Compounds for fast and efficient click release

Abstract

To provide a compound capable of achieving quick and efficient click release.SOLUTION: A compound satisfying the following formula (19) and a pharmaceutically acceptable salt thereof are provided.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The invention disclosed herein relates to compounds, combinations, kits for use in bioorthogonal release reactions and methods of using same. [Background technology]

[0002] Selective chemical reactions that are orthogonal to the diverse functions of biological systems are called bio-orthogonal reactions and occur between two abiotic groups with exclusive mutual reactivity. These can be used to selectively modify biochemical structures, such as proteins or nuclei. They typically proceed in water and at near-ambient temperatures and can be applied to complex chemical environments, such as those found in living organisms.

[0003] Bioorthogonal reactions are widely useful tools with applications spanning chemical synthesis, materials science, chemical biology, diagnostics, and medicine. Particularly prominent application areas for bioorthogonal reactions include drug delivery agents and prodrugs for pharmaceutical applications, as well as a variety of reversible bioconjugates and advanced spectroscopic bioprobes for applications in the field of bioanalysis.

[0004] One prominent bioorthogonal reaction is the inverse-electron-demand Diels Alder (IEDDA) reaction between trans-cyclooctene (TCO) and tetrazine (TZ). In a previous study, the IEDDA reaction was used for pretargeted radioimmunoimaging, in which tumor-bearing mice were treated with trans-cyclooctene (TCO)-tagged antibodies or antibody fragments. One or more days later, a radiolabeled tetrazine probe was administered and selectively bound to the TCO tag of the tumor-bound antibody [R. Rossin, MS Robillard, Curr. Opin. Chem. Biol. 2014, 21, 161-169].

[0005] Based on IEDDA conjugation, a release reaction has been developed. This release reaction, called IEDDA pyridazine elimination, is a "click-to-release" approach that allows instantaneous and selective release depending on the conjugation [RM Versteegen, R. Rossin, W. ten Hoeve, HM Janssen, MS Robillard, Angew. Chem. Int. Ed. 2013, 52, 14112-14116]. The IEDDA reaction between a tetrazine (i.e., a diene) and an alkene (i.e., a dienophile) gives 4,5-dihydropyridazine, which typically tautomerizes to 1,4- and 2,5-dihydropyridazine. The 1,4-dihydropyridazine product derived from a TCO containing tetrazine and allylic carbamate-linked doxorubicin (Dox) has been demonstrated to be prone to CO2 and Dox elimination via an electronic cascade mechanism, ultimately producing an aromatic pyridazine. Triggered release has been demonstrated in phosphate-buffered saline (PBS), serum, cell culture, and mice, and is promising for a wide range of applications in medicine, chemical biology, and synthetic chemistry, including triggered drug release, biomolecule uncaging, and capture and release strategies.

[0006] In general, removal of the IEDDA pyridazine allows for controlled manipulation of a variety of substrates in relatively complex environments in the presence of a variety of other chemical functional groups. This control can be temporal, random, and spatial. The manipulation can be versatile, for a variety of purposes, including, but not limited to, activating, deactivating, releasing, capturing, or otherwise altering constructs bound to the chemically cleavable group.

[0007] Removal of the IEDDA pyridazine has been applied to triggered drug release from antibody-drug conjugates (ADCs) capable of participating in the IEDDA reaction (Figure 1). ADCs are a promising class of biopharmaceuticals that combine the targeting specificity of monoclonal antibodies (mAbs) or mAb fragments with the potency of small molecule toxins. Classical ADCs are designed to bind to internalizing cancer cell receptors, leading to ADC uptake and subsequent intracellular release of the drug via enzymes, thiols, or lysosomal pH. Delivering toxins to tumors while minimizing peripheral damage to healthy tissues allows the use of highly potent drugs, resulting in improved therapeutic outcomes. The use of IEDDA pyridazine removal for ADC activation allows targeting of non-internalizing receptors because the drug is cleaved chemically rather than biologically.

[0008] In general, prodrugs, which may include ADCs, are of interest for IEDDA pyridazine elimination reactions in which a drug is deactivated, conjugated, or masked by a moiety and reactivated, released, or unmasked after the IEDDA reaction has occurred.

[0009] Further background art to the aforementioned technology includes International Publication No. WO 2012 / 156919, International Publication No. WO 2012156918A1, International Publication No. WO 2014 / 081303, and US Patent Application Publication No. 20150297741. In this specification, the dienophile, TCO, described above, is used as a chemically cleavable group, for example, a chemically cleavable group in a protecting group in synthetic chemistry, a cleavable linker or mask in chemical biology, in vitro diagnostics, and in vivo prodrug activation. The group is attached to a construct (e.g., a molecule, protein, peptide, polymer, dye, surface) such that release of the dienophile from the construct can be triggered by allowing the dienophile to react with a diene, TZ, described above. The dienophile is an eight-membered non-aromatic cyclic alkenylene or alkenyl group, particularly a TCO group.

[0010] In some applications, the TCO may be part of a prodrug that is first injected into the subject's bloodstream and targeted to a certain part of the body, such as a tumor. A certain percentage of the prodrug is then immobilized at the target site, while another percentage is removed by the body. After several hours or days, an active agent containing tetrazine is administered to release the drug from the prodrug, preferably only at the target site. The tetrazine itself is also subject to clearance by the body at a certain clearance rate.

[0011] Generally, in a first step, a tetrazine reacts with a dienophile-containing construct (e.g., a dienophile-containing prodrug) to form a conjugate. This is called a click conjugation process. The construct is then preferably released from the construct-dienophile (e.g., prodrug) via one or more mechanisms. It will be understood that a high yield in the click conjugation process, i.e., a high click conjugation yield, does not necessarily result in a high yield of the released construct, i.e., a high drug release yield.

[0012] From a bioorthogonal perspective, the chemistry works well.

[0013] However, it is desirable to develop better IEDDA reactions.

[0014] Achieving high construct release yields in IEDDA reactions remains a challenge both in vivo and in vitro. In particular, it is desirable for the reaction between a construct-bearing dienophile and a diene to result in high construct release yields in vitro and / or in vivo.

[0015] Furthermore, achieving fast release of constructs in IEDDA reactions remains a challenge both in vivo and in vitro. In particular, it is desirable for the reaction between a construct-bearing dienophile and a diene to result in high construct release yields in vitro and / or in vivo.

[0016] The motivation for these tetrazines is that they typically have lower reactivity than tetrazines that have been successfully used for in vivo click conjugation. These more reactive tetrazines provide good click conjugation yields but poor click release yields. The aforementioned study [RM Versteegen, R. Rossin, W. ten Hoeve, HM Janssen, MS Robillard, Angew. Chem. Int. Ed. 2013, 52, 14112-14116] showed, for example, that 3,6-bis-(2-pyridyl)-1,2,4,5-tetrazine provided high click conjugation rates and yields, but very poor click release yields of only 7% in PBS / MeCN (3:1) or only 12% in serum.

[0017] Additionally, for tetrazines that provide high release yields, such as 3,6-bis-methyl-1,2,4,5-tetrazine, this release typically takes several hours and typically exhibits release yields of up to 80-90% [RM Versteegen, R. Rossin, W. ten Hoeve, HM Janssen, MS Robillard, Angew. Chem. Int. Ed. 2013, 52, 14112-14116].

[0018] For at least these reasons, it is desirable to improve the click release rate and click release yield of tetrazine motifs that have relatively high reactivity (i.e., high click conjugation yield) toward dienophiles in vitro and / or in vivo.

[0019] Another desire is to improve the click release rate and possible click release yield of tetrazines that have relatively good click conjugation and / or release yield in vitro and / or in vivo.

[0020] Another desire is to improve the click release rate and preferably also the click release yield of tetrazines that have low reactivity towards dienophiles in vitro and / or in vivo.

[0021] Another desire is to achieve a combination of high click conjugation yields of construct-bearing dienophiles and high construct release yields both in vitro and in vivo.

[0022] Another desire is to achieve a combination of high click conjugation reaction rates with dienophiles, high click conjugation yields between the dienophiles and tetrazines bearing constructs, and high construct release yields, preferably both in vitro and in vivo.

[0023] Previous studies have attempted to address this desire by engineering dienes rather than dienophiles to provide conjugation yields, or increased release yields / rates, or both.

[0024] Previous work ([R. Rossin, SM J van Duijnhoven, W. ten Hoeve, HM Janssen, LH J Kleijn, FJ M Hoeben, RM Versteegen, MS Robillard, Bioconj. Chem., 2016, 27, 1697-1706]) has shown that conjugation of 10 kDa dextran to 3-methyl-6-(2-pyridyl)-tetrazine or 3-methyl-6-(methylene)-tetrazine resulted in high click conjugation yields with TCO in vivo, but suboptimal drug release yields both in vitro and in vivo.

[0025] Another publication aimed at in vitro reactions ([Fan et al., Angew. Chem. Int. Ed., 2016, 55, 14046-14050]) showed that small, asymmetric tetrazines conjugated more efficiently with TCO than symmetric bis-alkyltetrazines, but the click-release yields were not quantitative.

[0026] Two other publications [Carlson et al., J. Am. Chem. Soc. 2018, 140, 3603-3612] and [Sarris et al., Chemistry 2018, 24, 18075-18081] showed that bis-alkyltetrazines bearing carboxylate or amine groups can enhance click release rates, but that these tetrazines remain relatively unreactive in click conjugation with TCO. Summary of the Invention [Problem to be solved by the invention]

[0027] It would be desirable to develop compounds that address one or more of the above-mentioned problems and / or needs. [Means for solving the problem]

[0028] In one aspect, the present invention relates to compounds satisfying the following formula (19) and pharmaceutically acceptable salts thereof: [ka] where: R 48 is -OH, -OC(O)Cl, -OC(O)ON-succinimidyl, -OC(O)O-4-nitrophenyl, -OC(O)O-tetrafluorophenyl, -OC(O)O-pentafluorophenyl, -OC(O)-(S P ) k C A , -OC(S)-(S P ) k C A , -SC(O)-(SP ) k C A , -SC(S)-(S P ) k C A , -O-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A , -S-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A , and -(S P ) k C A selected from the group consisting of: r is an integer ranging from 0 to 2; each s is independently 0 or 1; each i is independently an integer ranging from 0 to 4, preferably 0 or 1; j is an integer ranging from 0 to 4, preferably 0 or 1; each k is independently 0 or 1; L C is a self-immolative linker, and S P is a spacer; Each C A and C B are independently selected from the group consisting of organic molecules and inorganic molecules; Here, at least one of the following conditions (a) to (c) is satisfied: (a)X 1 , X 2 , X 3 , X 4 , X 5 At least one of the 47 Y T1 and Y T1 is H a positioned in cis relative to ; (b)X 3 is Y T3 and (c)X 1 , X 2 , X 3 , X 4 , X 5 are part of a fused ring satisfying one of the following formulas (20a) to (20g): [ka] Y T2 is positioned syn relative to the eight-membered dienophile ring; where X a and X b is X a -X b or X b -X a is X 1 -X 2 , X 2 -X 3 , X 3 -X 4 , or X 4 -X 5 is part of the eight-membered ring of formula (19) such that Regarding formulas (20a) to (20c), X a and X b is CR 47 , preferably CH; For formula (20d) to formula (20g), X a and X b independently, CR 47 or N, preferably CR 47 , more preferably CH; X 6 and X 8 are each independently, Y T3 , C(R 47 )Y T2 、 C(R 47 )2, O, S, C(O), C(S), and S(O)2; X 7 is Y T3 and Z T selected from the group consisting of: X 6 Y T3 If X 7 is Z T and; X 7 Y T3 If X 6 and X 8 are each independently C(R 47 )Y T2 , C(R 47 )2, O, and S, preferably C(R 47 )Y T2 or C(R 47 )2; Here, the fused ring satisfying formula (20g) is at least one Y T2 or Y T3 Includes parts; Regarding formula (20b) and formula (20f), two R 47 are combined, =C-(R 47 )2, =S, or =O; However, X 1 -X 2 , X 2 -X 3 , X 3 -X 4 , and X 4 -X 5 is not -OO-, -NN-, -ON- or -NO-; X to the remainder of the ring fused to the 8-membered dienophile ring of formula (20a) to formula (20g) a and X b The direct bonds from are cis relative to each other, and H ais cis relative to; Preferably, the adjacent pair of atoms -OO- is not part of a fused ring of formula (20a) to formula (20g); Y T1 are OH, SH, N(R 38 )2, C(O)OH, C(S)OH, C(O)SH, C(S)SH, ON(R 38 )2, SO4H, SO3H, SO2H, PO4H2, PO3H, PO2H, and C(N)N(R 38 )2 is selected from the group consisting of; Y T2 are OH, SH, and N(R 38 )2 is selected from the group consisting of; Y T3 is NR 38 and is not adjacent to C(O), C(S), S(O), or S(O)2; Remaining X 1 , X 2 , X 3 , X 4 , X 5 are independent of each other, and X 1 , X 2 , X 3 , X 4 , X 5 Two or fewer of these are O, NR 38 or N, so that C(R 47 )2 and O; where R 48 -OC(O)-(S P ) k C A , -OC(S)-(S P ) k C A , -SC(O)-(S P ) k C A , or -SC(S)-(S P ) k C A If S P (if k>0) or C A (when k=0) is connected to R via an atom selected from the group consisting of O, C, S and N, preferably secondary or tertiary N. 48is bonded to -OC(O)-, -OC(S)-, -SC(O)- or -SC(S)-, where this atom is S P or C A is part of; preferably, R 48 is OC(O)-(S P ) k C A and S P or C A is connected to R via the N atom 48 bonded to -OC(O); where R 48 -O-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A , or -S-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A and r is 0, then S P (if k>0) or C A (when k=0) is R at the allylic position of the trans-cyclooctene ring of formula (19) via a group selected from the group consisting of —C(O)— and —C(S)—. 48 is bonded to the -O- or -S- moiety of P or C A is part of where R 48 -O-(LC ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A , or -S-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A and r is 1, L C is -C(Y C2 )Y C1 - and a carbon atom, preferably an aromatic carbon, bonded to the -O- or -S- moiety in the allylic position of the trans-cyclooctene ring of formula (19) through a group selected from the group consisting of L C is part of where Y C1 -O-, -S-, and -NR 36 - selected from the group consisting of where Y C2 is selected from the group consisting of O and S; where R 48 -O-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B) j ) r -(S P ) k C A , or -S-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A and r is 1, then S P (if k>0) or C A (when k=0) is connected to L through a moiety selected from the group consisting of -O-, -S-, and -N-, preferably secondary N or tertiary N. C where this moiety is S P or C A is part of where R 48 -(S P ) k C A If S P (if k>0) or C A (when k=0) is bonded to the allylic position of the trans-cyclooctene of formula (19) through an -O- or -S- atom, where this atom is S P or C A is part of where Z T However, C1~C 12 Alkylene group, C2-C 12 Alkenylene group, C7-C 12 Alkynylene group, C6 arylene group, C4-C5 heteroarylene group, C3-C8 cycloalkylene group, C5-C8 cycloalkenylene group, C5-C 12 Alkyl(hetero)arylene groups, C5-C 12 (Hetero)arylalkylene group, C4-C 12 Alkylcycloalkylene group, C4-C12 and cycloalkylalkylene groups, wherein the alkylene group, alkenylene group, alkynylene group, (hetero)arylene group, cycloalkylene group, cycloalkenylene group, alkyl(hetero)arylene group, (hetero)arylalkylene group, alkylcycloalkylene group, and cycloalkylalkylene group are selected from the group consisting of -(S P ) i -C B (wherein i is independently a number ranging from 0 to 4, and preferably i is 0 or 1), -Cl, -F, -Br, -I, -OR 37 , -N(R 37 )2, =O, =NR 37 , -SR 37 , -SO3H, -PO3H 、 -PO4H2, -NO2, and -Si(R 37 )3, and may be substituted by a moiety selected from the group consisting of —O—, —S—, —NR 37 may contain one or more heteroatoms selected from the group consisting of -, -P- and -Si-, wherein the N, S and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized; Here, each R 37 and R 36 are independently hydrogen atoms, -(S P ) i -C B , C1~C 24 Alkyl groups, C2-C 24 Alkenyl groups, C2-C 24 Alkynyl groups, C6-C 24 Aryl groups, C2-C 24 Heteroaryl groups, C3-C 24 Cycloalkyl groups, C5-C 24 Cycloalkenyl group, C 12 ~C 24 Cycloalkynyl group, C3-C 24 (Cyclo)alkyl(hetero)aryl groups, C3-C 24 (Hetero)aryl(cyclo)alkyl, C4-C 24 (Cyclo)alkenyl(hetero)aryl group, C4-C 24(Hetero)aryl(cyclo)alkenyl groups, C4-C 24 (Cyclo)alkynyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkynyl group, C4-C 24 Alkylcycloalkyl groups and C4-C 24 cycloalkylalkyl groups; where i is an integer ranging from 0 to 4, preferably i is 1; where R 37 and R 36 is not a hydrogen atom, but -Cl, -F, -Br, -I, -OH, -NH2, -SO3H, -PO3H 、 -PO4H 2、 optionally substituted with a moiety selected from the group consisting of -NO2, -CF3, =O, =NH, and -SH, and optionally containing one or more heteroatoms selected from the group consisting of O, S, NH, P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized; where R 38 are independent, R 37 and R 36 is selected from the groups listed for 38 is not attached to the rest of the molecule through a C(O), C(S), S(O), or S(O)2; Here, each R 47 are independently hydrogen atoms, -(S P ) i -C B , -F, -Cl, -Br, -I, -OR 37 , -N(R 37 )2, -SO3, -PO3 - 、 -NO2, -CF3, -SR 37 , S(=O)2N(R 37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R 37 , SC(=S)R 37 , N.R. 37 C(=O)-R 37 , N.R. 37 C(=S)-R 37 , N.R.37 C(=O)OR 37 , N.R. 37 C(=S)OR 37 , N.R. 37 C(=O)SR 37 , N.R. 37 C(=S)SR 37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R 37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR 37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37 )2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , S(O)R 37 , -S(O)2R 37 , N.R. 37 S(O)2R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , C1~C 24 Alkyl groups, C2-C 24 Alkenyl groups, C2-C 24 Alkynyl groups, C6-C 24 Aryl groups, C2-C 24 Heteroaryl groups, C3-C 24 Cycloalkyl groups, C5-C 24 Cycloalkenyl group, C 12 ~C 24 Cycloalkynyl group, C3-C 24 (Cyclo)alkyl(hetero)aryl groups, C3-C 24 (Hetero)aryl(cyclo)alkyl, C4-C 24 (Cyclo)alkenyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkenyl groups, C4-C24 (Cyclo)alkynyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkynyl group, C4-C 24 Alkylcycloalkyl groups and C4-C 24 cycloalkylalkyl groups; wherein i is independently an integer ranging from 0 to 4, preferably i is an integer ranging from 0 to 1; Here, the above alkyl group, alkenyl group, alkynyl group, aryl, heteroaryl, cycloalkyl group, cycloalkenyl group, cycloalkynyl group, (cyclo)alkyl(hetero)aryl group, (hetero)aryl(cyclo)alkyl group, (cyclo)alkenyl(hetero)aryl group, (hetero)aryl(cyclo)alkenyl group, (cyclo)alkynyl(hetero)aryl group, (hetero)aryl(cyclo)alkynyl group, alkylcycloalkyl group, cycloalkylalkyl group are -Cl, -F, -Br, -I, -OR 37 , -N(R 37 )2, -SO3R 37 , -PO3(R 37 ) 2、 -PO4(R 37 ) 2、 -NO2, -CF3, =O, =NR 37 , and -SR 37 and may be substituted with a moiety selected from the group consisting of O, S, NR 37 , and one or more heteroatoms selected from the group consisting of N, S, and P atoms, which may be oxidized, and wherein a plurality of N atoms may be quaternized; Here, two R 37、 R 38、 R 47 The group may be contained within a ring, Here, two R 37、 R 38、 R 47 Groups may also be contained within a ring to form a ring fused to the 8-membered trans ring.

[0029] In a further aspect, the present invention relates to a combination comprising a compound according to the invention and a diene, preferably a tetrazine.

[0030] In yet another aspect, the present invention relates to a compound of the invention, or a combination according to the invention, for use as a medicament.

[0031] In yet another aspect, the present invention relates to a compound of the invention or a combination according to the invention for use in the treatment of a disease in a subject, preferably a human, wherein the disease is selected from the group consisting of cancer, a central nervous system (CNS) disease, an infectious disease, inflammation, and a cardiovascular disease.

[0032] In yet another aspect, the present invention relates to a non-therapeutic method for releasing a molecule from a compound according to the present invention, said non-therapeutic method comprising contacting a compound according to formula (19), as defined herein, with a diene, as defined herein.

[0033] In another aspect, the present invention provides a non-therapeutic method for imaging a compound according to the present invention in a subject, preferably a human, comprising: (a) administering to said subject a compound according to Formula (19), as defined herein, including a label; (b) imaging a compound according to formula (19) present in said subject. The process includes the steps of: wherein said label is selected from the group consisting of radionuclides, fluorescent dyes, and phosphorescent dyes.

[0034] In another aspect, the present invention relates to a non-therapeutic use of a compound according to the invention or a combination according to the invention for imaging in a subject, preferably a human, wherein the compound or the combination of at least one compound according to formula (19) with a diene comprises a label selected from the group consisting of a radionuclide, a fluorescent dye, and a phosphorescent dye.

[0035] In yet another aspect, the present invention relates to a non-therapeutic method of use of a compound according to the invention or a combination according to the invention for releasing a molecule, preferably in vitro. [Brief explanation of the drawings]

[0036] [Figure 1] Figure 1 shows a preferred embodiment of the present invention. In both panels, the ADC is administered to a cancer patient and is allowed to circulate and bind to targets on cancer cells. After the free-circulating ADC has sufficiently cleared from the circulation, e.g., two days after injection, an active agent is administered and distributed systemically, allowing the prodrug attached to the cancer, i.e., the ADC, to react with a trigger, releasing the drug, which can then penetrate and kill neighboring cancer cells. Panel A shows cleavage of a carbamate-linked drug, and panel B shows cleavage of an ether-linked drug. [Figure 2] Figure 2 shows a preferred embodiment of the present invention. An antibody construct comprising a bispecific (anti-tumor and anti-CD3) antibody and a masking moiety (blocking protein) is administered to a cancer patient and is able to circulate and bind to targets on cancer cells. After the free-circulating construct has sufficiently cleared from the circulation, e.g., two days after injection, an active agent is administered and distributed systemically, allowing it to react with the trigger of the cancer-linked prodrug, i.e., ADC, unmasking it, after which T cells bind to the bispecific antibody, resulting in tumor killing. [Figure 3] Figure 3 shows the in vivo assembly of a functional cell penetration peptide (CPP) at the target site, resulting in CPP-induced drug internalization. [Figure 4] Figure 4 shows the in vivo unmasking of a functional cell penetration peptide (CPP) at the target site, resulting in CPP-induced drug internalization. [Figure 5]Figure 5 illustrates the use of the present invention in radioimmunotherapy: a radiolabeled antibody is administered, circulates, and binds an internalized cancer receptor, and after sufficient internalization has occurred, an activator is administered that cleaves the radiolabel (e.g., a moiety containing a radiometal chelate complex) from the antibody, resulting in rapid renal clearance of radioactivity from the blood and non-target tissues (but not radioactivity internalized by tumor cells). [Figure 6] FIG. 6 illustrates the use of compounds of the invention for site-specific antibody conjugation, e.g., with a drug, for the production of ADCs. [Figure 7] Figure 7 shows the results of in vitro experiments performed with TCO constructs containing a quenched fluorophore and activator objects of the present invention. The constructs and activators react very rapidly in PBS, cell culture medium, and human plasma to release and quench the fluorophore, resulting in a detectable increase in fluorescence in solution. DETAILED DESCRIPTION OF THE INVENTION

[0037] The present invention is based on the enlightened insight that the compounds, combinations, and kits according to the invention better address one or more of the needs mentioned above, which is surprising at least because these needs are met by the specific characteristics of the dienophile, and not by the specific characteristics of the diene, on which the background art focuses.

[0038] In one aspect, the use of the compounds, combinations, and kits according to the present invention results in high release yields and / or fast release.

[0039] In another aspect, the use of the compounds, combinations and kits according to the present invention results in fast release.

[0040] In another aspect, the use of the compounds, combinations, and kits according to the present invention results in fast click conjugation and high release yields and / or release rates.

[0041] The present invention is broadly based on the enlightened insight that compounds according to formula (19) described herein fulfill one or more of the above-mentioned needs and / or solve one or more of the above-mentioned problems.

[0042] In another aspect, it has been found that the combination of a compound according to formula (19) described herein with a diene as defined herein satisfies one or more of the above-mentioned needs and / or solves one or more of the above-mentioned problems.

[0043] In yet another aspect, it has been found that a kit containing a combination according to the present invention satisfies one or more of the above-mentioned needs and / or solves one or more of the above-mentioned problems.

[0044] In a still further aspect, the combinations according to the invention, and the kits according to the invention, have been found to be useful for the treatment or imaging of patients or animals.

[0045] In a still further aspect, compounds according to the invention, combinations according to the invention, and kits according to the invention have been found to be useful in in vitro and / or in vivo bioorthogonal reactions.

[0046] Without wishing to be bound by theory, the inventors believe that in the structure according to formula (19), Y as defined herein T1 , Y T2 and / or Y T3 The presence of the Y group, when contacted with a diene, is particularly advantageous compared to known TCOs. T1 , Y T2 and / or Y T3This is believed to result in a higher click release yield and / or click release rate compared to the same TCO lacking the group. Still not wishing to be bound by theory, the inventors presently believe this is the result of a destabilizing effect on dihydropyridazine tautomer intermediates, particularly the 4,5- and / or 1,4-dihydropyridazine tautomer intermediates formed upon conjugation of the TCO to the tetrazine.

[0047] Moreover, compounds according to formula (19) described herein have been found to provide high click release yields and click release rates when contacted with dienes that provide high click conjugation yields and rates.

[0048] Referring to the example in Scheme 1A below, without wishing to be bound by theory, the inventors believe that upon formation of 4,5-dihydropyridazine tautomer 3, Y T1 H with cis-positioned moieties (e.g., OH, NH2) a and hydrogen bonding with , inducing deprotonation and tautomerization to, among other things, the 1,4-dihydropyridazine intermediate, followed by C via pathway A and / or pathway B. A Pathway A involves 1,4-elimination, leaving free C A , 5, and subsequently 6. Route B provides C via a carbamate linker. A Y at the carbon to which T1 It involves nucleophilic attack of C A This results in the release of 7 and the formation of 8. T1 , Y T2 and Y T3 It is believed that the moiety results in rapid and high-yield release via one or both of these pathways.

[0049] [ka]

[0050] The present invention may be configured as follows. [Section 1] Compounds satisfying the following formula (19) and pharmaceutically acceptable salts thereof: [ka] where: R 48 is -OH, -OC(O)Cl, -OC(O)ON-succinimidyl, -OC(O)O-4-nitrophenyl, -OC(O)O-tetrafluorophenyl, -OC(O)O-pentafluorophenyl, -OC(O)-(S P ) k C A , -OC(S)-(S P ) k C A , -SC(O)-(S P ) k C A , -SC(S)-(S P ) k C A , -O-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A , -S-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A , and (S P )k C A selected from the group consisting of: r is an integer ranging from 0 to 2; each s is independently 0 or 1; each i is independently an integer ranging from 0 to 4, preferably 0 or 1; j is an integer ranging from 0 to 4, preferably 0 or 1; each k is independently 0 or 1; L C is a self-immolative linker, and S P is a spacer; Each C A and C B are independently selected from the group consisting of organic molecules and inorganic molecules; Here, at least one of the following conditions (a) to (c) is satisfied: (a)X 1 , X 2 , X 3 , X 4 , X 5 At least one of the 47 Y T1 and;Y T1 is H a positioned in cis relative to ; (b)X 3 is Y T3 and (c)X 1 , X 2 , X 3 , X 4 , X 5 are part of a fused ring satisfying one of the following formulas (20a) to (20g): [ka] Y T2 is positioned syn relative to the eight-membered dienophile ring; where X a and X b is X a -X b or X b -X a is X1 -X 2 , X 2 -X 3 , X 3 -X 4 , or X 4 -X 5 is part of the eight-membered ring of formula (19) such that Regarding formulas (20a) to (20c), X a and X b is CR 47 , preferably CH; For formula (20d) to formula (20g), X a and X b independently, CR 47 or N, preferably CR 47 , more preferably CH; X 6 and X 8 are each independently, Y T3 , C(R 47 )Y T2 、 C(R 47 )2, O, S, C(O), C(S), and S(O)2; X 7 is Y T3 and Z T selected from the group consisting of: X 6 Y T3 If X 7 is Z T and; X 7 Y T3 If X 6 and X 8 are each independently C(R 47 )Y T2 , C(R 47 )2, O, and S, preferably C(R 47 )Y T2 or C(R 47 )2; Here, the fused ring satisfying formula (20g) is at least one Y T2 or Y T3 Includes parts; Regarding formula (20b) and formula (20f), two R 47 are combined, =C-(R 47 )2, =S, or =O; However, X 1 -X 2 , X 2 -X 3 , X 3 -X 4 , and X 4 -X 5 is not -OO-, -NN-, -ON- or -NO-; X to the remainder of the ring fused to the 8-membered dienophile ring of formula (20a) to formula (20g) a and X b The direct bonds from are cis relative to each other, and H a is cis relative to; Preferably, the adjacent pair of atoms -OO- is not part of a fused ring of formula (20a) to formula (20g); Y T1 are OH, SH, N(R 38 )2, C(O)OH, C(S)OH, C(O)SH, C(S)SH, ON(R 38 )2, SO4H, SO3H, SO2H, PO4H2, PO3H, PO2H, and C(N)N(R 38 )2 is selected from the group consisting of; Y T2 are OH, SH, and N(R 38 )2 is selected from the group consisting of; Y T3 is NR 38 and is not adjacent to C(O), C(S), S(O), or S(O)2; Remaining X 1 , X 2 , X 3 , X 4 , X 5 are independent of each other, and X 1 , X 2 , X 3 , X 4 , X 5 Two or fewer of these are O, NR 38 or N, so that C(R 47)2 and O; where R 48 -OC(O)-(S P ) k C A , -OC(S)-(S P ) k C A , -SC(O)-(S P ) k C A , or -SC(S)-(S P ) k C A If S P (if k>0) or C A (when k=0) is connected to R via an atom selected from the group consisting of O, C, S and N, preferably secondary or tertiary N. 48 is bonded to -OC(O)-, -OC(S)-, -SC(O)- or -SC(S)-, where this atom is S P or C A is part of where R 48 -O-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A , or -S-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C Aand r is 0, then S P (if k>0) or C A (when k=0) is R at the allylic position of the trans-cyclooctene ring of formula (19) via a group selected from the group consisting of —C(O)— and —C(S)—. 48 is bonded to the -O- or -S- moiety of P or C A is part of where R 48 -O-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A , or -S-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A and r is 1, L C is -C(Y C2 )Y C1 - and a carbon atom, preferably an aromatic carbon, bonded to the -O- or -S- moiety in the allylic position of the trans-cyclooctene ring of formula (19) through a group selected from the group consisting of L C is part of where Y C1 -O-, -S-, and -NR 36 - selected from the group consisting of where YC2 is selected from the group consisting of O and S; where R 48 -O-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A , or -S-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A and r is 1, then S P (if k>0) or C A (when k=0) is connected to L through a moiety selected from the group consisting of -O-, -S-, and -N-, preferably secondary N or tertiary N. C where this moiety is S P or C A is part of where R 48 -(S P ) k C A If S P (if k>0) or C A (when k=0) is bonded to the allylic position of the trans-cyclooctene of formula (19) through an -O- or -S- atom, where this atom is S P or C A is part of where Z THowever, C1~C 12 Alkylene group, C2-C 12 Alkenylene group, C7-C 12 Alkynylene group, C6 arylene group, C4-C5 heteroarylene group, C3-C8 cycloalkylene group, C5-C8 cycloalkenylene group, C5-C 12 Alkyl(hetero)arylene groups, C5-C 12 (Hetero)arylalkylene group, C4-C 12 Alkylcycloalkylene group, C4-C 12 and a cycloalkylalkylene group, wherein the alkylene group, alkenylene group, alkynylene group, (hetero)arylene group, cycloalkylene group, cycloalkenylene group, alkyl(hetero)arylene group, (hetero)arylalkylene group, alkylcycloalkylene group, and cycloalkylalkylene group are selected from the group consisting of (S P ) i -C B (wherein i is independently an integer ranging from 0 to 4), -Cl, -F, -Br, -I, -OR 37 , -N(R 37 )2, =O, =NR 37 , -SR 37 , -SO3H, -PO3H 、 -PO4H2, -NO2, and -Si(R 37 )3, and may be substituted by a moiety selected from the group consisting of —O—, —S—, —NR 37 may contain one or more heteroatoms selected from the group consisting of -, -P- and -Si-, wherein the N, S and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized; Here, each R 37 and R 36 are independently hydrogen atoms, -(S P ) i -C B (i is independently an integer ranging from 0 to 4), C1 to C 24 Alkyl groups, C2-C 24 Alkenyl groups, C2-C 24 Alkynyl groups, C6-C 24 Aryl groups, C2-C 24 Heteroaryl groups, C3-C24 Cycloalkyl groups, C5-C 24 Cycloalkenyl group, C 12 ~C 24 Cycloalkynyl group, C3-C 24 (Cyclo)alkyl(hetero)aryl groups, C3-C 24 (Hetero)aryl(cyclo)alkyl, C4-C 24 (Cyclo)alkenyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkenyl groups, C4-C 24 (Cyclo)alkynyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkynyl group, C4-C 24 Alkylcycloalkyl groups and C4-C 24 cycloalkylalkyl groups; where i is an integer ranging from 0 to 4, preferably i is 1; where R 37 and R 36 is not a hydrogen atom, but -Cl, -F, -Br, -I, -OH, -NH2, -SO3H, -PO3H 、 -PO4H 2、 optionally substituted with a moiety selected from the group consisting of -NO2, -CF3, =O, =NH, and -SH, and optionally containing one or more heteroatoms selected from the group consisting of O, S, NH, P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized; where R 38 are independent, R 37 and R 36 is selected from the groups listed for 38 is not attached to the rest of the molecule through a C(O), C(S), S(O), or S(O)2; Here, each R 47 are independently hydrogen atoms, -(S P ) i -C B , -F, -Cl, -Br, -I, -OR 37 , -N(R 37 )2, -SO3, -PO3 -、 -NO2, -CF3, -SR 37 , -S(=O)2N(R 37 )2, - OC(=O)R 37 , -SC(=O)R 37 , -OC(=S)R 37 , -SC(=S)R 37 , -NR 37 C(=O)-R 37 , -NR 37 C(=S)-R 37 , -NR 37 C(=O)OR 37 , -NR 37 C(=S)OR 37 , -NR 37 C(=O)SR 37 , -NR 37 C(=S)SR 37 , -OC(=O)N(R 37 )2, -SC(=O)N(R 37 )2, -OC(=S)N(R 37 )2, -SC(=S)N(R 37 )2, -NR 37 C(=O)N(R 37 )2, -NR 37 C(=S)N(R 37 )2, -C(=O)R 37、 -C(=S)R 37、 -C(=O)N(R 37 )2, -C(=S)N(R 37 )2, -C(=O)OR 37 , -C(=O)SR 37 , -C(=S)OR 37 , -C(=S)SR 37 , -S(O)R 37 , -S(O)2R 37 , -NR 37 S(O)2R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , C1~C 24 Alkyl groups, C2-C 24 Alkenyl groups, C2-C 24 Alkynyl groups, C6-C 24 Aryl groups, C2-C24 Heteroaryl groups, C3-C 24 Cycloalkyl groups, C5-C 24 Cycloalkenyl group, C 12 ~C 24 Cycloalkynyl group, C3-C 24 (Cyclo)alkyl(hetero)aryl groups, C3-C 24 (Hetero)aryl(cyclo)alkyl, C4-C 24 (Cyclo)alkenyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkenyl groups, C4-C 24 (Cyclo)alkynyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkynyl group, C4-C 24 Alkylcycloalkyl groups and C4-C 24 cycloalkylalkyl groups; wherein preferably, i is an integer ranging from 0 to 1; Here, the alkyl group, alkenyl group, alkynyl group, aryl, heteroaryl, cycloalkyl group, cycloalkenyl group, cycloalkynyl group, (cyclo)alkyl(hetero)aryl group, (hetero)aryl(cyclo)alkyl group, (cyclo)alkenyl(hetero)aryl group, (hetero)aryl(cyclo)alkenyl group, (cyclo)alkynyl(hetero)aryl group, (hetero)aryl(cyclo)alkynyl group, alkylcycloalkyl group, cycloalkylalkyl group are -Cl, -F, -Br, -I, -OR 37 , -N(R 37 )2, -SO3R 37 , -PO3(R 37 ) 2、 -PO4(R 37 ) 2、 -NO2, -CF3, =O, =NR 37 , and -SR 37 and may be substituted with a moiety selected from the group consisting of O, S, NR 37 , and one or more heteroatoms selected from the group consisting of N, S, and P atoms, which may be oxidized, and wherein a plurality of N atoms may be quaternized; Here, two R 37、 R 38、 R 47 The group may be contained within a ring, Here, two R 37、 R 38、 R 47 Groups may also be contained within a ring to form a ring fused to the 8-membered trans ring. [Section 2] Item 1. The compound according to item 1, wherein at most one of the conditions (a) to (c) is satisfied. [Section 3] Item 1, a compound selected from the group consisting of: and their enantiomers according to formula (19): [ka] [Section 4] The compound according to any one of items 1 to 3, which is selected from the group consisting of: [ka] [ka] [ka] [Section 5] A combination comprising the compound according to any one of items 1 to 4 and a diene, preferably tetrazine. [Section 6] Item 6. The combination according to Item 5, wherein the diene is a tetrazine satisfying the following formula (4), preferably a pharmaceutically acceptable salt thereof: [ka] where: Each moiety Q1 and Q2 is independently a hydrogen atom, -F, -Cl, -Br, -I, -OR 37 , -N(R 37 )2, -SO3, -PO3 - 、 -NO2, -CF3, -SR 37 , -S(=O)2N(R37 )2、-OC(=O)R 37 、-SC(=O)R 37 、-OC(=S)R 37 、-SC(=S)R 37 、-NR 37 C(=O)-R 37 、-NR 37 C(=S)-R 37 、-NR 37 C(=O)O-R 37 、-NR 37 C(=S)O-R 37 、-NR 37 C(=O)S-R 37 、-NR 37 C(=S)S-R 37 、-OC(=O)N(R 37 )2、-SC(=O)N(R 37 )2、-OC(=S)N(R 37 )2、-SC(=S)N(R 37 )2、-NR 37 C(=O)N(R 37 )2、-NR 37 C(=S)N(R 37 )2、-C(=O)R 37、 -C(=S)R 37、 -C(=O)N(R 37 )2、-C(=S)N(R 37 )2、-C(=O)O-R 37 、-C(=O)S-R 37 、-C(=S)O-R 37 、-C(=S)S-R 37 、-S(O)R 37 、-S(O)2R 37 、-NR 37 S(O)2R 37 、-ON(R 37 ) 2、 -NR 37 OR 37, alkyl groups, alkenyl groups, alkynyl groups, aryl groups, heteroaryl groups, cycloalkyl groups, cycloalkenyl groups, cycloalkynyl groups, (cyclo)alkyl(hetero)aryl groups, (hetero)aryl(cyclo)alkyl, (cyclo)alkenyl(hetero)aryl groups, (hetero)aryl(cyclo)alkenyl groups, (cyclo)alkynyl(hetero)aryl groups, (hetero)aryl(cyclo)alkynyl groups, alkylcycloalkyl groups, and cycloalkylalkyl groups; Here, H, -F, -Cl, -Br, -I, -OH, -NH2, -SO3, -PO3 - 、 The Q1 and Q2 groups that are not -NO2 or -CF3 are preferably -Cl, -F, -Br, -I, -OR 37 , -N(R 37 )2, -SO3R 37 , -PO3(R 37 ) 2、 -PO4(R 37 ) 2、 -NO2, -CF3, =O, =NR 37 , and -SR 37 and may be substituted with a moiety selected from the group consisting of O, S, NR 37 , and one or more heteroatoms selected from the group consisting of N, S, and P atoms, which may be oxidized, and wherein a plurality of N atoms may be quaternized; where Q1 and Q2 groups are -(S P ) D -R 87 may be bound to; where D is 0 or 1, and each R 87 are individually selected from the group consisting of a biomolecule, a polymer, a peptoid, a dendrimer, a lipid, a micelle, a liposome, a polymersome, a particle, a bead, a gel, a metal complex, an organic molecule, an organometallic moiety, an albumin-binding moiety, a radionuclide-containing moiety, a dye moiety, a chelating moiety, and an imaging probe; And preferably, at least one of the moieties Q1 and Q2 is not a hydrogen atom. [Section 7] Item 7. The combination according to item 6, wherein Q1 and Q2 are selected from the group consisting of a hydrogen atom, phenyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2,6-pyrimidyl, 2,5-pyrimidyl, 3,5-pyrimidyl, and 2,4-pyrimidyl; and any Q1 or Q2 that is not a hydrogen atom may be substituted as defined in claim 6. [Section 8] In equation (4), (a) Q1 and Q2 are selected from the group consisting of 2-pyridyl, 3-pyridyl, and 4-pyridyl; or (b) Q1 is selected from the group consisting of 2,6-pyrimidyl, 2,5-pyrimidyl, 3,5-pyrimidyl, and 2,4-pyrimidyl; and Q2 is (hetero)alkyl; or (c) Q1 is phenyl and Q2 is a hydrogen atom; and in (a) to (c), all Q1 and Q2 that are not hydrogen atoms may be substituted as defined in claim 6. Item 6 or 7. The combination according to item 6 or 7. [Section 9] A compound according to any one of items 1 to 4 or a combination according to any one of items 5 to 8 for use as a pharmaceutical. [Section 10] The compound according to any one of items 1 to 4 or the combination according to any one of items 5 to 8, for use in treating a disease in a subject, preferably a human, wherein the disease is selected from the group consisting of cancer, central nervous system (CNS) disease, infectious disease, inflammation, and cardiovascular disease. [Section 11] A non-therapeutic method for releasing a molecule from a compound according to any one of items 1 to 4, comprising contacting a compound according to formula (19) according to any one of items 1 to 4 with a diene according to any one of items 5 to 8. [Section 12] A non-therapeutic method for imaging a compound according to any one of items 1 to 4 in a subject, preferably a human, comprising: (a) administering to the subject a compound according to formula (19) described in any one of items 1 to 4, including a label; (b) imaging a compound according to formula (19) present in said subject. Including, the label is selected from the group consisting of a radionuclide, a fluorescent dye, and a phosphorescent dye; The non-therapeutic method. [Section 13] A non-therapeutic use of a compound according to any one of items 1 to 4 or a combination according to any one of items 5 to 8 for imaging in a subject, preferably a human, wherein the compound or the combination of at least one compound according to formula (19) with a diene comprises a label selected from the group consisting of a radionuclide, a fluorescent dye, and a phosphorescent dye. [Section 14] A non-therapeutic use of a compound according to any one of items 1 to 4 or a combination according to any one of items 5 to 8 for releasing a molecule, preferably in vitro. definition

[0051] The present invention will be further described with respect to particular embodiments and with reference to certain drawings but the invention is not limited thereto and will be described only by the claims. Any reference signs in the claims should not be construed as limiting the scope of the invention. The drawings described are schematic and not limiting. In the drawings, the size of some of the elements may be exaggerated and not drawn to scale for illustrative purposes. Where an indefinite or definite article is used when referring to a singular noun, such as "a" or "an," "the," this includes the plural of that noun unless otherwise stated.

[0052] When used in the detailed description and claims of the present invention, the verb "comprise" and its conjugations are used in an open-ended sense meaning that items following the word are included, but items not specifically mentioned are not excluded.

[0053] In addition, reference to an element by the indefinite article "a" or "an" does not exclude the possibility that a plurality of elements is present, unless the context clearly requires that there is only one element. Thus, the indefinite article "a" or "an" usually means "at least one."

[0054] Therefore, the scope of the expression "a device comprising means A and B" should not be limited to a device consisting only of components A and B. It means that, in the context of the present invention, the only relevant components of the device are A and B.

[0055] The compounds disclosed in the detailed description and claims of the present invention may contain one or more asymmetric centers, and different diastereomers and / or enantiomers may exist in the compound. The description of any compound in the detailed description and claims of the present invention is meant to encompass all diastereomers and mixtures thereof, unless otherwise specified. Therefore, when a specific stereoisomer is shown in this specification, it will be understood that the compound is limited to this specific stereoisomer. For example, in formula (19), Y T2 is arranged syn relative to the eight-membered dienophile ring. This still includes the syn enantiomer, but Y T2 is positioned anti relative to the eight-membered dienophile ring.

[0056] Additionally, the description of any compound in the detailed description of the invention and in the claims is meant to encompass both each enantiomer and any mixture of the enantiomers, racemic or otherwise, unless otherwise specified. When the structure of a compound is shown as a particular enantiomer, it is to be understood that the invention of the present application is not limited to that particular enantiomer, unless otherwise specified. When the structure of a compound is shown as a particular diastereomer, it is to be understood that the invention of the present application is not limited to that particular diastereomer, unless otherwise specified.

[0057] The compounds can occur in various tautomeric forms. The compounds according to the present invention are meant to encompass all tautomeric forms unless otherwise specified. When the structure of a compound is shown as a specific tautomer, it should be understood that the invention of this application is not limited to that specific tautomer unless otherwise specified.

[0058] The compounds disclosed in the detailed description and claims of the present invention may further exist as exo and endo diastereomers. Unless otherwise specified, the description of any compound in the detailed description and claims of the present invention is meant to encompass both the respective exo and endo diastereomers of the compound, as well as mixtures thereof. When the structure of a compound is shown as a particular endo or exo diastereomer, it should be understood that the invention of this application is not limited to that particular endo or exo diastereomer, unless otherwise specified.

[0059] The compounds disclosed in the detailed description and claims of the present invention may further exist as anti- and syn-diastereomers. Unless otherwise specified, the description of any compound in the detailed description and claims of the present invention is meant to encompass both the respective anti- and syn-diastereomers of the compound, as well as mixtures thereof. When the structure of a compound is shown as a specific syn- or anti-diastereomer, it should be understood that the invention of this application is not limited to that specific syn- or anti-diastereomer, unless otherwise specified.

[0060] Regarding equation (19), where Y T2 is positioned syn relative to the 8-membered dienophile ring; "syn" means that Y is on the same side of the ring fused to the 8-membered dienophile ring as the 8-membered dienophile ring. T2 Those skilled in the art will recognize that there is a Y T2 It will be understood that Y can be defined as facing, rather than facing away from, the eight-membered dienophile ring, and also as being oriented end-on with respect to the eight-membered dienophile ring. T2 is X a and X b Adjacent to and X 1 , X 2 , X 3 , X 4 , X 5 For clarity, "facing" will be understood as "near to." Furthermore, for clarity, "located close to" means facing the X moiety, which is selected from Y T2 But X a and X b Adjacent to and X 1 , X 2 , X 3 , X 4 , X 5 For clarity, Y is meant to be cis-oriented relative to the X moiety, which is selected from T2 is Xa and X b is meant to be oriented cis relative to the direct bond from to the remaining 8-membered dienophile ring.

[0061] Unless otherwise specified, the compounds of the present invention and / or groups thereof may be protonated or deprotonated. It will be understood that compounds may carry multiple charges, which may be of opposite sign. For example, in a compound containing an amine and a carboxylic acid, the amine may be protonated while the carboxylic acid is simultaneously deprotonated.

[0062] In some formulas, groups or substituents are indicated by reference to letters, such as "A," "B," "X," "Y," and various (numbered) "R" groups. Additionally, the number of repeat units is indicated by reference to letters, such as -(CH) n The definitions of these letters should be read with reference to each formula, i.e., in different formulas, these letters can each independently have different meanings unless otherwise specified.

[0063] In some of the formulas and text that follow, references are made to "alkyl," "heteroalkyl," "aryl," "heteroaryl," "alkenyl," "alkynyl," "alkylene," "alkenylene," "alkynylene," "arylene," "cycloalkyl," "cycloalkenyl," "cycloakynyl," "arenetriyl," and the like. The number of carbon atoms contained in these groups, excluding carbon atoms contained in any substituents defined below, can be indicated by a designation preceding such term (e.g., "C1-C8 alkyl" means that the alkyl can have from 1 to 8 carbon atoms). For the avoidance of doubt, a butyl group substituted with -OCH3 will be referred to as a C4 alkyl because the carbon atoms in the substituent are not included in the carbon number count.

[0064] The unsubstituted alkyl group has the general formula C n H 2n+1and may be linear or branched. The alkyl group may be substituted with one or more substituents as further specified herein. Examples of alkyl groups include methyl, ethyl, propyl, 2-propyl, t-butyl, 1-hexyl, 1-dodecyl, etc. Unless otherwise specified, the alkyl group may have one or more heteroatoms independently selected from the group consisting of O, NR5, S, P, and Si, where N, S, and P atoms may be oxidized, and multiple N atoms may be quaternized. In preferred embodiments, up to two heteroatoms may be consecutive, such as in -CH2-NH-OCH3 and -CH2-O-Si(CH3)3. In some preferred embodiments, the heteroatoms are not directly bonded to each other. Examples of heteroalkyl include CH2CH2-O-CH3, -CH2CH2-NH-CH3, -CH2CH2-S(O)-CH3, -CH = It includes CH-O-CH3, -Si(CH3)3. In a preferred embodiment, the C1-C4 alkyl has no more than two heteroatoms.

[0065] A cycloalkyl group is a cyclic alkyl group. Unsubstituted cycloalkyl groups have at least 3 carbon atoms and have the general formula C n H 2n-1 The cycloalkyl group is substituted by one or more substituents as further specified herein. Examples of cyclopropyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Unless otherwise specified, the cycloalkyl group may have one or more heteroatoms independently selected from the group consisting of O, NR5, S, P, and Si, where N, S, and P atoms may be optionally oxidized, and multiple N atoms may be optionally quaternized.

[0066] Alkenyl groups have one or more carbon-carbon double bonds and can be linear or branched. Unsubstituted alkenyl groups having one C—C double bond have the general formula C n H 2n-1 Unsubstituted alkenyl groups having two C—C double bonds have the general formula Cn H 2n-3 The alkenyl group may have a terminal carbon-carbon double bond and / or an internal carbon-carbon double bond. A terminal alkenyl group is an alkenyl group in which the carbon-carbon double bond is located at the terminal position of the carbon chain. The alkenyl group may also have two or more carbon-carbon double bonds. Examples of alkenyl groups include ethenyl, propenyl, isopropenyl, t-butenyl, 1,3-butadienyl, 1,3-pentadienyl, and the like. Unless otherwise specified, the alkenyl group may be optionally substituted with one or more independently selected substituents as defined below. Unless otherwise specified, the alkenyl group may have one or more heteroatoms independently selected from the group consisting of O, NR5, S, P, and Si, where the N, S, and P atoms may be optionally oxidized and multiple N atoms may be optionally quaternized.

[0067] Alkynyl groups have one or more carbon-carbon triple bonds and can be linear or branched. Unsubstituted alkynyl groups having one CC triple bond have the general formula C n H 2n-3 The alkynyl group may have a terminal carbon-carbon triple bond and / or an internal carbon-carbon triple bond. A terminal alkynyl group is an alkynyl group in which the carbon-carbon triple bond is located at the terminal position of the carbon chain. The alkynyl group may also have two or more carbon-carbon triple bonds. Unless otherwise specified, the alkynyl group may be substituted with one or more independently selected substituents defined below. Examples of alkynyl groups include ethynyl, propynyl, isopropynyl, t-butynyl, and the like. Unless otherwise specified, the alkynyl group may have one or more heteroatoms independently selected from the group consisting of O, NR5, S, P, and Si, where the N, S, and P atoms may be optionally oxidized and multiple N atoms may be optionally quaternized.

[0068] An aryl group refers to an aromatic hydrocarbon ring system having 6 to 24 carbon atoms, more preferably 6 to 12 carbon atoms, and may have a monocyclic or polycyclic structure. When the aryl group has a polycyclic structure, it is preferably a bicyclic structure. The aryl group may be substituted with one or more substituents further specified herein. Examples of the aryl group are phenyl and naphthyl.

[0069] The arylalkyl group and the alkylaryl group have at least 7 carbon atoms and can have a monocyclic or polycyclic structure. The arylalkyl group and the alkylaryl group can be substituted with one or more substituents as further specified herein. An example of an arylalkyl group is benzyl. An example of an alkylaryl group is 4-tert-butylphenyl.

[0070] Preferably, the heteroaryl group has 5 to 16 carbon atoms and 1 to 5 heteroatoms. The heteroaryl group has at least two carbon atoms (i.e., at least C2) and one or more heteroatoms N, O, P, or S. The heteroaryl group can have a monocyclic or polycyclic structure. The heteroaryl group can be optionally substituted with one or more substituents further specified herein. Examples of suitable heteroaryl groups include pyridinyl, quinolinyl, pyrimidinyl, pyrazinyl, pyrazolyl, imidazolyl, thiazolyl, pyrrolyl, furanyl, triazolyl, benzofuranyl, indolyl, purinyl, benzoxazolyl, thienyl, phosphoryl, and oxazolyl.

[0071] Heteroarylalkyl and alkylheteroaryl groups have at least three carbon atoms (i.e., at least C3) and can have monocyclic and bicyclic structures. The heteroaryl groups may be optionally substituted with one or more substituents as further specified herein.

[0072] When an aryl group is designated as a (hetero)aryl group, the designation is meant to encompass aryl and heteroaryl groups. Similarly, alkyl(hetero)aryl groups are meant to encompass alkylaryl and alkylheteroaryl groups, and alkyl(hetero)aryl groups are meant to encompass arylalkyl and heteroarylalkyl groups. Thus, C2-C 24 The (hetero)aryl group is C2-C 24 Heteroaryl groups and C6-C 24 It should be construed to include aryl groups. 24 The alkyl(hetero)aryl group is C7-C 24 Alkylaryl groups and C3-C 24 It is meant to include alkylheteroaryl groups, as well as C3-C 24 (Hetero)arylalkyl is C7-C 24 Aryl alkyl groups and C3-C 24 Heteroarylalkyl groups are meant to be included.

[0073] A cycloalkenyl group is a cyclic alkenyl group. Unsubstituted cycloalkenyl groups having one double bond have the general formula C n H 2n-3 The cycloalkenyl group may be further substituted with one or more substituents as further specified herein. An example of a cycloalkenyl group is cyclopentenyl. Unless otherwise specified, the cycloalkenyl group may contain one or more heteroatoms independently selected from the group consisting of O, NR5, S, P, and Si, where N, S, and P atoms may be optionally oxidized, and multiple N atoms may be optionally quaternized.

[0074] A cycloalkynyl group is a cyclic alkynyl group. An unsubstituted cycloalkynyl group having one triple bond has the general formula C n H 2n-5The cycloalkynyl group may be further substituted by one or more substituents as further specified herein. An example of a cycloalkynyl group is cyclooctynyl. Unless otherwise specified, the cycloalkynyl group may contain one or more heteroatoms independently selected from the group consisting of O, NR5, S, P and Si, where N, S and P atoms may be oxidized and multiple N atoms may be quaternized.

[0075] When referring to a (hetero)aryl group, the designation is meant to encompass both aryl and heteroaryl groups. An alkyl(hetero)aryl group refers to an alkylaryl group and an alkylheteroaryl group. A (hetero)arylalkyl group refers to an arylalkyl group and a heteroarylalkyl group. In general, when (hetero) is placed before a group, it refers to both versions of the group without the hetero- prefix as well as the group with the hetero- prefix.

[0076] As used herein, the prefix hetero- indicates that the group contains one or more heteroatoms selected from the group consisting of O, N, S, P, and Si. It will be understood that a group containing the prefix hetero- contains heteroatoms by definition. Thus, if a group containing the prefix hetero- is part of a list of groups that are defined as optionally containing heteroatoms, then it is not optional for the group containing the prefix hetero- to contain heteroatoms, but rather it is by definition the case.

[0077] As used herein, when the prefix hetero- is used to refer to a combination of groups, it will be understood that the prefix hetero- refers only to the group immediately preceding it. For example, heteroarylalkyl refers to a combination of a heteroaryl group and an alkyl group, not a combination of a heteroaryl group and a heteroalkyl group. Thus, when the prefix hetero- is used to refer to a combination of groups that are part of a list of groups that indicate that heteroatoms may be included, it will be understood that it is optional only for groups in the combination without the prefix hetero-, as it is not optional by definition for groups in the combination with the prefix hetero- (see above). For example, when heteroarylalkyl is part of a list of groups that indicate that heteroatoms may be included, the heteroaryl portion is considered to have a heteroatom by definition, while having a heteroatom is optional for the alkyl portion.

[0078] As used herein, the prefix cyclo- indicates that a group is cyclic. When the prefix cyclo- is used in combination with multiple groups, it will be understood that the prefix cyclo- refers to only the group directly adjacent to it. For example, cycloalkylalkenylene refers to the combination of a cycloalkylene group (see the definition of the suffix -ene below) and an alkenylene group, rather than the combination of a cycloalkylene and a cycloalkenylene group.

[0079] In general, when (cyclo) is placed before a group, it refers to both versions of the group without the prefix cyclo- as well as the group with the prefix cyclo-.

[0080] As used herein, the suffix -ene indicates a divalent group, i.e., the group is bonded to at least two other moieties. An example of an alkylene is propylene (-CH-CH-CH-), which is bonded to another moiety at both ends. When a group having the suffix -ene is substituted with -H at one position, it is understood that the group is the same as the group without the suffix. For example, an alkylene substituted with -H is the same as an alkyl group. That is, propylene substituted with -H at one end, i.e., -CH-CH-CH-, is logically the same as propyl, i.e., -CH-CH-CH-.

[0081] In this specification, when a combination of multiple groups is listed with the suffix -ene, it indicates a divalent group, i.e., the group is bonded to at least two other moieties, where each group in the combination has one bond to one of these two moieties. Thus, for example, alkylarylene is understood to be a combination of an arylene group and an alkylene group. An example of an alkylarylene group is -phenyl-CH-, and an example of an arylalkylene group is -CH-phenyl-.

[0082] As used herein, the suffix -triyl indicates a trivalent group, i.e., the group is bonded to at least three other moieties. An example of an arenetriyl is shown below:

[0083] [ka]

[0084] Here, the wavy lines indicate bonds to different groups on the main compound.

[0085] When a group with the suffix -triyl is substituted with -H at one position, it is understood to be the same as a divalent group with the suffix -ene. For example, an arenetriyl substituted with -H is the same as an arylene group. Similarly, when a group with the suffix -triyl is substituted with -H at two positions, it is understood to be the same as a monovalent group. For example, an arenetriyl substituted with two -H is the same as an aryl group.

[0086] When a group, such as an alkyl group, has a heteroatom, it is understood that this group is the same as the hetero variant of this group. For example, when an alkyl group has a heteroatom, this group is the same as a heteroalkyl group. Similarly, when an aryl group has a heteroatom, this group is the same as a heteroaryl group. In this specification, "having" and its conjugations are understood to mean that when a group has a heteroatom, this heteroatom is part of the backbone of the group. For example, C alkylene containing N refers to -NH-CH-CH-, -CH-NH-CH-, and -CH-CH-NH-.

[0087] Unless otherwise specified, a group may have heteroatoms at non-terminal positions or at one or more terminal positions. In this case, "terminal" refers to a terminal position within the group, not necessarily a terminal position within the compound as a whole. For example, if an ethylene group has a nitrogen atom, this could refer to -NH-CH-CH-, -CH-NH-CH-, and -CH-CH-NH-. For example, if an ethyl group has a nitrogen atom, this could refer to -NH-CH-CH, -CH-NH-CH, and -CH-CH-NH.

[0088] In this specification, cyclic compounds (i.e., aryl, cycloalkyl, cycloalkenyl, etc.) are understood to be monocyclic, polycyclic, or branched. The number of carbon atoms in a cyclic compound refers not only to the number of carbon atoms in one ring, but also to the fact that carbon atoms can be contained in multiple rings. These rings can be fused to the main ring or substituted on the main ring. For example, C , which may contain heteroatoms, can be used. 10 Aryl refers inter alia to a naphthyl group (fused ring) or, for example, a bipyridyl group (substituted rings, both containing an N atom).

[0089] Unless otherwise specified, the term "hetero" refers to a (hetero)alkyl group, a (hetero)alkenyl group, a (hetero)alkynyl group, a (hetero)cycloalkyl group, a (hetero)cycloalkenyl group, a (hetero)cycloalkynyl group, a (hetero)alkylcycloalkyl group, a (hetero)alkylcycloalkenyl group, a (hetero)alkylcycloalkynyl group, a (hetero)cycloalkylalkyl group, a (hetero)cycloalkenylalkyl group, a (hetero)cycloalkynylalkyl group, a (hetero)alkenylcycloalkyl group, a (hetero)alkenylcycloalkenyl a (hetero)alkenylcycloalkynyl group, a (hetero)cycloalkylalkenyl group, a (hetero)cycloalkenylalkenyl group, a (hetero)cycloalkynylalkenyl group, a (hetero)alkynylcycloalkyl group, a (hetero)alkynylcycloalkenyl group, a (hetero)alkynylcycloalkynyl group, a (hetero)cycloalkylalkynyl group, a (hetero)cycloalkenylalkynyl group, a (hetero)cycloalkynylalkynyl group, a (hetero)aryl group, a (hetero)arylalkyl group, a (hetero)arylalkenyl a (hetero)arylalkynyl group, an alkyl(hetero)aryl group, an alkenyl(hetero)aryl group, an alkynyl(hetero)aryl group, a cycloalkyl(hetero)aryl group, a cycloalkenyl(hetero)aryl group, a cycloalkynyl(hetero)aryl group, a (hetero)arylcycloalkyl group, a (hetero)arylcycloalkenyl group, a (hetero)arylcycloalkynyl group, a (hetero)alkylene group, a (hetero)alkenylene group, a (hetero)alkynylene group, a (hetero)cycloalkylene group, a (hetero)cyclo The alkenylene group, (hetero)cycloalkynylene group, (hetero)arylene group, alkyl(hetero)arylene group, (hetero)arylalkylene group, (hetero)arylalkenylene group, (hetero)arylalkynylene group, alkenyl(hetero)arylene, alkynyl(hetero)arylene, (hetero)arenetriyl group, (hetero)cycloalkanetriyl group, (hetero)cycloalkenetriyl, and (hetero)cycloalkyntriyl groups include -Cl, -F, -Br, -I, -OH, -NH2, -SO3H, -PO3H 、-PO4H2, -NO2, -CF3, =O, =NR5, -SR5, C1~C 24 Alkyl groups, C2-C 24 Alkenyl groups, C2-C 24 Alkynyl groups, C6-C 24 Aryl groups, C2-C 24 Heteroaryl groups, C3-C 24 Cycloalkyl groups, C5-C 24 Cycloalkenyl group, C 12 ~C 24 Cycloalkynyl group, C3-C 24 Alkyl(hetero)aryl groups, C3-C 24 (Hetero)arylalkyl groups, C4-C 24 (Hetero)arylalkenyl groups, C4-C 24 (Hetero)arylalkynyl group, C4-C 24 Alkenyl(hetero)aryl groups, C4-C 24 Alkynyl(hetero)aryl groups, C4-C 24 Alkylcycloalkyl groups, C6-C 24 Alkylcycloalkenyl group, C 13 ~C 24 Alkylcycloalkynyl group, C4-C 24 Cycloalkylalkyl groups, C6-C 24 Cycloalkenylalkyl groups, C 13 ~C 24 Cycloalkynylalkyl groups, C5-C 24 Alkenylcycloalkyl groups, C7-C 24 Alkenylcycloalkenyl group, C 14 ~C 24 Alkenylcycloalkynyl group, C5-C 24 Cycloalkylalkenyl group, C7-C 24 Cycloalkenyl alkenyl group, C 14 ~C 24 Cycloalkynylalkenyl group, C5-C 24 Alkynylcycloalkyl groups, C7-C 24 Alkynylcycloalkenyl group, C 14 ~C 24 Alkynylcycloalkynyl group, C5-C 24Cycloalkylalkynyl group, C7-C 24 Cycloalkenylalkynyl group, C 14 ~C 24 Cycloalkynylalkynyl group, C5-C 24 Cycloalkyl(hetero)aryl groups, C7-C 24 Cycloalkenyl(hetero)aryl group, C 14 ~C 24 Cycloalkynyl(hetero)aryl groups, C5-C 24 (Hetero)arylcycloalkyl groups, C7-C 24 (Hetero)arylcycloalkenyl groups, and C 14 ~C 24 (Hetero)arylcycloalkynyl groups may be substituted with one or more substituents independently selected from the group consisting of (hetero)arylcycloalkynyl groups. Unless otherwise specified, the substituents disclosed herein may contain one or more heteroatoms selected from the group consisting of O, S, NR5, P, and Si, where N, S, and P atoms may be oxidized, and where multiple N atoms may be quaternized. Preferably, these substituents may contain one or more heteroatoms selected from the group consisting of O, S, and NR5.

[0090] In a preferred embodiment, the substituents are -Cl, -F, -Br, -I, -OH, -NH2, -SO3H, -PO3H 、 -PO4H2, -NO2, -CF3, =O, =NR5, -SR5, C1~C 12 Alkyl groups, C2-C 12 Alkenyl groups, C2-C 12 Alkynyl groups, C6-C 12 Aryl groups, C2-C 12 Heteroaryl groups, C3-C 12 Cycloalkyl groups, C5-C 12 Cycloalkenyl group, C 12 Cycloalkynyl group, C3-C 12 Alkyl(hetero)aryl groups, C3-C 12 (Hetero)arylalkyl groups, C4-C 12 (Hetero)arylalkenyl groups, C4-C 12(Hetero)arylalkynyl group, C4-C 12 Alkenyl(hetero)aryl groups, C4-C 12 Alkynyl(hetero)aryl groups, C4-C 12 Alkylcycloalkyl groups, C6-C 12 Alkylcycloalkenyl group, C 13 ~C 16 Alkylcycloalkynyl group, C4-C 12 Cycloalkylalkyl groups, C6-C 12 Cycloalkenylalkyl groups, C 13 ~C 16 Cycloalkynylalkyl groups, C5-C 12 Alkenylcycloalkyl groups, C7-C 12 Alkenylcycloalkenyl group, C 14 ~C 16 Alkenylcycloalkynyl group, C5-C 12 Cycloalkylalkenyl group, C7-C 12 Cycloalkenyl alkenyl group, C 14 ~C 16 Cycloalkynylalkenyl group, C5-C 12 Alkynylcycloalkyl groups, C7-C 12 Alkynylcycloalkenyl group, C 14 ~C 16 Alkynylcycloalkynyl group, C5-C 12 Cycloalkylalkynyl group, C7-C 12 Cycloalkenylalkynyl group, C 14 ~C 16 Cycloalkynylalkynyl group, C5-C 12 Cycloalkyl(hetero)aryl groups, C7-C 12 Cycloalkenyl(hetero)aryl group, C 14 ~C 16 Cycloalkynyl(hetero)aryl groups, C5-C 12 (Hetero)arylcycloalkyl groups, C7-C 12 (Hetero)arylcycloalkenyl groups, and C 14 ~C 16 (hetero)arylcycloalkynyl groups.

[0091] In a preferred embodiment, the substituents are -Cl, -F, -Br, -I, -OH, -NH2, -SO3H, -PO3H 、 -PO4H2, -NO2, -CF3, =O, =NR5, -SR5, C1-C7 alkyl group, C2-C7 alkenyl group, C2-C7 alkynyl group, C6-C7 aryl group, C2-C7 heteroaryl group, C3-C7 cycloalkyl group, C5-C7 cycloalkenyl group, C 12 Cycloalkynyl group, C3-C7 alkyl(hetero)aryl group, C3-C7 (hetero)arylalkyl group, C4-C7 (hetero)arylalkenyl group, C4-C7 (hetero)arylalkynyl group, C4-C7 alkenyl(hetero)aryl group, C4-C7 alkynyl(hetero)aryl group, C4-C7 alkylcycloalkyl group, C6-C7 alkylcycloalkenyl group, C 13 ~C 16 Alkylcycloalkynyl group, C4-C7 cycloalkylalkyl group, C6-C7 cycloalkenylalkyl group, C 13 ~C 16 Cycloalkynylalkyl group, C5-C7 alkenylcycloalkyl group, C7-C7 alkenylcycloalkenyl group, C 14 ~C 16 Alkenylcycloalkynyl group, C5-C7 cycloalkylalkenyl group, C7-C8 cycloalkenylalkenyl group, C 14 ~C 16 Cycloalkynylalkenyl group, C5-C7 alkynylcycloalkyl group, C7-C8 alkynylcycloalkenyl group, C 14 ~C 16 Alkynylcycloalkynyl group, C5-C7 cycloalkylalkynyl group, C7-C8 cycloalkenylalkynyl group, C 14 ~C 16 Cycloalkynylalkynyl group, C5-C7 cycloalkyl(hetero)aryl group, C7-C8 cycloalkenyl(hetero)aryl group, C 14 ~C 16 Cycloalkynyl(hetero)aryl groups, C5-C7 (hetero)arylcycloalkyl groups, C7-C8 (hetero)arylcycloalkenyl groups, and C 14 ~C16 (hetero)arylcycloalkynyl group, C4-C8 (hetero)arylalkenyl group, C4-C8 (hetero)arylalkynyl group, C4-C8 alkenyl(hetero)aryl group, C4-C8 alkynyl(hetero)aryl group, C5-C9 cycloalkyl(hetero)aryl group, C7-C 11 Cycloalkenyl(hetero)aryl group, C 14 ~C 18 Cycloalkynyl(hetero)aryl group, C5-C9(hetero)arylcycloalkyl group, C7-C 11 (Hetero)arylcycloalkenyl groups, and C 14 ~C 18 (hetero)arylcycloalkynyl groups.

[0092] Unless otherwise specified, any non-cyclic group disclosed herein should be understood to be linear or branched. In particular, (hetero)alkyl groups, (hetero)alkenyl groups, (hetero)alkynyl groups, (hetero)alkylene groups, (hetero)alkenylene groups, (hetero)alkynylene groups, etc., are linear or branched unless otherwise specified.

[0093] The general term "sugar" is used herein to refer to monosaccharides such as glucose (Glc), galactose (Gal), mannose (Man), and fucose (Fuc). The term "sugar derivative" is used herein to refer to derivatives of monosaccharide sugars, i.e., monosaccharide sugars having substituents and / or functional groups. Examples of sugar derivatives include amino sugars and sugar acids such as glucosamine (GlcNH), galactosamine (GalNH), N-acetylglucosamine (GlcNAc), N-acetylgalactosamine (GalNAc), sialic acid (Sia), which is also called N-acetylneuraminic acid (NeuNAc) and N-acetylmuramic acid (MurNAc), glucuronic acid (GlcA), and iduronic acid (IdoA).

[0094] A sugar may have no further substitutions and is understood to be a monosaccharide. A sugar may be further substituted at one or more of its hydroxyl groups, in which case it is understood to be a disaccharide or an oligosaccharide. A disaccharide has two monosaccharide moieties linked together. An oligosaccharide chain may be linear or branched and may contain 3 to 10 monosaccharide moieties.

[0095] The term "protein" is used herein in its ordinary scientific sense. Polypeptides having about 10 or more amino acids are considered proteins herein. Proteins may have naturally occurring amino acids, but may also have unnatural amino acids. The term "protein" as used herein is understood to include antibodies and antibody fragments.

[0096] The term "peptide" is used herein in its ordinary scientific sense. As used herein, a peptide is considered to have a number of amino acids ranging from 2 to 9.

[0097] The term "peptoid" is used herein in its ordinary scientific sense.

[0098] An antibody is a protein, typically produced by the immune system, that can recognize and bind to a specific antigen. While antibodies or immunoglobulins derived from IgG antibodies are particularly well suited for use in the present invention, immunoglobulins from any of several classes or subclasses can be selected, such as IgG, IgA, IgM, IgD, and IgE. Suitably, the immunoglobulin is of the IgG subclass (IgG1, IgG2, IgG3, and IgG4) or class IgG, including, but not limited to, class IgM, which can specifically bind to a specific epitope on an antigen. Antibodies can be intact immunoglobulins derived from natural or recombinant sources, and can be immunoreactive portions of intact immunoglobulins.Antibodies can be in a variety of forms, such as polyclonal antibodies, monoclonal antibodies, camelized single domain antibodies, recombinant antibodies, anti-idiotype antibodies, antibody fusions, multispecific antibodies, antibody fragments such as Fv, VHH, Fab, F(ab)2, Fab', Fab'-SH, F(ab')2, single chain variable fragment antibodies (scFv), tandem / bis-scFv, Fc, pFc', scFv-Fc, disulfide Fv (dsFv), bispecific antibodies (bc-scFv), such as BiTE antibodies, trispecific antibody derivatives such as tribodies, camelid antibodies, minibodies, nanobodies, resurfaced antibodies, etc. The antibodies may exist in a variety of forms, including, but not limited to, monoclonal antibodies, humanized antibodies, fully human antibodies, single domain antibodies (sdAb, also known as Nanobodies™), dual-affinity antibodies, e.g., chimeric antibodies, chimeric antibodies with at least one human constant region, dual-affinity retargeting proteins (DART™, dual-affinity retargeting proteins), and multimers and derivatives thereof, such as bivalent or multivalent single-chain variable fragments (e.g., di-scFvs, tri-scFvs), bivalent or multivalent single-chain variable fragments including, e.g., minibodies, diabodies, triabodies, tribodies, tetrabodies, etc., and multivalent antibodies.Reference is made to [Trends in Biotechnology 2015, 33, 2, 65], [Trends Biotechnol. 2012, 30, 575-582], and [Canc. Gen. Prot. 2013 10, 1-18] and [BioDrugs 2014, 28, 331-343], the contents of which are incorporated herein by reference. An "antibody fragment" refers to at least a portion of the variable region of an immunoglobulin that binds to its target, i.e., the antigen-binding region. Other embodiments are drugs or targeting agents. T and the use of antibody mimetics, such as, but not limited to, Affimers, Anticalins, Avimers, Alphabodies, Affibodies, DARPins, and multimers, and derivatives thereof; see [Trends in Biotechnology 2015, 33, 2, 65], the contents of which are incorporated herein by reference. For the avoidance of doubt, in the context of the present invention, the term "antibody" is meant to encompass all of the antibody variants, fragments, derivatives, fusions, analogs, and mimetics outlined in this paragraph, unless otherwise specified.

[0099] A linker is defined herein as a moiety that connects two or more elements of a compound, for example, in a bioconjugate, a biomolecule and a targeting moiety are covalently linked to each other via a linker.

[0100] A biomolecule is defined herein as any molecule that can be isolated from nature or that is composed of smaller molecular building blocks that are components of naturally occurring macromolecular structures, in particular nucleic acids, proteins, glycans, and lipids. Examples of biomolecules include enzymes, (non-catalytic) proteins, polypeptides, peptides, amino acids, oligonucleotides, monosaccharides, oligosaccharides, polysaccharides, glycans, lipids, and hormones.

[0101] As used herein, organic molecule is defined as a molecule that has C-H bond.As used herein, "organic molecule" is understood to include biomolecules, such as nucleic acid (oligonucleotide, polynucleotide, DNA, RNA), peptide, protein (particularly antibody), carbohydrate (monosaccharide, oligosaccharide and polysaccharide), aptamer, hormone, toxin, steroid, cytokine and lipid; small organic molecules as defined herein; polymers (particularly polyethylene glycol); LNA and PNA; amino acid; peptoid; molecule containing radionuclide; fluorescent dye; drug; resin (particularly polystyrene and agarose); beads; particle (particularly polymersome, liposome and bead); gel; surface; organometallic compound; metal complex; cell; and combinations thereof.

[0102] As used herein, inorganic molecule is defined as any molecule that is not an organic molecule, i.e., does not have C-H bond.As used herein, "inorganic molecule" is understood to include surfaces (particularly chips, wafers, gold, metal, silica-based surfaces, such as glass), particles, such as beads (particularly magnetic beads, gold beads), silica-based particles, polymer-based materials, iron oxide particles; carbon nanotubes; carbon allotropes (particularly fullerenes, such as Buckminsterfullerene; graphite; graphene, diamond, Lonsdaleite, Q carbon, linear acetylenic carbon, amorphous carbon, and carbon nanotubes); drugs (particularly cisplatin); metal complexes; and combinations thereof.

[0103] As used herein, a "particle" is preferably defined as a microparticle or a nanoparticle.

[0104] The term "salts thereof" refers to compounds formed when an acidic proton, typically an acid proton, is replaced by a cation, such as a metal cation or an organic cation. The term "salts thereof" also refers to compounds formed when an amine is protonated. Where applicable, the salt is a pharmaceutically acceptable salt, although this is not required for salts that are not intended for administration to patients. For example, in the salts of a compound, the compound may be protonated with an inorganic or organic acid to form a cation, where the conjugate base of the inorganic or organic acid is the anionic component of the salt.

[0105] The term "pharmaceutically acceptable salt" means a salt that is acceptable for administration to a patient, e.g., a mammal (a salt having a counterion that has acceptable mammalian safety for a given dosing regimen). Such salts can be derived from pharmaceutically acceptable inorganic or organic bases and pharmaceutically acceptable inorganic or organic acids.

[0106] "Pharmaceutically acceptable salt" refers to pharmaceutically acceptable salts of a compound, which salts are derived from a variety of organic and inorganic counterions known in the art and include, for example, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, etc., as well as, where the molecule possesses a basic functional group, salts of organic or inorganic acids, such as hydrochloride, hydrobromide, formate, tartrate, besylate, mesylate, acetate, maleate, oxalate, etc.

[0107] The logarithm of the partition-coefficient, or Log P, is used herein as a measure of the hydrophobicity of a compound. Typically, Log P is defined as follows:

[0108]

number

[0109] Those skilled in the art will know how to determine the partition coefficient of a compound without undue experimentation. Alternatively, those skilled in the art will know that software is available to reliably estimate Log P values ​​as a function of, for example, within ChemDraw® software or tools available online.

[0110] The unified atomic mass unit, or Dalton, is abbreviated herein as Da. Those skilled in the art know that the Dalton is a common unit of molecular weight and that 1 Da corresponds to 1 g / mole (gram per mole).

[0111] It will be understood that the words "moiety" and "group" are used interchangeably herein when referring to a portion of a molecule.

[0112] It will be understood that when the heteroatom is -(R')2-, where X is a heteroatom and R' is a moiety, this means that two moieties R' are bonded to the heteroatom.

[0113] The group may be, for example, -((R 51 )2-R 52 )2- or similar notation, where R 51 and R 52 is a part, this means that first the part R 51 and R 52 Instead of selecting parts and writing out the formula, 51 and R 52 -R before the part is selected 51 -R 51 -R 52 -R 51 -R 51 -R 52 - It will be understood that this indicates that the item should be written as

[0114] Inverse Electron Demand Diels-Alder Reaction (IEDDA) Established IEDDA conjugation chemistry generally involves a pair of reactants, including a suitable diene, such as a tetrazine derivative, e.g., an electron-deficient tetrazine, as one reactant (i.e., one bioorthogonal reactive group), and a suitable dienophile, e.g., trans-cyclooctene (TCO), as the other reactant (i.e., the other bioorthogonal reactive group). The extremely fast reaction of a (substituted) tetrazine, particularly an electron-deficient tetrazine, with the TCO moiety results in an intermediate that rearranges to a dihydropyridazine Diels-Alder adduct by eliminating N as the only byproduct. The initially formed 4,5-dihydropyridazine product can tautomerize to either the 1,4- or 2,5-dihydropyridazine product, especially in an aqueous environment. The following reaction scheme shows the [4 + 2] IEDDA reaction between (3,6)-di-(2-pyridyl)-s-tetrazine diene and trans-cyclooctene dienophile, followed by the retro Diels-Alder reaction to form the product and dinitrogen. Because the trans-cyclooctene derivative does not have an electron-withdrawing group like in the classical Diels-Alder reaction, this type of Diels-Alder reaction is distinguished from the classical reaction and is often referred to as the "inverse electron-demand Diels-Alder reaction (IEDDA)." In the following text, both reaction step sequences, i.e., the initial Diels-Alder cycloaddition reaction (typically an inverse electron-demand Diels-Alder cycloaddition reaction) and the subsequent retro Diels-Alder reaction, will be referred to as the "inverse electron-demand Diels-Alder reaction" or "inverse electron-demand Diels-Alder conjugation" or "IEDDA" for short. In this case, the product of the reaction is the IEDDA adduct or conjugate. This is illustrated in Scheme 1 below.

[0115] [ka]

[0116] The two reactive species are abiotic and do not undergo rapid metabolism or side reactions in vitro or in vivo. They are bioorthogonal, i.e., they react selectively with each other in physiological media. Therefore, the compounds and methods of the present invention can be used in living organisms. Moreover, the reactive groups are relatively small and can be introduced into biological samples or organisms without significantly altering the size of the biomolecules in the biological sample or organism. The literature on the inverse electron demand Diels-Alder reaction and the behavior of the pair of reactants includes the following: [Thalhammer et al., Tetrahedron Lett., 1990, 31, 47, 6851-6854], [Wijnen et al., J. Org. Chem., 1996, 61, 2001-2005], [Blackman et al., J. Am. Chem. Soc., 2008, 130, 41, 13518-19], Rossin et al., Angew. Chem. Int. Ed. 2010, 49, 3375], [Devaraj et al., Angew. Chem. Int. Ed. 2009, 48, 7013], [Devaraj et al., Angew. Chem. Int. Ed., 2009, 48, 1-5].

[0117] IEDDA pyridazine elimination reaction Hereinafter, the dienophile, TCO, contained in the combinations and kits of the present invention may be referred to as the "trigger." The dienophile is bound to Construct A at the allylic position. Furthermore, the tetrazine used in the IEDDA pyridazine elimination reaction may be referred to as the "activator." In the present invention, the term Construct A is used to indicate any substance, carrier, biological, or chemical group that is desired to initially place it in a bound (or masked) state and be able to trigger its release from that state.

[0118] The inventors have demonstrated that the dihydropyridazine product derived from a tetrazine (active agent) and a TCO containing a carbamate-linked drug (doxorubicin, Construct-A) at the allylic position tends to eliminate CO and the amine-containing drug, ultimately affording an aromatic pyridazine.

[0119] Without wishing to be bound by theory, the inventors believe that the activator triggers the release of Construct-A via a cascade mechanism within the IEDDA adduct, i.e., dihydropyridazine. The cascade mechanism may be a simple one-step reaction or may consist of multiple steps involving one or more intermediate structures. These intermediates may be stable for some time or may quickly decompose into a thermodynamic end product or the next intermediate structure. In either case, whether it is a simple step or a multi-step process, the result of the cascade mechanism is the release of Construct-A from the IEDDA adduct. Without wishing to be bound by theory, the design of the diene results in an unfavorable distribution of electrons within the IEDDA adduct, and therefore, a rearrangement of these electrons must occur. This situation initiates the cascade mechanism and, therefore, triggers the release of Construct-A. Specifically, without wishing to be bound by theory, the inventors believe that the NH moieties contained in various dihydropyridazine tautomers of the IEDDA adduct, such as the 1,4-dihydropyridazine tautomer, can initiate an electronic cascade reaction, a concerted or sequential shift of electrons across several bonds, resulting in the release of Construct-A. Because the Trigger-Construct-A conjugate itself is relatively stable, the cascade reaction within the Trigger and / or the release of Construct-A from the Trigger cannot occur efficiently or before the IEDDA reaction. The cascade occurs only after the Activator and Trigger-Construct conjugate react and are incorporated into the IEDDA adduct.

[0120] Referring to Scheme 2 below, and without wishing to be bound by theory, the inventors believe that pyridazine elimination occurs from the 1,4-dihydropyridazine tautomer 4. Upon formation of 4,5-dihydropyridazine 3, tautomerization affords intermediates 4 and 7, of which 2,5-dihydropyridazine 7 is C A Instead, it can be slowly converted to aromatic 8, which also A cannot be removed or it can tautomerize back to intermediate 3. Upon formation of 4, C A is removed almost instantly, leaving free C as an amine. A This gives 8, and the pyridazine elimination products 5 and 6. This elimination reaction has been shown to work equally well for cleaving carbonates, esters, and ethers from the TCO trigger [Versteegen et al., Angew. Chem. Int. Ed., 2018, 57, 10494]. The trigger in Scheme 2 can also be used to cleave construct-B (C B ), in which case it cannot be released from the trigger, thereby allowing Construct-A to be separated from Construct-B by IEDDA pyridazine elimination.

[0121] [ka]

[0122] In a preferred embodiment, the dienophile trigger moiety used in the present invention has a trans-cyclooctene ring. In this specification, for ease of reading, this 8-membered ring moiety is defined as a trans-cyclooctene moiety or abbreviated as a "TCO" moiety. It will be understood that the 8-membered ring essentially acts as a dienophile and can be released from its conjugated construct-A upon reaction.

[0123] The dienophiles of the present invention can react with tetrazines in an inverse electron demand Diels-Alder reaction (i.e., DDA). The IEDDA reaction of a trigger with an activator results in the release of Construct A via an electron cascade-based elimination, termed "pyridazine elimination." When an activator reacts with a trigger capable of removing Construct A, the combined process of reaction and removal of Construct A is termed "IEDDA pyridazine elimination."

[0124] The present invention provides a trigger conjugated to Construct-A that reacts with an activator, resulting in cleavage of the trigger from Construct-A. In one notable embodiment, this results in cleavage of Construct-A from Construct-B. In other embodiments, the trigger cleavage results in cleavage of one Construct-A from another Construct-A, where both can be released from the self-immolative linker attached to the trigger. In other embodiments, the trigger cleavage results in cleavage of one or more Construct-A from one or more Construct-B. Construct-B is a construct that is bound to a dienophile and cannot be released from the dienophile unless it is bound at the allylic position via a spacer or automimetic linker that is also attached to Construct-A. In a preferred embodiment, the trigger is used as a reversible covalent bond between two molecular species.

[0125] Scheme 3a below is a general scheme for construct release according to the present invention, where the released construct is Construct-A (C A ), and another construct, Construct-B (C B ), may be attached to the dienophile but not through an allylic position, where Construct-B cannot be released from the dienophile, and where either Construct-A or -B is the administration agent and the other is the label. Formula 1

[0126] JPEG2026004404000015.jpg119170

[0127] Scheme 3b below is a general scheme for releasing a construct according to another embodiment of the present invention, wherein Construct-B (C B ) is attached to the dienophile via a spacer or self-immolative linker that also attaches to Construct-A, and wherein when the spacer or self-immolative linker is released from the allylic position, Construct-B and Construct-A are released from the Trigger and from each other.

[0128] JPEG2026004404000016.jpg122170

[0129] Release of the construct occurs through a potent abiotic, bioorthogonal reaction between a dienophile (trigger) and a diene (activator), i.e., IEDDA, ​​as previously described. The masked or bound construct is a construct-dienophile conjugate. Construct A is likely linked to one or more additional constructs A via self-immolative linkers. It will be understood that the dienophile and diene groups shown in Scheme 3 in the IEDDA adduct and in the final product after release are residues of the dienophile and diene groups, respectively, after these groups are converted in the IEDDA reaction.

[0130] C A and C B The difference is C B and C B The bond between the moiety holding the trigger and the diene is not cleaved upon reaction with the diene, while the bond between the trigger and the diene holding the trigger and the diene is not cleaved upon reaction with the diene. A and C A The bond between the trigger and the moiety holding the C is cleaved upon reaction with the diene. A and C B The portion holding the trigger is a self-immolative linker L CTo clarify, C B is connected to the trigger C If it is connected to C B L holding C is released from the trigger upon reaction with the diene, while C B is released L C Similarly, C B If is directly connected to the trigger, C B will not be released from the trigger upon reaction with the diene. Those skilled in the art will understand that if it is necessary to separate one construct (1) from another construct (2), one of the following requirements must be met: 1) One C A is construct 1, and other C A is construct 2, 2) Construct 1 is C A and construct 2 is C B That is, 3) Constructs 1 and 2 are both C B However, one C B is directly connected to the trigger, and the other is L C or, provided that one of the C B The part is the other C B Partially different L C is connected to the part.

[0131] In one aspect, the present invention provides the use of a tetrazine as an activator for the release of a TCO-linked construct in a chemical, biological, or physiological environment. In this regard, the present invention also relates to a tetrazine as an activator for the release of a TCO-linked substance in a chemical, biological, or physiological environment. The fact that the reaction is bioorthogonal and that many structural options exist for reaction pairs will be apparent to those skilled in the art. For example, the IEDDA reaction is known in the fields of bioconjugation, diagnostics, and pre-targeted medicine. See, for example, WO 2010 / 119382, WO 2010 / 119389, and WO 2010 / 051530. While the present invention presents an entirely different use of the reaction, it will be understood that the various structural possibilities available for IEDDA reaction pairs, such as those used in pre-targeting, are also applicable in the field of the present invention.

[0132] For example, in medically active substances, whose in vitro or in vivo actions often change with small structural changes, the present invention requires, first and foremost, appropriate chemical reactivity combined with sufficient stability for the intended use. Thus, possible structures extend to those known by those skilled in the art to be reactive as dienophiles.

[0133] Compounds according to formula (19) In formula (19), r is an integer ranging from 0 to 2. In a preferred embodiment, r is 0. In a preferred embodiment, r is 1. In a preferred embodiment, r is 2. In formula (19), each s is independently 0 or 1. In a preferred embodiment, s is 0. In a preferred embodiment, s is 1. In formula (19), each i is independently an integer ranging from 0 to 4, preferably 0 or 1. In formula (19), j is an integer ranging from 0 to 4, preferably 0 to 2, more preferably 0 or 1. In formula (19), each k is independently 0 or 1.

[0134] In a preferred embodiment, in formula (19), only condition (a) is satisfied.

[0135] In a preferred embodiment, in equation (19), only condition (b) is satisfied.

[0136] In a preferred embodiment, in formula (19), only condition (c) is satisfied.

[0137] In a preferred embodiment, two R 37 , R 38 , R 47 may be included in a ring to form a ring fused to an 8-membered trans ring, the ring fused to the 8-membered trans ring is as defined in WO 2012 / 156919 A1, which is incorporated herein by reference in its entirety. More preferably, the ring fused to the 8-membered trans ring is as defined in WO 2012 / 156919 A1, page 15, line 25 to page 18, line 9.

[0138] Z T Z T is C1~C 12 Alkylene group, C2-C 12 Alkenylene group, C7-C 12Alkynylene group, C6 arylene group, C4-C5 heteroarylene group, C3-C8 cycloalkylene group, C5-C8 cycloalkenylene group, C5-C 12 Alkyl(hetero)arylene groups, C5-C 12 (Hetero)arylalkylene group, C4-C 12 Alkylcycloalkylene group, C4-C 12 and a cycloalkylalkylene group, wherein the above alkylene group, alkenylene group, alkynylene group, (hetero)arylene group, cycloalkylene group, cycloalkenylene group, alkyl(hetero)arylene group, (hetero)arylalkylene group, alkylcycloalkylene group, and cycloalkylalkylene group are selected from the group consisting of (S P ) i -C B (wherein i is independently a number ranging from 0 to 4, and preferably i is 0 or 1), -Cl, -F, -Br, -I, -OR 37 , -N(R 37 )2, =O, =NR 37 , -SR 37 , -SO3H, -PO3H 、 -PO4H2, -NO2, and -Si(R 37 )3, and is optionally substituted with a moiety selected from the group consisting of —O—, —S—, —NR 37 It may contain one or more heteroatoms selected from the group consisting of -, -P- and -Si-, wherein the N, S and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0139] Preferably, Z Tis selected from the group consisting of a C1-C6 alkylene group, a C2-C6 alkenylene group, a C7 alkynylene group, a C6 arylene group, a C4-C5 heteroarylene group, a C3-C6 cycloalkylene group, a C5-C8 cycloalkenylene group, a C5-C8 alkyl(hetero)arylene group, a C5-C8 (hetero)arylalkylene group, a C4-C8 alkylcycloalkylene group, and a C4-C8 cycloalkylalkylene group, wherein the above alkylene group, alkenylene group, alkynylene group, (hetero)arylene group, cycloalkylene group, cycloalkenylene group, alkyl(hetero)arylene group, (hetero)arylalkylene group, alkylcycloalkylene group, and cycloalkylalkylene group are each independently selected from the group consisting of (S P ) i -C B (wherein i is independently a number ranging from 0 to 4, and preferably i is 0 or 1), -Cl, -F, -Br, -I, -OR 37 , -N(R 37 )2, =O, =NR 37 , -SR 37 , -SO3H, -PO3H 、 -PO4H2, -NO2, and -Si(R 37 )3, and is optionally substituted with a moiety selected from the group consisting of —O—, —S—, —NR 37 It may contain one or more heteroatoms selected from the group consisting of -, -P- and -Si-, wherein the N, S and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0140] Preferably, Z T is selected from the group consisting of a C1 to C3 alkylene group, a C2 to C3 alkenylene group, a C3 alkynylene group, a C3 heteroarylene group, and a C3 cycloalkylene group, wherein the alkylene group, the alkenylene group, the alkynylene group, the heteroarylene group, and the cycloalkylene group are each selected from the group consisting of (S P ) i -C B (wherein i is independently a number ranging from 0 to 4, and preferably i is 0 or 1), -Cl, -F, -Br, -I, -OR 37 , -N(R 37)2, =O, =NR 37 , -SR 37 , -SO3H, -PO3H 、 -PO4H2, -NO2, and -Si(R 37 )3, and is optionally substituted with a moiety selected from the group consisting of —O—, —S—, —NR 37 In a preferred embodiment, the Z may contain one or more heteroatoms selected from the group consisting of -, -P-, and -Si-, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized. T In a preferred embodiment, the Z group is unsubstituted. T The group does not have heteroatoms.

[0141] R 48 Preferably, R 48 -OC(O)-(S P ) k C A If and S P (if k>0) or C A (when k=0) is bonded to -OC(O)- through an atom selected from the group consisting of O, C, S and N, preferably secondary or tertiary N, where this atom is S P or C A Preferably, R 48 Ga-OL C -(S P ) k C A If and S P (if k>0) or C A (when k=0) is connected to L through an atom selected from the group consisting of -O-, -S-, and -N-, preferably secondary N or tertiary N. C wherein the moiety is S P or C A It is part of R 48 is preferably arranged in the axial direction of the eight-membered dienophile ring. 48 is H a Preferably, the transformer is disposed relatively to the

[0142] Other Preferred Embodiments In a preferred embodiment, Y T1 is OH, N(R 38 )2, C(O)OH, ON(R 38 )2, SH, more preferably OH, N(R 38 )2, ON(R 38 )2, more preferably OH, N(R 38 )2, and most preferably Y T1 is OH. In another preferred embodiment, Y T1 is N(R 38 )2, more preferably NR 38 H, most preferably NH. In a preferred embodiment, Y T2 and Y T3 does not exist and Y T1 is OH, N(R 38 )2, C(O)OH, ON(R 38 )2, more preferably OH, N(R 38 )2, and most preferably Y T1 is OH. In a preferred embodiment, Y T2 is OH. In a preferred embodiment, Y T1 and Y T3 does not exist and Y T2 is OH. X 1 and X 5 is preferably not O.

[0143] In a preferred embodiment, when a fused ring satisfying any of formulas (20a) through (20f) is present, there are no other rings fused to the 8-membered dienophile ring.

[0144] In a preferred embodiment, R 37、 R 38、 R 47 The groups are OH, SH, N(R 38 )2, ON(R 38 )2, C(N)N(R 38 )2. In a preferred embodiment, R 37、 R 38、 R47 The base is 、 OH, SH, N(R 38 )2, C(O)OH, C(S)OH, C(O)SH, C(S)SH, ON(R 38 )2, SO4H, SO3H, SO2H, PO4H2, PO3H, PO2H, C(N)N(R 38 )2.

[0145] In a preferred embodiment, the compound of formula (19) is Y T1 , Y T2 , Y T3 and more preferably, no more than two moieties, and even more preferably, one moiety, each independently selected from the group consisting of:

[0146] In a preferred embodiment, X 2 , X 3 , X 4 One of them is CR 47 Y T1 and most preferably X 3 or X 4 is CR 47 Y T1 In a preferred embodiment, X 2 , X 3 , X 4 One of them is CR 47 Y T1 and most preferably X 3 or X 4 is CR 47 Y T1 and the remaining X 1 , X 2 , X 3 , X 4 , X 5 is C(R 47 )2, preferably CH 2、 In a preferred embodiment, X 2 , X 3 , X 4 Two of them are CR 47 Y T1 and the remaining X 1 , X 2 , X 3 , X 4 , X5 is C(R 47 )2, preferably CH 2、 In a preferred embodiment, X 3 is Y T3 and the remaining X 1 , X 2 , X 3 , X 4 , X 5 is C(R 47 )2, preferably CH 2、 In a preferred embodiment, X 2 and X 3 is a part of a fused ring satisfying one of formulas (20a) to (20g), and the remaining X 1 , X 2 , X 3 , X 4 , X 5 is C(R 47 )2, preferably CH 2、 is.

[0147] In a preferred embodiment, X 3 and X 4 is a part of a fused ring satisfying one of formulas (20a) to (20g), and the remaining X 1 , X 2 , X 3 , X 4 , X 5 is C(R 47 )2, preferably CH 2、 is.

[0148] In a preferred embodiment, the fused ring satisfies formula (20a): In a preferred embodiment, the fused ring satisfies formula (20b): In a preferred embodiment, the fused ring satisfies formula (20c):

[0149] In the case of formula (20g), X 6 and X 8 Both are Y T3 If X 7 is preferably not a C1 alkylene. 6 and X 8 Both are Y T3, preferably NR 38 and X 7 is preferably a C2 alkylene. In the case of formula (20g), X 6 and X 8 are both C(R 47 ) 2、 Preferably CH 2、 and X 7 is Y T3 , preferably NR 38、 It is preferable that:

[0150] In a preferred embodiment, X a and X b In a preferred embodiment, the fused ring satisfies formula (20a) and X a and X b is CH.

[0151] In a preferred embodiment, X 1 , X 2 , X 3 , X 4 , and X 5 R included in 37、 R 38、 R 47 4 or less of the X parts (i.e., not per part X) 1 ~X 5 (total for each) are not H, preferably no more than 3 are not H, more preferably no more than 2 are not H, and most preferably no more than 1 is not H.

[0152] X 1 , X 2 , X 3 , X 4 , X 5 The two R's included in 37、 R 38、 R 47 When groups are included in a ring to form rings fused to an 8-membered trans ring, the rings fused to the 8-membered trans ring are R 47 Preferably, the cycloalkylene group is a C3-C7 cycloalkylene group or a C4-C7 cycloalkenylene group, which may be substituted with or contain a heteroatom, as described above.

[0153] In a preferred embodiment, C A and / or C. B is R as defined herein 32 is attached to the rest of the molecule via a residue of 32 is equal to or is included in the spacer.

[0154] Those skilled in the art will recognize that "R 32 The residue of "C A and / or C. B and trigger, spacer or L C R for forming a conjugate between 32 It will be understood that the term "conjugated" refers to the conjugated reaction product of a hydroxyl group with another chemical group.

[0155] In other embodiments, C A and / or C. B is C as defined herein M2 is attached to the rest of the molecule via M2 is equal to or included in the spacer.

[0156] In still other embodiments, C A and / or C. B is C as defined herein X is attached to the rest of the molecule via X is equal to or included in the spacer.

[0157] In a preferred embodiment, moiety C X , C M2 and R 32 The residue is C A and / or C. B is included in.

[0158] In a preferred embodiment, C M2is selected from the group consisting of amine, amide, thioamide, aminooxy, ether, carbamate, thiocarbamate, urea, thiourea, sulfonamide, and sulfonecarbamate.

[0159] In a preferred embodiment, C M2 is R 10 is equal to.

[0160] In a preferred embodiment, C M2 is C X In a preferred embodiment, C M2 is as follows:

[0161] [ka]

[0162] [ka]

[0163] Here, the dashed line indicates C A Or C B Bond to, or C A Or C B and the wavy line indicates the bond to the rest of the dienophile. A Or C B Bond to, or C A Or C B and the dashed line indicates the bond to the rest of the dienophile.

[0164] In a preferred embodiment, C X is as follows:

[0165] [ka]

[0166] Here, the dashed line indicates C A Or C BBond to, or C A Or C B and the wavy line indicates the bond to the rest of the dienophile. A Or C B Bond to, or C A Or C B and the dashed line indicates the bond to the rest of the dienophile.

[0167] C M2 and C X Referring to the scheme above with examples of A Or C B is a protein, e.g., an antibody, the dashed line A Or C B Bond to, or C A Or C B The bond to

[0168] In a preferred embodiment, when k or i is 0, C A and / or C. B -O-, -C(R 6 )2-, -NR 6 is linked to the remainder of Formula 19 through a moiety selected from the group consisting of -, -C(O)-, and -S-, wherein the moiety is C A and / or C. B is part of.

[0169] In a preferred embodiment, when k or i is at least 1, C A and / or C. B -O-, -C(R 6 )2-, -NR 6 -S- via a moiety selected from the group consisting of -, -C(O)-, and -S- P wherein the moiety is linked to C A and / or C. B is part of, and S P is -O-, -C(R 6 )2-, -NR 6is linked to the remainder of Formula 19 through a moiety selected from the group consisting of -, -C(O)-, and -S-, wherein the moiety is S P is part of.

[0170] In a preferred embodiment, no more than three C B is included in the structure of formula (19), more preferably two or less, most preferably one or less, C B is included in the structure of formula (19).

[0171] In a preferred embodiment, C prior to conjugation to the remainder of the compound of formula (19) A and / or C. B -OH, -NHR', -CO2H, -SH, -SS-, -SCH3-, -N3, terminal alkynyl, terminal alkenyl, -C(O)R', C8 to C 12 and at least one moiety selected from the group consisting of (hetero)cycloalkynyl, C-C cycloalkenyl, nitrone, nitrile oxide, (imino)sydnone, isonitrile, (oxa)norbornene, and tetrazine, wherein the moiety satisfies formula (19) and is C M2 or C X To form a compound containing the moiety, a dienophile and an R 32 is used for conjugation to a moiety containing

[0172] In a preferred embodiment, the C A and / or C. B is selected from the group consisting of amines, amides, thioamides, aminooxys, carbamates, thiocarbamates, ureas, thioureas, sulfonamides, and sulfonecarbamates. M2 is attached to the remainder of the compound of formula (19) via

[0173] In a preferred embodiment, C M2 is R 10 is equal to.

[0174] In a preferred embodiment, C A or C BWhen is conjugated via -SH or -SS-, C M2 is selected from the group consisting of:

[0175] [ka]

[0176] where the wavy line indicates the bond to the rest of the molecule and the dashed line indicates the C A or C B The bond to

[0177] In a preferred embodiment, moiety C A , C B or C when the Administration Agent is conjugated via -SMe M2 is as follows:

[0178] [ka]

[0179] where the wavy line indicates the bond to the rest of the molecule and the dashed line indicates the C A , C B or exhibits binding to the administered agent.

[0180] In a preferred embodiment, C A or C B When is conjugated via -NR'-, C M2 is selected from the group consisting of:

[0181] [ka]

[0182] where the wavy line indicates the bond to the rest of the molecule and the dashed line indicates the C A or C B The bond to

[0183] In a preferred embodiment, CA or C B is conjugated via -C- derived from a moiety that was -C(O)R' or -C(O)R'-, C M2 is selected from the group consisting of:

[0184] [ka]

[0185] where the wavy line indicates the bond to the rest of the molecule and the dashed line indicates the C A or C B The bond to

[0186] In a preferred embodiment, C A or C B is conjugated via —C(O)—, which is derived from the moiety that was —C(O)OH, M2 is selected from the group consisting of:

[0187] [ka]

[0188] where the wavy line indicates the bond to the rest of the molecule and the dashed line indicates the C A or C B The bond to

[0189] In a preferred embodiment, C A or C B When is conjugated via -O-, C M2 is selected from the group consisting of:

[0190] [ka]

[0191] where the wavy line indicates the bond to the rest of the molecule and the dashed line indicates the C A or C B The bond to

[0192] In a preferred embodiment, C A or C B R containing an alkyne group 32 When conjugated via -N3, the resulting C X has a triazole ring, wherein each C X is independently selected from the group consisting of:

[0193] [ka]

[0194] where the wavy line indicates the bond to the rest of the molecule and the dashed line indicates the C A or C B The bond to

[0195] R 6 Preferably, each R 6 are independently hydrogen atoms, -(S P ) i -C B , C1~C 24 Alkyl groups, C2-C 24 Alkenyl groups, C2-C 24 Alkynyl groups, C6-C 24 Aryl groups, C2-C 24 Heteroaryl groups, C3-C 24 Cycloalkyl groups, C5-C 24 Cycloalkenyl group, C 12 ~C 24 Cycloalkynyl group, C3-C 24 (Cyclo)alkyl(hetero)aryl groups, C3-C 24 (Hetero)aryl(cyclo)alkyl, C4-C 24 (Cyclo)alkenyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkenyl groups, C4-C 24 (Cyclo)alkynyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkynyl group, C4-C24 An alkylcycloalkyl group, and C4 - C 24 Selected from the group consisting of cycloalkylalkyl groups, where i is an integer in the range of 0 - 4, preferably, i is 0 or 1; Here, R where it is not a hydrogen atom 6 The group may be substituted with a moiety selected from the group consisting of -Cl, -F, -Br, -I, -OH, -NH2, -SO3H, -PO3H 、 -PO4H 2、 -NO2, -CF3, =O, =NH, and -SH, and may contain one or more heteroatoms selected from the group consisting of O, S, NH, P, and Si, where the N atom, S atom, and P atom may be oxidized, and where multiple N atoms may be quaternized.

[0196] In a preferred embodiment, each R 6 Is individually a hydrogen atom, a C1 - C8 alkyl group, a C2 - C8 alkenyl group, a C2 - C8 alkynyl group, a C6 - C 12 Aryl, C2 - C 12 Heteroaryl, a C3 - C8 cycloalkyl group, a C5 - C8 cycloalkenyl group, a C3 - C 12 Alkyl(hetero)aryl group, a C3 - C 12 (Hetero)arylalkyl group, a C4 - C 12 Alkylcycloalkyl group, a C4 - C 12 Cycloalkylalkyl group, a C5 - C 12 Cycloalkyl(hetero)aryl group, and a C5 - C 12 (Hetero)arylcycloalkyl group, and where R where it is not a hydrogen atom 6 The group may be substituted with a moiety selected from the group consisting of -Cl, -F, -Br, -I, -OH, -NH2, -SO3H, -PO3H 、 -PO4H 2、 -NO2, -CF3, =O, =NH, and -SH, and may contain one or more heteroatoms selected from the group consisting of O, S, NH, P, and Si, where the N atom, S atom, and P atom may be oxidized, and where multiple N atoms may be quaternized.

[0197] In a preferred embodiment, R 6 is a hydrogen atom, a C1-C4 alkyl group, a C2-C4 alkenyl group, a C2-C4 alkynyl group, a C6-C8 aryl, a C2-C8 heteroaryl, a C3-C6 cycloalkyl group, a C5-C6 cycloalkenyl group, a C3-C 10 Alkyl(hetero)aryl groups, C3-C 10 (Hetero)arylalkyl group, C4-C8 alkylcycloalkyl group, C4-C8 cycloalkylalkyl group, C5-C 10 Cycloalkyl(hetero)aryl groups and C5-C 10 (hetero)arylcycloalkyl groups, wherein any R that is not a hydrogen atom is selected from the group consisting of 6 Groups include -Cl, -F, -Br, -I, -OH, -NH2, -SO3H, and -PO3H 、 -PO4H 2、 It may be substituted with a moiety selected from the group consisting of -NO2, -CF3, =O, =NH, and -SH, and may contain one or more heteroatoms selected from the group consisting of O, S, NH, P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0198] In a preferred embodiment, R 6 represents a hydrogen atom, a C1-C4 alkyl group, a C2-C4 alkenyl group, and C 4~6 (hetero)aryl groups, where R 6 In the above, the alkyl group, alkenyl group, and (hetero)aryl group are -Cl, -F, -Br, -I, -OH, -NH2, ═O, -SH, -SO3H, -PO3H 、 It may be substituted with a moiety selected from the group consisting of -PO4H2 and -NO2, and may contain up to two heteroatoms selected from -O-, -S-, -NH-, -P- and -Si-, wherein the N, S and P atoms are optionally oxidized.

[0199] In a preferred embodiment, R6 represents a hydrogen atom, a C1-C3 alkyl group, a C2-C3 alkenyl group, and C 4~6 (hetero)aryl groups, where R 6 In the above, the alkyl group, alkenyl group, and (hetero)aryl group are -Cl, -F, -Br, -I, -OH, -NH2, ═O, -SH, -SO3H, -PO3H 、 It may be substituted with a moiety selected from the group consisting of -PO4H2 and -NO2, and may contain up to two heteroatoms selected from -O-, -S-, -NH-, -P- and -Si-, wherein the N, S and P atoms are optionally oxidized.

[0200] In a preferred embodiment, R is not a hydrogen atom. 6 The group is unsubstituted. In a preferred embodiment, R is not a hydrogen atom. 6 The group does not contain heteroatoms. In a preferred embodiment, R 6 The group is a hydrogen atom.

[0201] R 7 In a preferred embodiment, each R 7 are independently hydrogen atoms, -(S P ) i -C B , -F, -Cl, -Br, -I, -OR 37 , -N(R 37 )2, -SO3, -PO3 - 、 -NO2, -CF3, -SR 37 , S(=O)2N(R 37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R 37 , SC(=S)R 37 , N.R. 37 C(=O)-R 37 , N.R. 37 C(=S)-R 37 , N.R. 37 C(=O)OR 37 , N.R. 37C(=S)OR 37 , N.R. 37 C(=O)SR 37 , N.R. 37 C(=S)SR 37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R 37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR 37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37 )2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , S(O)R 37 , -S(O)2R 37 , N.R. 37 S(O)2R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , C1~C 24 Alkyl groups, C2-C 24 Alkenyl groups, C2-C 24 Alkynyl groups, C6-C 24 Aryl groups, C2-C 24 Heteroaryl groups, C3-C 24 Cycloalkyl groups, C5-C 24 Cycloalkenyl group, C 12 ~C 24 Cycloalkynyl group, C3-C 24 (Cyclo)alkyl(hetero)aryl groups, C3-C 24 (Hetero)aryl(cyclo)alkyl, C4-C 24 (Cyclo)alkenyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkenyl groups, C4-C 24 (Cyclo)alkynyl(hetero)aryl group, C4-C 24(Hetero)aryl(cyclo)alkynyl group, C4-C 24 Alkylcycloalkyl groups and C4-C 24 cycloalkylalkyl groups, where i is an integer ranging from 0 to 4, preferably i is 0 or 1; Here, the above alkyl group, alkenyl group, alkynyl group, aryl, heteroaryl, cycloalkyl group, cycloalkenyl group, cycloalkynyl group, (cyclo)alkyl(hetero)aryl group, (hetero)aryl(cyclo)alkyl group, (cyclo)alkenyl(hetero)aryl group, (hetero)aryl(cyclo)alkenyl group, (cyclo)alkynyl(hetero)aryl group, (hetero)aryl(cyclo)alkynyl group, alkylcycloalkyl group, cycloalkylalkyl group are -Cl, -F, -Br, -I, -OR 37 , -N(R 37 )2, -SO3R 37 , -PO3(R 37 ) 2、 -PO4(R 37 ) 2、 -NO2, -CF3, =O, =NR 37 , and -SR 37 and may be substituted with a moiety selected from the group consisting of O, S, NR 37 , P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0202] In a preferred embodiment, each R 7 are independently a hydrogen atom, -F, -Cl, -Br, -I, or -OR 37 , -N(R 37 )2, -SO3, -PO3 - 、 -NO2, -CF3, -SR 37 , S(=O)2N(R 37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R 37 , SC(=S)R 37 , N.R. 37C(=O)-R 37 , N.R. 37 C(=S)-R 37 , N.R. 37 C(=O)OR 37 , N.R. 37 C(=S)OR 37 , N.R. 37 C(=O)SR 37 , N.R. 37 C(=S)SR 37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R 37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR 37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37 )2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , S(O)R 37 , -S(O)2R 37 , N.R. 37 S(O)2R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , C1-C8 alkyl group, C2-C8 alkenyl group, C2-C8 alkynyl group, C6-C 12 Aryl groups, C2-C 12 Heteroaryl group, C3-C8 cycloalkyl group, C5-C8 cycloalkenyl group, C3-C 12 Alkyl(hetero)aryl groups, C3-C 12 (Hetero)arylalkyl groups, C4-C 12 Alkylcycloalkyl groups, C4-C 12 Cycloalkylalkyl groups, C5-C 12 Cycloalkyl(hetero)aryl groups and C5-C 12and (hetero)arylcycloalkyl groups, wherein the alkyl group, alkenyl group, alkynyl group, aryl, heteroaryl, cycloalkyl group, cycloalkenyl group, alkyl(hetero)aryl group, (hetero)arylalkyl group, alkylcycloalkyl group, cycloalkylalkyl group, cycloalkyl(hetero)aryl group, and (hetero)arylcycloalkyl group are selected from the group consisting of -Cl, -F, -Br, -I, -OR 37 , -N(R 37 )2, -SO3R 37 , -PO3(R 37 ) 2、 -PO4(R 37 ) 2、 -NO2, -CF3, =O, =NR 37 , and -SR 37 and may be substituted with a moiety selected from the group consisting of O, S, NR 37 , P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0203] In a preferred embodiment, each R 7 are independently a hydrogen atom, -F, -Cl, -Br, -I, or -OR 37 , -N(R 37 )2, -SO3, -PO3 - 、 -NO2, -CF3, -SR 37 , S(=O)2N(R 37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R 37 , SC(=S)R 37 , N.R. 37 C(=O)-R 37 , N.R. 37 C(=S)-R 37 , N.R. 37 C(=O)OR 37 , N.R. 37 C(=S)OR 37 , N.R. 37 C(=O)SR 37 , N.R. 37C(=S)SR 37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R 37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR 37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37 )2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , S(O)R 37 , -S(O)2R 37 , N.R. 37 S(O)2R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , C1-C4 alkyl group, C2-C4 alkenyl group, C2-C4 alkynyl group, C6-C8 aryl group, C2-C8 heteroaryl group, C3-C6 cycloalkyl group, C5-C6 cycloalkenyl group, C3-C 10 Alkyl(hetero)aryl groups, C3-C 10 (Hetero)arylalkyl groups, C4-C 10 Alkylcycloalkyl groups, C4-C 10 Cycloalkylalkyl groups, C5-C 10 Cycloalkyl(hetero)aryl groups and C5-C 10 and (hetero)arylcycloalkyl groups, wherein the alkyl group, alkenyl group, alkynyl group, aryl, heteroaryl, cycloalkyl group, cycloalkenyl group, alkyl(hetero)aryl group, (hetero)arylalkyl group, alkylcycloalkyl group, cycloalkylalkyl group, cycloalkyl(hetero)aryl group, and (hetero)arylcycloalkyl group are selected from the group consisting of -Cl, -F, -Br, -I, -OR 37 , -N(R37 )2, -SO3R 37 , -PO3(R 37 ) 2、 -PO4(R 37 ) 2、 -NO2, -CF3, =O, =NR 37 , and -SR 37 and may be substituted with a moiety selected from the group consisting of O, S, NR 37 , P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0204] In a preferred embodiment, each R 7 are independently a hydrogen atom, a C1-C3 alkyl group, a C2-C3 alkenyl group, and C 4~6 and (hetero)aryl groups, wherein the alkyl, alkenyl, and (hetero)aryl groups are optionally substituted with a moiety selected from the group consisting of -Cl, -F, -Br, -I, -OH, -NH, =O, =NH, -N(CH), -S(O)CH, and -SH, and may contain up to one heteroatom selected from the group consisting of -O-, -S-, -NH-, -P-, and -Si-, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0205] In a preferred embodiment, R 7 is preferably selected from the group consisting of a hydrogen atom, methyl, —CH—CH—N(CH) and —CH—CH—S(O)—CH. In a preferred embodiment, R that is not a hydrogen atom 7 The group is unsubstituted. In a preferred embodiment, R is not a hydrogen atom. 7 The group does not contain heteroatoms. In a preferred embodiment, R 7 The group is a hydrogen atom.

[0206] R 8 and R9 Preferably, R 8 and R 9 is R 7 In a preferred embodiment, at least one or all of R 8 is —H. In a preferred embodiment, at least one or all of R 8 is —CH3. In a preferred embodiment, at least one or all of R 9 is —H. In a preferred embodiment, at least one or all of R 9 is -CH3.

[0207] R 32 R 32 is a conjugation moiety, which is a part of a construct, such as construct-B, or a spacer, linker, L C A conjugate or coupling is a chemical group that can be used for the conjugation or coupling of a ligand, a trigger, or other molecule or construct of interest. Those skilled in the art are aware of the myriad strategies available for chemoselective, non-selective, or enzymatic coupling or conjugation of one molecule or construct to another.

[0208] In a preferred embodiment, R 32 is a moiety that allows conjugation to proteins containing natural and / or unnatural amino acids. Moieties suitable for conjugation are known to those skilled in the art. Conjugation strategies can be found, for example, in [O. Boutureira, GJL Bernardes, Chem. Rev., 2015, 115, 2174-2195].

[0209] In a particularly preferred embodiment, R 32is an N-maleimidyl group, a halogenated N-alkylamide group, a sulfonyloxy N-alkylamide group, a vinyl sulfone group, an (activated) carboxylic acid, a benzenesulfonyl halide, an ester group, a carbonate group, a sulfonyl halide group, a thiol group, or a derivative thereof, C 2~6 Alkenyl group, C 2~6 Alkynyl group, C 7~18 Cycloalkynyl group, C 5~18 Heterocycloalkynyl group, bicyclo[6.1.0]non-4-yn-9-yl] group, C 3~12 Cycloalkenyl groups, azide groups, phosphine groups, nitrile oxide groups, nitrone groups, nitrile imine groups, isonitrile groups, diazo groups, ketone groups, (O-alkyl)hydroxylamino groups, hydrazine groups, N-maleimidyl halide groups, aryloxymaleimides, dithiophenolmaleimides, bromo- and dibromo-pyridazinediones, 2,5-dibromohexanediamide groups, alkynone groups, 3-arylpropiolonitrile groups, 1,1-bis(sulfonylmethyl)-methylcarbonyl groups, or elimination derivatives thereof. derivatives), carbonyl halide groups, allenamide groups, 1,2-quinone groups, isothiocyanate groups, isocyanate groups, aldehyde groups, triazine groups, tetrazine groups, squaric acid, 2-imino-2-methoxyethyl groups, (oxa)norbornene groups, (imino)sydnone, methylsulfonylphenyloxadiazole groups, aminoxy groups, 2-aminobenzamidoxime groups, ethynylphosphonamidate, groups reactive in Pictet-Spengler ligation and hydrazino-Pictet-Spengler (HIPS) ligation, DNA intercalators, and photocross linkers.

[0210] In a preferred embodiment, R 32 is an N-maleimidyl group connected to the remainder of the compound according to formula (19) via the N atom of the N-maleimidyl group.

[0211] In another preferred embodiment, R32 is selected from the group consisting of a hydroxyl group, an amine group, a halogen, a vinylpyridine group, a disulfide group, a pyridyl disulfide group, a sulfonyloxy group, a mercaptoacetamide group, an anhydride group, a sulfonylated hydroxyacetamide group, a sulfonyl chloride, a thiosemicarbazone, a hydrazine carboxylate, and an aryl hydrazide.

[0212] In other embodiments, R 32 is a group that can be attached to another group by an enzyme, such as a sortase or tubulin tyrosine ligase.

[0213] R 36 In a preferred embodiment, R 36 is R 37 is as defined for

[0214] In equation (19), R 36 is preferably a hydrogen atom, a C1 to C8 alkyl group, a C2 to C8 alkenyl group, and C 4~6 and (hetero)aryl groups, wherein the alkyl, alkenyl, and (hetero)aryl groups are selected from the group consisting of -Cl, -F, -Br, -I, -OH, -NH2, =O, -SH, -SO3H, -PO3H. 、 It may be substituted with a moiety selected from the group consisting of -PO4H2 and -NO2, and may contain up to two heteroatoms selected from the group consisting of -O-, -S-, -NH-, -P- and -Si-, where the N, S and P atoms are optionally oxidized.

[0215] In preferred embodiments, R36 groups that are not hydrogen atoms are unsubstituted. In preferred embodiments, R36 groups that are not hydrogen atoms do not contain heteroatoms.

[0216] R 37 Preferably, each R37 are independently hydrogen atoms, -(S P ) i -C B , C1-C8 alkyl group, C2-C8 alkenyl group, C2-C8 alkynyl group, C6-C 12 Aryl, C2-C 12 Heteroaryl, C3-C8 cycloalkyl group, C5-C8 cycloalkenyl group, C3-C 12 Alkyl(hetero)aryl groups, C3-C 12 (Hetero)arylalkyl groups, C4-C 12 Alkylcycloalkyl groups, C4-C 12 Cycloalkylalkyl groups, C5-C 12 Cycloalkyl(hetero)aryl groups and C5-C 12 (hetero)arylcycloalkyl groups, where i is in the range of 0 to 4, preferably 1, and where any R that is not a hydrogen atom is selected from the group consisting of: 37 Groups include -Cl, -F, -Br, -I, -OH, -NH2, -SO3H, and -PO3H 、 -PO4H 2、 It may be substituted with a moiety selected from the group consisting of -NO2, -CF3, =O, =NH, and -SH, and may contain one or more heteroatoms selected from the group consisting of O, S, NH, P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0217] Preferably, each R 37 are independently hydrogen atoms, -(S P ) i -C B , C1-C4 alkyl group, C2-C4 alkenyl group, C2-C4 alkynyl group, C6-C8 aryl, C2-C8 heteroaryl, C3-C6 cycloalkyl group, C5-C6 cycloalkenyl group, C3-C 10 Alkyl(hetero)aryl groups, C3-C 10 (Hetero)arylalkyl group, C4-C8 alkylcycloalkyl group, C4-C8 cycloalkylalkyl group, C5-C 10 Cycloalkyl(hetero)aryl groups and C5-C10 (hetero)arylcycloalkyl groups, where i is in the range of 0 to 4, preferably 1, and where any R that is not a hydrogen atom is selected from the group consisting of: 37 Groups include -Cl, -F, -Br, -I, -OH, -NH2, -SO3H, and -PO3H 、 -PO4H 2、 It may be substituted with a moiety selected from the group consisting of -NO2, -CF3, =O, =NH, and -SH, and may contain one or more heteroatoms selected from the group consisting of O, S, NH, P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0218] In a preferred embodiment, R is not a hydrogen atom. 37 The group is unsubstituted. In a preferred embodiment, R is not a hydrogen atom. 37 The group does not contain heteroatoms.

[0219] R 47 In a preferred embodiment, each R 47 are independently a hydrogen atom, -F, -Cl, -Br, -I, or -OR 37 , -N(R 37 )2, -SO3, -PO3 - 、 -NO2, -CF3, -SR 37 , S(=O)2N(R 37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R 37 , SC(=S)R 37 , N.R. 37 C(=O)-R 37 , N.R. 37 C(=S)-R 37 , N.R. 37 C(=O)OR 37 , N.R. 37 C(=S)OR 37 , N.R. 37 C(=O)SR 37 , N.R. 37 C(=S)SR37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R 37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR 37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37 )2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , S(O)R 37 , -S(O)2R 37 , N.R. 37 S(O)2R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , -(S P ) i -C B , C1-C8 alkyl group, C2-C8 alkenyl group, C2-C8 alkynyl group, C6-C 12 Aryl groups, C2-C 12 Heteroaryl group, C3-C8 cycloalkyl group, C5-C8 cycloalkenyl group, C3-C 12 Alkyl(hetero)aryl groups, C3-C 12 (Hetero)arylalkyl groups, C4-C 12 Alkylcycloalkyl groups, C4-C 12 Cycloalkylalkyl groups, C5-C 12 Cycloalkyl(hetero)aryl groups and C5-C 12(hetero)arylcycloalkyl groups, where i is a number ranging from 0 to 4, preferably 1, and the above alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, alkyl(hetero)aryl, (hetero)arylalkyl, alkylcycloalkyl, cycloalkylalkyl, cycloalkyl(hetero)aryl, and (hetero)arylcycloalkyl groups are selected from the group consisting of -Cl, -F, -Br, -I, -OR 37 , -N(R 37 )2, -SO3R 37 , -PO3(R 37 ) 2、 -PO4(R 37 ) 2、 -NO2, -CF3, =O, =NR 37 , and -SR 37 and may be substituted with a moiety selected from the group consisting of O, S, NR 37 , P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0220] In a preferred embodiment, each R 47 are independently a hydrogen atom, -F, -Cl, -Br, -I, or -OR 37 , -N(R 37 )2, -SO3, -PO3 - 、 -NO2, -CF3, -SR 37 , S(=O)2N(R 37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R 37 , SC(=S)R 37 , N.R. 37 C(=O)-R 37 , N.R. 37 C(=S)-R 37 , N.R. 37 C(=O)OR 37 , N.R. 37 C(=S)OR 37 , N.R. 37 C(=O)SR37 , N.R. 37 C(=S)SR 37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R 37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR 37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37 )2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , S(O)R 37 , -S(O)2R 37 , N.R. 37 S(O)2R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , -(S P ) i -C B , C1-C4 alkyl group, C2-C4 alkenyl group, C2-C4 alkynyl group, C6-C8 aryl group, C2-C8 heteroaryl group, C3-C6 cycloalkyl group, C5-C6 cycloalkenyl group, C3-C 10 Alkyl(hetero)aryl groups, C3-C 10 (Hetero)arylalkyl groups, C4-C 10 Alkylcycloalkyl groups, C4-C 10 Cycloalkylalkyl groups, C5-C 10 Cycloalkyl(hetero)aryl groups and C5-C 10(hetero)arylcycloalkyl groups, where i is a number ranging from 0 to 4, preferably 1, and the above alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, alkyl(hetero)aryl, (hetero)arylalkyl, alkylcycloalkyl, cycloalkylalkyl, cycloalkyl(hetero)aryl, and (hetero)arylcycloalkyl groups are selected from the group consisting of -Cl, -F, -Br, -I, -OR 37 , -N(R 37 )2, -SO3R 37 , -PO3(R 37 ) 2、 -PO4(R 37 ) 2、 -NO2, -CF3, =O, =NR 37 , and -SR 37 and may be substituted with a moiety selected from the group consisting of O, S, NR 37 , P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0221] In a preferred embodiment, R 47 is a portion that satisfies either one of the following formulas (2x) and (2y).

[0222] [ka]

[0223] [ka]

[0224] Here, in both formula (2x) and formula (2y), the wavy line indicates the bond to the rest of the molecule. M2 Construct-B(C B), preferably conjugated to a targeting agent, preferably selected from the group consisting of proteins, antibodies, peptoids and peptides.

[0225] In a preferred embodiment, in formula (19), X 3 is CR 47 Y T1 and X 1 , X 2 , X 4 , and X 5 If is CH2, X 3 R in 47 does not satisfy equation (2x) or equation (2y).

[0226] R 33 In a preferred embodiment, each respective R 33 is C1~C 12 Alkylene group, C2-C 12 Alkenylene group, C2-C 12 Alkynylene group, C6 arylene group, C4-C5 heteroarylene group, C3-C8 cycloalkylene group, C5-C8 cycloalkenylene group, C5-C 12 Alkyl(hetero)arylene groups, C5-C 12 (Hetero)arylalkylene group, C4-C 12 Alkylcycloalkylene group, C4-C 12 The alkylene group, alkenylene group, alkynylene group, (hetero)arylene group, cycloalkylene group, cycloalkenylene group, alkyl(hetero)arylene group, (hetero)arylalkylene group, alkylcycloalkylene group, and cycloalkylalkylene group are selected from the group consisting of -Cl, -F, -Br, -I, -OR 37 , -N(R')2, =O, =NR', -SR', -SO3H, -PO3H 、and optionally substituted with a moiety selected from the group consisting of -PO4H2, -NO2, and -Si(R')3, and optionally containing one or more heteroatoms selected from the group consisting of -O-, -S-, -NR'-, -P-, and -Si-, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0227] In a particularly preferred embodiment, each respective R 33 is selected from the group consisting of a C1 to C6 alkylene group, a C2 to C6 alkenylene group, and a C2 to C6 alkynylene group, more preferably selected from the group consisting of a C1 to C3 alkylene group, a C2 to C3 alkenylene group, and a C2 to C3 alkynylene group, wherein preferably the above alkylene group, alkenylene group, alkynylene group, cycloalkylene group, cycloalkenylene group, and cycloalkynylene group may contain one or more heteroatoms selected from the group consisting of O, S, NR5, P, and Si, wherein the N atom, S atom, and P atom are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0228] R 35 In a preferred embodiment, each respective R 35 is a C1-C8 alkylene group, a C2-C8 alkenylene group, a C2-C8 alkynylene group, a C6 arylene group, a C4-C5 heteroarylene group, a C3-C6 cycloalkylene group, a C5-C8 cycloalkenylene group, a C5-C 12 Alkyl(hetero)arylene groups, C5-C 12 (Hetero)arylalkylene group, C4-C 12 Alkylcycloalkylene group, C4-C 12and cycloalkylalkylene groups, wherein the above alkylene group, alkenylene group, alkynylene group, (hetero)arylene group, cycloalkylene group, cycloalkenylene group, alkyl(hetero)arylene group, (hetero)arylalkylene group, alkylcycloalkylene group, and cycloalkylalkylene group are selected from the group consisting of -Cl, -F, -Br, -I, -OR', -N(R')2, ═O, ═NR', -SR', -SO3H, and -PO3H. 、 and optionally substituted with a moiety selected from the group consisting of -PO4H2, -NO2, and -Si(R')3, and optionally containing one or more heteroatoms selected from the group consisting of -O-, -S-, -NR'-, -P-, and -Si-, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0229] In a preferred embodiment, each respective R 35 is selected from the group consisting of a C1 to C4 alkylene group, a C2 to C4 alkenylene group, a C2 to C4 alkynylene group, a C6 arylene group, a C4 to C5 heteroarylene group, and a C3 to C6 cycloalkylene group, wherein the alkylene group, alkenylene group, alkynylene group, (hetero)arylene group, and cycloalkylene group are selected from the group consisting of -Cl, -F, -Br, -I, -OR', -N(R')2, ═O, ═NR', -SR', -SO3H, -PO3H 、 and optionally substituted with a moiety selected from the group consisting of -PO4H2, -NO2, and -Si(R')3, and optionally containing one or more heteroatoms selected from the group consisting of -O-, -S-, -NR'-, -P-, and -Si-, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0230] R’ In a preferred embodiment, each R' is independently a hydrogen atom, a C1 to C6 alkylene group, a C2 to C6 alkenylene group, a C2 to C6 alkynylene group, a C6 arylene, a C4 to C5 heteroarylene, a C3 to C6 cycloalkylene group, a C5 to C8 cycloalkenylene group, a C5 to C12 Alkyl(hetero)arylene groups, C5-C 12 (Hetero)arylalkylene group, C4-C 12 Alkylcycloalkylene groups and C4-C 12 cycloalkyl alkylene groups.

[0231] In a preferred embodiment, each R' is independently a hydrogen atom, a C1-C4 alkylene group, a C2-C4 alkenylene group, a C2-C4 alkynylene group, a C6 arylene, a C4-C5 heteroarylene, a C3-C6 cycloalkylene group, a C5-C8 cycloalkenylene group, a C5-C8 alkyl(hetero)arylene group, a C5-C8 (hetero)arylalkylene group, a C4-C 12 It is selected from the group consisting of alkylcycloalkylene groups, and C4 to C8 cycloalkylalkylene groups.

[0232] Unless otherwise specified, for R', the above alkylene group, alkenylene group, alkynylene group, (hetero)arylene group, cycloalkylene group, cycloalkenylene group, alkyl(hetero)arylene group, (hetero)arylalkylene group, alkylcycloalkylene group, and cycloalkylalkylene group include -Cl, -F, -Br, -I, -OH, -NH2, ═O, -SH, -SO3H, and -PO3H. 、 It may be substituted with a moiety selected from the group consisting of -PO4H2, -NO2, and may contain one or more heteroatoms selected from the group consisting of -O-, -S-, -NH-, -P-, and -Si, wherein the N, S, and P atoms are optionally oxidized.

[0233] In a preferred embodiment, R' is R 37 is as defined for

[0234] In a preferred embodiment, C M2 , C X , and C A and C B For conjugation with R', R 37 is as defined for

[0235] R” In a preferred embodiment, each R″ is independently selected from the group consisting of:

[0236] [ka]

[0237] where the wavy line represents the bond to the ethylene glycol group, or optionally, R 32 R adjacent to 33 The dashed line indicates the bond to R 33 or binding to G.

[0238] In a preferred embodiment, R" is -CH2-C(O)NR'- or -CH2-NR', N, C5-C6 arenetriyl, C4-C5 heteroarenetriyl, C3-C6 cycloalkanetriyl, and C4-C6 cycloalkenetriyl, wherein the above arenetriyl, heteroarenetriyl, cycloalkanetriyl, and cycloalkenetriyl are not limited to -Cl, -F, -Br, -I, -OR', -N(R')2, -SR', -SO3H, -PO3H 、 It may be further substituted with a group selected from the group consisting of -PO4H2, -NO2, and -CF3, and may contain one or more heteroatoms selected from the group consisting of -O-, -S-, -NR'-, -P-, and -Si-, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized. Preferably, G is CR'.

[0239] L In a preferred embodiment, L is selected from the group consisting of -CH2-OCH3, -CH2-OH, -CH2-C(O)OH, -C(O)OH. In a preferred embodiment, L is preferably -CH2-OCH3.

[0240] t 1、t 2 、t 3 、t 4 、t 5 In a preferred embodiment, t1 is 0. In another embodiment, t1 is 1. In a preferred embodiment, t2 is 0. In another embodiment, t2 is 1. In a preferred embodiment, t3 is an integer in the range of 0 to 12. Preferably, t3 is an integer in the range of 1 to 10, more preferably an integer in the range of 2 to 8. In a particularly preferred embodiment, t3 is 4, and y is 1. In a preferred embodiment, t4 is 0. In another embodiment, t4 is 1. In a preferred embodiment, t5 is an integer in the range of 6 to 48, preferably 15 to 40, more preferably 17 to 35, even more preferably 20 to 30, and most preferably 22 to 28. In a particularly preferred embodiment, t5 is 23.

[0241] trans-cyclooctene In a preferred embodiment, the dienophile trigger moiety used in the present invention has a trans-cyclooctene ring, and in particular refers to a structure satisfying formula (19), where the ring may contain one or more heteroatoms. Because heterocyclic monoalkenylene 8-membered rings are also known to have dienophile activity, those skilled in the art are familiar with the fact that dienophile activity does not necessarily depend on the presence of all carbon atoms in the ring.

[0242] Therefore, in general, the present invention is not strictly limited to trans-cyclooctene. Those skilled in the art of organic chemistry will recognize that there are other 8-membered ring-based dienophiles that contain the same endocyclic double bond as trans-cyclooctene, but may have one or more heteroatoms elsewhere in the ring. That is, the present invention generally relates to 8-membered non-aromatic cyclic alkene moieties, preferably cyclooctene moieties, more preferably trans-cyclooctene moieties.

[0243] It should be noted that depending on the choice of nomenclature, the TCO dienophile can also be referred to as E-cyclooctene. Referring to conventional nomenclature, it will be understood that as a result of substitution on the cyclooctene ring, the same cyclooctene isomer can be formally designated as a Z-isomer, depending on the position and molecular weight of the substituent. In the present invention, any substitution variant of the present invention, regardless of whether it is formally an "E" or "Z" isomer, or a "cis" or "trans" isomer, will be considered a derivative of unsubstituted trans-cyclooctene or unsubstituted E-cyclooctene. The terms "trans-cyclooctene" (TCO) and E-cyclooctene are used interchangeably, and are maintained for all dienophiles according to the present invention, even when the substituent formally requires the opposite nomenclature. That is, the present invention relates to cyclooctene in which carbon atoms 1 and 6, numbered below in formula 4b, are in the E (entgegen), i.e., trans, position.

[0244] [ka]

[0245] The TCO may consist of multiple isomers, and the substituents in the TCO, such as R 48 , also including the equatorial versus axial position of R. In this respect, reference is made to Whitham et al. J. Chem. Soc. (C), 1971, 883-896, which describes the synthesis and characterization of the equatorial and axial isomers of trans-cyclooct-2-en-ol, defined as (1RS,2RS) and (1SR,2RS), respectively. In these isomers, the OH substituent is in either the equatorial or axial position. In a preferred embodiment, R 48 For a TCO structure where R can be in either an axial or equatorial position, 48 is in the axial position.

[0246] Trans-cyclooctene or E-cyclooctene derivatives are particularly suitable as triggers, given their high reactivity. The trans-cyclooctene (TCO) moiety may include at least two exocyclic bonds fixed in substantially the same plane, and / or the trans-cyclooctene (TCO) moiety may include at least one substituent in an axial position, rather than an equatorial position. Those skilled in the art of organic chemistry will understand that the term "fixed in substantially the same plane" refers to a bonding theory in which bonds are typically considered to be fixed in the same plane. Typical examples of such fixation in the same plane include double bonds and strained fused rings; for example, the at least two exocyclic bonds can be two double bonds to an oxygen atom (i.e., C=O). The at least two exocyclic bonds can also be single bonds between two adjacent carbon atoms, provided that these bonds are part of a fused ring (i.e., fused to a TCO ring) that has a substantially planar structure, thereby fixing the two single bonds in substantially the same plane. Examples of the latter include strained rings, such as cyclopropyl and cyclobutyl. Without wishing to be bound by theory, the inventors believe that the presence of at least two exocyclic bonds in the same plane will result in at least partial planarization of the TCO ring, which can result in higher reactivity in the IEDDA reaction. Background material providing further guidance is WO 2013 / 153254.

[0247] The dienophiles for use in the present invention can be synthesized by those skilled in the art based on known synthetic routes to cyclooctene and the corresponding ring containing one or more heteroatoms. Those skilled in the art are also aware of the abundance of cyclooctene derivatives that can be synthesized via ring-closing metathesis reactions using Grubbs catalysts. As mentioned above, TCOs may contain one or more heteroatoms in the ring. This is itself well accessible to those skilled in the art (e.g., WO 2016025480). For example, the presence of thioethers in TCOs has been mentioned (Cere et al. J. Org. Chem. 1980, 45, 261). Also, for example, the presence of -O-SiR2-O moieties in TCOs has been mentioned (Prevost et al. J. Am. Chem. Soc. 2009, 131, 14182). The leaving group (R) located at the allylic position has also been mentioned (Cere et al. J. Org. Chem. Soc. 2009, 131, 14182). 48 References have been made to TCO syntheses in which Z is an ether, ester, carbonate, carbamate, or thiocarbamate: [Versteegen et al Angew.Chem.Int.Ed.2018,57,10494] and [Steiger et al Chem Comm 2017,53,1378]. Exemplary compounds include the following structures, shown with multiple literature references below: When a cyclooctene derivative is depicted as Z-cyclooctene, it is believed that it can be converted to the E-cyclooctene analog.

[0248] [ka]

[0249] Preferred triggers of the present invention include the following:

[0250] [ka]

[0251] Further preferred triggers of the present invention include the following:

[0252] [ka]

[0253] Construct-A(C A ) and Construct-B (C B ) Construct A and Construct B include small molecules, organic molecules, metal coordination compounds, molecules containing radionuclides, chelates containing radioactive metals, inorganic molecules, organometallic molecules, biomolecules, polymers, resins, particles (e.g., microparticles and nanoparticles), liposomes, micelles, polymersomes, gels, surfaces, cells, biological tissues, and pathogens.

[0254] In a preferred prodrug or in vivo embodiment, C A is selected from the group consisting of a drug, a targeting agent, a label, an administration agent, and a masking moiety. A is an agent, preferably an agent as defined herein.

[0255] In some preferred prodrug or in vivo embodiments, C B is selected from the group consisting of a drug, a targeting agent, a label, an administration agent, and a masking moiety. B is selected from the group consisting of a targeting agent and a masking moiety.

[0256] In a preferred embodiment, the compound of formula (19) comprises at least one label and at least one administration agent, and preferably satisfies at least one of the following conditions (i) to (iii): (i) At least one C A is a label, and at least one C A is the administered agent; (ii) at least one C A is a label, and at least one CB is the administered agent; (iii) at least one C A is a dosage form, and at least one C B is a sign; For all conditions (i) to (iii), X 1 ~X 5 contains a dosage agent, and at least one C A If is a sign, X 1 ~X 5 At least one C A does not contain the same sign as A is the label, and X 1 ~X 5 There is at least one C A If it contains the same indicator as X 1 ~X 5 does not include the administered agent; 48 At least one C in B If is the administered agent, R 48 Other C included in B is preferably not a label. In a preferred embodiment, only condition (i) is met. In a preferred embodiment, only condition (ii) is met. In a preferred embodiment, only condition (iii) is met. In a preferred embodiment, both conditions (i) and (ii) are met. In a preferred embodiment, both conditions (i) and (iii) are met. In a preferred embodiment, both conditions (ii) and (iii) are met. In a preferred embodiment, all three conditions (i) to (iii) are met.

[0257] In a preferred in vitro embodiment, C A and C BExamples of constructs include, but are not limited to, small molecules, organic molecules (including fluorescent dyes), metal coordination compounds, molecules containing radionuclides, chelates containing radioactive metals, inorganic molecules, organometallic molecules, biomolecules, drugs, polymers, resins (e.g., polystyrene, agarose), particles (e.g., beads, magnetic beads, gold, silica-based particles and materials, polymers and polymer-based materials, glass, iron oxide particles, microparticles and nanoparticles, such as liposomes and polymersomes), gels, surfaces (e.g., glass slides, chips, wafers, gold, metals, silica-based, polymers, plastics, resins), cells, biological tissues, and pathogens (viruses, bacteria, fungi, yeast). The constructs may, for example, include combinations of the constructs described above. Examples of biomolecules include carbohydrates, biotin, peptides, peptoids, lipids, proteins, enzymes, oligonucleotides, DNA, RNA, PNA, LNA, aptamers, hormones, toxins, steroids, cytokines, antibodies, antibody fragments (e.g., Fab2, Fab, scFV, diabodies, triabodies, VHH), antibody (fragment) fusions (e.g., bispecific and trispecific mAb fragments).

[0258] In a preferred in vitro embodiment, C A and C B is also defined herein as R 32 or R 32 where R 32 is a further C A and C B For example, C A Spacer S P via T R R is a maleimide or photocross linker attached to 32 The maleimide or photocross linker can be T R can be used to further conjugate to a protein. In this particular embodiment, C A and C Bis a biomolecule binding moiety.

[0259] In a preferred embodiment, each C A and C B are independently selected from the group consisting of organic molecules, inorganic molecules, organometallic molecules, resins, beads, glass, microparticles, nanoparticles, gels, surfaces, and cells. A and C B are independently selected from the group consisting of organic molecules and inorganic molecules.

[0260] In a preferred embodiment, each C A and C B are independently selected from the group consisting of small molecules, proteins, carbohydrates, peptides, peptoids, oligosaccharides, molecules including radionuclides, fluorescent dyes, inorganic molecules, organometallic molecules, polymers, lipids, oligonucleotides, DNA, RNA, PNA, LNA, drugs, resins, beads, glass, microparticles, nanoparticles, gels, surfaces, and cells.

[0261] Preferably, the small molecule is a small organic molecule. Preferably, the small molecule has a molecular weight of 2 kDa or less, more preferably 1 kDa or less, more preferably 750 Da or less, more preferably 500 Da or less, and most preferably 300 Da or less. Preferably, the small molecule has a molecular weight of at least 15 Da, more preferably at least 50 Da, more preferably at least 75 Da, and most preferably at least 100 Da.

[0262] It will be understood that "a molecule comprising a radionuclide" includes a chelating agent that chelates a radionuclide.

[0263] In another preferred embodiment, each C A and C B are independently moieties according to formula (5) as defined herein.

[0264] Construct - Trigger Assembly A construct trigger can be a single construct or multiple constructs C A and Trigger T R Optionally, the trigger further comprises a conjugate with one or more constructs C B is linked to.

[0265] The general formula for the construct trigger is shown below in equations (5a) and (5b). For the avoidance of doubt, Y C L C and C A Because it is part of Y C are not shown separately in equations (5a) and (5b).

[0266] [ka]

[0267] C A is construct A and C B is construct B, and S P is a spacer;T R is the trigger, and L C is a linker. Formula (5a): b, c, e, f, g, h≧0; a, d≧1. Formula (5b): c, e, f, g, h≧0; a, b, d≧1.

[0268] In the trigger-construct conjugate, construct C A and Trigger T R (TCO derivative) can be directly linked to each other. They can also be linked to each other via a self-immolative linker LC, which can consist of multiple (self-immolative or non-self-immolative) units. Referring to Formula 5a and Formula 5b, L C If contains a non-self-immolative unit, this unit is P and c≧1. The present invention provides a construct C having one or more dinucleotides. A It will be understood that this encompasses any conceivable manner in which one or more constructs C BTrigger or linker L C This applies when the spacer S is attached to the P The trigger or linker L C The same is true for the optional attachment of a spacer to a protein. Methods of affecting conjugation, for example, in the case of proteins, through reactive amino acids such as lysine or cysteine, are known to those skilled in the art. Exemplary conjugation methods are outlined herein below in the section on spacers.

[0269] Construct C A After the formation of the IEDDA adduct, construct C A It will be understood that the construct C is linked to the TCO in such a way that it can eventually be released. Generally, this is the case for construct C. A and the TCO, or a self-immolative linker L C In the case of , between the linker and TCO, and construct C A This means that the bond between the construct C and the linker should be cleavable. A and any linker is linked via a heteroatom, preferably via O, N, NH or S. The cleavable bond is preferably selected from the group consisting of carbamate, thiocarbamate, carbonate, ester, ether, thioether, amide, thioester bonds.

[0270] One C B It should be understood that the C can be modified with multiple triggers. For example, an antibody can be modified with four TCO-drug constructs by conjugation to four amino acid residues, where C A is a drug.

[0271] Similarly, one C AIt should be understood that the can be modified by multiple triggers. For example, a protein drug can be masked by conjugating four amino acid residues to four TCO-polyethylene glycol constructs, where the polyethylene glycol is C B is.

[0272] Furthermore, one C A can be modified with two or more triggers, where at least one trigger is C B and at least one trigger is linked to a targeting agent which is B where C A can be a drug, preferably a protein.

[0273] Spacer S P As used herein, "each individual S P When it is stated that "is linked at all ends to the rest of the structure," this refers to the spacer S P It will be understood that this refers to the fact that the spacer connects multiple moieties within a structure and, therefore, the spacer, by definition, has multiple termini. P can be linked to each individual moiety via different or identical moieties, each of which can be individually selected. Typically, these linking moieties are spacers S P It should be considered as part of itself. Spacer S P connects two moieties within a structure, "all ends" should be interpreted as "both ends." For example, if the spacer connects a transcyclooctene moiety to Construct A, the "rest of the molecule" refers to the transcyclooctene moiety and Construct A, while the connecting moiety between the spacer and the transcyclooctene moiety, Construct A (i.e., both ends), can be selected independently.

[0274] In a preferred embodiment, the spacer S Pis one or more spacer units S arranged linearly and / or branchedly. U and one or more C B Part and / or one or more L C or T R The spacer can be attached to a moiety such as C B One T R (See Example A below; Equation 5a and Equation 5b: f, e, a=1) or multiple T R (See examples B and C below; Equations 5a and 5b: f, e=1, a≧1), but it can also be used to connect C B -T R -C A The spacer unit can be used to adjust the properties, e.g., pharmacokinetic properties, of the conjugate (see Example D below; Formula 5a and Formula 5b: one or more of c, e, g, h > 1). Thus, the spacer unit does not necessarily connect two entities to each other; it can be used to adjust the properties, e.g., pharmacokinetic properties, of the conjugate (see Example D below; Formula 5a and Formula 5b: one or more of c, e, g, h > 1). R or L C Alternatively, the spacer may be C B T R The spacer may also contain a spacer unit that links to the spacer and, in addition, may contain another spacer unit that binds only to the spacer and serves to adjust the properties of the conjugate (see Example F below; Formula 5a and Formula 5b: e≧1). The spacer may also contain two different types of S U Constructs such as PEG linked to a peptide or PEG linked to an alkylene moiety (see Example E below; Formulas 5a and 5b: e≧1). For clarity, Example B illustrates the use of a multivalent branched S U Branched S by using U Example C shows a linear S U Branched polymers, such as peptides, can be used to U Formula 3

[0275] JPEG2026004404000035.jpg82170

[0276] The spacer can be bound to the active agent in similar designs as shown in Examples A-F above.

[0277] The spacer units include, but are not limited to, amino acids, nucleosides, nucleotides, and biopolymer fragments, such as oligopeptides or polypeptides, oligopeptoids or polypeptoids, or oligolactides or polylactides, or oligo- or polycarbohydrates, varying from 2 to 200, particularly 2 to 113, preferably 2 to 50, more preferably 2 to 24, more preferably 2 to 12 repeating units. Exemplary preferred biopolymers S U is a peptide. Still other examples are alkyl, alkylene, alkenyl, alkenylene, alkynyl, alkynylene, cycloalkyl, cycloalkylene, cycloalkenyl, cycloalkenylene, cycloalkynyl, cycloalkynylene, aryl, arylene, alkylaryl, alkylarylene, arylalkyl, arylalkylene, arylalkenyl, arylalkenylene, arylalkynyl, arylalkynylene, polyethyleneamino, polyamine, which may be substituted or unsubstituted, linear or branched, and may contain additional cyclic moieties and / or heteroatoms, preferably O, N and S, more preferably O; wherein in preferred embodiments, S in these examples is U contains 50 or fewer carbon atoms, more preferably 25 or fewer carbon atoms, more preferably 10 or fewer carbon atoms. U are independent, (CH2) r , (C3-C8 carbocyclo), O-(CH2) r , arylene, (CH2) r -Arylene, arylene-(CH2) r , (CH2) r -(C3-C8 carbocyclo), (C3-C8 carbocyclo)-(CH2) r , (C3-C8 heterocyclo), (CH2) r-(C3-C8 heterocyclo), (C3-C8 heterocyclo)-(CH2) r , -(CH2) r C(O)NR4(CH2) r , (CH2CH2O) r , (CH2CH2O) r CH2, (CH2) r C(O)NR4(CH2CH2O) r , (CH2) r C(O)NR4(CH2CH2O) r CH 2、 (CH2CH2O) r C(O)NR4(CH2CH2O) r , (CH2CH2O) r C(O)NR4(CH2CH2O) r CH2, (CH2CH2O) r C(O)NR4CH2; where r is independently an integer from 1 to 10, and R4 is as defined herein.

[0278] Spacer Unit S U Other examples are linear or branched polyalkylene glycols, such as polyethylene glycol (PEG) chains or polypropylene glycol (PPG) chains, with repeat units varying from 2 to 200, particularly from 2 to 113, preferably from 2 to 50, more preferably from 2 to 24, and more preferably from 2 to 12. When the polyalkylene glycol, such as PEG and PPG polymers, is attached via only one end of the polymer chain, the other end is preferably terminated with -OCH3, -OCH2CH3, or -OCH2CH2CO2H.

[0279] Other polymer spacer units are polymers and copolymers such as poly-(2-oxazoline), poly(N-(2-hydroxypropyl)methacrylamide) (HPMA), polylactic acid (PLA), polylactic-co-glycolic acid (PLGA), polyglutamic acid (PG), dextran, polyvinylpyrrolidone (PVP), poly(1-hydroxymethylethylenehydroxymethyl-formal) (PHF). Other exemplary polymers are polysaccharides, glycopolysaccharides, glycolipids, polyglycosides, polyacetals, polyketals, polyamides, polyethers, and polyesters. U Examples of naturally occurring polysaccharides that can be used as the polymer S include cellulose, amylose, dextran, dextrin, levan, fucoidan, carrageenan, inulin, pectin, amylopectin, glycogen, lyxenan, agarose, hyaluronan, chondroitin sulfate, dermatan sulfate, keratan sulfate, alginic acid, and heparin. In yet another exemplary embodiment, the polymer S U comprises copolymers of polyacetals / polyketals and hydrophilic polymers selected from the group consisting of polyacrylates, polyvinyl polymers, polyesters, polyorthoesters, polyamides, oligopeptides, polypeptides and derivatives thereof. Exemplary preferred polymers S U are PEG, HPMA, PLA, PLGA, PVP, PHF, dextran, oligopeptides, and polypeptides.

[0280] In some aspects of the invention, S U The polymers used in have molecular weights in the range of 2 to 200 kDa, 2 to 100 kDa, 2 to 80 kDa, 2 to 60 kDa, 2 to 40 kDa, 2 to 20 kDa, 3 to 15 kDa, 5 to 10 kDa, 500 daltons to 5 kDa.

[0281] Other exemplary S U are dendrimers, such as poly(propyleneimine) (PPI) dendrimers, PAMAM dendrimers, and glycol-based dendrimers.

[0282] S of the present invention U expressly encompasses, but is not limited to, conjugates prepared using commercially available crosslinker reagents such as BMPEO, BMPS, EMCS, GMBS, HBVS, LC-SMCC, MBS, MPBH, SBAP, SIA, SIAB, SMCC, SMPB, SMPH, sulfo-EMCS, sulfo-GMBS, sulfo-KMUS, sulfo-MBS, sulfo-SIAB, sulfo-SMCC, sulfo-SMPB, and SVSB, DTME, BMB, BMDB, BMH, BMOE, BM(PEO)3, and BM(PEO)4.

[0283] To construct a branched spacer, S-type amines based on one or more natural or unnatural amino acid, amino alcohol, amino aldehyde, or polyamine residues, or combinations thereof, which collectively provide the required functionality for branching, are used. U For example, serine has three functional groups, namely, an acid, an amino, and a hydroxyl group, and can be branched S U The attached amino acid may be viewed as an amino alcohol residue for the purpose of acting as a carboxyl group. Other exemplary amino acids are lysine and tyrosine.

[0284] In preferred embodiments, the spacer consists of one spacer unit, and therefore in those cases, S P is S U In a preferred embodiment, the spacer consists of 2, 3 or 4 spacer units.

[0285] In some aspects of the invention, S P has a molecular weight in the range of 2 to 200 kDa, 2 to 100 kDa, 2 to 80 kDa, 2 to 60 kDa, 2 to 40 kDa, 2 to 20 kDa, 3 to 15 kDa, 5 to 10 kDa, 500 daltons to 5 kDa. Phas a mass of 5000 daltons or less, 4000 daltons or less, 3000 daltons or less, 2000 daltons or less, 1000 daltons or less, 800 daltons or less, 500 daltons or less, 300 daltons or less, or 200 daltons or less. P has a mass from 100 daltons, from 200 daltons, from 300 daltons, to 5000 daltons. P has a mass of from 30, 50 or 100 daltons to 1000 daltons, from about 30, 50 or 100 daltons to 500 daltons.

[0286] In a preferred embodiment, S P is C1~C 12 Alkylene group, C2-C 12 Alkenylene group, C2-C 12 Alkynylene group, C6 arylene group, C4-C5 heteroarylene group, C3-C8 cycloalkylene group, C5-C8 cycloalkenylene group, C5-C 12 Alkyl(hetero)arylene groups, C5-C 12 (Hetero)arylalkylene group, C4-C 12 Alkylcycloalkylene group, C4-C 12 a spacer selected from the group consisting of cycloalkyl alkylene groups, wherein S P With regard to the above, the alkylene group, alkenylene group, alkynylene group, (hetero)arylene group, cycloalkylene group, cycloalkenylene group, alkyl(hetero)arylene group, (hetero)arylalkylene group, alkylcycloalkylene group, and cycloalkylalkylene group are not limited to -Cl, -F, -Br, -I, -OR', -N(R 37 )2, =O, =NR 37 , -SR 37 , and -Si(R 37 )3, and is optionally substituted with a moiety selected from the group consisting of —O—, —S—, —NR 37It may contain one or more heteroatoms selected from the group consisting of -, -P- and -Si-, wherein the N, S and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0287] In a preferred embodiment, S P is a compound having one or more moieties C as described herein. M2 , C X , or R 32 In a preferred embodiment, the above-mentioned C M2 , C X , or R 32 The residue of S P C B , C A , L C , T R Or other S P Combine with.

[0288] Linker L C L C is any self-immolative linker, which may be composed of multiple units arranged in a linear and / or branched manner, and which may comprise one or more C A A portion may be released.

[0289] To be more clear, if r is 0, then species C A directly constitutes the leaving group of the release reaction, and when r>0, the self-immolative linker L C constitutes the leaving group for the release reaction. C Structures, their uses, linkers L C Position and attachment method of construct C A and C B , and T R are known to those skilled in the art, see for example [Papot et al., Anticancer Agents Med. Chem., 2008, 8, 618-637]. Nevertheless, the self-immolative linker L CPreferred, but non-limiting examples of are benzyl derivatives, such as the benzyl derivative depicted below. There are two main self-immolation mechanisms: electron cascade elimination and cyclization-mediated elimination. The preferred example on the left below functions via a cascade mechanism, in which the bond between the allylic carbon of the trigger and the -O- or -S- attached to that carbon is cleaved, and Y C1 electron pairs, e.g., NR 6 The electron pair of , shifts to the benzyl moiety, resulting in an electron cascade and the formation of 4-hydroxybenzyl alcohol, CO2, and the liberated C A The preferred example in the center below works via a cyclization mechanism, where NR 6 Cleavage of the bond to C results in nucleophilic attack of the amine on the carbonyl to form the pentacyclic 1,3-dimethylimidazolidin-2-one, and A The preferred example on the right combines both mechanisms, where the linker binds CO and one unit of 4-hydroxybenzyl alcohol (Y C1 is O), but will also decompose into a single 1,3-dimethylimidazolidin-2-one unit.

[0290] [ka]

[0291] Here, the wavy line indicates a bond to -O- or -S- at the allylic position of trans-cyclooctene, and the dashed double-dashed line indicates C A The bond to

[0292] The self-immolative linker L C By substituting the benzyl group ofA It is possible to adjust the release rate of . Synthetic procedures for preparing such substituted benzyl derivatives are known to those skilled in the art (see, for example, [Greenwald et al., J. Med. Chem., 1999, 42, 3657-3667] and [Thornthwaite et al., Polym. Chem., 2011, 2, 773-790]). Some preferred substituted benzyl derivatives with different release rates are depicted below.

[0293] Self-immolative linkers that undergo cyclization include, but are not limited to, substituted and unsubstituted aminobutyric acid amides, appropriately substituted bicyclo[2.2.1] and bicyclo[2.2.2] ring systems, 2-aminophenylpropionic acid amides, and trimethyl lock-based linkers (see, e.g., [Chem. Biol. 1995, 2, 223], [J. Am. Chem. Soc. 1972, 94, 5815], [J. Org. Chem. 1990, 55, 5867], the contents of which are incorporated herein by reference). Preferably, cyclization leads to C A Emits L C So, C A The remainder of the sulfur atom is connected to L through the aromatic oxygen of the sulfur atom. C For example, an aromatic oxygen will be understood to mean an oxygen that is directly attached to an aromatic group.

[0294] L C Further preferred examples can be found in WO 2009017394(A1), U.S. Pat. No. 7,375,078, WO 2015038426A1, WO 2004043493, Angew. Chem. Int. Ed. 2015, 54, 7492-7509, the contents of which are incorporated herein by reference.

[0295] In some aspects of the invention, L Chas a mass of 1000 daltons or less, 500 daltons or less, 400 daltons or less, 300 daltons or less, or 10, 50 or 100 to 1000 daltons, 10, 50, 100 to 400 daltons, 10, 50, 100 to 300 daltons, 10, 50, 100 to 200 daltons, for example 10 to 1000 daltons, for example 50 to 500 daltons, for example 100 to 400 daltons.

[0296] Those skilled in the art will appreciate that one L C But C A Another L that is bound to C where the disconnecting agent and trigger T R Depending on the reaction with L C -L C -C A T R Emitted from multiple L C Part and C A It will be appreciated that the self-immolative release of both the hydroxyl group and the hydroxyl group will occur. C Regarding the formula, next, T R Other L C L to be connected to C is C A does not emit Y C1 and C A L, which is further connected to C is released.

[0297] In a preferred embodiment, L C is selected from the group consisting of linkers according to Group I, Group II, and Group III.

[0298] Linkers according to Group I are:

[0299] [ka]

[0300] where the wavy line may also represent a bond to -S- at the allylic position of trans-cyclooctene; where U, V, W, and Z are independently -CR 7 - and -N-; where e is either 0 or 1; and where X is -O-, -S-, and -NR 6 -; wherein preferably, independently, each R 8 and R 9 represents a hydrogen atom, a C1-C4 alkyl group, a C2-C4 alkenyl group, and C 4~6 (hetero)aryl groups; where R 8 and R 9 In the above, the alkyl group, alkenyl group, and (hetero)aryl group are -Cl, -F, -Br, -I, -OH, -NH2, ═O, -SH, -SO3H, -PO3H 、 and optionally substituted with a moiety selected from the group consisting of -PO4H2, and -NO2, and containing up to two heteroatoms selected from the group consisting of -O-, -S-, -NH-, -P-, and -Si-, wherein the N, S, and P atoms are optionally oxidized; wherein, in the case of a linker according to Group I, C A is linked to LC through a moiety selected from the group consisting of -O-, -N-, -C- and -S-, preferably from the group consisting of secondary amines and tertiary amines, wherein the moiety A Preferably, for Group I linkers, R 8 and R 9 are both hydrogen atoms.

[0301] Linkers according to Group II are:

[0302] [ka]

[0303] Here, the wavy line may also represent a bond to -S- at the allylic position of trans-cyclooctene; where m is an integer from 0 to 2, preferably m is 0; where e is either 0 or 1; where, in the case of a linker according to Group II, C A is connected to L through a moiety selected from the group consisting of -O-, -N-, -C- and -S-, preferably from the group consisting of secondary amines and tertiary amines. C wherein the moiety is linked to C A Preferably, for Group II linkers, R 8 and R 9 and R are hydrogen atoms. Preferably, for Group II linkers, R 7 is methyl or isopropyl.

[0304] Linkers according to Group III are:

[0305] [ka]

[0306] Here, the wavy line may also represent the bond to -S- at the allylic position of trans-cyclooctene; where, in the case of a linker according to Group III, C A is selected from -O- and -S-, preferably C 4~6 L through a moiety selected from the group consisting of -O- or -S- attached to a (hetero)aryl group C wherein the moiety is linked to C A where preferably, each R 6 are independently a hydrogen atom, a C1-C4 alkyl group, a C2-C4 alkenyl group, and C 4~6 (hetero)aryl groups, where R 6 In the above, the alkyl group, alkenyl group, and (hetero)aryl group are -Cl, -F, -Br, -I, -OH, -NH2, ═O, -SH, -SO3H, -PO3H 、and -NO2, and may contain up to two heteroatoms selected from the group consisting of -O-, -S-, -NH-, -P- and -Si-, wherein the N, S and P atoms may be oxidized, and wherein preferably each R 7 are independently a hydrogen atom, a C1-C3 alkyl group, a C2-C3 alkenyl group, and C 4~6 (hetero)aryl groups, where R 7 wherein the alkyl, alkenyl, and (hetero)aryl groups are optionally substituted with a moiety selected from the group consisting of -Cl, -F, -Br, -I, -OH, -NH, =O, =NH, -N(CH), -S(O)CH, and -SH, and are optionally flanked by up to one heteroatom selected from the group consisting of -O-, -S-, -NH-, -P-, and -Si-, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized, and wherein R 7 is preferably selected from the group consisting of a hydrogen atom, methyl, —CH—CH—N(CH) and —CH—CH—S(O)—CH.

[0307] Preferably, for Group III linkers, R 6 is a hydrogen atom. Preferably, for Group III linkers, R 6 is methyl.

[0308] R included in Groups I, II and III 6 , R 7 , R 8 , R 9 is also -(S P ) i -C B It can be.

[0309] For all linkers according to Group I and Group II, Y C1 is -O-, -S- and -NR 6 -, preferably -NR 6For all linkers according to Group III, Y C1 Ha-NR 6 For all linkers according to Group I, Group II and Group III, Y C2 is selected from the group consisting of O and S, preferably O.

[0310] Two L's C are linked to each other, L attached to -O- or -S- at the allylic position of trans-cyclooctene C is selected from the group consisting of linkers according to Group I and Group II, and is attached to -O- or -S- at the allylic position of trans-cyclooctene; C and C A L between C is selected from Group III, and the wavy line in the Group III structures represents L attached to -O- or -S- at the allylic position in the trans-cyclooctene ring, instead of a bond to the allylic -O- or -S- in the trans-cyclooctene ring. C The double dashed lines in the structures of Groups I and II indicate bonds to C A Instead of a bond to -O- or -S- at the allylic position of trans-cyclooctene, L C and C A L between C The bond to

[0311] In a preferred embodiment, L C is selected from the group consisting of linkers according to Group IV, Group V, Group VI, and Group VII, where linkers according to Group IV are:

[0312] [ka]

[0313] where the wavy line may also represent a bond to -S- in the allylic position of trans-cyclooctene; Ais selected from -O- and -S-, preferably -OC 5-8 -Arylene- and -SC 5~8 -arylene-, C wherein the moiety is linked to C A is part of.

[0314] Linkers according to Group V are:

[0315] [ka]

[0316] where the wavy line may also represent a bond to -S- in the allylic position of trans-cyclooctene; A is connected to L through a moiety selected from the group consisting of -O- and -S- C wherein the moiety is linked to C A In the first linker of group V, R 7 is preferably —(CH 2 ) 2 —N(CH 3 ) 2 .

[0317] Linkers according to Group VI are:

[0318] [ka]

[0319] where the wavy line may represent the bond to -S- in the allylic position of trans-cyclooctene; A is connected to L through a moiety selected from the group consisting of -O-, -N- and -S-, preferably a secondary or tertiary amine C wherein the moiety is linked to C A is part of.

[0320] Linkers according to Group VII are:

[0321] [ka]

[0322] Here, the wavy line may also represent a bond to the -S- in the allylic position of trans-cyclooctene; A is connected to L through a moiety selected from the group consisting of -O-, -N- and -S-, preferably from the group consisting of secondary amines and tertiary amines. C wherein the moiety is linked to C A where multiple double dashed lines are part of one L C If indicated within, each C A The portions are independently selected.

[0323] For all linkers according to Group IV, Group V, Group VI and Group VII, Y C1 is -O-, -S- and -NR 6 - is selected from the group consisting of

[0324] For groups IV to VII, preferably R 6 and R 7 is as defined herein, more preferably as defined for Groups I to III. For Groups I to VII, i is an integer ranging from 0 to 4, preferably 0 or 1, where j is 0 or 1. Preferably, R used in any one of Groups I to VII 6 , R 7 , R 8 , R 9 is not substituted. R 6 , R 7 , R 8 , R 9 is as defined herein. Preferably, R 6 is a hydrogen atom. Preferably, R 7 is a hydrogen atom. Preferably, R 8 is a hydrogen atom. Preferably, R 9 is a hydrogen atom.

[0325] Targeting The kits of the present invention are highly suitable for use in targeted imaging, targeted delivery of drugs and therapeutic radiation, and for selective biomolecule or tissue binding in vitro.

[0326] The term "primary target" as used herein preferably refers to a target for a therapeutic targeting agent. In other embodiments, the primary target refers to a target for imaging theranostics, diagnostics, or in vitro research. For example, the primary target can be any molecule present in an organism, tissue, or cell. Targets include cell surface targets, such as receptors and glycoproteins; structural proteins, such as amyloid plaques; abundant extracellular targets, such as stromal targets and tumor microenvironment targets; extracellular matrix targets, such as growth factors and proteases; intracellular targets, such as the surface of the Golgi apparatus, the surface of mitochondria, RNA, DNA, enzymes, components of cell signaling pathways; and / or foreign substances, such as pathogens, such as viruses, bacteria, fungi, yeast, or parts thereof. Examples of primary targets include compounds, such as proteins, whose presence or expression levels correlate with certain tissue or cell types, or whose expression levels are up- or down-regulated in certain disorders. According to a particular embodiment of the invention, the primary target is a protein, such as a receptor (internalizing or non-internalizing).

[0327] Furthermore, a preferred primary target is blood, i.e., to prolong the blood circulation of the compound and / or reduce extravasation into other tissues. For example, blood can be targeted by attaching polyethylene glycol (PEG) to the compound, which typically increases blood circulation time. For example, by attaching the compound to 500 nm PLGA particles, the conjugate will not easily flow out of the blood into other tissues (e.g., muscle), except for its rapid uptake and clearance via the liver and spleen. Organs, such as the liver, can also be a primary target. For example, a hexose can be attached to the compound to specifically target the liver.

[0328] A further major target is the general immune system. According to the present invention, the primary target may be any suitable target in the human or animal body or on a pathogen or parasite, for example a cell, such as the cell membrane and cell wall; a receptor, such as a cell membrane receptor; an intracellular structure, such as the Golgi apparatus or mitochondria; an enzyme, a receptor, DNA, RNA, a virus or virus particle, an antibody, a protein, a carbohydrate, a monosaccharide, a polysaccharide, a cytokine, a hormone, a steroid, a somatostatin receptor, a monoamine oxidase, a muscarinic receptor, a cardiomyopathy, a nervous system, a leukotriene receptor, e.g., on leukocytes, a urokinase plasminogen activator receptor (uPAR), a folate receptor, an apoptosis marker, an (anti-)angiogenesis marker, a gastrin receptor, a dopaminergic system, a serotonergic system, a GABAergic system, an adrenergic system, a cholinergic system, an opoid receptor, a GPIIb / IIIa receptor and other thrombosis-related receptors, fibrin, a calcitonin receptor, a vasoconstrictor ... receptor), tuftsin receptor, integrin receptor, fibronectin, VEGF / EGF and VEGF / EGF receptor, TAG72, CEA, A33, CD19, CD20,CD22, CD25, CD30, CD33, CD40, CD45, CD56, CD74, CD79, CD105, CD123, CD138, CD163, CD174, CD184, CD227, CD269, CD326, CD340, CD352, MUC1, MUC16, GPNMB, PSMA, Cripto, tenascin C, melanocortin-1 receptor, CD44v6, G250, HLA DR, ED-A, ED-B, TMEFF2, EphB2, EphA2, FAP, mesothelin, GD2, CAIX, 5T4, matrix metalloproteinase (MMP) metalloproteinase), ADAM-9, P / E / L-selectin receptor, LDL receptor, P-glycoprotein, neurotensin receptor, neuropeptide receptor, substance P receptor, NK receptor, CCK receptor, sigma receptor, interleukin receptor, herpes simplex virus tyrosine kinase, human tyrosine kinase, MSR1, FAP, CXCR, tumor endothelial marker (TEM), cMET, IGFR, FGFR, GPA33, hCG, HER2, HER3, CA19, TAM, LGALS3BP, nectin-4, IFGR, PD1, PDL1, AGS-5, AGS-16, endosialin, ETBR, TM4SF1, BCMA, GPC2, TROP-2, AXL, HLA-DR, B7-H3, MTX3, MTX5, EFNA4, NOTCH, tissue factor (TF) factor), PDGFR, GITR, OX40, RIG, MDA-5, NLRP1, NLRP3, AIM2, IDO, MEK, cGAS, and NKG2A.

[0329] According to a further specific embodiment of the present invention, the primary target and targeting agent are selected to result in specific or increased targeting of tissues or diseases, such as cancer, inflammation, infectious diseases, cardiovascular diseases such as thrombosis, atherosclerosis, hypoxic sites such as stroke, tumors, cardiovascular disorders, brain disorders, apoptosis, angiogenesis, organs, and reporter genes / enzymes. This can be achieved by selecting primary targets with tissue-, cell-, or disease-specific expression. For example, membrane folate receptors mediate the intracellular accumulation of folic acid and its analogs, such as methotrexate. While expression is restricted in normal tissues, the receptor is overexpressed in various tumor cell types.

[0330] In a preferred embodiment, the primary target is equivalent to a therapeutic target, which should be understood to be the entity that is targeted by a drug to have a therapeutic effect.

[0331] Targeting agent T T Targeting agent T T binds to the primary target. To allow specific targeting of the primary targets listed above, the targeting agent T TThese include, but are not limited to, antibodies, antibody derivatives, antibody fragments, antibody (fragment) fusions (e.g., bispecific and trispecific mAb fragments or derivatives thereof), proteins, peptides such as octreotide and its derivatives, VIP, MSH, LHRH, chemotactic peptides, cell-penetrating peptides, membrane-translocating moieties, bombesin, elastin, peptidomimetics, organic compounds, inorganic compounds, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, oligonucleotides, aptamers, viruses, whole cells, phages, drugs, polymers, liposomes, chemotherapeutic agents, receptor agonists and antagonists, cytokines, hormones, steroids, and toxins. Examples of organic compounds contemplated within the context of the present invention are or are derived from dyes, compounds targeting CAIX and PSMA, estrogens such as estradiol, androgens, progestins, corticosteroids, methotrexate, folic acid, and cholesterol.

[0332] According to a specific embodiment of the present invention, the primary target is a receptor, and a targeting agent capable of specifically binding to the primary target is used. Suitable targeting agents include, but are not limited to, ligands of such receptors or portions thereof that still bind to the receptor, such as receptor-binding peptides in the case of receptor-binding protein ligands. Other examples of protein targeting agents include insulin, transferrin, fibrinogen-gamma fragment, thrombospondin, claudins, apolipoprotein E, affibody molecules such as ABY-025, ankyrin repeat proteins, ankyrin-like repeat proteins, interferons such as alpha, beta, and gamma interferons, interleukins, lymphokines, colony-stimulating factors, and protein growth factors such as tumor growth factors, such as alpha, beta, tumor growth factors, platelet-derived growth factor (PDGF), uPAR targeting proteins, apolipoproteins, LDL, annexin V, endostatin, and angiostatin. Alternative examples of targeting agents include DNA, RNA, PNA and LNA, which are complementary to the primary target.

[0333] Examples of targeting peptides include LHRH receptor targeting peptides, EC-1 peptides, RGD peptides, HER2-targeting peptides, PSMA-targeting peptides, somatostatin-targeting peptides, and bombesin. Other examples of targeting agents include lipocalins, such as anticalins. One particular embodiment uses Affibodies™, as well as multimers and derivatives thereof.

[0334] In a preferred embodiment, T T is an antibody. In a preferred embodiment, T T is selected from antibodies and antibody derivatives, such as antibody fragments, fragment fusions, proteins, peptides, peptidomimetics, organic molecules, dyes, fluorescent molecules, enzyme substrates.

[0335] In a preferred embodiment, the T is an organic molecule. T has a molecular weight of less than 2000 Da, more preferably less than 1500 Da, more preferably less than 1000 Da, and even more preferably less than 500 Da.

[0336] In another preferred embodiment, the T T is selected from antibody fragments, fragment fusions, and other antibody derivatives that do not contain an Fc domain.

[0337] In other embodiments, the T T are polymers and accumulate as primary targets due to the EPR effect. Exemplary polymers for use in this embodiment include, but are not limited to, polyethylene glycol (PEG), poly(N-(2-hydroxypropyl)methacrylamide) (HPMA), polylactic acid (PLA), polylactic-glycolic acid (PLGA), polyglutamic acid (PG), polyvinylpyrrolidone (PVP), poly(1-hydroxymethylethylenehydroxymethyl-formal) (PHF). Other examples are copolymers of polyacetals / polyketals with hydrophilic polymers selected from the group consisting of polyacrylates, polyvinyl polymers, polyesters, polyorthoesters, polyamides, oligopeptides, polypeptides, and derivatives thereof. Other examples are oligopeptides, polypeptides, glycopolysaccharides, and polysaccharides, such as dextran and hyaluronan.

[0338] Typically, a suitable polymer is polyethylene glycol (PEG), preferably having a number of repeat units in the range of 2 to 4000 and a molecular weight in the range of 200 Da to 100,000 Da.

[0339] In addition, reference is made to [G. Pasut, FM Veronese, Prog. Polym. Sci. 2007, 32, 933-961].

[0340] Other TT's are nanoparticles, microparticles, liposomes, micelles, polymersomes, dendrimers, biomolecules, peptides, peptoids, proteins, carbohydrates, oligonucleotides, oligosaccharides, lipids, liposomes, albumin, albumin-binding moieties, dyes, fluorescent molecules, and enzyme substrates.

[0341] In other embodiments, the T T are selected from amino acids, nucleosides, nucleotides, carbohydrates, and biopolymer fragments, such as oligopeptides or polypeptides, oligopeptoids or polypeptoids, or oligolactides or polylactides, or oligo- or polycarbohydrates, oligonucleotides, varying in 2 to 200, in particular 2 to 113, preferably 2 to 50, more preferably 2 to 24, more preferably 2 to 12 repeating units.

[0342] In some aspects of the invention, the polymeric T T The moieties have molecular weights ranging from 2 to 200 kDa, 2 to 100 kDa, 2 to 80 kDa, 2 to 60 kDa, 2 to 40 kDa, 2 to 20 kDa, 3 to 15 kDa, 5 to 10 kDa, 500 daltons to 5 kDa.

[0343] Other exemplary T T The moieties are dendrimers, such as poly(propyleneimine) (PPI) dendrimers, PAMAM dendrimers, and glycol-based dendrimers.

[0344] In a preferred embodiment, the targeting agent T T are localized or retained in a particular system, tissue, or organ in the body, such as the blood circulation, lymphatic system, nervous system, digestive system, RES system, or organs such as the heart or kidney. For example, microparticles will localize in the liver, and large hydrophilic polymers will be retained in the circulation. Similarly, T T The use of an albumin binding moiety as a medicament will result in long-term retention in the circulation.

[0345] In a preferred embodiment, TT is used to alter the pharmacokinetics of the moiety to which it is attached. This can include, but is not limited to, slowing the blood clearance of the moiety, affecting the volume of distribution of the moiety (e.g., decreasing or increasing the volume of distribution), affecting the metabolism of the moiety, and / or affecting (preferably avoiding) the deposition or uptake of the moiety into non-target tissues. Exemplary T in this regard are: T are polymers, peptides, peptoids, dendrimers, proteins, carbohydrates, oligonucleotides, oligosaccharides, lipids, liposomes, micelles, nanoparticles, microparticles, albumin, albumin-binding moieties, and small to medium-sized organic molecules such as steroids and dyes. Typically, suitable polymers are polyethylene glycol (PEG) or polypropylene glycol (PPG).

[0346] According to a further specific embodiment of the present invention, the primary target and targeting agent are selected to result in specific or increased targeting of tissues or diseases, such as cancer, inflammation, infectious diseases, cardiovascular diseases such as thrombosis, atherosclerosis, hypoxic sites such as stroke, tumors, cardiovascular disorders, brain disorders, apoptosis, angiogenesis, organs, and reporter genes / enzymes. This can be achieved by selecting a primary target with tissue-, cell-, or disease-specific expression. For example, the CC49 antibody targets TAG72, whose expression is restricted in normal tissues, but the receptor is overexpressed in various solid tumor cell types.

[0347] In one embodiment, the targeting agent specifically binds to or complexes with a cell surface molecule, e.g., a cell surface receptor or antigen, for a given cell population. TFollowing specific binding or complex formation with the receptor, the cell allows uptake of the prodrug, which is then internalized within the cell. A subsequently administered active agent enters the cell and activates the prodrug, releasing the drug within the cell. In other embodiments, the targeting agent specifically binds to or complexes with a cell surface molecule, e.g., a cell surface receptor or antigen, for a given cell population. T Following specific binding or complex formation between the prodrug and the receptor, the cell does not permit uptake of the prodrug. A subsequently administered active agent will activate the prodrug outside the cell, after which the released drug will enter the cell.

[0348] As used herein, a T cell is a molecule that "specifically binds to or complexes with," or "targets," a cell surface molecule, an extracellular matrix target, or another target. T The ligand preferentially associates with the target through intermolecular forces. For example, the ligand has a dissociation constant (K) of less than about 50 nM, less than about 5 nM, or less than about 500 pM. d or K D ) can preferentially associate with the target.

[0349] In some embodiments, T T can be a cell-penetrating moiety, e.g., a cell-penetrating peptide. In a preferred embodiment, a T that becomes functional upon reaction of the trigger with the activator. T In a particularly preferred embodiment, the non-functional T T is a portion of the cell-penetrating peptide that is attached to another portion of the cell-penetrating peptide upon reaction of the trigger with the activator. T A peptide, preferably a cell-penetrating peptide, is unmasked upon reaction of the trigger with the activator.

[0350] In other embodiments, T T may be polymers, particles, gels, biomolecules, or other T listed above. TThe moiety is injected locally to create a local depot of prodrug or active agent, which can subsequently react with the active agent or prodrug, respectively.

[0351] In other embodiments, the targeting agent T T is a solid material, such as, but not limited to, a polymer, metal, or ceramic, wherein the solid material is or is contained in a cartridge, reservoir, or depot, wherein preferably the cartridge, reservoir, or depot is used for drug release in vivo.

[0352] In some embodiments, the targeting agent T T Also, D D In a particularly preferred embodiment, the T T inhibits drug D by binding to the primary target D In another preferred embodiment, the T T After the trigger is released, D It functions as:

[0353] In an embodiment of the present invention, T If it contains C B is preferably equal to

[0354] Masking part As discussed above, to avoid the drawbacks of current prodrug activation, such as low release yields and / or slow reactions, IEDDA pyridazine removal using compounds of the present invention can be used to trigger the release of a masking moiety from a masked drug. In this type of prodrug, the masking moiety is attached to the drug via a trigger, and this trigger is not intrinsically activated, for example, by an enzyme or a specific pH, but is activated by controlled administration of an active agent, i.e., a species that reacts with the trigger moiety in the prodrug to induce release of the masking moiety or drug from the trigger (or vice versa, i.e., release of the trigger or drug from the masking moiety; however, people can see this release process), resulting in activation of the drug.

[0355] The present invention provides kits for the administration and activation of prodrugs, wherein the prodrugs are covalently attached, directly or indirectly, to a trigger moiety, which in turn is a drug (D D and a masking moiety (M M wherein the activator comprises a tetrazine satisfying any one of formulas (4), (4a), or (6)-(14).

[0356] In another aspect, the present invention provides a masking moiety, M, linked directly or indirectly to a dienophile moiety satisfying formula (19) above. M , a prodrug comprising

[0357] In yet another aspect, the present invention provides a compound comprising one or more masking moieties M that provide a prodrug that can be activated by an abiotic, bioorthogonal reaction. M Drug D D The present invention provides a method for modifying a compound represented by the formula (19), comprising providing a masking moiety and a drug, and chemically linking the masking moiety and the drug to a dienophile moiety satisfying formula (19).

[0358] In a still further aspect, the present invention provides a masking moiety M M and Drug D D to a patient suffering from a disease that can be modulated by a drug, wherein a prodrug comprising a trigger moiety linked to

[0359] In a still further aspect, the invention is a compound comprising a dienophile moiety for use in prodrug therapy in an animal or human, the moiety being linked to a masking moiety M M The compound includes a bond to

[0360] In another aspect, the invention is the use of a diene as an activator for releasing, in a physiological environment, a substance covalently bound to a compound satisfying formula 19. In this regard, the invention also relates to a diene for use as an activator for releasing, in a physiological environment, a substance bound to a compound satisfying formula 19, and to a method for activating the release, in a physiological environment, of a substance bound to a compound satisfying formula 19, wherein a tetrazine is used as the activator.

[0361] In another aspect, the present invention provides the use of an inverse electron demand Diels-Alder reaction between a compound satisfying formula (19) and a dienophile, preferably trans-cyclooctene, as a chemical tool for releasing a substance administered in covalently bound form in a physiological environment, wherein the substance is bound to a compound satisfying formula (19).

[0362] For the avoidance of doubt, M M In the context of the present invention, where is removed from the antibody (i.e., drug), the terms "activatable antibody" and "prodrug" have the same meaning.

[0363] For the avoidance of doubt, M M In the context of the present invention, where is removed from the drug, the drug itself may contain an additional targeting agent T T The primary target may be bound to one or more primary targets without the use of a nucleotide. In this context, the primary target is preferably a therapeutic target.

[0364] In a preferred embodiment, the drug is coupled to a targeting agent T such that the prodrug can bind to the primary target. T Including activation and M M Following removal of the agonist, the drug binds to another primary target, which may be a therapeutic target.

[0365] In a preferred embodiment, the agent comprises one or more T T Includes parts.

[0366] For the avoidance of doubt, in the context of the use of masking moieties, primary target and therapeutic target are used interchangeably.

[0367] For the avoidance of doubt, one drug construct may be modified with multiple masking moieties.

[0368] In a preferred embodiment, an activatable antibody or prodrug of the invention is used in the treatment of cancer. In a preferred embodiment, an activatable antibody or prodrug of the invention is used in the treatment of an autoimmune disease or an inflammatory disease, such as rheumatoid arthritis. In a preferred embodiment, an activatable antibody or prodrug of the invention is used in the treatment of a fibrotic disease, such as idiopathic pulmonary fibrosis.

[0369] Exemplary classes of primary targets for activatable antibodies or prodrugs of the invention include, but are not limited to, cell surface receptors and secreted proteins (e.g., growth factors), soluble enzymes, structural proteins (e.g., collagen, fibronectin), etc. In preferred embodiments, the primary target is an extracellular target. In preferred embodiments, the primary target is an intracellular target.

[0370] In other embodiments, the agent is a bispecific or trispecific antibody derivative that binds to tumor cells and serves to recruit and activate immune effector cells (e.g., T-cells, NK cells), where the binding function of the immune effector cells is masked and inactivated by linking to the dienophile moiety described above, again serving to enable activation of bioorthogonal chemically activated agents.

[0371] D D C B If D D is preferably not attached to the remainder of the prodrug via its antigen binding domain. D is C A is.

[0372] Masking part M M For example, the conjugated drug D D or prodrugs, which further shield the prodrug, can be antibodies, proteins, peptides, polymers, polyethylene glycol, polypropylene glycol carbohydrates, aptamers, oligopeptides, oligonucleotides, oligosaccharides, carbohydrates, as well as peptides, peptoids, steroids, organic molecules, or combinations thereof. This shielding can be based, for example, on steric hindrance, but the shielding can also be based on the drug D D Such masking moieties can also be based on non-covalent interactions with the drug D D or to influence the in vivo properties of the prodrug (e.g., blood clearance; biodistribution; recognition by the immune system).

[0373] In a preferred embodiment, the masking moiety is an albumin binding moiety. In a preferred embodiment, the masking moiety is equal to a targeting agent. In a preferred embodiment, the masking moiety is bound to a targeting agent. In a preferred embodiment, C A Drug D D is C B Multiple M M wherein the attached M M At least one of them is T T In a preferred embodiment, C A D D If D D Spacer S P via T R is not bound to

[0374] In a preferred embodiment, T R It can itself act as a masking part, provided that C A is D D For clarity, in these embodiments, M M T without adhesion R The size of the drug D D is sufficient to protect the immune system from its primary target, which is preferably the therapeutic target in this context.

[0375] Modified D D M M binds its target allosterically or sterically. D This may reduce the ability of

[0376] In certain embodiments, M M is a peptide and antibody-based D D does not have more than 50% amino acid sequence similarity with its natural protein-based binding partner.

[0377] In a preferred embodiment, M M is a peptide between 2 and 40 amino acids in length.

[0378] In one embodiment, the M M is aimed at the target M D when combined with D The dissociation constant of M M D when not bonded to D D binds to its target in a manner that is at least 100 times greater than the dissociation constant for that target. D In another embodiment, M M D D The bond to D D reduces the ability of the target to bind to the target by at least 90%.

[0379] In a preferred embodiment, a masked D D M in M D binds the target D The ability of the unmasked D D by at least 50%, by at least 60%, by at least 70%, by at least 75%, by at least 80%, by at least 85%, by at least 90%, by at least 95%, by at least 96%, by at least 97%, by at least 98%, by at least 99%, or by 100% compared to the function of D. D The reduction in the ability of a target to bind can be measured, for example, by in vitro displacement assays, e.g., as in WO 2009 / 025846 and WO 2010 / 081173. D This can be determined by using an in vitro displacement assay, which has been described.

[0380] In a preferred embodiment, a masked D D D included in D is an antibody, which expressly encompasses full-length antibodies, antigen-binding fragments thereof, antibody derivatives, antibody analogs, antibody mimetics, and fusions of antibodies or antibody derivatives.

[0381] In some embodiments, the MM is not a natural binding partner of the antibody. M has no or substantially no homology to any natural binding partner of the antibody. M is no more than 5%, no more than 10%, no more than 15%, no more than 20%, no more than 25%, no more than 30%, no more than 35%, no more than 40%, no more than 45%, no more than 50%, no more than 55%, no more than 60%, no more than 65%, no more than 70%, no more than 75%, or no more than 80% similar to any natural binding partner of the antibody. M is no more than 5%, no more than 10%, no more than 15%, no more than 20%, no more than 25%, no more than 30%, no more than 35%, no more than 40%, no more than 45%, no more than 50%, no more than 55%, no more than 60%, no more than 65%, no more than 70%, no more than 75%, or no more than 80% identical to any natural binding partner of the antibody. M is 50% or less identical to any natural binding partner of the antibody. M is 25% or less identical to any natural binding partner of the antibody. M is 20% or less identical to any natural binding partner of the antibody. M is 10% or less identical to any natural binding partner of the antibody.

[0382] In the prodrug, M M and Trigger T R (dienophile derivatives) can be directly linked to each other. They can also be linked to each other via a spacer S P or a self-immolative linker L C The present invention relates to a dimer containing a dimer of M. M It will be understood that this encompasses any conceivable manner of attachment to M. M After the formation of the IEDDA adduct, M D DIt will be understood that the dienophile is linked to a dienophile such that it can ultimately be released from D D and the dienophile, or the self-immolative linker L C In the case of L C Between and dienophile and D D and L C Alternatively, this means that the bond between M M and the bond between the dienophile, or the self-immolative linker L C In the case of L C Between and dienophile and M M and L C This means that the bond between should be cleavable.

[0383] In a preferred embodiment, the antibody included in the masked antibody is a multi-antigen targeting antibody, comprising at least a first antibody or antigen-binding fragment or mimic thereof that binds to a first primary target, and a second antibody or antigen-binding fragment or mimic thereof that binds to a second primary target. In a preferred embodiment, the antibody included in the masked antibody is a multi-antigen targeting antibody, comprising a first antibody or antigen-binding fragment or mimic thereof that binds to a first primary target, a second antibody or antigen-binding fragment or mimic thereof that binds to a second primary target, and a third antibody or antigen-binding fragment or mimic thereof that binds to a third primary target. In a preferred embodiment, the multi-antigen targeting antibody binds to two or more different primary targets. In a preferred embodiment, the multi-antigen targeting antibody binds to two or more different epitopes on the same primary target. In a preferred embodiment, the multi-antigen targeting antibody binds to a combination of two or more different targets and two or more different epitopes on the same primary target. In a preferred embodiment, the masked multi-antigen targeting antibody comprises one M M group, or two or more M M It should be appreciated that preferably, at least one of the primary targets is a therapeutic target.

[0384] In a preferred embodiment of a multispecific activatable antibody, an scFv can be fused to the carboxyl terminus of the heavy chain of an IgG activatable antibody, to the carboxyl terminus of the light chain of an IgG activatable antibody, or to the carboxyl terminus of both the light and heavy chains of an IgG activatable antibody. In a preferred embodiment of a multispecific activatable antibody, an scFv can be fused to the amino terminus of the heavy chain of an IgG activatable antibody, to the amino terminus of the light chain of an IgG activatable antibody, or to the amino terminus of both the light and heavy chains of an IgG activatable antibody. In a preferred embodiment of a multispecific activatable antibody, an scFv can be fused to any combination of one or more carboxyl termini and one or more amino termini of an IgG activatable antibody. Methods for preparing multispecific antibodies are known to those skilled in the art. In addition, reference is made to [Weilde et al., Cancer Genomics & Proteomics 2013, 10, 1-18], [Weidle et al., Seminars in Oncology 2014, 41, 5, 653-660], and [Jachimowicz et al., BioDrugs (2014) 28:331-343], the contents of which are incorporated herein by reference.

[0385] In a preferred embodiment, T R M connected to M is attached to and masked by the antigen binding domain of IgG. R M connected to M is attached to and masked by the antigen-binding domain of at least one scFv. R M connected to M is attached to the antigen binding domain of IgG and masked, and T R M connected to M is attached to and masked by the antigen-binding domain of at least one scFv.

[0386] In a preferred embodiment, M Mhas a dissociation constant, i.e., the equilibrium Kd for binding to an antibody, which is the equilibrium K for binding the antibody to its primary target. d In a preferred embodiment, M M is the K for binding to the antibody d and K d is the K for binding the antibody to its primary target d In a preferred embodiment, M M is the K for binding to the antibody d and K d is the K for binding the antibody to its primary target d In a preferred embodiment, M M is the K for binding to the antibody d and K d is the K for binding the antibody to the primary target d In a preferred embodiment, M M is the K for binding to the antibody d and K d is the K for binding the antibody to the primary target d 1 to 5 times, 2 to 5 times, 2 to 10 times, 5 to 10 times, 5 to 20 times, 5 to 50 times, 5 to 100 times, 10 to 100 times, 10 to 1,000 times, 20 to 100 times, 20 to 1,000 times, or 100 to 1,000 times larger than

[0387] In a preferred embodiment, M M has a higher affinity for binding to the antibody than the affinity of the antibody for its primary target. M has an affinity for binding to the antibody that is approximately equal to the affinity of binding of the antibody to its primary target. Mhas a binding affinity to an antibody that is lower than the binding affinity of the antibody to its primary target. In a preferred embodiment, has a binding affinity to an antibody that is 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 25-fold, 50-fold, 100-fold, 250-fold, 500-fold, or 1,000-fold lower than the binding affinity of the antibody to its primary target. has a binding affinity to an antibody that is 1 to 5-fold, 2 to 5-fold, 2 to 10-fold, 5 to 10-fold, 5 to 20-fold, 5 to 50-fold, 5 to 100-fold, 10 to 100-fold, 10 to 1,000-fold, 20 to 100-fold, 20 to 1,000-fold, or 100 to 1,000-fold lower than the binding affinity of the antibody to its primary target. In a preferred embodiment, it has an affinity of binding to the antibody that is 2 to 20 times lower than the affinity of binding of the antibody to its primary target.

[0388] In a preferred embodiment, M M does not inhibit the binding of the antibody to its primary target. M does not prevent the prodrug, when in its cleaved state, from competing with the antibody for binding to the primary target.

[0389] In a preferred embodiment, the antibody has a dissociation constant of about 100 nM or less for binding to its primary target. In a preferred embodiment, the antibody has a dissociation constant of about 10 nM or less for binding to its primary target. In a preferred embodiment, the antibody has a dissociation constant of about 1 nM or less for binding to its primary target. In a preferred embodiment, the antibody has a dissociation constant of about 1 nM or less for binding to its primary target. M Binding of the antibody reduces the ability of the antibody to bind to its primary target, resulting in M M The dissociation constant (K d ) to the primary target, M M The K of the antibody when not bound to d In a preferred embodiment, M MBinding of the antibody reduces the ability of the antibody to bind to its primary target, resulting in M M The dissociation constant (K d ) to the primary target, M M The K of the antibody when not bound to d In a preferred embodiment, M M Binding of the antibody reduces the ability of the antibody to bind to its primary target, resulting in M M The dissociation constant (K d ) to the primary target, M M The K of the antibody when not bound to d In a preferred embodiment, M M Binding of the antibody reduces the ability of the antibody to bind to its primary target, resulting in M M The dissociation constant (K d ) to the primary target, M M The K of the antibody when not bound to d In a preferred embodiment, M M Binding of the antibody reduces the ability of the antibody to bind to its primary target, resulting in M M The dissociation constant (K d ) to the primary target, M M The K of the antibody when not bound to d In a preferred embodiment, for example, the non-binding steric M M When using M M Binding of the antibody reduces the ability of the antibody to bind to its primary target, resulting in M M The dissociation constant (K d ) to the primary target, M M The K of the antibody when not bound to dIn a preferred embodiment, for example, the non-bonded stereoisomer M M When using M M Binding of the antibody reduces the ability of the antibody to bind to its primary target, resulting in M M The dissociation constant (K d ) to the primary target, M M The K of the antibody when not bound to d In a preferred embodiment, for example, the non-bonded stereoisomer M M When using M M Binding of the antibody reduces the ability of the antibody to bind to its primary target, resulting in M M The dissociation constant (K d ) to the primary target, M M The K of the antibody when not bound to d At least 10,000,000 times larger than

[0390] Exemplary drugs that can be used in prodrugs related to the present invention using a masking moiety include, but are not limited to, antibodies, antibody derivatives, antibody fragments, proteins, aptamers, oligopeptides, oligonucleotides, oligosaccharides, carbohydrates, as well as peptides, peptoids, steroids, toxins, hormones, viruses, whole cells, and phages. In a preferred embodiment, the drug is a low- to medium-molecular-weight compound, preferably an organic compound (e.g., about 200 to about 2500 Da, preferably about 300 to about 1750 Da, more preferably about 300 to about 1000 Da).

[0391] In one embodiment, antibodies are used as drugs. Immunoglobulins from any class or subclass can be selected, such as IgG, IgA, IgM, IgD, and IgE, although antibodies or immunoglobulins derived from IgG antibodies are particularly well suited for use in the present invention. Preferably, the immunoglobulin is of the IgG subclass (IgG1, 2, 3, and 4) or class IgG, including, but not limited to, class IgM, which can specifically bind to a specific epitope on an antigen. Antibodies can be intact immunoglobulins derived from natural or recombinant sources, and can be immunoreactive portions of intact immunoglobulins.Antibodies include, for example, polyclonal antibodies, monoclonal antibodies, camelized single domain antibodies, recombinant antibodies, anti-idiotype antibodies, multispecific antibodies, antibody fragments such as Fv, VHH, Fab, F(ab)2, Fab', Fab'-SH, F(ab')2, single chain variable fragment antibodies (scFv), tandem / bis-scFv, Fc, pFc', scFv-Fc, disulfide Fv (dsFv), bispecific antibodies (bc-scFv), e.g., BiTE antibodies, camelized antibodies, minibodies, nanobodies, resurfaced antibodies, humanized antibodies, antibodies), fully human antibodies, single domain antibodies (sdAbs, also known as Nanobodies™), chimeric antibodies, chimeric antibodies comprising at least one human constant region, dual affinity antibodies, e.g., dual-affinity retargeting proteins (DART™), and multimers and derivatives thereof, e.g., bivalent or multivalent single-chain variable fragments, including bivalent or multivalent single-chain variable fragments (e.g., di-scFvs, tri-scFvs), e.g., minibodies, diabodies, triabodies, tribodies, tetrabodies, etc., and multivalent antibodies. Reference is made to [Trends in Biotechnology 2015, 33, 2, 65], [Trends Biotechnol. 2012, 30, 575-582] and [Canc. Gen. Prot. 2013 10, 1-18] and [BioDrugs 2014, 28, 331-343], the contents of which are incorporated herein by reference.Other embodiments use antibody mimetics as agents, such as affimers, anticalins, avimers, alphabodies, affibodies, DARPins, and multimers, and derivatives thereof; see Trends in Biotechnology 2015, 33, 2, 65, the contents of which are incorporated herein by reference. An "antibody fragment" refers to at least a portion of the variable region of an immunoglobulin that binds to its target, i.e., the antigen-binding region. Multimers may be linearly linked or branched, and may be derived from a single vector, or may be chemically or non-covalently linked. Methods for producing the above-listed constructs are known in the art. For the avoidance of doubt, in the context of the present invention, the term "antibody" is meant to encompass all antibody variants, fragments, derivatives, fusions, analogs, and mimetics outlined in this paragraph, unless otherwise specified.

[0392] Exemplary agents for which the present invention is suitable include, but are not limited to, proteins, peptides, oligosaccharides, oligonucleotides, mono-, bi- and tri-specific antibodies, as well as antibody fragments or protein fusions, preferably bi- and tri-specific antibodies. In a preferred embodiment, the activatable antibody or derivative is formulated as part of a pro-bispecific T cell engager (BITE) molecule.

[0393] Other embodiments utilize immunotoxins that are fusions or conjugates between a toxin and an antibody. Typical toxins included in immunotoxins are cholera toxin, lysine A, gelonin, saporin, bouganin, lysine, abrin, diphtheria toxin, Staphylococcus aureus enterotoxin, Bacillus Cyt2Aa1 toxin, Pseudomonas exotoxin PE38, Pseudomonas exotoxin PE38KDEL, granule-associated serine protease granzyme B, human ribonuclease (RNase), or other pro-apoptotic human proteins. Other exemplary cytotoxic human proteins that can be incorporated into fusion constructs are caspase 3, caspase 6, and BH3-interacting domain death agonist (BID). Current immunotoxins suffer from immunogenicity and toxicity issues, particularly for vascular endothelial cells. Targeting toxins with M M PEG or a peptide, and once the masked immunotoxin binds to its target, M It is expected that removing the .alpha.-amino acid will significantly reduce toxicity and immunogenicity problems.

[0394] In other embodiments, immunocytokines that are fusions or conjugates of cytokines and antibodies are used. Typical cytokines used in cancer therapy include IL-2, IL-7, IL-12, IL-15, IL-21, and TNF. A typical cytokine used in autoimmune disease is the anti-inflammatory IL-10. Target cytokines can be administered via M M PEG or a peptide, and once the masked immunocytokine binds to its target, M It is expected that the removal of the .alpha.-hydroxybenzoates will significantly reduce toxicity and immunogenicity problems. In other embodiments, small to medium sized organic agents are used.

[0395] In a preferred embodiment, the unmasked drug is multispecific and binds to two or more of the same or different primary targets. In a preferred embodiment, the multispecific drug comprises one or more (masked) antibodies (also referred to as binding moieties) designed to engage immune effector cells. In a preferred embodiment, the masked multispecific prodrug comprises one or more (masked) antibodies designed to engage leukocytes. In a preferred embodiment, the masked multispecific prodrug comprises one or more (masked) antibodies designed to engage T cells. In a preferred embodiment, the masked multispecific prodrug comprises one or more (masked) antibodies that engage surface antigens on leukocytes, such as T cells, natural killer (NK) cells, myelomonocytic cells, macrophages, and / or other immune effector cells. In a preferred embodiment, the immune effector cells are leukocytes, T cells, NK cells, or monocytes.

[0396] In an exemplary multispecific masked prodrug, the prodrug comprises an antibody (i.e., a targeting agent) for a cancer receptor, such as TAG72, an antibody for CD3 on T cells and an antibody for CD28 on T cells, wherein the antibody for CD3 or the antibody for CD28, or both antibodies, are M M Another example is an activatable antibody that includes an antibody to a cancer receptor and an antibody to CD3 on T cells, where the antibody to CD3 is masked by M M Another example is a prodrug with an antibody for a cancer receptor and an antibody for CD28 on T cells, where the antibody for CD28 is M M Another example is a prodrug with an antibody for a cancer receptor and an antibody for CD16a on NK cells, where the antibody for CD16a is M MIn yet another embodiment, the unmasked agent may bind to two different immune cells and further bind to tumor cells. The above multispecific antibody derivatives can be prepared, for example, by fusing or conjugating antibodies, antibody fragments, such as Fab, Fabs, scFv, camelid antibody heavy chain fragments, and proteins.

[0397] In some preferred embodiments, M M reduces binding of the agent to a primary target equivalent to the therapeutic target, selected from CD3, CD28, PD-L1, PD-1, LAG-3, TIGIT, TIM-3, B7H4, Vista, CTLA-4 polysialic acid and corresponding lectins. M masks T cell agonists, NK cell agonists, and DC cell agonists.

[0398] In a preferred embodiment of an immune effector cell engaging masked multispecific prodrug, e.g., a T cell engaging multispecific activatable antibody, at least one antibody included in the prodrug is a targeting agent and binds to a primary target, which is typically an antigen present on the surface of a tumor cell or other cell type associated with the disease, such as, but not limited to, EGFR, erbB2, EpCAM, PD-L1, B7H3, or CD71 (transferrin receptor), and at least one other antibody included in the prodrug binds to the primary target. The primary target typically binds to a stimulatory or inhibitory antigen present on the surface of T-cells, natural killer (NK) cells, myelomononuclear cells, macrophages, and / or other immune effector cells, such as, but not limited to, B7-H4, BTLA, CD3, CD4, CD8, CD16a, CD25, CD27, CD28, CD32, CD56, CD137, CTLA-4, GITR, HVEM, ICOS, LAG3, NKG2D, OX40, PD-1, TIGIT, TIM3, or VISTA. In a preferred embodiment, the targeted CD3 antigen is preferably CD3ε or CD3 epsilon.

[0399] One embodiment of the present disclosure is a multispecific activatable antibody that includes an antibody targeting agent directed against a tumor target and another agent that is an agonist antibody directed against a costimulatory receptor expressed on the surface of activated T cells or NK cells, where the agonist antibody is masked. Examples of costimulatory receptors include, but are not limited to, CD27, CD137, GITR, HVEM, NKG2D, and OX40. In this embodiment, when the prodrug binds to the tumor and is activated, it effectively crosslinks and activates the costimulatory receptors expressed by T cells or NK cells in a tumor-dependent manner, enhancing the activity of T cells or NK cells responding to any tumor antigens via their endogenous T cell or NK cell activating receptors. The activation dependency of these T cell or NK cell costimulatory receptors focuses the activity of the activated multispecific prodrug on tumor-specific T cells without activating all T cells, regardless of their antigen specificity.

[0400] One embodiment of the present disclosure is a multispecific activatable antibody that targets diseases characterized by T cell overstimulation, including, but not limited to, the microenvironment of autoimmune or inflammatory diseases. Such prodrugs include antibodies, e.g., IgG or scFv, directed to targets including surface antigens expressed in tissues targeted by T cells in autoimmune or inflammatory diseases, and antibodies, e.g., IgG or scFv, directed to inhibitory receptors expressed on the surface of T cells or NK cells, where the inhibitory antibodies on the T cells or NK cells are masked. Examples of inhibitory receptors include, but are not limited to, BTLA, CTLA-4, LAG3, PD-1, TIGIT, TIM3, and NK-expressed KIR. Examples of tissue antigens targeted by T cells in autoimmune diseases include, but are not limited to, surface antigens expressed on myelin or nerve cells in multiple sclerosis, or surface antigens expressed on pancreatic islet cells in type 1 diabetes. In this embodiment, the prodrug localizes to tissues under autoimmune attack or inflammation, is activated by an activator, and co-binds with T cell or NK cell inhibitory receptors to suppress the activity of autoreactive T cells that respond to any disease tissue target antigens via their endogenous TCRs or activating receptors.

[0401] Other non-limiting exemplary primary targets for binding moieties comprised in the agents of the invention are described in WO 2015 / 013671, the contents of which are incorporated herein by reference.

[0402] In other embodiments, the agent is a masked vaccine that can be unmasked at a desired time and / or at a selected location in the body, e.g., subcutaneously and / or near a lymph node. In other embodiments, the agent is a masked antigen, e.g., a masked peptide, that may be present within the Major Histocompatibility Complex (MHC), and that can be unmasked at a desired time and / or at a selected location in the body, e.g., subcutaneously and / or near a lymph node.

[0403] The prodrug may further comprise another linked drug, which is released upon target binding, either by proteases, pH, thiols, or by catabolism. Examples are provided in the review of antibody-drug conjugates in [Polakis, Pharmacol. Rev. 2016, 68, 3-19]. The present invention further contemplates that the prodrug can induce antibody-dependent cellular toxicity (ADCC) or complement-dependent cytotoxicity (CDC) upon unmasking one or more moieties of the prodrug. The present invention also contemplates that the prodrug can induce antibody-dependent cellular cytotoxicity (ADCC) or complement-dependent cytotoxicity (CDC) independently of unmasking one or more moieties of the prodrug.

[0404] Some embodiments use as the additional agent antiproliferative / antitumor agents, antibiotics, cytokines, anti-inflammatory agents, antivirals, antihypertensive agents, chemosensitizers, radiosensitizers, DNA damaging agents, antimetabolites, natural products, and analogs thereof, preferably proteins or antibodies.

[0405] Drugs Drugs that can be used in prodrugs (D D) or the agent that can be inactivated in the context of the present invention is a pharmaceutically active compound. In a preferred embodiment, the pharmaceutically active compound is selected from the group consisting of cytotoxins, antiproliferative / antitumor agents, antivirals, antibiotics, anti-inflammatory agents, chemosensitizers, radiosensitizers, immunomodulators, immunosuppressants, immunostimulators, antiangiogenic factors, enzyme inhibitors.

[0406] In preferred embodiments, these pharmaceutically active compounds are selected from the group consisting of antibodies, antibody derivatives, antibody fragments, proteins, aptamers, oligopeptides, oligonucleotides, oligosaccharides, carbohydrates, as well as peptides, peptoids, steroids, toxins, hormones, cytokines, and chemokines.

[0407] In a preferred embodiment, these agents are low to medium molecular weight compounds, preferably organic compounds (eg, about 200 to about 2500 Da, preferably about 300 to about 1750 Da, more preferably about 300 to about 1000 Da).

[0408] Exemplary types of cytotoxic agents for use as conjugates to TCOs and released upon IEDDA reaction with an active agent, e.g., for use in cancer therapy, include, but are not limited to, DNA damaging agents, DNA crosslinking agents, DNA binders, DNA alkylating agents, DNA intercalators, DNA cleaving agents, microtubule stabilizing and destabilizing agents, topoisomerase inhibitors, radiosensitizers, antimetabolites, natural products and analogs thereof, peptides, oligonucleotides, enzyme inhibitors, e.g., dihydrofolate reductase inhibitors and thymidylate synthase inhibitors.

[0409] Examples include colchicine, vinca alkaloids, anthracyclines (e.g., doxorubicin, epirubicin, idarubicin, daunorubicin), camptothecins, taxanes, taxol, vinblastine, vincristine, vindesine, calicheamicin, tubulysin, tubulysin M, cryptophycin, methotrexate, methopterin, aminopterin, dichloromethotrexate, irinotecan, enediynes, amanitin, deBouganin, dactinomycin, CC1065 and its analogs, duocarmycins, maytansine, maytansinoids, dolastatins, auristatins, pyridinobenzodiazepines and dimers (PBDs). dimers), indolinobenzodiazepines and dimers, pyridinobenzodiazepines and dimers, mitomycins (e.g., mitomycin C, mitomycin A, caminomycin), melphalan, leurosine, leurosideine, actinomycin, tallysomycin, lexitropsin, bleomycin, podophyllotoxin, etoposide, etoposide phosphate, staurosporine, esperamicin, the pteridine family of drugs, SN-38 and its analogs, platinum-based drugs, and cytotoxic nucleosides.

[0410] Other exemplary drug classes are angiogenesis inhibitors, cell cycle progression inhibitors, P13K / m-TOR / AKT pathway inhibitors, MAPK signaling pathway inhibitors, kinase inhibitors, protein chaperone inhibitors, HDAC inhibitors, PARP inhibitors, Wnt / Hedgehog signaling pathway inhibitors, and RNA polymerase inhibitors.

[0411] Examples of auristatins include dolastatin 10, monomethyl auristatin E (MMAE), auristatin F, monomethyl auristatin F (MMAF), auristatin F hydroxypropylamide (AF HPA), auristatin F phenylenediamine (AFP), monomethyl auristatin D (MMAD), auristatin PE, auristatin EB, auristatin EFP, auristatin TP, and auristatin AQ. Suitable auristatins are also described in U.S. Patent Application Publication Nos. 2003 / 0083263, 2011 / 0020343, and 2011 / 0070248; WO 09 / 117531, WO 2005 / 081711, WO 04 / 010957; WO 02 / 088172; FRET, and WO 01 / 24763, as well as U.S. Pat. Nos. 7,498,298; 6,884,869; 6,323,315; 6,239,104; 6,124,431; 6,034,065; 5,780,588; and 5,767,237. Nos.; U.S. Pat. Nos. 5,665,860; 5,663,149; 5,635,483; 5,599,902; 5,554,725; 5,530,097; 5,521,284; 5,504,191; 5,410,024; 5,138,036; 5,076,973; 4,986,988; 4,978,744; 4,879,278; 4,879,278; 4,816,444; and 4,486,414, the disclosures of which are incorporated herein by reference in their entireties.

[0412] Exemplary agents include dolastatins and their analogs, including dolastatin A (U.S. Pat. No. 4,486,414), dolastatin B (U.S. Pat. No. 4,486,414), dolastatin 10 (U.S. Pat. Nos. 4,486,444, 5,410,024, 5,504,191, 5,521,284, 5,530,097, 5,599,902, 5,635,483, 5,663,149, 5,665,860, 5,666,861, 5,666,862, 5,666,863, 5,666,864, 5,666,865 ... Nos. 5,780,588, 6,034,065, 6,323,315), dolastatin 13 (U.S. Pat. No. 4,986,988), dolastatin 14 (U.S. Pat. No. 5,138,036), dolastatin 15 (U.S. Pat. No. 4,879,278), dolastatin 16 (U.S. Pat. No. 6,239,104), dolastatin 17 (U.S. Pat. No. 6,239,104), and dolastatin 18 (U.S. Pat. No. 6,239,104), each of which is incorporated herein by reference in its entirety.

[0413] Exemplary maytansine maytansinoids, such as DM-1 and DM-4, ​​or maytansinoid analogs, including maytansinol and maytansinol analogs, see U.S. Pat. Nos. 4,424,219; 4,256,746; 4,294,757; 4,307,016; 4,313,946; 4,315,929; 4,331,598; 4,361,650; 4,362,6 No. 63; U.S. Patent No. 4,364,866; U.S. Patent No. 4,450,254; U.S. Patent No. 4,322,348; U.S. Patent No. 4,371,533; U.S. Patent No. 5,208,020; U.S. Patent No. 5,416,064; U.S. Patent No. 5,475,092; U.S. Patent No. 5,585,499; U.S. Patent No. 5,846,545; U.S. Patent No. 6,333,410; U.S. Patent No. 6,441,163; U.S. Patent No. 6,716,821 and U.S. Patent No. 7,276,497. Other examples include mertansine and ansamitocin.

[0414] Pyrrolobenzodiazepines (PBDs), explicitly including dimers and analogs, include, but are not limited to, pyrrolobenzodiazepines described in [Denny, Exp. Opin. Ther. Patents, 10(4):459-474 (2000)], [Hartley et al., Expert Opin Investig Drugs. 2011, 20(6):733-44], and [Antonow et al., Chem Rev. 2011, 111(4), 2815-64]. Exemplary indolinobenzodiazepines are described in the literature. Exemplary pyridinobenzodiazepines are described in the literature.

[0415] Calicheamicins include, for example, enediynes, esperamicins, and those described in US Pat. Nos. 5,714,586 and 5,739,116.

[0416] Examples of duocarmycins and analogs include CC1065, duocarmycin SA, duocarmycin A, duocarmycin B1, duocarmycin B2, duocarmycin C1, duocarmycin C2, duocarmycin D, DU-86, KW-2189, adozelesin, bizelesin, carzelesin, seco-adozelesin, CPI, and CBI. Other examples are described in, e.g., U.S. Pat. Nos. 5,070,092; 5,101,092; 5,187,186; 5,475,092; 5,595,499; 5,846,545; 6,534,660; 6,548,530; 6,586,618; 6,660,742; and U.S. Pat. Nos. 56,397; 7,049,316; 7,553,816; 8,815,226; U.S. Patent Application Publication No. 20150104407; U.S. Patent Application No. 61 / 988,011, filed May 2, 2014, and U.S. Patent Application No. 62 / 010,972, filed June 11, 2014, the disclosures of each of which are incorporated herein by reference in their entirety.

[0417] Exemplary vinca alkaloids include vincristine, vinblastine, vindesine, and navelbine, and those disclosed in U.S. Patent Application Publication Nos. 2002 / 0103136 and 2010 / 0305149, and in U.S. Patent No. 7,303,749, the disclosures of which are incorporated herein by reference in their entireties.

[0418] Exemplary epothilone compounds include epothilone A, B, C, D, E, and F, and derivatives thereof. Suitable epothilone compounds and derivatives thereof are described, for example, in U.S. Pat. Nos. 6,956,036; 6,989,450; 6,121,029; 6,117,659; 6,096,757; 6,043,372; 5,969,145; and 5,886,026; and WO 97 / 19086; WO 98 / 08849. No. WO 98 / 22461; WO 98 / 25929; WO 98 / 38192; WO 99 / 01124; WO 99 / 02514; WO 99 / 03848; WO 99 / 07692; WO 99 / 27890; and WO 99 / 28324, the disclosures of which are incorporated herein by reference in their entireties.

[0419] Exemplary cryptophycin compounds are described in U.S. Patent Nos. 6,680,311 and 6,747,021, the disclosures of which are incorporated herein by reference in their entireties. Exemplary platinum compounds include cisplatin, carboplatin, oxaliplatin, iproplatin, ormaplatin, and tetraplatin. Exemplary DNA-binding or alkylating agents include CC-1065 and its analogs, anthracyclines, calicheamicin, dactinomycin, mithromycin, pyrrolobenzodiazepines, indolinobenzodiazepines, pyridinobenzodiazepines, and the like. Exemplary microtubule-stabilizing and microtubule-destabilizing agents include taxane compounds, such as paclitaxel, docetaxel, tesetaxel, and carbazitaxel; maytansinoids, auristatins and their analogs, vinca alkaloid derivatives, epothilones, and cryptophycins. Exemplary topoisomerase inhibitors include camptothecin and camptothecin derivatives, camptothecin analogs, and unnatural camptothecins, such as CPT-11, SN-38, topotecan, 9-aminocamptothecin, rubitecan, gimatecan, karenitecin, ciratecan, raltotecan, exatecan, diflometotecan, belotecan, raltotecan, and S39625. Other camptothecin compounds that can be used in the present invention include, for example, the camptothecin compounds described in J. Med. Chem., 29:2358-2363 (1986); J. Med. Chem., 23:554 (1980); J. Med. Chem., 30:1774 (1987). Angiogenesis inhibitors include, but are not limited to, MetAP2 inhibitors, VEGF inhibitors, PIGF inhibitors, VGFR inhibitors, PDGFR inhibitors, and MetAP2 inhibitors. Exemplary VGFR and PDGFR inhibitors include sorafenib, sunitinib, and vatalanib. Exemplary MetAP2 inhibitors include fumagillol analogs, which refer to compounds that include the fumagillin core structure.Exemplary cell cycle progression inhibitors include, for example, CDK inhibitors such as BMS-387032 and PD0332991; Rho kinase inhibitors such as AZD7762; Aurora kinase inhibitors such as AZD1152, MLN8054 and MLN8237; PLK inhibitors such as BI2536, BI6727, GSK461364, ON-01910; and KSP inhibitors such as SB743921, SB715992, MK-0731, AZD8477, AZ3146, and ARRY-520. Exemplary P13K / m-TOR / AKT signaling pathway inhibitors include phosphoinositide 3-kinase (P13K) inhibitors, GSK-3 inhibitors, ATM inhibitors, DNA-PK inhibitors, and PDK-1 inhibitors. Exemplary P13 kinase inhibitors are disclosed in U.S. Patent No. 6,608,053 and include BEZ235, BGT226, BKM120, CAL263, demethoxyviridine, GDC-0941, GSK615, IC87114, LY294002, Palomid 529, perifosine, PF-04691502, PX-866, SAR245408, SAR245409, SF1126, wortmannin, XL147, and XL765. Exemplary AKT inhibitors include, but are not limited to, AT7867. Exemplary MAPK signaling pathway inhibitors include MEK inhibitors, Ras inhibitors, JNK inhibitors, B-Raf inhibitors, and B-p38 MAPK. Exemplary MEK inhibitors are disclosed in U.S. Patent No. 7,517,944 and include GDC-0973, GSK1120212, MSC1936369B, AS703026, RO5126766 and RO4987655, PD0325901, AZD6244, AZD8330, and GDC-0973. Exemplary B-raf inhibitors include CDC-0879, PLX-4032, and SB590885. Exemplary B-p38 MAPK inhibitors include BIRB 796, LY2228820, and SB 202190.Exemplary receptor tyrosine kinase inhibitors include, but are not limited to, AEE788 (NVP-AEE 788), BIBW2992 (Afatinib), lapatinib, erlotinib (Tarceva), gefitinib (Iressa), AP24534 (Ponatinib), ABT-869 (Linifanib), AZD2171, CHR-258 (Dovitinib), sunitinib (Sutent), sorafenib (Nexavar), and vatalinib. Exemplary protein chaperone inhibitors include HSP90 inhibitors. Exemplary inhibitors include 17AAG derivatives, BIIB021, BIIB028, SNX-5422, NVP-AUY-922, and KW-2478. Exemplary HDAC inhibitors include belinostat (PXD101), CUDC-101, droxinostat, ITF2357 (gibinostat, gabinostat), JNJ-26481585, LAQ824 (NVP-LAQ824, dacinostat), LBH-589 (panobinostat), MC1568, MGCD0103 (mosetinostat), MS-275 (entinostat), PCI-24781, pyroxamide (NSC 696085), SB939, trichostatin A, and vorinostat (SAHA). Exemplary PARP inhibitors include iniparib (BSI 201), olaparib (AZD-2281), ABT-888 (veliparib), AG014699, CEP9722, MK 4827, KU-0059436 (AZD2281), LT-673, 3-aminobenzamide, A-966492, and AZD2461. Exemplary Wnt / Hedgehog signaling pathway inhibitors include vismodegib, cyclopamine, and XAV-939. Exemplary RNA polymerase inhibitors include amatoxins. Exemplary amatoxins include alpha-amanitin, beta-amanitin, gamma-amanitin, eta-amanitin, amanulin, amanuric acid, amanisamide, amanone, and pro-amanulin.Exemplary immune modulators are APRIL, cytokines including, for example, IL-2, IL-7, IL-10, IL-12, IL-15, IL-21, TNF, interferon gamma, GMCSF, NDV-GMCSF, and agonists and antagonists of STING, agonists and antagonists of TLRs including, for example, TLR1 / 2, TLR3, TLR4, TLR7 / 8, TLR9, TLR12, GITR, CD3, CD28, CD40, CD74, CTLA4, OX40, PD1, PDL1, RIG, MDA-5, NLRP1, NLRP3, AIM2, IDO, MEK, cGAS, and CD25, agonists and antagonists of NKG2A. Other exemplary agents include puromycin, topetecan, rhizoxin, echinomycin, combretastatin, netropsin, estramustine, cemadotin, discodermolide, eleutherobin, mitoxantrone, pyrrolobenzimidazole (PBI), gamma interferon, tiaranostatin (A) and analogs, CDK11, immunotoxins, including, for example, lysine A, diphtheria toxin, and cholera toxin.

[0420] In exemplary embodiments of the invention, the drug moiety is a mitomycin compound, a vinca alkaloid compound, taxol or an analog, an anthracycline compound, a calicheamicin compound, a maytansinoid compound, an auristatin compound, a duocarmycin compound, SN38 or an analog, a pyrrolobenzodiazepine compound, an indolinobenzodiazepine compound, a pyridinobenzodiazepine compound, a tubulysin compound, a non-naturally occurring camptothecin compound, a DNA binding drug, a kinase inhibitor, a MEK inhibitor, a KSP inhibitor, a P13 kinase inhibitor, a topoisomerase inhibitor, or an analog thereof.

[0421] In one preferred embodiment, the agent is a non-naturally occurring camptothecin compound, a vinca alkaloid, a kinase inhibitor (e.g., P13 kinase inhibitors: GDC-0941 and PI-103), a MEK inhibitor, a KSP inhibitor, an RNA polymerase inhibitor, a PARP inhibitor, docetaxel, paclitaxel, doxorubicin, a dolastatin, a calicheamicin, SN38, a pyrrolobenzodiazepine, a pyridinobenzodiazepine, an indolinobenzodiazepine, a DNA binding agent, maytansinoids DM1 and DM4, auristatin MMAE, CC1065 and analogs thereof, camptothecin and analogs thereof, or SN-38 and analogs thereof.

[0422] In another preferred embodiment, the agent is selected from DNA binding agents and microtubule agents, including pyrrolobenzodiazepines, indolinobenzodiazepines, pyridinobenzodiazepines, maytansinoids, maytansines, auristatins, tubulysins, duocarmycins, anthracyclines, and taxanes. In another preferred embodiment, the agent is selected from colchicine, vinca alkaloids, tubulysins, irinotecan, inhibitory peptides, amanitin, and debouganin.

[0423] In another preferred embodiment, the agent is a radioactive moiety, which comprises a radioisotope for radiotherapy. The radionuclide used for the treatment is preferably: 24 Na, 32 P, 33 P, 47 Sc, 59 Fe, 67 Cu, 76 As, 77 As, 80 Br, 82 Br, 89 Sr, 90 Nb, 90 Y, 103 Ru, 105 Rh, 109 Pd, 111 Ag, 111 In, 121 Sn,127 Te, 131 I, 140 La, 141 Ce, 142 Pr, 143 Pr, 144 Pr, 149 Pm, 149 Tb, 151 Pm, 153 Sm, 159 Gd, 161 Tb, 165 Dy, 166 Dy, 166 Ho, 169 Er, 172 Tm, 175 Yb, 177 Lu, 186 Re, 188 Re, 198 Au, 199 Au, 211 At, 211 Bi, 212 Bi, 212 Pb, 213 Bi, 214 Bi, 223 Ra, 224 Ra, 225 Ac, and 227 The isotope is selected from the group consisting of Th.

[0424] The radioactive moiety is a metal, e.g. 177 When it is intended to include Lu, such radiometals are preferably provided in the form of a chelate. In such cases, the radioactive moiety preferably comprises a structural moiety capable of forming a coordination complex with such metal. A good example of this is the macrocyclic lanthanide(III) chelate derived from 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (H4dota).

[0425] In a preferred embodiment, the moiety is DTPA (diethylenetriaminepentaacetic acid), DOTA (1,4,7,10-tetraazacyclododecane-N,N',N",N"-tetraacetic acid), NOTA (1,4,7-triazacyclononane-N,N',N"-triacetic acid), TETA (1,4,8,11-tetraazacyclotetradecane-N,N',N",N'-tetraacetic acid), OTTA (N1-(p-isothiocyanatobenzyl)-diethylenetriamineN1, N2,N3,N3-tetraacetic acid), deferoxamine or DFO (N'[5-[[4-[[5-(acetylhydroxyamino)pentyl]amino]-1,4-dioxobutyl]hydroxyamino]pentyl]-N-(5-aminopentyl)-N-hydroxybutanediamide), and a chelating moiety selected from the group consisting of HYNIC (hydrazinonicotinamide), DOTAM, TACN, saclofazine, and 3,4-HOPO-based chelators.

[0426] In other embodiments, the radioactive moiety is a non-metallic radionuclide, e.g. 131 I, containing a prostethic group (i.e., phenol) bound by

[0427] In other embodiments, a combination of two or more different drugs is used. In preferred embodiments, the drug to be released is itself a prodrug designed to release an additional drug. In preferred embodiments, the drug to be released is itself a prodrug designed to be bound to another moiety (e.g., another prodrug) to form an active drug. In preferred embodiments, the drug has increased therapeutic efficacy after reaction of the trigger with the activator. In some embodiments, the drug has decreased therapeutic efficacy after reaction of the trigger with the activator.

[0428] The agent may comprise a membrane-translocating moiety (e.g., adamantine, polylysine / arginine, TAT, human lactoferrin) and / or a targeting agent (e.g., directed against a tumor cell receptor), which may be linked via a stable or unstable linker. Exemplary references include: Trends in Biochemical Sciences, 2015, 40, 12, 749; J. Am. Chem. Soc. 2015, 137, 12153-12160; Pharmaceutical Research, 2007, 24, 11, 1977.

[0429] Trigger or Linker L C one or more targeting agents (or C B ) plus the targeting agent T T But, spacer S P It will be further understood that the T may be attached to the drug, optionally via a T The targeting efficacy of increases after reaction of the trigger with an activator, wherein preferably the agent is C B wherein preferably the active agent is T T wherein, optionally, the active agent contains the above T T The targeting efficacy of is increased after reaction of the trigger with the activator.

[0430] Alternatively, the targeting agent (or C B It will further be understood that a linker such as a nucleotide sequence may include one or more additional agents attached to the targeting agent by other types of linkers, e.g., a linker that is cleavable by a protease, pH, a thiol, or by catabolism.

[0431] The present invention further contemplates that when the targeting agent is an appropriately selected antibody or antibody derivative, such targeting agent can induce antibody-dependent cellular toxicity (ADCC) or complement dependent cytotoxicity (CDC).

[0432] Some agents may contain or be replaced by an imageable label to measure the targeting and release of the agent.

[0433] It will be appreciated that chemical modifications may also be made to the desired compound to make reactions of that compound more convenient for the purpose of preparing the conjugates of the present invention.

[0434] Drugs containing an amine functionality for coupling to a TCO include mitomycin-C, mitomycin-A, daunorubicin, doxorubicin, aminopterin, actinomycin, bleomycin, 9-aminocamptothecin, N8-acetylspermidine, 1-(2-chloroethyl)-1,2-dimethanesulfonylhydrazide, tallysomycin, cytarabine, dolastatins (including auristatin), and their derivatives.

[0435] Drugs containing a hydroxyl functionality for coupling to TCOs include etoposide, camptothecin, taxol, esperamicin, 1,8-dihydroxy-bicyclo[7.3.1]trideca-4-9-diene-2,6-diyn-13-one (U.S. Pat. No. 5,198,560), podophyllotoxin, anguidine, vincristine, vinblastine, morpholine-doxorubicin, n-(5,5-diacetoxy-pentyl)doxorubicin, and derivatives thereof.

[0436] Agents containing sulfhydryl functional groups for coupling to TCOs include esperamicin and 6-mercaptopurine, and their derivatives.

[0437] The drug may optionally be linked to a self-immolative linker L C , or a combination thereof, and it will be understood that it may consist of multiple (self-immolative or non-self-immolative) units.

[0438] According to further particular embodiments of the present invention, the prodrug is selected to target and / or address diseases such as cancer, inflammation, infectious diseases, cardiovascular diseases such as thrombosis, atherosclerosis, hypoxic sites such as stroke, tumors, cardiovascular disorders, brain disorders, apoptosis, angiogenesis, organs, and reporter genes / enzymes.

[0439] In the prodrug, Construct-A and the TCO derivative can be directly linked to each other. They can also be linked to each other via a linker or a self-immolative linker L. C The dienophile TCOs can be bonded to each other via a dienophile TCO. It will be understood that the present invention encompasses any conceivable manner in which the dienophile TCO is attached to Construct-A. In a preferred embodiment, Construct-A is a drug. For example, in the case of proteins, methods for affecting conjugation to these drugs via reactive amino acids, such as lysine or cysteine, are known to those skilled in the art.

[0440] sign The compound of formula (19) preferably includes a label capable of providing a desired diagnostic, imaging, and / or radiotherapeutic effect.

[0441] In preferred embodiments, the label is selected from the group consisting of an MRI-imageable construct, a moiety comprising a spin label, a moiety comprising an optical label, an ultrasound-responsive construct, an X-ray-responsive moiety, a moiety comprising a radionuclide, a fluorescent dye, a luminescent dye, a FRET dye, a paramagnetic ion, a superparamagnetic particle, and a peptide.

[0442] Preferably, the label is a detectable label, particularly for imaging applications. As used herein, a "detectable label" refers to a moiety of a compound of formula (19) that allows the compound of formula (19) to be detected when present in a cell, tissue, or organism. One type of detectable label contemplated within the context of the present invention is a contrast-providing label. Different types of detectable labels are contemplated within the context of the present invention and are described herein below.

[0443] Thus, according to a particular embodiment of the present invention, the compounds, combinations, kits and methods of the present invention are used in imaging, particularly medical imaging, where a detectable label is used to identify the primary target and / or to assess the biodistribution of the compound of formula (19).

[0444] Preferred detectable labels for imaging are contrast-providing moieties used in classical imaging systems, e.g., MRI-imageable constructs, moieties containing spin labels, moieties containing optical labels, ultrasound-responsive constructs, X-ray-responsive moieties, moieties containing radionuclides, (bio)luminescence, and FRET-type dyes.

[0445] Furthermore, preferred detectable labels contemplated within the context of the present invention include, but are not necessarily limited to, fluorescent molecules, e.g., autofluorescent molecules, molecules that fluoresce in response to contact with a reagent, radioactive moieties or conjugates, biotin, e.g., biotin detected through binding of biotin with avidin, fluorescent tags, imaging constructs for MRI including paramagnetic metals, imaging reagents, e.g., those described in U.S. Pat. Nos. 4,741,900 and 5,326,856.

[0446] Preferably, the radionuclide contained in the label for imaging is 3 H, 11 C. 13 N,15 O. 18 F, 19 F, 44 Sc, 51 Cr, 52 Fe, 52 Mn, 55 Co, 60 Cu, 61 Cu, 62 Zn, 62 Cu, 63 Zn, 64 Cu, 66 Ga, 67 Ga, 68 Ga, 70 As, 71 As, 72 As, 74 As, 75 Se, 75 Br, 76 Br, 77 Br, 8O Br, 82 Br, 82 Rb, 86 Y, 88 Y, 89 Sr, 89 Zr, 97 Ru, 99m Tc, 110 In, 111 In, 113 In, 114 In, 117 Sn, 120 I, 122 Xe, 123 I, 124 I, 125 I, 166 Ho, 167 Tm, 169 Yb, 193 Pt, 195 Pt, 201 Tl, and 203 Pb. More preferably, the radionuclide contained in the label for imaging is an isotope selected from the group consisting of: 18 F, 44 Sc, 64 Cu, 68 Ga, 89 Zr, 99m Tc, 111 In, 123 I, 124I is an isotope selected from the group consisting of:

[0447] Other elements and isotopes, such as those used for therapy, may also be adapted for imaging in certain applications.

[0448] In a preferred embodiment, the MRI-imageable moiety is a paramagnetic ion or a superparamagnetic particle. The paramagnetic ion is preferably an element selected from the group consisting of Gd, Fe, Mn, Cr, Co, Ni, Cu, Pr, Nd, Yb, Tb, Dy, Ho, Er, Sm, Eu, Ti, Pa, La, Sc, V, Mo, Ru, Ce, Dy, and Tl. The ultrasound-responsive moiety can comprise a microbubble, the shell of which is made of phospholipids, and / or (biodegradable) polymers, and / or human serum albumin. The microbubbles can be filled with a fluorinated gas or liquid.

[0449] X-ray responsive moieties include, but are not limited to, iodine, barium, barium sulfate, gastrografin, or may comprise vesicles, liposomes, or polymer capsules filled with iodine compounds and / or barium sulfate.

[0450] Moreover, detectable labels contemplated within the context of the present invention also include peptides or polypeptides that can be detected by antibody binding, for example, by binding of a detectably labeled antibody or by detecting the bound antibody through a sandwich-type assay. In one embodiment, the detectable label is a small-sized organic PET and SPECT radioisotope, e.g., 18 F, 11 C. 123 I or 124 I. Due to their small size, organic PET or SPECT radioisotopes do not significantly affect the properties of targeting agents in general, particularly their membrane trafficking, making them ideally suited for monitoring intracellular events.

[0451] In a preferred embodiment, particularly when the compound of formula (19) is used in a therapeutic application, the label is a therapeutic label, wherein the label comprises a radioisotope for radiotherapy. The radionuclide used for therapy is preferably 24 Na, 32 P, 33 P, 47 Sc, 59 Fe, 67 Cu, 76 As, 77 As, 80 Br, 82 Br, 89 Sr, 90 Nb, 90 Y, 103 Ru, 105 Rh, 109 Pd, 111 Ag, 111 In, 121 Sn, 127 Te, 131 I, 140 La, 141 Ce, 142 Pr, 143 Pr, 144 Pr, 149 Pm, 149 Tb, 151 Pm, 153 Sm, 159 Gd, 161 Tb, 165 Dy, 166 Dy, 166 Ho, 169 Er, 172 Tm, 175 Yb, 177 Lu, 186 Re, 188 Re, 198 Au, 199 Au, 211 At, 211 Bi, 212 Bi, 212 Pb, 213 Bi, 214 Bi, 223 Ra, 224 Ra, 225 Ac, and 227More preferably, the radionuclide contained in the therapeutic label is an isotope selected from the group consisting of: 90 Y, 111 In, 131 I, 177 Lu, 186 Re, 188 Re, 211 At, 212 Pb, 213 Bi, 225 Ac, and 227 The isotope is selected from the group consisting of Th.

[0452] The label may be a metal, e.g., a 111 When it is intended to include In, it is preferably provided in the form of a chelate. In such cases, the label preferably contains a structural moiety capable of forming a coordination complex with such a metal. Good examples thereof are the macrocyclic lanthanide(III) chelates derived from 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (H4dota).

[0453] In a preferred embodiment, the label is -OR 37 , -N(R 37 )2, -CF3, -SR 37 , S(=O)2N(R 37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R 37 , SC(=S)R 37 , N.R. 37 C(=O)-R 37 , N.R. 37 C(=S)-R 37 , N.R. 37 C(=O)OR 37 , N.R. 37 C(=S)OR 37 , N.R. 37 C(=O)SR 37 , N.R. 37 C(=S)SR 37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR 37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37 )2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , C1~C 24 Alkyl groups, C2-C 24 Alkenyl groups, C2-C 24 Alkynyl groups, C6-C 24 Aryl groups, C2-C 24 Heteroaryl groups, C3-C 24 Cycloalkyl groups, C5-C 24 Cycloalkenyl group, C 12 ~C 24 Cycloalkynyl group, C3-C 24 (Cyclo)alkyl(hetero)aryl groups, C3-C 24 (Hetero)aryl(cyclo)alkyl, C4-C 24 (Cyclo)alkenyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkenyl groups, C4-C 24 (Cyclo)alkynyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkynyl group, C4-C 24 Alkylcycloalkyl groups and C4-C 24 cycloalkylalkyl groups; 37Groups, alkyl groups, alkenyl groups, alkynyl groups, aryl groups, heteroaryl groups, cycloalkyl groups, cycloalkenyl groups, cycloalkynyl groups, (cyclo)alkyl(hetero)aryl groups, (hetero)aryl(cyclo)alkyl groups, (cyclo)alkenyl(hetero)aryl groups, (hetero)aryl(cyclo)alkenyl groups, (cyclo)alkynyl(hetero)aryl groups, (hetero)aryl(cyclo)alkynyl groups, alkylcycloalkyl groups, cycloalkylalkyl groups are 3 H, 11 C. 13 N, 15 O. 18 F, 19 F, 44 Sc, 51 Cr, 52 Fe, 52 Mn, 55 Co, 60 Cu, 61 Cu, 62 Zn, 62 Cu, 63 Zn, 64 Cu, 66 Ga, 67 Ga, 68 Ga, 70 As, 71 As, 72 As, 74 As, 75 Se, 75 Br, 76 Br, 77 Br, 8O Br, 82 Br, 82 Rb, 86 Y, 88 Y, 89 Sr, 89 Zr, 97 Ru, 99m Tc, 110 In, 111 In, 113 In, 114 In, 117 Sn, 120 I, 122 Xe, 123 I, 124 I, 125 I, 166 Ho, 167 Tm, 169Yb, 193 Pt, 195 Pt, 201 Tl, 203 Pb, 24 Na, 32 P, 33 P, 47 Sc, 59 Fe, 67 Cu, 76 As, 77 As, 80 Br, 82 Br, 89 Sr, 90 Nb, 90 Y, 103 Ru, 105 Rh, 109 Pd, 111 Ag, 111 In, 121 Sn, 127 Te, 131 I, 140 La, 141 Ce, 142 Pr, 143 Pr, 144 Pr, 149 Pm, 149 Tb, 151 Pm, 153 Sm, 159 Gd, 161 Tb, 165 Dy, 166 Dy, 166 Ho, 169 Er, 172 Tm, 175 Yb, 177 Lu, 186 Re, 188 Re, 198 Au, 199 Au, 211 At, 211 Bi, 212 Bi, 212 Pb, 213 Bi, 214 Bi, 223 Ra, 224 Ra, 225 Ac, and 227 -substituted with and / or chelated with at least one isotope selected from the group consisting of -Th; and -Cl, -F, -Br, -I, -OR 37, -N(R 37 )2, -SO3R 37 , -PO3(R 37 ) 2、 -PO4(R 37 ) 2、 -NO2, -CF3, =O, =NR 37 , and -SR 37 and may be further substituted with a moiety selected from the group consisting of O, S, NR 37 , P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0454] In another preferred embodiment, the label is -OR 37 , -N(R 37 )2, -CF3, -SR 37 , S(=O)2N(R 37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R 37 , SC(=S)R 37 , N.R. 37 C(=O)-R 37 , N.R. 37 C(=S)-R 37 , N.R. 37 C(=O)OR 37 , N.R. 37 C(=S)OR 37 , N.R. 37 C(=O)SR 37 , N.R. 37 C(=S)SR 37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R 37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR 37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37)2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , C1~C 12 Alkyl groups, C2-C 12 Alkenyl groups, C2-C 12 Alkynyl groups, C6-C 12 Aryl groups, C2-C 12 Heteroaryl groups, C3-C 12 Cycloalkyl groups, C5-C 12 Cycloalkenyl group, C 12 ~C 12 Cycloalkynyl group, C3-C 12 (Cyclo)alkyl(hetero)aryl groups, C3-C 12 (Hetero)aryl(cyclo)alkyl, C4-C 12 (Cyclo)alkenyl(hetero)aryl group, C4-C 12 (Hetero)aryl(cyclo)alkenyl groups, C4-C 12 (Cyclo)alkynyl(hetero)aryl group, C4-C 12 (Hetero)aryl(cyclo)alkynyl group, C4-C 12 Alkylcycloalkyl groups and C4-C 12 cycloalkylalkyl groups; 37 Groups, alkyl groups, alkenyl groups, alkynyl groups, aryl groups, heteroaryl groups, cycloalkyl groups, cycloalkenyl groups, cycloalkynyl groups, (cyclo)alkyl(hetero)aryl groups, (hetero)aryl(cyclo)alkyl groups, (cyclo)alkenyl(hetero)aryl groups, (hetero)aryl(cyclo)alkenyl groups, (cyclo)alkynyl(hetero)aryl groups, (hetero)aryl(cyclo)alkynyl groups, alkylcycloalkyl groups, cycloalkylalkyl groups are 3 H, 11 C. 13 N, 15 O. 18 F, 19 F, 44 Sc, 51 Cr, 52 Fe, 52 Mn, 55Co、 60 Cu、 61 Cu、 62 Zn、 62 Cu、 63 Zn、 64 Cu、 66 Ga、 67 Ga、 68 Ga、 70 As、 71 As、 72 As、 74 As、 75 Se、 75 Br、 76 Br、 77 Br、 8O Br、 82 Br、 82 Rb、 86 Y、 88 Y、 89 Sr、 89 Zr、 97 Ru、 99m Tc、 110 In、 111 In、 113 In、 114 In、 117 Sn、 120 I、 122 Xe、 123 I、 124 I、 125 I、 166 Ho、 167 Tm、 169 Yb、 193 Pt、 195 Pt、 201 Tl、 203 Pb、 24 Na、 32 P、 33 P、 47 Sc、 59 Fe、 67 Cu、 76 As、 77 As、 80 Br、 82 Br、 89 Sr、 90 Nb、 90 Y、 103 Ru、 105 Rh、 109 Pd、 111 Ag、 111 In、 121 Sn、 127Te, 131 I, 140 La, 141 Ce, 142 Pr, 143 Pr, 144 Pr, 149 Pm, 149 Tb, 151 Pm, 153 Sm, 159 Gd, 161 Tb, 165 Dy, 166 Dy, 166 Ho, 169 Er, 172 Tm, 175 Yb, 177 Lu, 186 Re, 188 Re, 198 Au, 199 Au, 211 At, 211 Bi, 212 Bi, 212 Pb, 213 Bi, 214 Bi, 223 Ra, 224 Ra, 225 Ac, and 227 -substituted with and / or chelated with at least one isotope selected from the group consisting of -Th; and -Cl, -F, -Br, -I, -OR 37 , -N(R 37 )2, -SO3R 37 , -PO3(R 37 ) 2、 -PO4(R 37 ) 2、 -NO2, -CF3, =O, =NR 37 , and -SR 37 and may be further substituted with a moiety selected from the group consisting of O, S, NR 37 , P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0455] In a preferred embodiment, each label is independently -OR 37 , -N(R37 )2、-CF3、-SR 37 、S(=O)2N(R 37 )2、OC(=O)R 37 、SC(=O)R 37 、OC(=S)R 37 、SC(=S)R 37 、NR 37 C(=O)-R 37 、NR 37 C(=S)-R 37 、NR 37 C(=O)O-R 37 、NR 37 C(=S)O-R 37 、NR 37 C(=O)S-R 37 、NR 37 C(=S)S-R 37 、OC(=O)N(R 37 )2、SC(=O)N(R 37 )2、OC(=S)N(R 37 )2、SC(=S)N(R 37 )2、NR 37 C(=O)N(R 37 )2、NR 37 C(=S)N(R 37 )2、C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2、C(=S)N(R 37 )2、C(=O)O-R 37 、C(=O)S-R 37 、C(=S)O-R 37 、C(=S)S-R 37, a C1-C6 alkyl group, a C2-C6 alkenyl group, a C2-C6 alkynyl group, a C6 aryl group, a C2-C6 heteroaryl group, a C3-C6 cycloalkyl group, a C5-C6 cycloalkenyl group, a C8 cycloalkynyl group, a C3-C6 (cyclo)alkyl(hetero)aryl group, a C3-C6 (hetero)aryl(cyclo)alkyl, a C4-C6 (cyclo)alkenyl(hetero)aryl group, a C4-C6 (hetero)aryl(cyclo)alkenyl group, a C4-C6 (cyclo)alkynyl(hetero)aryl group, a C4-C6 (hetero)aryl(cyclo)alkynyl group, a C4-C6 alkylcycloalkyl group, and a C4-C6 cycloalkylalkyl group; 37 Groups, alkyl groups, alkenyl groups, alkynyl groups, aryl groups, heteroaryl groups, cycloalkyl groups, cycloalkenyl groups, cycloalkynyl groups, (cyclo)alkyl(hetero)aryl groups, (hetero)aryl(cyclo)alkyl groups, (cyclo)alkenyl(hetero)aryl groups, (hetero)aryl(cyclo)alkenyl groups, (cyclo)alkynyl(hetero)aryl groups, (hetero)aryl(cyclo)alkynyl groups, alkylcycloalkyl groups, cycloalkylalkyl groups are 3 H, 11 C. 13 N, 15 O. 18 F, 19 F, 44 Sc, 51 Cr, 52 Fe, 52 Mn, 55 Co, 60 Cu, 61 Cu, 62 Zn, 62 Cu, 63 Zn, 64 Cu, 66 Ga, 67 Ga, 68 Ga, 70 As, 71 As, 72 As, 74 As, 75 Se, 75 Br, 76 Br, 77 Br, 8O Br,82 Br、 82 Rb、 86 Y、 88 Y、 89 Sr、 89 Zr、 97 Ru、 99m Tc、 110 In、 111 In、 113 In、 114 In、 117 Sn、 120 I、 122 Xe、 123 I、 124 I、 125 I、 166 Ho、 167 Tm、 169 Yb、 193 Pt、 195 Pt、 201 Tl、 203 Pb、 24 Na、 32 P、 33 P、 47 Sc、 59 Fe、 67 Cu、 76 As、 77 As、 80 Br、 82 Br、 89 Sr、 90 Nb、 90 Y、 103 Ru、 105 Rh、 109 Pd、 111 Ag、 111 In、 121 Sn、 127 Te、 131 I、 140 La、 141 Ce、 142 Pr、 143 Pr、 144 Pr、 149 Pm、 149 Tb、 151 Pm、 153 Sm、 159 Gd、 161 Tb、 165 Dy、 166 Dy、 166 Ho、 169 Er、 172 Tm、 175 Yb、177 Lu, 186 Re, 188 Re, 198 Au, 199 Au, 211 At, 211 Bi, 212 Bi, 212 Pb, 213 Bi, 214 Bi, 223 Ra, 224 Ra, 225 Ac, and 227 -substituted with and / or chelated with at least one isotope selected from the group consisting of -Th; and -Cl, -F, -Br, -I, -OR 37 , -N(R 37 )2, -SO3R 37 , -PO3(R 37 ) 2、 -PO4(R 37 ) 2、 -NO2, -CF3, =O, =NR 37 , and -SR 37 and may be further substituted with a moiety selected from the group consisting of O, S, NR 37 , P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0456] In a preferred embodiment, the label is derived from a prosthetic group. Those skilled in the art will appreciate that a prosthetic group may be a radionuclide, e.g. 131 It will be appreciated that I is a precursor that can be radiolabeled to form a label.

[0457] In another preferred embodiment, the label is C1-C 12 Alkyl groups, C2-C 12 Alkenyl groups, C2-C 12 Alkynyl groups, C6-C 12 Aryl groups, C2-C 12 Heteroaryl groups, C3-C 12 Cycloalkyl groups, C5-C 12Cycloalkenyl group, C 12 ~C 12 Cycloalkynyl group, C3-C 12 (Cyclo)alkyl(hetero)aryl groups, C3-C 12 (Hetero)aryl(cyclo)alkyl, C4-C 12 (Cyclo)alkenyl(hetero)aryl group, C4-C 12 (Hetero)aryl(cyclo)alkenyl groups, C4-C 12 (Cyclo)alkynyl(hetero)aryl group, C4-C 12 (Hetero)aryl(cyclo)alkynyl group, C4-C 12 Alkylcycloalkyl groups and C4-C 12 cycloalkylalkyl groups; 3 H, 11 C. 13 N, 15 O. 18 F, 19 F, 75 Br, 76 Br, 77 Br, 8O Br, 82 Br, 120 I, 123 I, 124 I, 125 I, 32 P, 33 P, 131 I, 211 At least one isotope selected from the group consisting of At, ... 37 , -N(R 37 )2, -SO3R 37 , -PO3(R 37 ) 2、 -PO4(R 37 ) 2、 -NO2, -CF3, =O, =NR 37 , and -SR 37 and may be further substituted with a moiety selected from the group consisting of O, S, NR 37, P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0458] In a preferred embodiment, the label comprises a chelating moiety. In a preferred embodiment, the label is selected from the group consisting of DTPA (diethylenetriaminepentaacetic acid), DOTA (1,4,7,10-tetraazacyclododecane-N,N',N",N"-tetraacetic acid), DOTAGA anhydride (2,2',2"-(10-(2,6-dioxotetrahydro-2H-pyran-3-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid), NOTA (1,4,7-triazacyclononane-N,N',N"-triacetic acid), TE TA (1,4,8,11-tetraazacyclotetradecane-N,N',N",N'-tetraacetic acid), OTTA (N1-(p-isothiocyanatobenzyl)-diethylenetriamine N1,N2,N3,N3-tetraacetic acid), deferoxamine, or DFO (N'[5-[[4-[[5-(acetylhydroxyamino)pentyl]amino]-1,4-dioxobutyl]hydroxyamino]pentyl]-N-(5-aminopentyl)-N-hydroxybutanediamide), and DFO * and a conjugate of HYNIC (hydrazinonicotinamide); and the chelating moiety is selected from the group consisting of: a DFO derivative called 44 Sc, 51 Cr, 52 Fe, 52 Mn, 55 Co, 60 Cu, 61 Cu, 62 Zn, 62 Cu, 63 Zn, 64 Cu, 66 Ga, 67 Ga, 68 Ga, 70 As, 71 As, 72 As, 74 As, 75 Se, 82 Rb,86 Y、 88 Y、 89 Sr、 89 Zr、 97 Ru、 99m Tc、 110 In、 111 In、 113 In、 114 In、 117 Sn、 122 It is、 166 Ho、 167 Tm、 169 Yb、 193 Pt、 195 Pt、 201 Tl、 203 Pb、 24 Ankle, 47 Scr、 59 Faith、 67 Cu、 76 As、 77 As、 89 Sr、 90 Nb、 90 Y、 103 Ru、 105 Rh、 109 Pd、 111 Ag、 111 In、 121 Sn、 127 For, 140 The、 141 What、 142 Pr、 143 Pr、 144 Pr、 149 Pm、 149 Tb、 151 Pm、 153 Sm、 159 Gd、 161 Tb、 165 Dy、 166 Dy、 166 Ho、 169 Er、 172 Tm、 175 Yb、 177 Lu、 186 Re, 188 Re, 198 I、 199 I、 211 Assets, 211 Wind、 212 Wind、 212 Pb、 213 Wind、 214Bi, 223 Ra, 224 Ra, 225 Ac, and 227 It chelates with a metal selected from the group consisting of Th.

[0459] In a preferred embodiment, the metal chelate comprises an acyclic derivative of ethylenediaminotetraacetic acid (EDTA) or diethylenediaminotetraacetic acid (DTPA).

[0460] [ka]

[0461] Here, the dashed line indicates the bond to the rest of the molecule.

[0462] In another preferred embodiment, the metal chelate comprises an acyclic chelator containing a carboxy-pyridine group as shown below.

[0463] [ka]

[0464] Here, the dashed line indicates the bond to the rest of the molecule.

[0465] In another preferred embodiment, the metal chelate comprises a cyclic derivative of 1,4,7,10-tetraazadodecane (cyclen).

[0466] [ka]

[0467] Here, the dashed line indicates the bond to the rest of the molecule.

[0468] In another preferred embodiment, the metal chelate comprises a derivative of 1,4,7-triazacyclononane (TACN):

[0469] [ka]

[0470] Here, the dashed line indicates the bond to the rest of the molecule.

[0471] In yet another preferred embodiment, the metal chelate comprises a macrocyclic chelator containing N and O heteroatoms, as described below.

[0472] [ka]

[0473] Here, the dashed line indicates the bond to the rest of the molecule.

[0474] In another preferred embodiment, the metal chelate comprises a derivative of the cryptand agent sarcofazine (Sar), described below.

[0475] [ka]

[0476] Here, the dashed line indicates the bond to the rest of the molecule.

[0477] In another preferred embodiment, the metal chelate comprises a linear or cyclic chelating agent containing a hydroxamate group, as described below.

[0478] [ka]

[0479] Here, the dashed line indicates the bond to the rest of the molecule.

[0480] In yet another preferred embodiment, the metal chelate comprises a linear or cyclic chelating agent containing a 3-hydroxy-4-pyridinone (3,4-HOPO) group, as described below, and derivatives thereof.

[0481] [ka]

[0482] Here, the dashed line indicates the bond to the rest of the molecule.

[0483] In other preferred embodiments, the metal chelate comprises a linear or cyclic chelating agent containing N, S, and P heteroatoms, as described below.

[0484] [ka]

[0485] where the dashed line indicates the bond to the rest of the molecule and M is 99m Tc, 186 Re, and 188 Re indicates a radionuclide selected from the group consisting of:

[0486] In other preferred embodiments, the metal chelates contain the following residues: glycine, serine, cysteine, lysine, and alanine.

[0487] [ka]

[0488] where the dashed line indicates the bond to the rest of the molecule and M is 99m Tc, 186 Re, and 188 Re indicates a radionuclide selected from the group consisting of:

[0489] In another preferred embodiment, the metal chelate comprises a hydrazinonicotinic acid derivative (HYNIC) and a co-ligand, as described below.

[0490] [ka]

[0491] where the dashed line indicates the bond to the rest of the molecule and M is 99m Tc, 186 Re, and 188 Re indicates a radionuclide selected from the group consisting of:

[0492] In another preferred embodiment, the chelate comprises a carbonyl group and a chelator containing N, O, and S heteroatoms or cyclopentadienyl, as described below.

[0493] [ka]

[0494] where the dashed line indicates the bond to the rest of the molecule and M is 99m Tc, 186 Re, and 188 Re indicates a radionuclide selected from the group consisting of:

[0495] In some preferred embodiments of the invention, the label is 18 They contain F and can be prepared by those skilled in the art based on known synthetic routes using known labeled synthons or prosthetic groups. 18 Some non-limiting examples of labels containing F are shown below.

[0496] [ka]

[0497] Here, the dashed line indicates the bond to the rest of the molecule.

[0498] In a preferred embodiment of the invention, the label is 123 I, 124 I, 125 I, 131 I, and 211and are synthesized by one skilled in the art based on known synthetic routes using prosthetic groups. Some preferred embodiments of such labels are set forth below.

[0499] [ka]

[0500] where the dashed line indicates the bond to the rest of the molecule and X is 123 I, 124 I, 125 I, 131 I, or 211 Indicates At.

[0501] In yet another preferred embodiment of the present invention, the label comprises: 123 I, 124 I, 125 I, 131 I, and 211 At; and is synthesized by one skilled in the art based on known synthetic routes using the closo-decaborate (2-) group.

[0502] [ka]

[0503] where the dashed line indicates the bond to the rest of the molecule and X is 123 I, 124 I, 125 I, 131 I, or 211 Indicates At.

[0504] Administered drug The agent can be any construct where it is desired to modify the construct with a label for imaging or radiotherapy and to remove the imaging or radiotherapy label at a specific time after injection. This is particularly true for targeted imaging and radiotherapy to a site, e.g., a tumor, within the body of a subject, particularly a human. The only requirement is an additional linker to the label, a Trigger T. R It should be noted that the exact attachment of the trigger to the administered agent will depend on the molecular structure of both molecules, but this usually does not pose a particular challenge to those skilled in the art, since many proven conjugation methods and attachment moieties exist for a variety of biomolecules. The attachment may also be via a spacer, such as a polyethylene glycol (PEG) chain.

[0505] Typically, the administered agent can bind to a primary target, as defined herein. The primary target can be a target to which a targeting agent binds, or the administered agent can be a therapeutic target depending on the effect of the drug. In a preferred embodiment, the primary target is a therapeutic target, and the targeting agent is a drug and binds to the primary target.

[0506] In a preferred embodiment, the administration agent is a targeted agent as defined herein.

[0507] Preferably, the agent is selected from the group consisting of proteins, peptoids and peptides. Most preferably, the agent is an antibody.

[0508] In other preferred embodiments, the agent is selected from the group consisting of antibodies, antibody-drug conjugates, antibody derivatives, antibody fragments, proteins, polymers, polymer-drug conjugates, drugs including liposomes and polymersomes, nanoparticles, microparticles, aptamers, oligopeptides, oligonucleotides, oligosaccharides, carbohydrates, as well as peptides, peptoids, steroids, toxins, hormones, cytokines, and chemokines.

[0509] In another preferred embodiment, the administration agent is equivalent to a targeting agent, and the targeting agent is radiolabeled with a therapeutic radioisotope to target the therapeutic radiation to tissues expressing the primary target.

[0510] In another preferred embodiment, the administered agent is equivalent to a targeting agent, and the targeting agent is radiolabeled with a therapeutic radioisotope for imaging tissues expressing the primary target.

[0511] In a preferred embodiment, the agent is an antibody, more preferably an antibody comprising an FcRn-binding domain, more preferably an intact IgG antibody.

[0512] In another preferred embodiment, the agent is an antibody comprising an albumin-binding moiety. In another preferred embodiment, the agent is a protein comprising an albumin-binding moiety. In another preferred embodiment, the agent is equivalent to a drug. In another preferred embodiment, the agent is equivalent to a drug, and the drug is labeled using the present invention for purposes of imaging drug distribution in vivo.

[0513] Drugs that can be used in the administration preparations related to the present invention include pharmaceutically active compounds such as antibodies, antibody-drug conjugates, antibody derivatives, antibody fragments, proteins, biomolecules, polymer-drug conjugates, drug-containing liposomes and polymersomes, aptamers, oligopeptides, oligonucleotides, oligosaccharides, carbohydrates, as well as peptides, peptoids, steroids, toxins, hormones, cytokines, and chemokines. Other drugs that can be used are low- to medium-molecular-weight compounds, preferably organic compounds (e.g., about 200 to about 2500 Da, preferably about 300 to about 1750 Da, more preferably about 300 to about 1000 Da).

[0514] In preferred embodiments, the pharmaceutically active compound or agent is selected from the group consisting of cytotoxins, antiproliferative / antitumor agents, antivirals, antibiotics, anti-inflammatory agents, chemosensitizers, radiosensitizers, immunomodulators, immunosuppressants, immunostimulators, anti-angiogenic factors, and enzyme inhibitors.

[0515] In other preferred embodiments, the agent is designed to act in the central nervous system, for example in the context of Alzheimer's and Parkinson's disease, for example antibodies against beta amyloid and Tau proteins.

[0516] Exemplary classes of cytotoxic agents, e.g., for use in cancer therapy, include, but are not limited to, DNA damaging agents, DNA crosslinking agents, DNA binding agents, DNA alkylating agents, DNA intercalators, DNA cleaving agents, microtubule stabilizing and destabilizing agents, topoisomerase inhibitors, radiosensitizers, antimetabolites, natural products and their analogs, peptides, oligonucleotides, enzyme inhibitors, e.g., dihydrofolate reductase inhibitors and thymidylate synthase inhibitors.

[0517] Exemplary immunomodulatory agents include PD-L1, PD-1, LAG-3, OX40, TIGIT, TIM-3, B7H4, Vista, CTLA-4, APRIL, cytokines including IL-2, IL-7, IL-10, IL-12, IL-15, IL-21, TNF, interferon gamma, GMCSF, NDV-GMCSF, and STING agonists and antagonists. agonists and antagonists of TLRs, including TLR1 / 2, TLR3, TLR4, TLR7 / 8, TLR9, and TLR12; agonists and antagonists of GITR, CD3, CD28, CD40, CD74, CTLA4, OX40, PD1, PDL1, RIG, MDA-5, NLRP1, NLRP3, AIM2, IDO, MEK, cGAS, and CD25, and NKG2A.

[0518] It will be appreciated that chemical modifications may also be made to the agent to make it more convenient to react that compound for the purpose of preparing the conjugates of the invention.

[0519] In a preferred embodiment, the administered agent prior to conjugation to the remainder of the compound of Formula (19) is selected from the group consisting of -OH, -NHR', -COH, -SH, -SS-, -SCH-, -N, terminal alkynyl, terminal alkenyl, -C(O)R', C-C 12 and R' comprises at least one moiety selected from the group consisting of (hetero)cycloalkynyl, nitrone, nitrile oxide, (imino)sydnone, isonitrile, (oxa)norbornene, and tetrazine, wherein R' is R 37 where the moiety used for conjugation to the moiety satisfies formula (19) and is equal to C M2 or C X To form a compound containing the moiety, a dienophile, the label, and R 32 Includes:

[0520] In a preferred embodiment, the administration agent is a C selected from the group consisting of amines, amides, thioamides, aminooxys, carbamates, thiocarbamates, ureas, thioureas, sulfonamides, and sulfonecarbamates. M2 In a preferred embodiment, C M2 is R as defined herein 10 Preferably, the dosage form is equal to C M2 and C X are bonded as described above for

[0521] Preferably, the therapeutic agent, preferably an antibody, is modified with additional moieties equivalent to formula (19), except that these additional moieties do not include therapeutic agents (because the first-mentioned therapeutic agent is already attached to said moiety). In a preferred embodiment, the therapeutic agent, preferably an antibody, is attached to additional moieties at positions 1-8, more preferably positions 1-6, and even more preferably positions 1-4, as defined in this paragraph.

[0522] Further embodiments of compounds of formula (19) In a preferred embodiment, in the compounds of the present invention, the administered agent comprises an antibody, preferably, the administered agent is an antibody.

[0523] In a preferred embodiment, in the compounds of the invention, the label is a radiolabel, preferably a chelating moiety that chelates a radioisotope.

[0524] Jien The combination according to the present invention comprises a compound according to formula (19) and a diene, preferably a tetrazine. In a preferred embodiment, the combination is in the form of a kit.

[0525] The tetrazine compounds of the present invention may be referred to herein as "activators." Tetrazines typically react with other bioorthogonal reactive groups, i.e., dienophiles (see above). The diene of the activator is selected so that it can react with the dienophile of the TCO via a Diels-Alder cycloaddition followed by a retro-Diels-Alder reaction to give an IEDDA adduct. This intermediate adduct then releases Construct A, where this release can be triggered by various circumstances or conditions related to the specific molecular structure of the IEDDA adduct.

[0526] From the structure of formula (19), one or more moieties R 48 The compound used to release the compound is referred to herein as the active agent.

[0527] In a preferred embodiment, the activator is a tetrazine. Tetrazine is a diene and is highly reactive with dienophiles, particularly TCO constructs (see above). The diene of the activator is selected so that it can react with a dienophile, such as TCO, for example, via Diels-Alder cycloaddition followed by a retro-Diels-Alder reaction to give an IEDDA adduct. This intermediate adduct then releases construct-A.

[0528] In general, synthetic routes to tetrazines are readily available to those skilled in the art based on standard knowledge in the art. For example, Lions et al,J.Org.Chem.,1965,30,318-319;Horwitz et al,J.Am.Chem.Soc.,1958,80,3155-3159;Hapiot et al,New J.Chem.,2004,28,387-392,Kaim et al. al,Z.Naturforsch.,1995,50b,123-127;Yang et al.,Angew.Chem.2012,124,5312-5315;Mao et al.,Angew.Chem.Int.Ed.2019,58,1106-1109;Qu et al. al.Angew.Chem.Int.Ed.2018,57,12057-12061;Selvaraj et al.,Tetrahedron Lett. 2014, 55, 4795-4797; Fan et al., Angew. Chem. Int. Ed. 2016, 55, 14046-14050 contain references to tetrazine synthesis routes.

[0529] Preferably, the active agent is a tetrazine, preferably a pharmaceutically acceptable salt thereof, satisfying formula (4):

[0530] [ka]

[0531] wherein each moiety Q1 and Q2 is independently a hydrogen atom, -F, -Cl, -Br, -I, -OR 37 , -N(R 37 )2, -SO3, -PO3 - 、 -NO2, -CF3, -SR 37 , S(=O)2N(R 37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R 37 , SC(=S)R 37 , N.R. 37 C(=O)-R 37 , N.R. 37C(=S)-R 37 , N.R. 37 C(=O)OR 37 , N.R. 37 C(=S)OR 37 , N.R. 37 C(=O)SR 37 , N.R. 37 C(=S)SR 37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R 37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR 37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37 )2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , S(O)R 37 , -S(O)2R 37 , N.R. 37 S(O)2R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , alkyl groups, alkenyl groups, alkynyl groups, aryl groups, heteroaryl groups, cycloalkyl groups, cycloalkenyl groups, cycloalkynyl groups, (cyclo)alkyl(hetero)aryl groups, (hetero)aryl(cyclo)alkyl, (cyclo)alkenyl(hetero)aryl groups, (hetero)aryl(cyclo)alkenyl groups, (cyclo)alkynyl(hetero)aryl groups, (hetero)aryl(cyclo)alkynyl groups, alkylcycloalkyl groups, and cycloalkylalkyl groups.

[0532] In formula (4), H, -F, -Cl, -Br, -I, -OH, -NH2, -SO3, -PO3 - 、The Q1 and Q2 groups that are not -NO2 or -CF3 are preferably -Cl, -F, -Br, -I, -OR 37 , -N(R 37 )2, -SO3R 37 , -PO3(R 37 ) 2、 -PO4(R 37 ) 2、 -NO2, -CF3, =O, =NR 37 , and -SR 37 and optionally substituted with a moiety selected from the group consisting of O, S, NR 37 , P, and Si, where the N, S, and P atoms are optionally oxidized, and where multiple N atoms are optionally quaternized. Preferably, each Q1 and Q2 contains no more than 4 substituents, more preferably no more than 3 substituents, even more preferably no more than 2 substituents, and most preferably no more than 1 substituent.

[0533] In formula (4), the Q1 and Q2 groups are selected from the group consisting of polymers, particles, peptides, peptoids, dendrimers, proteins, aptamers, carbohydrates, oligonucleotides, oligosaccharides, lipids, steroids, liposomes, targeting agents, and the like. T , R 87 , albumin-binding moiety, and chelating moiety, radionuclide-containing moiety, or drug D D may be bonded to

[0534] In formula (4), preferably, at least one of the moieties Q1 and Q2 is not a hydrogen atom.

[0535] In formula (4), preferably, each moiety Q1 and Q2 independently represents a hydrogen atom, -F, -Cl, -Br, -I, -OR 37 , -N(R 37 )2, -SO3, -PO3 - 、 -NO2, -CF3, -SR 37 , S(=O)2N(R 37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R37 , SC(=S)R 37 , N.R. 37 C(=O)-R 37 , N.R. 37 C(=S)-R 37 , N.R. 37 C(=O)OR 37 , N.R. 37 C(=S)OR 37 , N.R. 37 C(=O)SR 37 , N.R. 37 C(=S)SR 37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R 37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR 37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37 )2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , S(O)R 37 , -S(O)2R 37 , N.R. 37 S(O)2R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , C1~C 24 Alkyl groups, C2-C 24 Alkenyl groups, C2-C 24 Alkynyl groups, C6-C 24 Aryl groups, C2-C 24 Heteroaryl groups, C3-C 24 Cycloalkyl groups, C5-C 24 Cycloalkenyl group, C 12 ~C 24 Cycloalkynyl group, C3-C 24 (Cyclo)alkyl(hetero)aryl groups, C3-C24 (Hetero)aryl(cyclo)alkyl, C4-C 24 (Cyclo)alkenyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkenyl groups, C4-C 24 (Cyclo)alkynyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkynyl group, C4-C 24 Alkylcycloalkyl groups and C4-C 24 cycloalkylalkyl groups.

[0536] In a preferred embodiment, the Q1 and Q2 groups that are not hydrogen atoms are unsubstituted.

[0537] In a preferred embodiment, Q1 and Q2 in formula (4) are each independently a hydrogen atom, -F, -Cl, -Br, -I, -OR 37 , -N(R 37 )2, -SO3, -PO3 - 、 -NO2, -CF3, -SR 37 , S(=O)2N(R 37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R 37 , SC(=S)R 37 , N.R. 37 C(=O)-R 37 , N.R. 37 C(=S)-R 37 , N.R. 37 C(=O)OR 37 , N.R. 37 C(=S)OR 37 , N.R. 37 C(=O)SR 37 , N.R. 37 C(=S)SR 37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R 37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37 )2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , S(O)R 37 , -S(O)2R 37 , N.R. 37 S(O)2R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , C1-C8 alkyl group, C2-C8 alkenyl group, C2-C8 alkynyl group, C6-C 12 Aryl groups, C2-C 12 Heteroaryl group, C3-C8 cycloalkyl group, C5-C8 cycloalkenyl group, C3-C 12 Alkyl(hetero)aryl groups, C3-C 12 (Hetero)arylalkyl groups, C4-C 12 Alkylcycloalkyl groups, C4-C 12 Cycloalkylalkyl groups, C5-C 12 Cycloalkyl(hetero)aryl groups and C5-C 12 (hetero)arylcycloalkyl groups.

[0538] In a preferred embodiment, Q1 and Q2 in formula (4) are each independently a hydrogen atom, -F, -Cl, -Br, -I, -OR 37 , -N(R 37 )2, -SO3, -PO3 - 、 -NO2, -CF3, -SR 37 , S(=O)2N(R 37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R 37 , SC(=S)R 37 , N.R. 37 C(=O)-R 37 , N.R.37 C(=S)-R 37 , N.R. 37 C(=O)OR 37 , N.R. 37 C(=S)OR 37 , N.R. 37 C(=O)SR 37 , N.R. 37 C(=S)SR 37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R 37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR 37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37 )2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , S(O)R 37 , -S(O)2R 37 , N.R. 37 S(O)2R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , C1-C4 alkyl group, C2-C4 alkenyl group, C2-C4 alkynyl group, C6-C8 aryl group, C2-C8 heteroaryl group, C3-C6 cycloalkyl group, C5-C6 cycloalkenyl group, C3-C 10 Alkyl(hetero)aryl groups, C3-C 10 (Hetero)arylalkyl groups, C4-C 10 Alkylcycloalkyl groups, C4-C 10 Cycloalkylalkyl groups, C5-C 10 Cycloalkyl(hetero)aryl groups and C5-C 10 (hetero)arylcycloalkyl groups.

[0539] In a preferred embodiment, Q1 and Q2 in formula (4) are selected from the group consisting of a hydrogen atom, a C1-C8 alkyl, a phenyl, a 2-pyridyl, a 3-pyridyl, a 4-pyridyl, a 2,6-pyrimidyl, a 2,5-pyrimidyl, a 3,5-pyrimidyl, and a 2,4-pyrimidyl; and Q1 and Q2 that are not hydrogen atoms are selected from the group consisting of -F, -Cl, -Br, -I, -OR 37 , -N(R 37 )2, -SO3, -PO3 - 、 -NO2, -CF3, -SR 37 , S(=O)2N(R 37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R 37 , SC(=S)R 37 , N.R. 37 C(=O)-R 37 , N.R. 37 C(=S)-R 37 , N.R. 37 C(=O)OR 37 , N.R. 37 C(=S)OR 37 , N.R. 37 C(=O)SR 37 , N.R. 37 C(=S)SR 37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R 37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR 37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37 )2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , S(O)R 37 , -S(O)2R 37 , N.R. 37S(O)2R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , C1~C 24 Alkyl groups, C2-C 24 Alkenyl groups, C2-C 24 Alkynyl groups, C6-C 24 Aryl groups, C2-C 24 Heteroaryl groups, C3-C 24 Cycloalkyl groups, C5-C 24 Cycloalkenyl group, C 12 ~C 24 Cycloalkynyl group, C3-C 24 (Cyclo)alkyl(hetero)aryl groups, C3-C 24 (Hetero)aryl(cyclo)alkyl, C4-C 24 (Cyclo)alkenyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkenyl groups, C4-C 24 (Cyclo)alkynyl(hetero)aryl group, C4-C 24 (Hetero)aryl(cyclo)alkynyl group, C4-C 24 Alkylcycloalkyl groups and C4-C 24 A moiety selected from the group consisting of cycloalkylalkyl groups, preferably -F, -Cl, -Br, -I, -OR 37 , -N(R 37 )2, -SO3, -PO3 - 、 -NO2, -CF3, -SR 37 , S(=O)2N(R 37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R 37 , SC(=S)R 37 , N.R. 37 C(=O)-R 37 , N.R. 37 C(=S)-R 37 , N.R. 37 C(=O)OR 37 , N.R. 37 C(=S)OR 37 , N.R. 37 C(=O)SR37 , N.R. 37 C(=S)SR 37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R 37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR 37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37 )2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , S(O)R 37 , -S(O)2R 37 , N.R. 37 S(O)2R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , C1-C8 alkyl group, C2-C8 alkenyl group, C2-C8 alkynyl group, C6-C 12 Aryl groups, C2-C 12 Heteroaryl group, C3-C8 cycloalkyl group, C5-C8 cycloalkenyl group, C3-C 12 Alkyl(hetero)aryl groups, C3-C 12 (Hetero)arylalkyl groups, C4-C 12 Alkylcycloalkyl groups, C4-C 12 Cycloalkylalkyl groups, C5-C 12 Cycloalkyl(hetero)aryl groups and C5-C 12 A moiety selected from the group consisting of (hetero)arylcycloalkyl groups, more preferably -F, -Cl, -Br, -I, -OR 37 , -N(R 37 )2, -SO3, -PO3 - 、 -NO2, -CF3, -SR 37 , S(=O)2N(R37 )2, OC(=O)R 37 , SC(=O)R 37 , OC(=S)R 37 , SC(=S)R 37 , N.R. 37 C(=O)-R 37 , N.R. 37 C(=S)-R 37 , N.R. 37 C(=O)OR 37 , N.R. 37 C(=S)OR 37 , N.R. 37 C(=O)SR 37 , N.R. 37 C(=S)SR 37 , OC(=O)N(R 37 )2, SC(=O)N(R 37 )2, OC(=S)N(R 37 )2, SC(=S)N(R 37 )2, NR 37 C(=O)N(R 37 )2, NR 37 C(=S)N(R 37 )2, C(=O)R 37、 C(=S)R 37、 C(=O)N(R 37 )2, C(=S)N(R 37 )2, C(=O)OR 37 , C(=O)SR 37 , C(=S)OR 37 , C(=S)SR 37 , S(O)R 37 , -S(O)2R 37 , N.R. 37 S(O)2R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , C1-C4 alkyl group, C2-C4 alkenyl group, C2-C4 alkynyl group, C6-C8 aryl group, C2-C8 heteroaryl group, C3-C6 cycloalkyl group, C5-C6 cycloalkenyl group, C3-C 10 Alkyl(hetero)aryl groups, C3-C 10 (Hetero)arylalkyl groups, C4-C 10 Alkylcycloalkyl groups, C4-C10 Cycloalkylalkyl groups, C5-C 10 Cycloalkyl(hetero)aryl groups and C5-C 10 It may be optionally substituted with a moiety selected from the group consisting of (hetero)arylcycloalkyl groups.

[0540] In a preferred embodiment, in formula (4), (a) Q1 and Q2 are independently selected from the group consisting of 2-pyridyl, 3-pyridyl, and 4-pyridyl; (b) Q1 is selected from the group consisting of 2,6-pyrimidyl, 2,5-pyrimidyl, 3,5-pyrimidyl, and 2,4-pyrimidyl; and Q2 is (hetero)alkyl; or (c) Q1 is phenyl and Q2 is a hydrogen atom; (d) Q1 is phenyl and Q2 is phenyl; (e) Q1 is phenyl and Q2 is C1-C8 alkyl; (f) Q1 and Q2 are C1-C8 alkyl; and In (a) to (f), all Q1 and Q2 that are not hydrogen atoms may be substituted as defined in the previous paragraph.

[0541] In a preferred embodiment, the active agent can be a multimeric compound containing multiple dienes, as defined herein, including, but not limited to, biomolecules, proteins, peptides, peptoids, polymers, dendrimers, liposomes, micelles, particles, polymeric particles, or other polymeric constructs.

[0542] Preferred tetrazines Formula (4a) Preferred tetrazines are those according to formula (4) below, preferably pharmaceutically acceptable salts thereof:

[0543] [ka]

[0544] wherein each moiety Q1 and Q2 is independently selected from the group consisting of a hydrogen atom and a moiety according to formula (5):

[0545] [ka]

[0546] where the dashed line indicates the bond to the rest of the molecule, where R 10 , R 11 , and R 12 is as defined herein.

[0547] In a preferred embodiment, each f in formula (5) is an integer independently selected from the range of 0 to 24, preferably from the range of 1 to 12, more preferably from the range of 2 to 6, and even more preferably from 1 to 3. In a preferred embodiment, f is 1. In another preferred embodiment, f is an integer in the range of 12 to 24. In a preferred embodiment, in formula (5), g is an integer in the range of 0 to 12, preferably from 1 to 6, and more preferably from 2 to 4. In a preferred embodiment, in formula (5), each h is independently 0 or 1. In a preferred embodiment, g is 0 and f is 1. In a preferred embodiment, g is 1 and f is 1.

[0548] When a compound according to the invention contains multiple moieties satisfying formula (5), each g, h, and f is independently selected.

[0549] In a preferred embodiment, the moiety according to formula (5) may be substituted with other moieties independently selected from the moieties according to formula (5). In a preferred embodiment, the moiety according to formula (5) may be unsubstituted with other moieties independently selected from the moieties according to formula (5).

[0550] In a preferred embodiment, the moiety according to formula (5) is R 87 is.

[0551] In a preferred embodiment, the moiety according to formula (5) is R M Fill the molecule from the group.

[0552] It is preferred that at least one of the moieties Q1 and Q2 in formula (4a) is not a hydrogen atom.

[0553] In a preferred embodiment, Q1 in formula (4a) is a C6-C 24 Aryl and C2-C 24 heteroaryl and optionally further substituted with a moiety according to formula (5), preferably with no more than two, more preferably no more than one moiety according to formula (5).

[0554] In a preferred embodiment, Q1 in formula (4a) is a C6-C 24 Aryl and C2-C 24 heteroaryl and optionally further substituted with a moiety according to formula (5), preferably with up to two, more preferably up to one moiety according to formula (5), and Q2 in formula (4a) is selected from the group consisting of C6-C 24 Aryl and C2-C 24 heteroaryl and optionally further substituted with a moiety according to formula (5), preferably with no more than two, more preferably no more than one moiety according to formula (5).

[0555] In a preferred embodiment, Q1 in formula (4a) is selected from the group consisting of C6 aryl and C3-C5 heteroaryl, and is optionally further substituted with at least one moiety according to formula (5), preferably with up to two, more preferably up to one moiety according to formula (5). Preferred heteroaryls herein are 2-pyridyl, 3-pyridyl, 4-pyridyl, 2,6-pyrimidyl, 3,5-pyrimidyl, 2,5-pyrimidyl, 2,4-pyrimidyl, 2,4-imidazyl, 2,5-imidazyl, phenyl, 2,3-pyrazyl, 3,4-pyrazyl, oxazole, isoxazole, thiazole, oxazoline, 2-pyrryl, 3-pyrryl, 2-thiophene, and 3-thiophene.

[0556] In a preferred embodiment, Q1 in formula (4a) is a C3-C5 heteroaryl and is optionally further substituted with at least one moiety according to formula (5), preferably with up to two, more preferably up to one moiety according to formula (5), and Q2 is a C3-C5 heteroaryl and is optionally further substituted with a moiety according to formula (5), preferably with up to two, more preferably up to one moiety according to formula (5). Preferred heteroaryls herein are 2-pyridyl, 3-pyridyl, 4-pyridyl, 2,6-pyrimidyl, 3,5-pyrimidyl, 2,5-pyrimidyl, 2,4-pyrimidyl, 2,4-imidazyl, 2,5-imidazyl, phenyl, 2,3-pyrazyl, 3,4-pyrazyl, pyrazyl, isoxazole, thiazole, oxazoline, 2-pyrryl, 3-pyrryl, 2-thiophene, and 3-thiophene.

[0557] In a preferred embodiment, Q1 in formula (4a) is C3-C5 heteroaryl and is optionally further substituted with at least one moiety according to formula (5), preferably no more than two, more preferably no more than one moiety according to formula (5), and Q2 is -H.

[0558] In a preferred embodiment, Q1 in formula (4a) is a phenyl ring and is optionally further substituted with at least one moiety according to formula (5), preferably no more than two, more preferably no more than one moiety according to formula (5), and Q2 is -H.

[0559] In a preferred embodiment, Q1 in formula (4a) is a phenyl ring and is optionally further substituted with at least one moiety according to formula (5), preferably with no more than two, more preferably no more than one moiety according to formula (5), and Q2 is a phenyl ring and is optionally further substituted with at least one moiety according to formula (5), preferably with no more than two, more preferably no more than one moiety according to formula (5).

[0560] In a preferred embodiment, Q1 in formula (4a) is a phenyl ring and is optionally further substituted with at least one moiety according to formula (5), preferably no more than two, more preferably no more than one moiety according to formula (5), and Q2 is selected from the group consisting of C6 aryl and C 3~5 heteroaryl, and optionally further substituted with at least one moiety according to formula (5), preferably no more than two, more preferably no more than one moiety according to formula (5).

[0561] In a preferred embodiment, Q1 in formula (4a) is C1 to C 12 alkyl and optionally further substituted with at least one moiety according to formula (5), preferably no more than two, more preferably no more than one moiety according to formula (5), and Q2 is C aryl and C 3~5 heteroaryl, and optionally further substituted with at least one moiety according to formula (5), preferably no more than two, more preferably no more than one moiety according to formula (5).

[0562] In a preferred embodiment, Q1 in formula (4a) is C1 to C 12alkyl and optionally further substituted with at least one moiety according to formula (5), preferably no more than two, more preferably no more than one moiety according to formula (5), and Q2 in formula (4a) is C1-C 12 It is alkyl and may be further substituted with at least one moiety according to formula (5), preferably with no more than two, more preferably no more than one moiety according to formula (5).

[0563] In a preferred embodiment, Q2 is equal to Q1.

[0564] R 10 In a preferred embodiment, each R 10are independently -O-, -S-, -SS-, -NR4-, -N=N-, -C(O)-, -C(O)NR4-, -OC(O)-, -C(O)O-, -OC(O)O-, -OC(O)NR4-, -NR4C(O)-, -NR4C(O)O-, -NR4C(O)NR4-, -SC(O)-, -C(O)S-, -SC(O)O-, -OC(O)S-, -SC(O)NR4-, -NR4C(O)S-, -S(O)-, -S(O)2-, -OS(O)2-, - S(O2)O-, -OS(O)2O-, -OS(O)2NR4-, -NR4S(O)2O-, -C(O)NR4S(O)2NR4-, -OC(O)NR4S(O)2NR4-, -OS(O)-, -OS(O)O-, -OS (O)NR4-, -ONR4C(O)-, -ONR4C(O)O-, -ONR4C(O)NR4-, -NR4OC(O)-, -NR4OC(O)O-, -NR4OC(O)NR4-, -ONR4C(S)-, -ONR4C( S)O-, -ONR4C(S)NR4-, -NR4OC(S)-, -NR4OC(S)O-, -NR4OC(S)NR4-, -OC(S)-, -C(S)O-, -OC(S)O-, -OC(S)NR4-, -NR4C(S )-, -NR4C(S)O-, -SS(O)2-, -S(O)2S-, -OS(O2)S-, -SS(O)2O-, -NR4OS(O)-, -NR4OS(O)O-, -NR4OS(O)NR4-, -NR4OS(O)2- , -NR4OS(O)2O-, -NR4OS(O)2NR4-, -ONR4S(O)-, -ONR4S(O)O-, -ONR4S(O)NR4-, -ONR4S(O)2O-, -ONR4S(O)2NR4-, -ONR4S(O)2-, -OP(O)(R4)2-, -SP(O)(R4)2-, -NR4P(O)(R4)2-, and combinations thereof, where R4 is defined as described herein. In a preferred embodiment, each R 10are independently -O-, -S-, -SS-, -NR4-, -N=N-, -C(O)-, -C(O)NR4-, -OC(O)-, -C(O)O-, -OC(O)NR4-, -NR4C(O)-, -NR4C(O)O-, -NR4C(O)NR4-, -SC(O)-, -C(O)S-, -SC(O)O-, -OC(O)S-, -SC(O )NR—, —NRC(O)S—, —S(O)—, —S(O)—, —C(O)NRS(O)NR—, —OC(O)NRS(O)NR—, —OC(S)—, —C(S)O—, —OC(S)NR—, —NRC(S)—, —NRC(S)O—, and —SS(O)—.

[0565] Preferably, R 10 and wherein each R4 is independently selected from the group consisting of a hydrogen atom and a C1-C4 alkyl.

[0566] R 11 In a preferred embodiment, each R 11 are independent, C1~C 24 Alkylene group, C2-C 24 Alkenylene group, C2-C 24 Alkynylene group, C6-C 24 Arylene, C2-C 24 Heteroarylene, C3-C 24 Cycloalkylene group, C5-C 24 Cycloalkenylene group, and C 12 ~C 24 and wherein the alkylene group, alkenylene group, alkynylene group, cycloalkylene group, cycloalkenylene group, and cycloalkynylene group are preferably selected from the group consisting of O, S, NR 36 , P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0567] In a preferred embodiment, each R 11 are independent, C1~C12 Alkylene group, C2-C 12 Alkenylene group, C2-C 12 Alkynylene group, C6-C 12 Arylene, C2-C 12 Heteroarylene, C3-C 12 Cycloalkylene group, C5-C 12 Cycloalkenylene group, and C 12 and wherein preferably the alkylene group, alkenylene group, alkynylene group, cycloalkylene group, cycloalkenylene group, and cycloalkynylene group are selected from the group consisting of O, S, NR 36 , P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0568] In a preferred embodiment, each R 11 are independently selected from the group consisting of a C1-C6 alkylene group, a C2-C6 alkenylene group, a C2-C6 alkynylene group, a C6-C6 arylene, a C2-C6 heteroarylene, a C3-C6 cycloalkylene group, and a C5-C6 cycloalkenylene group; and wherein the above alkylene group, alkenylene group, alkynylene group, cycloalkylene group, cycloalkenylene group, and cycloalkynylene group are preferably selected from the group consisting of O, S, NR 36 , P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0569] In a preferred embodiment, R 11 The groups further include -Cl, -F, -Br, -I, -OH, -NH2, -SO3H, and -PO3H 、 -PO4H2, -NO2, -CF3, =O, =NR 36 , -SR 36 , C1~C 24 Alkyl groups, C2-C 24 Alkenyl groups, C2-C 24Alkynyl groups, C6-C 24 Aryl groups, C2-C 24 Heteroaryl groups, C3-C 24 Cycloalkyl groups, C5-C 24 Cycloalkenyl group, C 12 ~C 24 Cycloalkynyl group, C3-C 24 Alkyl(hetero)aryl groups, C3-C 24 (Hetero)arylalkyl groups, C4-C 24 (Hetero)arylalkenyl groups, C4-C 24 (Hetero)arylalkynyl group, C4-C 24 Alkenyl(hetero)aryl groups, C4-C 24 Alkynyl(hetero)aryl groups, C4-C 24 Alkylcycloalkyl groups, C6-C 24 Alkylcycloalkenyl group, C 13 ~C 24 Alkylcycloalkynyl group, C4-C 24 Cycloalkylalkyl groups, C6-C 24 Cycloalkenylalkyl groups, C 13 ~C 24 Cycloalkynylalkyl groups, C5-C 24 Alkenylcycloalkyl groups, C7-C 24 Alkenylcycloalkenyl group, C 14 ~C 24 Alkenylcycloalkynyl group, C5-C 24 Cycloalkylalkenyl group, C7-C 24 Cycloalkenyl alkenyl group, C 14 ~C 24 Cycloalkynylalkenyl group, C5-C 24 Alkynylcycloalkyl groups, C7-C 24 Alkynylcycloalkenyl group, C 14 ~C 24 Alkynylcycloalkynyl group, C5-C 24 Cycloalkylalkynyl group, C7-C 24 Cycloalkenylalkynyl group, C 14 ~C 24 Cycloalkynylalkynyl group, C5-C 24Cycloalkyl(hetero)aryl groups, C7-C 24 Cycloalkenyl(hetero)aryl group, C 14 ~C 24 Cycloalkynyl(hetero)aryl groups, C5-C 24 (Hetero)arylcycloalkyl groups, C7-C 24 (Hetero)arylcycloalkenyl groups, and C 14 ~C 24 and (hetero)arylcycloalkynyl groups, wherein the substituents are O, S, NR 36 , P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0570] In a preferred embodiment, R 11 The groups further include -Cl, -F, -Br, -I, -OH, -NH2, -SO3H, and -PO3H 、 -PO4H2, -NO2, -CF3, =O, =NR 36 , -SR 36 , C1~C 12 Alkyl groups, C2-C 12 Alkenyl groups, C2-C 12 Alkynyl groups, C6-C 12 Aryl groups, C2-C 12 Heteroaryl groups, C3-C 12 Cycloalkyl groups, C5-C 12 Cycloalkenyl group, C 12 Cycloalkynyl group, C3-C 12 Alkyl(hetero)aryl groups, C3-C 12 (Hetero)arylalkyl groups, C4-C 12 (Hetero)arylalkenyl groups, C4-C 12 (Hetero)arylalkynyl group, C4-C 12 Alkenyl(hetero)aryl groups, C4-C 12 Alkynyl(hetero)aryl groups, C4-C 12 Alkylcycloalkyl groups, C6-C 12 Alkylcycloalkenyl group, C13 ~C 18 Alkylcycloalkynyl group, C4-C 12 Cycloalkylalkyl groups, C6-C 12 Cycloalkenylalkyl groups, C 13 ~C 18 Cycloalkynylalkyl groups, C5-C 12 Alkenylcycloalkyl groups, C7-C 12 Alkenylcycloalkenyl group, C 14 ~C 16 Alkenylcycloalkynyl group, C5-C 12 Cycloalkylalkenyl group, C7-C 12 Cycloalkenyl alkenyl group, C 14 ~C 16 Cycloalkynylalkenyl group, C5-C 12 Alkynylcycloalkyl groups, C7-C 12 Alkynylcycloalkenyl group, C 14 ~C 16 Alkynylcycloalkynyl group, C5-C 12 Cycloalkylalkynyl group, C7-C 12 Cycloalkenylalkynyl group, C 14 ~C 16 Cycloalkynylalkynyl group, C5-C 12 Cycloalkyl(hetero)aryl groups, C7-C 12 Cycloalkenyl(hetero)aryl group, C 14 ~C 16 Cycloalkynyl(hetero)aryl groups, C5-C 12 (Hetero)arylcycloalkyl groups, C7-C 12 (Hetero)arylcycloalkenyl groups, and C 14 ~C 16 and (hetero)arylcycloalkynyl groups, wherein the substituents are O, S, NR 36 , P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

[0571] In a preferred embodiment, R 11 The groups further include -Cl, -F, -Br, -I, -OH, -NH2, -SO3H, and -PO3H 、 -PO4H2, -NO2, -CF3, =O, =NR 36 , -SR 36 , C1-C6 alkyl group, C2-C6 alken...

Claims

1. A compound satisfying the following formula (19) or a pharmaceutically acceptable salt thereof: 【Chemistry 01】 where: The compound is 【Chemical 02】 is selected from the group consisting of wherein X 1 , X 2 , X 3 , X 4 , and X 5 are CH 2 ; Y T1 is selected from the group consisting of OH, SH, and NH 2 ; where the dashed line indicates the bond to the R 38 group; R 48 is -OC(O)-(S P ) k C A , -OC(S)-(S P ) k C A , -SC(O)-(S P ) k C A , -SC(S)-(S P ) k C A , -O-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A , and -S-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A , and -(S P ) k C A selected from the group consisting of: r is 0 or 1; each s is independently 0 or 1; each i is independently an integer ranging from 0 to 4; j is an integer ranging from 0 to 4; each k is independently 0 or 1; L C is a self-immolative linker, and S P is a spacer; Each C A and C B are independently selected from the group consisting of organic molecules and inorganic molecules; wherein an organic molecule is a molecule that contains a C-H bond; and wherein an inorganic molecule is a molecule that does not contain a C-H bond; where R 48 -OC(O)-(S P ) k C A , -OC(S)-(S P ) k C A , -SC(O)-(S P ) k C A , or -SC(S)-(S P ) k C A If S P (if k>0) or C A (when k=0) is connected to R via an atom selected from the group consisting of O, C, S, and N. 48 is bonded to -OC(O)-, -OC(S)-, -SC(O)- or -SC(S)-, where this atom is S P or C A is part of where R 48 -O-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A , or -S-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A and r is 0, then S P (if k>0) or C A (when k=0) is R at the allylic position of the trans-cyclooctene ring of formula (19) via a group selected from the group consisting of —C(O)— and —C(S)—. 48 is bonded to the -O- or -S- moiety of P or C A is part of where R 48 -O-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A , or -S-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A and r is 1, L C is -C(Y C2 )Y C1 - and a carbon atom, wherein the group is linked to the -O- or -S- moiety in the allylic position of the trans-cyclooctene ring of formula (19) through a group selected from the group consisting of L C is part of where Y C1 -O-, -S-, and -NR 36 - selected from the group consisting of where Y C2 is selected from the group consisting of O and S; where R 48 -O-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A , or -S-(L C ((S P ) k C A ) s ((S P ) k C A ) s ((S P ) i -C B ) j ) r -(S P ) k C A and r is 1, then S P (if k>0) or C A (when k=0) is connected to L through a moiety selected from the group consisting of —O—, —S—, and —N—. C where this moiety is S P or C A is part of where R 48 -(S P ) k C A If S P (if k>0) or C A (when k=0) is attached to the allylic position of the trans-cyclooctene of formula (19) through an -O- or -S- atom, where this atom is S P or C A is part of Here, each of R 38 , R 37 and R 36 are independently hydrogen atoms, -(S P ) i -C B (i is independently an integer ranging from 0 to 4), C 1 ~C 24 Alkyl group, C 2 ~C 24 Alkenyl group, C 2 ~C 24 Alkynyl group, C 6 ~C 24 Aryl group, C 2 ~C 24 Heteroaryl groups, C 3 ~C 24 Cycloalkyl groups, C 5 ~C 24 Cycloalkenyl group, C 12 ~C 24 Cycloalkynyl group, C 3 ~C 24 (Cyclo)alkyl(hetero)aryl groups, C 3 ~C 24 (Hetero)aryl(cyclo)alkyl, C 4 ~C 24 (Cyclo)alkenyl(hetero)aryl group, C 4 ~C 24 (Hetero)aryl(cyclo)alkenyl group, C 4 ~C 24 (Cyclo)alkynyl(hetero)aryl group, C 4 ~C 24 (Hetero)aryl(cyclo)alkynyl group, C 4 ~C 24 Alkylcycloalkyl groups, and C 4 ~C 24 cycloalkylalkyl groups; where SP and CB are as defined above; where R 37 and R 36 is not a hydrogen atom, but -Cl, -F, -Br, -I, -OH, -NH 2 , -SO 3 H, -PO 3 H 、 -PO 4 H 2、 -NO 2 , -CF 3 , ═O, ═NH, and —SH, and optionally containing one or more heteroatoms selected from the group consisting of O, S, NH, P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized; provided that when the nitrogen atom is substituted by an R 38 group, the R 38 group is not C(O), C(S), S(O), or S(O) 2 is not attached to the nitrogen via Here, each R 47 are independently hydrogen atoms, -(S P ) i -C B , -F, -Cl, -Br, -I, -OR 37 , -N(R 37 ) 2 , -SO 3 , -PO 3 - 、 -NO 2 , -CF 3 , -SR 37 , -S(=O) 2 N(R 37 ) 2 , - OC(=O)R 37 , -SC(=O)R 37 , -OC(=S)R 37 , -SC(=S)R 37 , -NR 37 C(=O)-R 37 , -NR 37 C(=S)-R 37 , -NR 37 C(=O)OR 37 , -NR 37 C(=S)OR 37 , -NR 37 C(=O)SR 37 , -NR 37 C(=S)SR 37 , -OC(=O)N(R 37 ) 2 , -SC(=O)N(R 37 ) 2 , -OC(=S)N(R 37 ) 2 , -SC(=S)N(R 37 ) 2 , -NR 37 C(=O)N(R 37 ) 2 , -NR 37 C(=S)N(R 37 ) 2 , -C(=O)R 37、 -C(=S)R 37、 -C(=O)N(R 37 ) 2 , -C(=S)N(R 37 ) 2 , -C(=O)OR 37 , -C(=O)SR 37 , -C(=S)OR 37 , -C(=S)SR 37 , -S(O)R 37 , -S(O) 2 R 37 , -NR 37 S(O) 2 R 37 , -ON(R 37 ) 2、 -NR 37 OR 37 , C 1 ~C 24 Alkyl group, C 2 ~C 24 Alkenyl group, C 2 ~C 24 Alkynyl group, C 6 ~C 24 Aryl group, C 2 ~C 24 Heteroaryl groups, C 3 ~C 24 Cycloalkyl groups, C 5 ~C 24 Cycloalkenyl group, C 12 ~C 24 Cycloalkynyl group, C 3 ~C 24 (Cyclo)alkyl(hetero)aryl groups, C 3 ~C 24 (Hetero)aryl(cyclo)alkyl, C 4 ~C 24 (Cyclo)alkenyl(hetero)aryl group, C 4 ~C 24 (Hetero)aryl(cyclo)alkenyl group, C 4 ~C 24 (Cyclo)alkynyl(hetero)aryl group, C 4 ~C 24 (Hetero)aryl(cyclo)alkynyl group, C 4 ~C 24 Alkylcycloalkyl groups, and C 4 ~C 24 wherein SP and CB are as defined above; and wherein, for R 47 , the alkyl group, alkenyl group, alkynyl group, aryl, heteroaryl, cycloalkyl group, cycloalkenyl group, cycloalkynyl group, (cyclo)alkyl(hetero)aryl group, (hetero)aryl(cyclo)alkyl group, (cyclo)alkenyl(hetero)aryl group, (hetero)aryl(cyclo)alkenyl group, (cyclo)alkynyl(hetero)aryl group, (hetero)aryl(cyclo)alkynyl group, alkylcycloalkyl group, and cycloalkylalkyl group are selected from the group consisting of -Cl, -F, -Br, -I, -OR 37 , -N(R 37 ) 2 , -SO 3 R 37 , -PO 3 (R 37 ) 2、 -PO 4 (R 37 ) 2、 -NO 2 , -CF 3 , =O, =NR 37 , and -SR 37 and may be substituted with a moiety selected from the group consisting of O, S, NR 37 , P, and Si, wherein the N, S, and P atoms are optionally oxidized, and wherein multiple N atoms are optionally quaternized.

2. The compound according to claim 1, wherein R 48 is -OC(O)-CA.

3. A compound described in claim 1 or 2, wherein CA is a drug.

4. The compound of claim 3, wherein the drug is selected from the group consisting of cytotoxins, antiproliferative / antitumor agents, antivirals, antibiotics, anti-inflammatory agents, chemosensitizers, radiosensitizers, immunomodulators, immunosuppressants, immunostimulants, anti-angiogenic factors, and enzyme inhibitors.

5. The compound described in claim 3, wherein the drug is selected from the group consisting of monomethylauristatin E, doxorubicin, pyrrolobenzodiazepines and dimers (PBDs), indolinobenzodiazepines and dimers, and pyridinobenzodiazepines and dimers.

6. The compound according to any one of claims 1 to 5, wherein R 47 is -(SP) i -CB, and CB is a peptide or protein.

7. The compound 【Chemistry 03】 and enantiomers thereof; A compound according to any one of claims 1 to 6, wherein X 1 , X 2 , X 3 , X 4 and X 5 are CH 2 and R 47 is -(SP) i -CB, where CB is a peptide or protein.

8. The compound of claim 7, wherein R 48 is -OC(O)-CA, and CA is selected from the group consisting of monomethylauristatin E, doxorubicin, pyrrolobenzodiazepines and dimers (PBDs), indolinobenzodiazepines and dimers, and pyridinobenzodiazepines and dimers.

9. The compound 【Chemical 04】 The compound according to any one of claims 1 to 5, 10. A kit comprising the compound according to any one of claims 1 to 9 and a diene.

11. A compound according to any one of claims 1 to 9 or a kit according to claim 10 for use as a pharmaceutical.

12. A compound described in any one of claims 1 to 9, or a kit described in claim 10, for use in treating a disease in a subject, wherein the disease is selected from the group consisting of cancer, central nervous system (CNS) disease, infectious disease, inflammation, and cardiovascular disease.

13. The compound of claim 12, wherein the compound is as defined in claim 7 and the disease is cancer.

14. The compound of claim 12, wherein the compound is as defined in claim 8 and the disease is cancer.

15. The compound described in claim 14, wherein CA is monomethyl auristatin E.

16. An agent for use in treating a subject, the agent comprising a compound described in any one of claims 1 to 9, the agent being administered to the subject, and a molecule being released from the compound when the compound comes into contact with a diene in the subject.

17. The agent described in claim 16, wherein the agent comprises a compound described in claim 7.

18. The agent described in claim 16, wherein the agent comprises a compound described in claim 8.

19. The agent described in claim 18, wherein CA is monomethyl auristatin E.

20. An agent for use in imaging, the agent comprising a compound described in any one of claims 1 to 9, the agent being administered to a subject, and the compound in the subject being imaged.

21. A kit for use in imaging a subject, the kit comprising a compound according to any one of claims 1 to 9, or the compound and a diene, wherein the compound, or in the case of the compound and the diene, at least one of the compound and the diene, comprises a label selected from the group consisting of a radionuclide, a fluorescent dye, and a phosphorescent dye.

22. A kit for use in the delivery of a molecule, comprising a compound according to any one of claims 1 to 9, or said compound and a diene.

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