil.

[0421]

[0422] 1H-NMR (360MHz, DMSO): δ = 13.26 (bs, 1H), 8.18 (bs, 1H), 7.84 (d, J = 1.5Hz, 1H), 6.59 (dd, J = 2.8Hz, 1.5Hz, 1H), 3.63-3 .59(m,2H),3.47-3.40(m,2H),2.46(s,3H),2.26(s,4H),1.73-1.58(m,4H),1.41(s,9H),1.21(s,9H),0.94(s,6H). 13 C-NMR (90MHz, DMSO): δ=196.3,172.8,156.6,155.9,151.7,143.6,130.4,10 9.6,106.8,81.9,81.5,52.1,47.7,42.1,29.7,27.8,27.5,27.3,21.7,17.2. Rf=0.50(DCM / EtOAc 2:1),HPLC(10-90%,7min):t R =4.50min,MS(ESI):m / z=568.1[M+Na] + ,546.1[M+H] + ,445.9[M-Boc+H] + .

[0423] tert-Butyl-(((tert-butyloxycarbonyl)imino)(1H-pyrazol-1-yl)methyl)(2-((1-(4,4-dimethyl-2, 6-Dioxocyclohexylidene)ethyl)amino)pentyl)carbamate (A9)

[0424] Reaction of N,N′-di-BOC-1H-pyrazole-1-carboximidamide with A4 according to GP13, followed by purification by flash chromatography (DCM / EtOAc 2:1) afforded A9 (1.41 g, 2.52 mmol, 79%) as a yellow viscous oil.

[0425]

[0426] 1 H-NMR (360MHz, DMSO): δ = 13.27 (bs, 1H), 8.17 (bs, 1H), 7.84 (d, J = 1.5Hz, 1H), 6.59 (dd, J = 2.8Hz, 1.5Hz, 1H), 3.60-3.55 (m, 2H ),3.43-3.37(m,2H),2.46(s,3H),2.26(s,4H),1.68-1.54(m,4H),1.42(s,9H),1.42-1.35(m,2H),1.20(s,9H),0.94(s,6H). 13C-NMR (90MHz, DMSO): δ=196.3,172.8,156.6,155.9,151.7,143.6,130.4,10 9.6,106.8,81.9,81.5,52.1,47.7,42.1,29.7,27.8,27.5,27.3,21.7,17.2. Rf=0.53(DCM / EtOAc 2:1),HPLC(10-90%,7min):t R =4.61min,MS(ESI):m / z=582.1[M+Na] + ,560.1[M+H] + ,460.1[M-Boc+H] + ,360.1[M-2Boc+H] + .

[0427] tert-Butyl-(((tert-butyloxycarbonyl)imino)(1H-pyrazol-1-yl)methyl)(2-((1-(4,4-dimethyl-2, 6-Dioxocyclohexylidene)ethyl)amino)hexyl)carbamate (A10)

[0428] Reaction of N,N′-di-BOC-1H-pyrazole-1-carboximidamide with A5 according to GP13, followed by purification by flash chromatography (DCM / EtOAc 2:1) afforded A10 (1.41 g, 2.46 mmol, 88%) as a yellow viscous oil.

[0429]

[0430] 1 H-NMR (360MHz, DMSO): δ = 13.26 (t, 1H), 8.18 (bs, 1H), 7.85 (d, 1H), 6.59 (dd, 1H), 3.56 (t, 2H), 3.39 (dt, 2H), 2.46 (s ,3H),2.26(s,4H),1.65-1.60(m,2H),1.57-1.53(m,2H),1.41(s,9H),1.36-1.31(m,2H),1.20(s,9H),0.94(s,6H). 13 C-NMR (90MHz, DMSO): δ=196.2,172.6,156.6,151.9,143.5,130.3,109.5,106. 8,82.0,81.6,52.3,48.1,42.5,29.7,28.2,27.8,27.5,27.3,25.7,25.5,17.3. Rf=0.66(DCM / EtOAc 2:1),HPLC(10-90%,15min):t R =10.36min,MS(ESI):m / z=574.1,[M+H] +,474.1[M-Boc+H] + ,374.2[M-2Boc+H] + .

[0431] tert-Butyl-(((tert-butoxycarbonyl)imino)(1H-pyrazol-1-yl)methyl)(hex-5-yn-1-yl)carbamic acid Ester (A11)

[0432] Reaction of N,N′-di-BOC-1H-pyrazole-1-carboximidamide with 5-hexin-1-ol according to GP13, followed by purification by flash chromatography (DCM / EtOAc 2:1) afforded A11 (0.94 g, 2.40 mmol, 75%) as a yellow viscous oil.

[0433]

[0434] Rf=0.53(DCM / EtOAc 2:1),HPLC(10-90%,7min):t R =4.73min,MS(ESI):m / z=413.1

[0435] [M+Na]+,391.1[M+H] + ,291.0[M-Boc+H] + ,189.9[M-2Boc+H] + .

[0436] 4. Synthesis of Cyclic Pentapeptide Analogs (P1-P6) with Alkylated Hexylguanidino Residues

[0437] Peptide intermediate c(y(OtBu)-(NHex-NH2)aR(Pbf)-Nal-G)(P0)

[0438] According to GP1, TCP-resin loaded Fmoc-Gly-OH, the linear peptide Hd-Ala-R (Pbf) -Nal-G was synthesized according to standard Fmoc-operation (GP2 and GP4, respectively). The peptide was subsequently alkylated with Dde-aminohexanol according to GP10, and Fmoc-d-Tyr (OtBu) -OH (GP4) was coupled to the Ns-deprotected peptide. After deprotection (GP2), cleavage (GP6) and backbone cyclization (GP7) from the resin were performed. After removing the Dde-protecting group (GP8), the crude peptide was precipitated in a saturated NaCl aqueous solution and lyophilized from an ACN / H2O solution. P0 (132 mg, 126 μM, 31%) was obtained as a light yellow powder (purity > 90%).

[0439]

[0440] Acetylated Alkylguanidine Derivatives (P1-P6)

[0441] P0 was guanylated with precursors A6-A10 according to GP11. After the guanylation reaction was completed, the compound was treated in situ with hydrazine (final concentration 2% v / v) for 5 minutes. Subsequently, an excess of Ac2O (20 equivalents) was added and the mixture was stirred for 30 minutes. Subsequently, the peptide was precipitated in a saturated NaCl aqueous solution. The acid-labile side chain protecting groups were removed according to GP9, and the crude peptides were purified by semi-preparative HPLC to obtain compounds P1-P5. By reacting P0 and A11 according to GP11, the crude peptide was precipitated from water, followed by side chain deprotection (GP9) and purification by semi-preparative HPLC to obtain P6.

[0442]

[0443]

[0444]

[0445] 5. Synthesis of Functionalized P5 Analogs

[0446] Cel-S-Hex-CPCR4.3(P5a)

[0447] H2N-Hex-CPCR4.3 (1 equivalent) and Cel(7Ac)-merc-OPfp 54 To a solution of 1.5 eq. in DMF was added DIPEA (3 eq.) and the solution was stirred at room temperature for 30 minutes. The crude product was then precipitated with ether and redissolved in MeOH. For deacetylation, a catalytic amount of KCN was added and the peptide methanol solution was stirred at room temperature overnight. Once RP-HPLC monitoring showed complete removal of all acetyl protecting groups, the peptide was precipitated with ether, washed, dried and purified by preparative RP-HPLC.

[0448]

[0449] HPLC (30-60% in 15 min): t R =7.08min.MS(ESI):m / z=649.6[M+2H] 2+

[0450] DOTA-Hex-CPCR4.3 (P5b) (and Ga / Lu reference compounds (P5c and P5d)

[0451] P0 was completely deprotected as in GP9 and solution-phase coupled with DOTA as in GP14.

[0452]

[0453] HPLC (15-45% in 15 minutes): t R=9.3min.MS(ESI):m / z=1272.2[M+H] + ,1294.1[M+Na] + ,636.9

[0454] [M+2H] 2+

[0455] Prepare the respective nat Ga-DOTA-(P5c) and nat Lu-DOTA-(P5d) reference compound.

[0456] nat Ga-P5b=P5c:MS(ESI):m / z=1339.7[M+H] + ,670.6[M+2H] 2+

[0457] nat Lu-P5b=P5d:MS(ESI):m / z=1443.8[M+H] + ,722.7[M+2H] 2+

[0458] Sulfo2-Cy5-Hex-CPCR4.3(P5e)

[0459] P0 was fully deprotected according to GP9 and solution phase coupling with Sulfo2-Cy5 (free acid) was performed according to GP16.

[0460]

[0461] HPLC (15-45% in 15 minutes): t R =13.2min.MS(ESI):m / z=1511.0[M+H] + ,756.3[M+2H] 2+ .

[0462] Sulfo2-Cy5(Hex-CPCR4.3)2(P5f)

[0463] P0 was completely deprotected according to GP9, and solution phase coupling with Sulfo2-Cy5 (symmetrical diacid) was performed according to GP16 using 2.5 equivalents of P0 and 1 equivalent of Sulfo2-Cy5 (symmetrical diacid).

[0464]

[0465] HPLC (15-45% in 15 minutes): t R =11.5min.MS(ESI):m / z=1239.3[M+2H] 2+,826.9[M+3H] 3+ .

[0466] Sulfo2-Cy5-PEG4-Hex-CPCR4.3(P5g)

[0467] After removing the acid-labile side chain protecting groups of P0 according to GP9, the peptide was dissolved in DMF and coupled with Fmoc-NH-PEG4-COOH (1.5 equivalents) using HOBt (1.5 equivalents) and TBTU (1.5 equivalents) as coupling agents and DIPEA (4.5 equivalents) as a base. After the coupling reaction was completed, the crude peptide was precipitated from ether and immediately redissolved in piperidine / DMF (20:80, v / v) to remove the Fmoc-protecting group. After 10 minutes at room temperature, the deprotected PEG4-conjugate was precipitated with ether and washed with ether. Coupling with Sulfo2-Cy5 (free acid) was performed according to GP16.

[0468]

[0469] HPLC (20-70% in 15 min): t R =7.15min.

[0470] MS (ESI): m / z = 1758.2 [M+H] + ,879.5[M+2H] 2+ ,890.8[M+H+Na] 2+ .

[0471] Cel-Sulfo2-Cy5-PEG4-Hex-CPCR4.3(P5h)

[0472] After removing the acid-labile side chain protecting groups of P0 according to GP9, the peptide was dissolved in DMF and coupled with Fmoc-NH-PEG4-COOH (1.5 equivalents) using HOBt (1.5 equivalents) and TBTU (1.5 equivalents) as coupling agents and DIPEA (4.5 equivalents) as a base. After the coupling reaction was completed, the crude peptide was precipitated from ether and immediately redissolved in piperidine / DMF (20:80, v / v) to remove the Fmoc-protecting group. After 10 minutes at room temperature, the deprotected PEG4-conjugate was precipitated with ether and washed with ether. Coupling with cellobiose-ethyl-Sulfo2-Cy5 (free acid) was performed according to GP16.

[0473]

[0474] HPLC (10-45% in 15 minutes): t R =11.3min.MS(ESI):m / z=1057.0[M+2H] 2+ .

[0475] Cel-Sulfo2-Cy5-arg-Hex-CPCR4.3(P5i)

[0476] After removing the acid-labile side chain protecting groups of P0 according to GP9, the peptide was dissolved in DMF and coupled with Fmoc-D-Arg(Pbf)-OH (1.5 equivalents) using HOBt (1.5 equivalents) and TBTU (1.5 equivalents) as coupling agents and DIPEA (4.5 equivalents) as a base. After the coupling reaction was completed, the crude peptide was precipitated from ether and immediately redissolved in piperidine / DMF (20:80, v / v) to remove the Fmoc-protecting group. After 10 minutes at room temperature, the deprotected arg-conjugate was precipitated with ether and washed with ether. Coupling with cellobiose-ethyl-Sulfo2-Cy5 (free acid) was performed according to GP16.

[0477]

[0478] HPLC (10-45% in 15 minutes): t R =11.1min.MS(ESI):m / z=1011.8[M+2H] 2+ .

[0479] 6-(6-(3-(di(benzyloxy)phosphoryl)propionamido)hexanamido)hexanoic acid (D1)

[0480] TCP resin was loaded with Fmoc-6-aminocaproic acid (Fmoc-Ahx-OH) according to GP1. Successive Fmoc-deprotection, coupling with Fmoc-Ahx-OH, Fmoc-deprotection and coupling with dibenzylphos-phonopropionic acid were carried out according to GP2 and GP4, respectively. D1 was obtained as a white solid (110 mg, 196 μmol, 25%) after cleavage from the resin (GP6) and freeze drying.

[0481]

[0482] HPLC (10-90% in 15 minutes): t R =6.83min,MS(ESI):m / z=1120.9[2M+H] + ,561.2[M+H] +

[0483] Phosphonate-Ahx2-Hex-CPCR4.3(P5k)

[0484] P0 (11.3 mg, 1.0 eq.) was coupled with D1 (5.9 mg, 1.3 eq.) using HOBt (1.5 eq.) and TBTU (1.5 eq.) as coupling agents and DIPEA (4.5 eq.) as a base. The fully protected peptide was purified by preparative RP-HPLC. The purified peptide was then deprotected and lyophilized according to GP9 to give P5k as a white solid (3.7 mg, 37%).

[0485]

[0486] HPLC (10-90% in 15 minutes): t R =4.30min,MS(ESI):m / z=1247.7[M+H] + ,624.7[M+2H] 2+ .

[0487] 21-(Bis(benzyloxy)phosphoryl)-10,19-dioxo-3,6,12,15-tetraoxa-9,18-diaza-eicosyl Monodecanoic acid (D2)

[0488] TCP resin was loaded with Fmoc-3,6-dioxooctanoic acid (Fmoc-O2Oc-OH) according to GP1. Successive Fmoc-deprotection, coupling with Fmoc-O2Oc-OH, Fmoc-deprotection and coupling with dibenzylphosphonic acid propionic acid were performed according to GP2 and GP4, respectively. After cleavage (GP6) from the resin and freeze drying, D1 was obtained as a white solid (87 mg, 18%).

[0489]

[0490] HPLC (10-90% in 15 minutes): t R =6.26min.MS(ESI):m / z=625.2[M+H] + .

[0491] Phosphonate-(O2Oc)2Hex-CPCR4.3(P5m)

[0492] P0 (10.5 mg, 1.0 eq.) was coupled with D2 (6.1 mg, 1.3 eq.) using HOBt (1.5 eq.) and TBTU (1.5 eq.) as coupling agents and DIPEA (4.5 eq.) as a base. The fully protected peptide was purified by preparative RP-HPLC. The purified peptide was then deprotected and freeze-dried according to GP9 to give P5m as a white solid (2.4 mg, 24%).

[0493]

[0494] HPLC (10-90% in 15 minutes): t R=5.05min.MS(ESI):m / z=1311.7[M+H] + ,656.7[M+2H] 2+ .

[0495] Carboxyl-QDs coated with P5 (P5-QD)

[0496] P0 was deprotected according to GP9, dried and lyophilized. Carboxyl quantum dots (Life technologies, 655ITK TM Carboxyl quantum dots) (0.8 nmol in 100 μl 50 mM borate buffer, (pH = 8.3)) to a total volume of 920 μl. Then, 1-ethyl-3-(3-dimethylamino-propyl) carbodiimide (800 equivalents, 640 nmol) in 12.2 μl 10 mM borate buffer and deprotected P0 (40 equivalents, 32 nmol) dissolved in 64 μl 10 mM borate buffer were added. The consumption of unreacted peptide in the reaction mixture was monitored by RP-HPLC. After the coupling reaction was completed (30-60 minutes at room temperature), the excess coupling agent was removed from the P5-QD suspension by repeating the buffer exchange using 50 mM borate buffer (pH = 8.3) and an Amicon Ultra-4 ultrafiltration unit (4 mL, Millipore) with a cutoff value of 100 kDa. Final reconstitution with 50 mM borate buffer yielded the peptide-coated quantum dots P5-QD as a 4 μM suspension. Based on the chosen stoichiometry, the coverage of the quantum dots was calculated to be approximately 40% of the reacted carboxylate.

[0497] 6. Synthesis of the hexylguanidinyl side chain acylation precursor in R

[0498] N-(tert-Butyloxycarbonyl)-S-methylisothiourea (B1)

[0499] S-methylisothiourea-hemisulfate (1.4 g, 10 mmol, 1.0 equiv) and di(tert-butyl)dicarbonate (2.2 g, 10 mmol, 1.0 equiv) were dissolved in a two-phase solution of vigorously stirred DCM and saturated NaHCO3 aqueous solution at room temperature for 1 day. After the two phases were separated, the organic layer was collected and the aqueous phase was extracted with DCM (3×). The organic layers were combined, dried over Na2SO4, and the solvent was removed in vacuo. After purification by flash chromatography (hexane / EtOAc 4:1), B1 (1.1 g, 5.8 mmol, 58%) was obtained as a colorless sticky solid.

[0500]

[0501] 1H-NMR (360MHz, DMSO): δ = 2.30 (s, 3H), 1.39 (s, 9H). 13 C-NMR (90MHz, DMSO): δ = 165.6, 154.1, 78.3, 28.4, 13.3. R f =0.30 (hexane / EtOAc 3:1), HPLC (10-90% in 15 min): t R =1.31min,MS(ESI):m / z=190.8[M+H] + .

[0502] N-acetyl glycine (B2)

[0503] Glycine (1.0g, 13.3mmol, 1.0 equivalent) is dissolved in 30ml DMF, 4.5ml DIPEA (26.6mmol, 2.0 equivalent) is added subsequently, and the solution is then cooled to 0 ℃. Afterwards, Ac2O (3.8ml, 39.9mmol, 3.0 equivalent) is added dropwise, and the solution is stirred at room temperature for 2h. After adding 20ml MeOH, the mixture is stirred for 30 minutes, and the solvent is removed in vacuo. The crude product is placed in 20ml EtOAc and washed with an equal volume of 1M HCl. The organic layer is dried with Na2SO4, and the solvent is removed in vacuo. B2 is obtained as a white solid (1.2g, 10.3mmol, 77%).

[0504]

[0505] 1 H-NMR (360MHz, DMSO): δ = 12.48 (s, 3H), 8.14 (t, J = 5.9Hz, 1H), 3.71 (d, J = 5.9, 2H), 1.84 (s, 3H). 13 C-NMR (90MHz, DMSO): δ = 171.61, 169.9, 40.8, 22.4. MS(ESI):m / z=140.0[M+Na] + ,117.9[M+H] + .

[0506] N-Acetyl-6-aminocaproic acid (B3)

[0507] 6-aminocaproic acid (2.0g, 15.3mmol, 1.0 equivalent) is dissolved in 50ml DMF, 5.2ml DIPEA (30.6mmol, 2.0 equivalent) is added subsequently, and the solution is then cooled to 0 ℃.Afterwards, Ac o (4.3ml, 45.9mmol, 3.0 equivalent) is added dropwise, and the solution is stirred at room temperature. After adding 20 milliliters of MeOH, the mixture is stirred for 30 minutes, and the solvent is removed in vacuo. The crude product is placed in 30ml EtOAc, washed with isopyknic 1M HCl. Organic layer Na sO dry, remove the solvent in vacuo. Obtain B3, it is a white solid (2.1g, 12.1mmol, 79%).

[0508]

[0509] 1 H-NMR (360MHz, DMSO): δ = 11.93 (bs, 1H), 7.79 (bs, 1H), 3.0 (dt, J = 6.9Hz, 5.5Hz, 2H), 2.1 9(t,J=7.2Hz,2H),1.78(s,3H),1.54-1.42(m,2H),1.40-1.33(m,2H),1.29-1.22(m,2H). 13 C-NMR (90MHz, DMSO): δ = 174.4, 168.94, 38.4, 33.6, 28.9, 26.0, 24.3, 22.6. HPLC(10-90%,7min):t R =0.70min,MS(ESI):m / z=196.0[M+Na] + ,173.9[M+H] + ,347.0[2M+H] + ,369.0[2M+Na] + ,385.1[2M+K] + .

[0510] Tert-butyl-(acetamido(methylthio)methylene)carbamate (B4)

[0511] B1 (250 mg, 1.3 mmol, 1.0 equiv) was dissolved in DMF, and DIPEA (440 μl, 2.6 mmol, 2.0 equiv) was added to the stirred solution. After Ac2O (245 μl, 5.2 mmol, 4.0 equiv) was added dropwise, the mixture was stirred at room temperature for 1 hour, then placed in MeOH (2 ml) and stirred for 15 minutes. The solvent was removed in vacuo, and the crude product was purified by flash chromatography (hexane / EtOAc 3: 1). B4 was obtained as a white solid (241 mg, 1.05 mmol, 80%).

[0512]

[0513] 1 H-NMR (360MHz, DMSO): δ = 2.27 (s, 3H), 2.05 (s, 3H), 1.42 (s, 9H). 13 C-NMR (90MHz, DMSO): δ = 168.44, 161.51, 157.84, 27.77, 23.72, 13.92. R f =0.62 (hexane / EtOAc 3:1). HPLC (10-90% in 15 min): t R =7.13min,MS(ESI):m / z=232.7[M+H] + ,176.8[M-tBu+H] + .

[0514] Tert-butyl-((2-acetamidoacetamido)(methylthio)methylene)carbamate (B5)

[0515] B1 (130 mg, 0.68 mmol, 1.0 eq) was dissolved in 4 ml DMF, 1 ml DMF solution containing B2 (95 mg, 0.82 mmol, 1.2 eq), HATU (312 mg, 0.82 mmol, 1.2 eq) and DIPEA (230 μl, 1.36 mmol, 2.0 eq) was added and stirred at room temperature for 1 hour. After evaporation of the solvent, the compound was purified by flash chromatography (EtOAc / MeOH 50:1). B5 (153 mg, 0.44 mmol, 78%) was obtained as a white solid.

[0516]

[0517] Rf = 0.28 (EtOAc / MeOH 50:1). HPLC (10-90% in 15 min): t R =5.84min.MS(ESI):m / z=289.8

[0518] [M+H] + ,233.8[M-tBu+H] + ,189.9[M-Boc+H] + .

[0519] Tert-butyl ((6-acetamidohexanamido)(methylthio)methylene)carbamate (B6)

[0520] B1 (150 mg, 0.8 mmol, 1.0 eq) was dissolved in 4 ml DMF, 1 ml DMF solution containing B3 (166 mg, 0.96 mmol, 1.2 eq), HATU (365 mg, 0.96 mmol, 1.2 eq) and DIPEA (270 μl, 1.6 mmol, 2.0 eq) was added and stirred at room temperature for 1 hour. After evaporation of the solvent, the compound was purified by flash chromatography (EtOAc / MeOH 50:1). B6 (210 mg, 0.56 mmol, 76%) was obtained as a white solid.

[0521]

[0522] 1 H-NMR (360MHz, DMSO): δ = 7.77 (t, 1H), 2.99 (dt, 2H), 2.33 (t, 2H), 2.28 (s, 3H), 1. 77(s,3H),1.54-1.48(m,2H),1.42(s,9H),1.37-1.34(m,2H),1.27-1.21(m,2H). 13 C-NMR (90MHz, DMSO): δ = 175.8, 169.4, 161.6, 157.7, 80.4, 38.5, 33.4, 28.9, 27.9, 24.8, 24.2, 22.7, 14.0. R f =0.40 (EtOAc / MeOH 50:1). HPLC (10-90% in 15 min): t R =7.00min,MS(ESI):m / z=368.0[M+Na] + ,346.0[M+H] + ,246.1[M-Boc+H] + .

[0523] Tert-butyl-((methylthio)(pent-4-ynylamino)methylene)carbamate (B7)

[0524] B1 (100 mg, 0.52 mmol, 1.0 equivalent) was dissolved in 4 ml DMF, 1 ml DMF solution containing 4-pentynoic acid (61 mg, 0.62 mmol, 1.2 equivalent), HATU (230 mg, 0.6 mmol, 1.2 equivalent) and DIPEA (170 μl, 1.0 mmol, 2.0 equivalent) was added, and stirred at room temperature for 1 h. After evaporation of the solvent, the compound was purified by flash chromatography (DCM / EtOAc 2:1). B7 (111 mg, 0.41 mmol, 79%) was obtained as a white solid.

[0525]

[0526] 1 H-NMR (360MHz, DMSO): δ = 11.17 (bs, 1H), 2.79-2.77 (m, 1H), 2.57 (t, J = 7.1Hz, 2H), 2.41-2.35 (m, 2H), 2.28 (s, 3H), 1.43 (s, 9H). 13 C-NMR (90 MHz, DMSO): δ = 170.0, 161.7, 158.3, 83.4, 80.6, 72.1, 35.3, 28.2, 14.4, 13.7. Rf = 0.87 (DCM / EtOAc 2:1). HPLC (10-90% in 15 minutes): t R =9.24min,MS(ESI):m / z=271.1[M+H] + ,171.0[M-Boc+H] + .

[0527] 7. Synthesis of Cyclic Pentapeptide Analogs with Acylated Hexylguanidino Residues (P7-P10)

[0528] Guanidinization of P0 was carried out according to GP11 with acylated precursors (B4-B7, 1.3 equivalents). After the reaction was completed, the peptide was precipitated in a saturated NaCl solution and treated with a solution containing 90% TFA, 5% DCM, 2.5% H2O and 2.5% TIPS to remove residual acid-labile side chain protecting groups. Compounds P7-P10 were subsequently purified by semi-preparative HPLC.

[0529]

[0530]

[0531] 8. Synthesis of D-Cys-for-Gly Substituted Peptide Scaffold Pc0

[0532] Peptide R was synthesized similarly to that described previously. 55 , cyclo(ya'(hexylguanidino)Rnalc) (Pc0) was prepared. After assembling the pentapeptide on HD-Cys-(2-Cl)Trt resin using solid phase peptide synthesis, including N-alkylation of the D-ala residue, the fully protected peptide was cleaved from the resin and cyclized using diphenylphosphoryl azide. After selective removal of the Dde protecting group on the hexylamino side chain according to GP8, solution phase guanidination was performed using 1-H-pyrazole-1-carboxamidine (10 equiv) in DMF and DIPEA (20 equiv). Subsequently, the peptide was treated with trifluoroacetic acid, triisopropylsilane (TIPS) and water (95, 2.5, 2.5; v / v / v) for 40 min and precipitated in diethyl ether to obtain crude Pc0 as a light yellow powder with sufficient purity for further functionalization.

[0533]

[0534] HPLC (10-90% B in 15 minutes): t R =11min.MS(ESI):m / z=832.7[M+H] + ,854.6[M+Na] + ,416.9[M+2H] 2+

[0535] 9. Synthesis of Linker Moieties for D-Cys-Functionalization

[0536] 1-(Fmoc),4-(2-chloroacetyl)-phenylenediamine (C1)

[0537] 1,4-(Boc)-phenylenediamine (460 mg, 1 eq) was dissolved in 2 mL of tetrahydrofuran (THF) and 5 mL of 10% Na2CO3 was added. The resulting suspension was gradually added to a solution of Fmoc-Cl (500 mg, 1 eq) in THF (2 mL) at 0°C. After the reaction was complete, the reaction mixture was diluted with water and THF. The organic solvent was then evaporated in vacuo to precipitate the product, which was separated by centrifugation, washed with water and dried in vacuo. To remove the Boc protecting group, the dried product was redissolved in TFA / CH2Cl2. After 10 minutes at room temperature, the solvent was evaporated in vacuo to give 1-(Fmoc),4-phenylenediamine as a yellow oil. The crude product was then dissolved in anhydrous CH2Cl2 and DIPEA (2.5 eq) and 2-chloroacetyl chloride (1 eq) were added. After 30 minutes at room temperature, the solvent was evaporated in vacuo and the product was recrystallized from ethyl acetate to give a white solid product.

[0538]

[0539] HPLC (40-100% in 15 minutes): t R =12.26min.MS(ESI):m / z=457.1[M+Na] + .

[0540] 1-(Fmoc)-3-(2-chloroacetyl)-phenylenediamine (C2)

[0541] The synthesis of C2 was carried out essentially as described for C1. However, purification of the final product was carried out using SiO2 column chromatography and a 1:1 mixture of ethyl acetate / petroleum ether (v / v) as eluent.

[0542]

[0543] HPLC (40-100% in 15 minutes): t R=12.42min.MS(ESI):m / z=457.1[M+Na] + .

[0544] Fmoc-(2-chloroacetyl)Lys-CO-NH2(C3)

[0545] Fmoc-Lys-CO-NH2 (125 mg, 1 eq) was dissolved in CH2Cl2, and DIPEA (2.5 eq) and 2-chloroacetyl chloride (1 eq) were added. After 30 min at room temperature, the product precipitated from the reaction mixture. After filtration, the crude product was recrystallized from ethyl acetate and lyophilized.

[0546]

[0547] HPLC (40-100% in 15 minutes): t R =9.68min.MS(ESI):m / z=443.9[M+H] + ,466.1[M+Na] + .

[0548] 1-Fmoc-6-(2-chloroacetyl)-hexanediamine (C4)

[0549] Fmoc-1,6-hexanediamine (200 mg, 1 eq.) was dissolved in acetonitrile / methanol, and DIPEA (2.5 eq.) and 2-chloroacetyl chloride (1.7 eq.) were added. After 60 min at room temperature and cooling to -20 °C for several hours, the crude product precipitated from the reaction mixture. After filtration, the white crystals were dried and used as is without further purification.

[0550]

[0551] HPLC (40-100in%15min):t R =10.02min.MS(ESI):m / z=437.2[M / 2+H] + .

[0552] 10. Synthesis of Acetylated D-Cys-for-Gly Substituted Peptides Pc1-Pc5

[0553] Cyclo[ya'(hexylguanidino)RNalc(CH2-CO-NH-(CH2)4-CH(CO-NH2)-NH-Ac)](Pc1)

[0554] Alkylation of Pc0 and C3 was performed according to GP17. The crude product was then redissolved in piperidine / DMF (80 / 20, v / v) to remove the Fmoc protecting group on the linker unit. After 10 minutes at room temperature, the deprotected peptide was precipitated in water, washed and dried. Acetylation of the free amino functional group was achieved by dissolving the peptide in a solution of acetic anhydride, DIPEA and DMF (10:5:85; v / v / v) and stirring at room temperature for 1 hour. After precipitation in water, Pc1 was purified using preparative HPLC.

[0555]

[0556] HPLC (15-45% in 15 minutes): t R =6.8min.MS(ESI):m / z=1059.8[M+H] + ,1081.8[M+Na] + ,1097.6

[0557] [M+K] +

[0558] Cyclo[ya'(hexylguanidino)RNalc(CH2-CO-NH-(CH2)6-NH-Ac)](Pc2)

[0559] Alkylation of Pc0 with C4 was performed according to GP17. Subsequent Fmoc-deprotection and N-acetylation were performed as described for Pc1.

[0560]

[0561] HPLC (15-45% in 15 minutes): t R =9min.MS(ESI):m / z=1030.7[M+H] + ,1052.6[M+Na] + .

[0562] Cyclo[ya'(hexylguanidino)RNalc(-(CH2)6-NH-Ac)](Pc3)

[0563] Alkylation of Pc0 with Boc-6-aminohexyl bromide was performed according to GP17. This was followed by Boc-deprotection according to GP9 and N-acetylation as described for PC1.

[0564]

[0565] HPLC (15-45% in 15 minutes): t R =10min.MS(ESI):m / z=973.6[M+H] + ,995.5[M+Na] +

[0566] Cyclo[ya'(hexylguanidino)RNalc(-CH2-CO-NH-CH2-(p-aryl)-CH2-NH-Ac)](Pc4)

[0567] Alkylation of Pc0 with C1 was performed according to GP17. Subsequent Fmoc-deprotection and N-acetylation were performed as described for Pc1.

[0568]

[0569] HPLC (15-45% in 15 minutes): t R=8.7min.MS(ESI):m / z=1050.6[M+H] + ,1072.6[M+Na] + ,525.8

[0570] [M+2H] 2+ .

[0571] Cyclo[ya'(hexylguanidino)RNalc(-CH2-CO-NH-CH2-(m-aryl)-CH2-NH-Ac)](Pc5)

[0572] Alkylation of Pc0 with C2 was performed according to GP17. Subsequent Fmoc-deprotection and N-acetylation were performed as described for Pc1.

[0573]

[0574] HPLC (15-45% in 15 minutes): t R =8.8min.MS(ESI):m / z=1050.8[M+H] + ,1072.7[M+Na] + ,525.9

[0575] [M+2H] 2+ .

[0576] Cyclo[ya'(hexylguanidino)RNalc(-CH2-CO-NH-CH2-(m-aryl)-CH2-NH-DOTA)](Pc5a)

[0577] Alkylation of Pc0 and C2 was performed according to GP17. The crude product was then redissolved in piperidine / DMF (80 / 20, v / v) to remove the Fmoc protecting group on the linker unit. After 10 minutes at room temperature, the deprotected peptide was precipitated in water, washed and dried. Coupling of DOTA to free amino functional groups was performed according to GP14.

[0578]

[0579] HPLC (15-35% in 15 minutes): t R =11.9min.MS(ESI):m / z=1396.1[M+H] + ,698.4[M+2H] 2+ .

[0580] Cyclo[a'(hexylguanidino)RNalc(-CH2-CO-NH-CH2-(m-aryl)-CH2-NH- nat Ga-DOTA)](Pc5b)

[0581] Preparation of Pc5a according to GP15 nat Ga-complex.

[0582] HPLC (15-35% in 15 minutes): t R =12.5min.MS(ESI):m / z=1463.0[M+H] +,732.1[M+2H] 2+ .

[0583] Cyclo[a'(hexylguanidino)RNalc(-CH2-CO-NH-CH2-(m-aryl)-CH2-NH- nat Lu-DOTA)](Pc5c)

[0584] Preparation of Pc5a according to GP15 nat Lu-complex.

[0585] HPLC (15-35% in 15 minutes): t R =11.7min.MS(ESI):m / z=1567.0[M+H] + ,784.5[M+2H] 2+ .

[0586] Cyclo[a'(hexylguanidino)RNalc(-CH2-CO-NH-CH2-(m-aryl)-CH2-NH- nat Cu-DOTA)](Pc5d)

[0587] Preparation of Pc5a according to GP15 nat Cu-complex.

[0588] HPLC (15-35% in 15 minutes): t R =12.9min.MS(ESI):m / z=1456.1[M+H] + ,729.1[M+2H] 2+ .

[0589] Coating of carboxyl-QDs with Pc5 (Pc5-QD)

[0590] Alkylation of Pc0 with C2 was performed according to GP17. The crude product was then redissolved in piperidine / DMF (80 / 20, v / v) to remove the Fmoc protecting group on the linker unit. After 10 min at room temperature, the deprotected peptide was precipitated in water, washed, dried and lyophilized. The carboxyl quantum dots (Life technologies, 655ITK TMCarboxyl quantum dots) (0.8 nmol in 100 μl 50 mM borate buffer (pH = 8.3)) to a total volume of 920 μl. Then, 1-ethyl-3-(3-dimethylamino-propyl) carbodiimide (800 equivalents, 640 nmol) in 12.2 μl 10 mM borate buffer and the deprotected peptide (40 equivalents, 32 nmol) dissolved in 64 μl 10 mM borate buffer were added. The consumption of unreacted peptide in the reaction mixture was monitored by RP-HPLC. After the coupling reaction was completed (30-60 minutes at room temperature), the excess coupling agent was removed from the Pc5-QD suspension by repeating the buffer exchange using 50 mM borate buffer (pH = 8.3) and an Amicon Ultra-4 ultrafiltration unit (4 mL, Millipore) with a cutoff value of 100 kDa. Final reconstitution with 50 mM borate buffer yielded the peptide-coated quantum dots Pc5-QD as a 4 μM suspension. Based on the chosen stoichiometry, the coverage of the quantum dots was calculated to be approximately 40% of the reacted carboxylate.

[0591] Example 3

[0592] Other Materials and Methods

[0593] 1. Radiolabeling

[0594] 1.1. Radioiodine labeling

[0595] use Methods All peptides were radioiodinated. Briefly, 100-200 μg of peptide was dissolved in 5-10 μL DMSO. The solution was diluted with 0.5 mL TRIS iodination buffer (25 mM Tris·HCl, 0.4 M NaCl, pH 7.5) and transferred to a 150 μg Coated Eppendorf reaction tube. Add [ 125 I]NaI (18-20 MBq, HartmannAnalytic, Braunschweig, Germany) or [ 123I] NaI (220MBq, GE Healthcare, Braunschweig, Germany), the reaction vessel was briefly vortexed, and the labeling reaction was allowed to proceed at room temperature for 15 minutes. The peptide solution was then removed from the insoluble oxidant. Gradient RP-HPLC was used to achieve separation of the labeled product from the unlabeled precursor. For in vitro binding studies, the HPLC product fractions were used as is and diluted to the desired concentration using their respective assay media. For biodistribution experiments, each product fraction was diluted with water and sent to a SepPak Plus C-18 column (Waters, Eschborn, Germany). The column was washed with water, and then 1ml of acetonitrile was used to elute the immobilized radioactive peptide. The solvent was removed by bubbling an argon stream through the radioligand solution for 20 minutes at 90°C. The radioiodine-labeled peptide was then reconstructed to an activity concentration of approximately 1Mbq / 100 μL using PBS and used in vivo animal studies.

[0596] 1.2. 68 Ga-labeling

[0597] The SnO2 matrix (iTHEMBA LABS, South Africa) was eluted with 1 M HCl (5.5 mL). 68 Ge / 68 Ga generator acquisition 68 Ga was immobilized on a strong cation exchange column (SCX-Chromafix, size M, Macherey-Nagel, Düren, Germany).

[0598] For animal studies, similar to previously published 68 Ga labeling operation 56 (SCINTOMICS GmbH, Germany) in Gallelut + System preparation 68 Ga-pentixafor. In short, 68 The Ga generator elution fraction (1.25 mL, 600-800 MBq, buffered to pH 3.3 with 900 μL of 14.4 g HEPES in 12 mL water) was reacted with 3.5 nmol of the respective DOTA-peptide (P5b, Pc5a) for 5 minutes. The radiochemical purity was always >99% as determined by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). 1 mL PBS was added and the mixture was concentrated in vacuo to a total volume of 1 mL to obtain a solvent-free preparation with a specific activity of 100-150 MBq / nmol.

[0599] 1.3. 177 Lu-labeled

[0600] for 177Lu-labeled, the respective DOTA peptides (P5b, Pc5a) were dissolved in water to obtain a 1 mM solution. The required volume of this solution was added to 177 LuCl3 in 0.04 M HCl (itg Isotope Technologies Garching, Garching, Germany; activity concentration: 370 Mbq / 500 μl) was used to allow the peptide to react with 177 The activity ratio of Lu is 1 nmol peptide / 225 MBq 177 LuCl3. 1M NH4OAc (calculated as 10% of the total reaction volume) was added to the mixture and the mixture was heated to 95°C for 20 minutes. After cooling, the radiochemical purity was determined using thin layer chromatography (TLC) (usually > 98%). For in vitro and in vivo studies, the reaction mixture was diluted to the desired activity concentration with PBS and used in the experiment as is.

[0601] 2. Lipophilicity Determination

[0602] 500 μL of octanol (n=6) was added to a solution of about 2 kBq radiolabeled peptide in 500 μL PBS (pH 7.4). The vial was vortexed vigorously for 3 minutes. In order to achieve quantitative phase separation, the vial was centrifuged at 14,600 × g for 6 minutes in Biofuge 15 (Heraeus Sepatech, Osterode, Germany). The activity concentration in 100 μL samples of both the aqueous phase and the organic phase was measured in a γ-counter. The distribution coefficient P was calculated. OW and log P OW , the distribution coefficient P OW Defined as the molar concentration ratio of a single species A between octanol and water at equilibrium, the log P OW It is an important parameter used to characterize the lipophilicity of a compound.

[0603] 3. In vitro evaluation

[0604] 3.1. Cell culture

[0605] Jurkat human T lymphocytes were maintained in RPMI 1640 medium supplemented with 10% fetal calf serum (FCS). Daudi cells (human Burkitt's B cell lymphoma) were grown in RPMI-1640 medium supplemented with 10% FCS, 2 mM L-glutamine, 1% non-essential amino acids (NEA), 50 μM β-mercaptoethanol, and 100 units / mL penicillin / streptomycin. Chem-1 cells stably transfected with hCXCR4 (HTS004C ChemiScreen TMCells (Merck Millipore, Darmstadt, Germany) were cultured in DMEM supplemented with 10% FCS, 1% NEA, and 1% HEPES (1 M). Eμ-Myc1080 mouse B cell lymphoma cells 57 C6 rat glioma cells (kindly provided by Prof. Rolf Mentlein, Institute of Anatomy, University of Kiel, Germany) were cultured in DMEM medium containing 10% FCS, 2 mM L-glutamine and 100 units / mL penicillin / streptomycin.

[0606] All cell lines were cultured at 37°C in a humidified 5% CO2 atmosphere. Culture media and supplements were obtained from Biochrom (Berlin, Germany) or Gibco (life technologies, Darmstadt, Germany). In the assay medium used for internalization studies, 5% bovine serum albumin (BSA; Sigma, St. Louis, USA) was used instead of FCS. Cells were counted using a Countesse automated cell counter (Invitrogen, Carlsbad, USA).

[0607] 3.2. IC 50 Determination of

[0608] To determine hCXCR4-affinity, Jurkat cells and [ 125 I]FC131 or 125 I]R1 was used as a radioligand for competition binding experiments. Eμ-Myc1080 mouse B cell lymphoma cells were used 57 or C6 rat glioma cells, and [ 125 I]R1 was used as a radioligand to measure its affinity to mCXCR4 and rCXCR4.

[0609] For IC with Jurkat or Eμ-Myc1080 cells 50 The determination was carried out at increasing concentrations (10 -11 Up to 10 -5 M) of each target peptide (n = 3 / concentration), 2×10 5 cells (or 4×10 5The cells; the exact number of cells used in each evaluation is given in the results section) were incubated with 100.000 cpm of each radioligand (about 0.1 nM). The total sample volume was 250 μL. After incubation for 120 minutes with gentle stirring (200 mot / min) at room temperature (RT), the tubes were centrifuged (5 min, 450 g, Megafuge 1.0, Heraeus Thermo Scientific) and the supernatant was carefully removed. After washing twice with 400 μL cold HBSS, the amount of cell-bound radioligand was quantified using a γ-counter.

[0610] In the case of adherent C6 glioma cells, cells were harvested with trypsin / EDTA (0.05% and 0.02%) in PBS the day before the experiment, centrifuged and resuspended in culture medium to a concentration of approximately 150.000 cells / mL. The suspension was transferred to a PLL-coated 24-well plate (1 mL / well, Greiner, Solingen, Germany) and placed in an incubator overnight. On the day of the experiment, the culture medium was removed, the cells were washed with 250 μL HBSS, and then equilibrated in 200 μL HBSS (1% BSA) at 37°C for a minimum of 15 minutes before the experiment. Then, at increasing concentrations (10 -11 Up to 10 -5 Cells were incubated with 100.000 cpm of each radioligand (about 0.1 nM) in the presence of 100.000 cpm of each target compound (n=3 / concentration). The total sample volume was 250 μL. After incubation for 120 minutes at room temperature (RT), the supernatant was removed and the cells were washed twice with 200 μL of cold HBSS. Then, the cells were lysed with 250 μL of 1N NaOH, the lysate was transferred to a vial and combined with 250 μL of HBSS for flushing the wells. Quantification of free and bound activity was performed in a γ-counter.

[0611] IC 50 Values ​​were calculated using PRISM 6 software (Graph Pad Software, San Diego, CA) using a three-parameter sigmoidal dose-response curve fit.

[0612] 3.3. Internalization study of hCXCR4 in Chem-1 cells

[0613] On the day of the experiment, the culture medium was removed, the cells were washed with 250 μL HBSS (1% BSA), and then balanced at least 15 minutes in 200 μL assay culture medium (RPMI+5% BSA) at 37°C before the experiment. Then, 25 μL / well assay culture medium (control) or 1 mM AMD3100 HBSS solution (non-specific binding assay) (n=3 for each time point), followed by 25 μL target radioligands in the assay culture medium. In all internalization tests, the final radioligand concentration was 0.2-1 nM.

[0614] At 37 ℃, incubate different time points until 60 minutes, remove incubation medium, rinse cells twice with 250 μ L HBSS.Washing medium is merged with the supernatant of the previous step.This part represents the amount of free radioligand.In order to remove the (acid releasable) radioactivity of surface binding, cells are then incubated with 250 μ L ice-cold acid wash buffer (0.02M NaOAc is buffered to pH=5 with AcOH).After removing the acid wash buffer, thoroughly rinse cells with another 250 μ L ice-cold acid wash buffer.The two acid wash parts are merged.Release internalization activity by incubation with 250 μ L 1N NaOH, and transfer to a vial and merge with 250 μ L PBS for flushing wells.The amount of free, acid releasable and internalized activity is quantified in a γ-counter.

[0615] 3.4. Cellular binding and internalization studies using fluorescent ligands and coated QDs

[0616] 24 hours before the experiment, Chem-1 cells (125.000 / well) were seeded on glass coverslips in well plates. Then, the culture medium was removed, the cells were washed with 250 μL HBSS (1% BSA), and then equilibrated in 200 μL assay medium (DMEM / Ham's F12+5% BSA) at 4°C or 37°C for a minimum of 15 minutes before the experiment. In the case of Daudi or Eμ-Myc 1080 cells, a suspension of each cell in assay medium (DMEM / Ham's+5% BSA, 100.000 cells / 200 μl) was prepared. Then, the cells were incubated with the respective fluorescent ligand alone, or with the fluorescent ligand (final ligand concentrations are specified in the figure legend), 2 μg / ml Hoechst (1 mg / ml, 0.5 μl) and 2 μM Lysotracker Green (1 mM, 0.5 μl) for 1 hour at 4°C, room temperature or 37°C, respectively. Nonspecific binding was determined in the presence of 100 μM AMD3100. After incubation, cells were washed twice with ice-cold PBS, immediately before fluorescence microscopy images were taken.

[0617] 4. In vivo evaluation

[0618] To determine general tracer pharmacokinetics, in vivo biodistribution studies were optionally performed in Black Six or CD-1 normal mice. For the most promising compounds, additional biodistribution studies were performed in CB-17-SCID mice bearing Daudi (human B cell lymphoma) xenografts.

[0619] In general, mice were injected intravenously (iv) with the respective radioligand (0.4-3.7 MBq in 100 μL PBS (pH 7.4)) under isoflurane anesthesia. Animals (n=3-5) were sacrificed 60 minutes after injection (pi) and the target organs were dissected. The radioactivity in the weighed tissue samples was measured using a γ-counter. Data are expressed as % ID / g tissue (mean ± SD).

[0620] In order to determine the CXCR4-specificity of tracer accumulation, competition studies were performed in parallel. The species-independent CXCR4 antagonist AMD-3100 (50 μg per mouse, dissolved in 10 μL PBS) was co-injected with the radioligand. Animals (n=3-4) were sacrificed 60 minutes after injection, and then the activity accumulation in the target organ was determined as described above.

[0621] Example 4

[0622] result

[0623] 1. Lipophilicity

[0624] Table 1: Lipophilicity (log P O / PBS )

[0625]

[0626] 2. Determination of CXCR4 Affinity

[0627] Table 2a: Binding affinities (IC50 in nM) of novel CXCR4 ligands for human CXCR4 (hCXCR4). Jurkat human T-cell leukemia cells (200.000 cells / sample) and [ 125 I] FC-131 was used as a radioligand to determine affinity. Each experiment was performed in triplicate, and the results are the mean ± SD from three independent experiments.

[0628]

[0629]

[0630] Table 2b: Binding affinities (IC50 in nM) of novel CXCR4 ligands for human CXCR4 (hCXCR4). Jurkat human T-cell leukemia cells (400.000 cells / sample) and [ 125 I] FC-131 was used as a radioligand to determine affinity. Each experiment was performed in triplicate, and the results are the mean ± SD from three independent experiments.

[0631]

[0632] Table 3: Binding affinities (IC50 in nM) of novel CXCR4 ligands for human, mouse and rat CXCR4 (hCXCR4, mCXCR4 and rCXCR4). Jurkat human T-cell leukemia (hCXCR4), Eμ-Myc1080 mouse B-cell lymphoma and C6 rat glioblastoma (200.000 cells / sample) and [ 125 I]R1 was used as the radioligand to determine affinity. Each experiment was performed in triplicate and the results are the mean ± SD from three independent experiments.

[0633]

[0634]

[0635] 3. Fluorescence Microscopy

[0636] Chem-1 cells stably transfected with hCXCR4 were incubated with the respective Sulfo-Cy5-conjugated peptides for 1 h at 4°C or 37°C. The ligand concentrations used in the respective experiments are given in the figure legends. Nonspecific ligand binding was determined in the presence of AMD3100 (100 μM). Nuclei were stained with Hoechst (2 μg / ml) and ligand internalization into endosomes and lysosomes was confirmed by co-staining with lysotracker green (2 μM). See Figures 2 to 8 .

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Claims

1. A compound of the following formula (IV) or a pharmaceutically acceptable salt thereof, in: R 1 It is methyl; R 4 is H; R 5 It is H; R 6 is represented by at least one selected from -NH-R B 、-NH-C(=NH)-NH-R B 、-NH-C(=NH)-NH-R 14 and -NH-C(=NH)-NH-C(=O)-R 15 An alkyl group substituted with a substituent; R 14 and R 15 are independently C1-C10 alkyl groups, which may be substituted by at least one substituent selected from -NH-C(=O)-CH3 and -C≡CH; R B is a group comprising at least one selected from the following: (i) a chelating moiety, (ii) a chelate formed by a chelating moiety and a chelated radioactive or non-radioactive cation, and (v) quantum dots, And R B It may also contain a group of formula (II) Its use The indicated bond is bound to the rest of the compound, where R 1 To R 5 and R 7 To R 10 As defined above, and wherein R 16 is an alkanediyl group, preferably a C1-C6 alkanediyl group, and R 17 is selected from -NH- and -NH-C(=NH)-NH-; And among them The compound of formula (I) comprises at least one selected from -SR A 、-NH-R B and -NH-C(=NH)-NH-R B substituent.

2. The compound or salt of claim 1, wherein the chelated cation is selected from the group consisting of Cu, Ga and Lu cations.

3. The compound or salt of claim 2, wherein the chelated cation is selected from 64 Cu, 67 Cu, 68 Ga and 177 The cation of Lu.

4. The compound or salt according to any one of claims 1 to 3, wherein the chelating moiety of (i) or (ii) is a residue obtainable by forming an amide bond from a carboxyl group contained in DOTA.

5. The compound or salt of any one of claims 1 to 4, wherein R B Is the formula -L B1 -R B1 A group selected from the group of formula (VIa) to (VIf): -R 18 -NH-R B1 (VIa), -R 18 -NH-(C(=O)-R 19 -NH) u -R B1 (VIb), -R 18 -NH-(C(=O)-R 20 -NH) v -R B1 (VIc), -(C=O)-R 21 -NH-R B1 (VId), -(C=O)-R 21 -NH-(C(=O)-R 22 -NH) w -R B1 (VIe), -(C=O)-R 21 -NH-(C(=O)-R 23 -NH) x -R B1 (VIf), in: R 18 is a C1-C10 alkanediyl group, preferably a straight-chain alkanediyl group, more preferably a straight-chain C2-C6 alkanediyl group, and most preferably a straight-chain C6 alkanediyl group, R 19 is a C1-C10 alkanediyl group, preferably a straight-chain alkanediyl group, more preferably a straight-chain C2-C6 alkanediyl group, u is 1, 2 or 3, preferably 1 or 2, R 20 is an oligoether moiety, preferably a linear oligoether moiety, more preferably a linear oligoether moiety having 5 to 10 carbon atoms and 2 or 3 oxygen atoms providing ether bonds, v is 1, 2 or 3, preferably 1 or 2, R 21 is a C1-C10 alkanediyl group, preferably a straight-chain alkanediyl group, more preferably a straight-chain C2-C6 alkanediyl group, and most preferably a straight-chain C6 alkanediyl group, R 22 is a C1-C10 alkanediyl group, preferably a straight-chain alkanediyl group, more preferably a straight-chain C2-C6 alkanediyl group, w is 1, 2 or 3, preferably 1 or 2, R 23 is an oligoether moiety, preferably a linear oligoether moiety, more preferably a linear oligoether moiety having 5 to 10 carbon atoms and 2 or 3 oxygen atoms providing ether bonds, x is 1, 2 or 3, preferably 1 or 2; And R B1 is a group selected from the following or contains a group selected from the following (i) a chelating moiety, (ii) a chelate formed by the chelating moiety (i) and a chelated radioactive or non-radioactive cation, and (v) quantum dots, And R B1 It may also contain a group of formula (II) as defined in claim 1.

6. A pharmaceutical composition for use in therapy or diagnosis, comprising or consisting of a compound or salt according to any one of claims 1 to 5.

7. Use of the compound or salt according to any one of claims 1 to 5 in the preparation of a medicament for nuclear medicine, nuclear molecular imaging, optical imaging, or targeted intracavitary radiotherapy.

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