Compounds for complexing rare earth elements and / or s-, p-, d-block metals, their coordination compounds, peptide conjugates, methods for their preparation and their uses - Patents.com

Abstract

Title: Compounds for complexing rare earth elements and / or s-, p-, d-block metals, their coordination compounds, peptide conjugates, methods for their preparation and their uses The present invention relates to compounds of general formula (I), [Formula 1] JPEG2024533430000465.jpg31169 where Y is nitrogen or N-oxide and R 1 is hydrogen; halogen; hydroxyl; azide; amine; trifluoromethyl; -COO-alkyl; -CH2CH(OMe)2; -SH; -SO3H; -SO2Ar (where Ar is phenyl); NO2; [Case 2] JPEG2024533430000466.jpg7169 ;C6~C 10 Aryl (optionally substituted with -NH2, -NO2, -N, -COOH, -CH2Cl and / or -CH2COOH); C7-C 10 arylalkyl (optionally substituted with -NH2, -NO2, -COOH, -CH2Cl, and / or -CH2COOH); R 2 is an azide group; or [C3] JPEG2024533430000467.jpg9169 (wherein n is an integer ranging from 1 to 3); A is H;-(CH2) n COOH; -CH(CH3)COOH; -CH2P(=O)(OR)2 (where R is H or alkyl); -CH((CH2) n COOH)COOH;-CH((CH2) n -NH2)COOH, where n is an integer from 1 to 3; -CH2C(=O)(NH2); -CH2C(=O)(NH)-CH2COOH; -CH2P(=O)(OH)(Ar), where Ar is phenyl optionally substituted with C1-C6 alkyl; Z is [C4] JPEG2024533430000468.jpg60169 selected from the group consisting of; R 3 teeth, [C5] JPEG2024533430000469.jpg6169 And R 4 is H; halogen; piperidinyl; trimethylsilyl; triisopropylsilyl; phenyl; C1-C6 alkyl; C3-C6 cycloalkyl; -CF3; -(CH2) n NH2;-(CH2) n NHFmoc;-(CH2) n NHBoc;adamantyl; [C6] JPEG2024533430000470.jpg7169 R 5 is R 1 ,or, [C7] JPEG2024533430000471.jpg6169 and / or R 2 and R 3 Both, forming a 1,2,3-triazole group; with the proviso that at most one A is H. The invention further relates to coordination compounds thereof, peptide conjugates thereof, methods for their preparation and their use.

Description

Detailed Description of the Invention

[0001] [Current state of the technology] The present invention relates to novel macrocyclic compounds suitable as bridging chelators for complexing rare earth elements and / or s-, p-, and d-block metals to form highly stable coordination compounds. These coordination compounds are suitable for use as labels for quantitative detection of peptide conjugates and / or as contrast agents for magnetic resonance imaging. The present invention further relates to methods for preparing said chelators and for tracking peptide / protein-containing pharmaceuticals.

[0002] [Background technology] Metal elements are used in biomedical applications such as imaging contrast agents, radiotherapeutic agents, or bioanalytical labels. Most of these applications require the metal to be bound to a stable chelate that can be covalently linked to other molecules, such as peptide- or antibody-based targeting vectors. The most common example of a chelator applicable to most metal elements is DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) and its derivatives. A wide range of specialized chelators for specific metal elements have also been developed. [Price EW, Orvig C. (2014), Chem. Soc. Rev. 43(1), 260-290]

[0003] The rare earth elements (scandium - Sc, yttrium - Y, lanthanum - La, cerium - Ce, praseodymium - Pr, neodymium - Nd, promethium - Pm, samarium - Sm, europium - Eu, gadolinium - Gd, terbium - Tb, dysprosium - Dy, holmium - Ho, erbium - Er, thulium - Tm, ytterbium - Yb, and lutetium - Lu) are a group of metals that offer a wide range of medical applications. 90 Y, 153 Sm and 177 Lu-based radiopharmaceuticals are FDA approved; 166Clinical trials are underway for Ho, and others are being investigated using positron emission tomography (PET), single photon emission computed tomography (SPECT), or other methods that show favorable therapeutic properties ( 44 Sc, 47 Sc, 86 Y, 149 Pm, 159 Gd, 149 Tb, 161 Tb, 165 Dy, 161 Ho, 169 Er and 175 Yb). Stable, non-radioactive Gd chelates are clinically used as contrast agents in magnetic resonance imaging (MRI). Stable isotopes of rare earth elements are also used as labels for analytical purposes. Compounds of interest labeled in this way can be visualized, tracked, and quantified based on the element's unique luminescence or isotopic mass, which has zero background in biological systems. The unique isotopic mass is particularly useful, as it allows simultaneous quantification of many compounds in a single analysis (multiplexing), typically using an inductively coupled plasma mass spectrometer (ICP-MS) [Bodenmiller B. et al. (2012), Nat. Biotechnol. 30(9), 858-867]. Other metallic elements belonging to the s-, p-, and d-blocks of the periodic system, such as 89 Sr, 223 Ra, 117m Sn, 212 Pb, 213 Bi, 64 Cu, 225 Ac has been used in radiopharmaceutical compounds for imaging or therapy.

[0004] For practical applications, metal chelates must be highly stable so that the metal ion does not easily escape from the chelator or dissociate from the carrier molecule. In this regard, thermodynamic stability constants are insufficient information, since most applications are performed under conditions quite different from thermodynamic equilibrium (e.g., in vivo). Instead, kinetic inertness, which characterizes the rate at which the metal ion dissociates from the chelate under given conditions, must be considered. Kinetic inertness is strongly correlated with the rigidity of the chelator. DTPA-type (diethylenetriaminepentaacetic acid) acyclic chelators have a more flexible structure and are less kinetically inert and more rigid than DOTA-type macrocyclic chelators. Chelates with high kinetic inertness are particularly desirable for in vivo applications where the metal chelate is exposed to an excess of competing biological chelators and metal ions. Therefore, rigid, kinetically inert macrocyclic chelators are preferred over flexible acyclic chelators. For example, for approved or developmental radionuclides, 177 Virtually all radiopharmaceuticals based on Lu (half-life 6.7 days) use the macrocyclic chelator DOTA. This is because the metal must remain bound to the target molecule in vivo for several weeks to achieve the desired therapeutic effect. Similarly, for MRI contrast agents, macrocyclic chelates of gadolinium(III) are preferred over acyclic DTPA types. Recently, it has been found that acyclic agents release large amounts of free gadolinium in vivo, resulting in long-term deposition in the human brain [Fur ML, Caravan P. (2019), Metallomics 11(2), 240-254]. Following this finding and to prevent potential harm to patients, the use of acyclic MRI contrast agents has been severely restricted worldwide by drug regulatory agencies. Therefore, the importance of kinetic inertness of metal chelates is increasing for medical and other applications, and chelators offering even greater inertness are needed.

[0005] A method to further enhance kinetic inertness is to reinforce and rigidify the macrocyclic chelator with additional rings. For example, the bridged macrocyclic chelator cb-TE2A (4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane) provides a much more inert chelate with copper(II) ion than its non-bridged analogue [Boswell CA et al. (2004), J. Med. Chem. 47(6), 1465-1474]. Another similar chelator, cb-TEDPA (6,6'-((1,4,8,11-tetraazabicyclo[6.6.2]hexadecane-4,11-diyl)bis(methylene))dipicolinic acid), has been shown to provide exceptionally inert lanthanide(III) chelates [Rodriguez-Rodriguez A. et al. (2016), Inorg. Chem. 55(5), 2227-2239]. Another cyclen-based bridged chelator provided an extremely inert copper(II) chelate [Esteves, CV et al. (2013), Inorg. Chem. 52(9), 5138-5153]. A common feature of these chelators is that the bridge is independent of the coordinating pendant arm and points away from the pendant arm in the chelate. Chelators have also been prepared in which the bridge links the pendant arm itself, but the impact on the stability of the chelate was strongly negative [Vipond, J. et al. (2007), Inorg. Chem. 46(7), 2584-2595].

[0006] Moreover, too high kinetic inertness can be counterproductive and limit practical application, since the energy barrier the metal ion must overcome to exit the chelate is similar to the barrier the metal ion must overcome to enter the chelate during chelate formation. Generally, the more inert the chelate, the more difficult its formation. Higher temperatures and longer reaction times accelerate the formation kinetics. However, these reaction conditions are often incompatible with the half-life of metal nuclides in the preparation of sensitive targeting vectors (e.g., antibodies) and / or radiopharmaceuticals.

[0007] There remains a strong need for new types of chelating agents that rapidly form kinetically inert metal complexes under mild synthetic conditions.

[0008] DISCLOSURE OF THE INVENTION We have overcome the problems of the background art by developing a new type of chelating agent that forms a bridge after complexation of the metal ion, rigidifying the structure and enhancing kinetic inertness. The bridge is formed between two coordinating pendant arms based on a cycloaddition reaction between an alkyne and an azide substituent on the opposite pendant arm. This reaction occurs spontaneously after the formation of the unbridged chelate, without the usual need for catalysis by Cu(I) ions. Thus, the chelating agent acts as a one-way trap for the metal ion. The initial formation of the unbridged chelate is relatively fast. Subsequently, a bridge forms, trapping the metal within the chelating agent. For rare earth elements, the resulting bridged chelate exhibits extremely high kinetic inertness, up to six orders of magnitude greater than that of analogous chelates using DOTA. This exceptional inertness enables the novel use of chelates as analytical labels, capable of withstanding harsh hydrolysis conditions in concentrated acid and allowing for the direct quantification of the chelate in lysates using liquid chromatography-mass spectrometry (LC-MS). The use of LC-MS is advantageous because it is a much more common instrument than ICP-MS. The extremely high inertness is also advantageous for future in vivo uses of the chelates, such as as MRI contrast agents or radiopharmaceuticals, since the free metal is not released from the coordination compound in vivo and therefore no deposition of the metal occurs in the human or animal body.

[0009] The compounds of the present invention can act as highly efficient molecular traps, coordinating metal ions in a highly stable, rigid, and well-defined manner. The ligands can be prepared in a relatively few steps of organic synthesis. After the metal ion is coordinated, a click reaction takes place between the azide group and the triple bond present in the opposing pendant arms of the macrocycle, forming a triazole bridge and trapping the metal ion within the cage (forming a so-called click-zip complex). The formation of the triazole is irreversible and is depicted in Scheme 1 below.

[0010] Scheme 1. Schematic representation of the click-zip principle illustrated by the example of the ligand TD647.

[0011] [ka]

[0012] In a first aspect, the subject of the present invention relates to compounds of general formula (I)

[0013] [ka]

[0014] where: Y is nitrogen (N); N-oxide (N + -O - selected from the group consisting of; R 1 is H; halogen; -OH; -N3; ​​-(CH2) n N3 (where n is an integer ranging from 1 to 3); -NR2 (where R is independently selected from H or C1-C6 alkyl, which may be branched or straight chain); -(CH2) n NR2 (where n and R are as defined above); C6 to C 10 aryl (optionally -NH2, -NO2, -N3,

[0015] [ka]

[0016] , -COOH, -CH2Cl and / or -CH2COOH); C7 to C 10 arylalkyl (optionally -NH2, -NO2, -N3,

[0017] [ka]

[0018] -COOH, -CH2Cl, and / or -CH2COOH); -CF3; -COOR (wherein R is as defined above); -(CH2) n COOR (where n and R are as defined above);

[0019] [ka]

[0020] ;-CH2CH(OMe)2;

[0021] [ka]

[0022] -SH; -SO3H; -SO2Ar (wherein Ar is phenyl); NO2; R 2 teeth,

[0023] [ka]

[0024] ;or

[0025] [ka]

[0026] (wherein n is an integer ranging from 1 to 3); and preferably, R 2 teeth

[0027] [ka]

[0028] or

[0029] [ka]

[0030] and more preferably, R 2 teeth

[0031] [ka]

[0032] and; A is H;-(CH2) n COOH (where n is an integer from 1 to 3); -CH(CH3)COOH; -CH((CH2) n CH3)COOH (wherein n is as defined above); -CH2P(=O)(OR)2 (wherein R is as defined above); -CH((CH2) n -COOH)COOH (where n is an integer from 1 to 3); -CH((CH) n -NH2)COOH (wherein n is an integer from 1 to 3); -CH2C(=O)(NH2); -CH2C(=O)(NH)-CH2COOH; -CH2P(=O)(OH)(Ar) (wherein Ar is phenyl optionally substituted with C1-C6 alkyl); Z is

[0033] [ka]

[0034] is selected from the group consisting of where: R 3 teeth,

[0035] [ka]

[0036] and R 4is H; halogen; piperidinyl; trimethylsilyl; triisopropylsilyl; phenyl; C1-C6 alkyl (which may be branched or linear); C3-C6 cycloalkyl; -CF3; -(CH2) n NHR 6 (where n is an integer ranging from 1 to 3, and R 6 is selected from H, fluorenylmethyloxycarbonyl, tert-butyloxycarbonyl and benzyloxycarbonyl; —C(CH3)2(NHR 6 ) (where R 6 is as defined above); adamantyl;

[0037] [ka]

[0038] ;-(CH2) n COOH (where n is as defined above); C-C 10 aryl (optionally -NH2, -NO2, -N3,

[0039] [ka]

[0040] , -COOH, -CH2Cl, and / or -CH2COOH); R 5 is R 1 ,or,

[0041] [ka]

[0042] (where R 4 is as defined above); preferably, R 5 H; -CF3; halogen; -OH; C7~C 10arylalkyl (optionally substituted with -NH, -NO, -COOH, -CHCl, and / or -CHCOOH); or

[0043] [ka]

[0044] and / or R 2 and R 3 are both expressions

[0045] [ka]

[0046] , or the expression

[0047] [ka]

[0048] a 1,2,3-triazole group (where R 4 is as defined above); However, at most one A may be H.

[0049] If the compounds of the invention contain a chiral center, all enantiomers, mixtures of enantiomers and racemates fall within the scope of the invention. The invention further includes the compounds of formula (I) in the form of their salts with alkali metals, ammonium or amines, as well as the compounds of formula (I) in the form of addition salts with acids.

[0050] The term "halogen" means any iozotop of F, Cl, Br, and I. In particular, this term refers to 19 F, 35 Cl, 37 Cl, 79 Br, 81 Br, 127non-radioactive izotopes such as I; and 18 F, 36 Cl, 77 Br, 83 Br, 123 I, 124 I, 125 I, 131 It contains radioisotopes such as I.

[0051] In one embodiment, A is H; -(CH) n COOH (where n is an integer from 1 to 3); -CH(CH3)COOH; -CH((CH2) n -CH3)COOH (where n is an integer from 1 to 3); -CH2P(=O)(OR)2 (where R is as defined above); -CH2C(=O)(NH2); -CH2C(=O)(NH)-CH2COOH; -CH2P(=O)(OH)(Ar) (where Ar is phenyl which may be optionally substituted with C1-C6 alkyl).

[0052] In one embodiment, R 1 is H; halogen; -OH; -N; -CHN; -NR (wherein R is independently selected from H or C1-C6 alkyl, which may be branched or straight chain); -CHNR (wherein R is as defined above); C6-C 10 Aryl (optionally substituted with -NH, -NO, -COOH and / or -CHCOOH); C to C 10 Arylalkyl (optionally substituted with -NH, -NO, -COOH, -CHCl and / or -CHCOOH; -CF; -COOR (wherein R is as defined above); -(CH) n COOR (where n and R are as defined above);

[0053] [ka]

[0054] ;-CH2CH(OMe)2;

[0055] [ka]

[0056] -SH; -SO3H; -SO2Ar (wherein Ar is phenyl); NO2; Preferably, R 1 -H; halogen; -OH; -N; -CHN; -N(CH); phenyl; carboxyphenyl; -CF; -COOR (where R is as defined above); and

[0057] [ka]

[0058] is selected from.

[0059] In one embodiment, R 4 is H; halogen; piperidinyl; trimethylsilyl; triisopropylsilyl; phenyl; C1-C6 alkyl, which may be branched or straight-chain; C3-C6 cycloalkyl; -CF3; -CH2NHR 6 (where R 6 is selected from H, fluorenylmethyloxycarbonyl, tert-butyloxycarbonyl and benzyloxycarbonyl; —C(CH3)2(NHR 6 ) (where R 6 is as defined above); adamantyl;

[0060] [ka]

[0061] is selected from the group consisting of Preferably, R 4is selected from the group consisting of H; trifluoromethyl; 4-piperidinyl; -CH-NH; phenyl; cyclopropyl; adamantyl; terc-butyl; trimethylsilyl (TMS); triisopropylsilyl (TIPS); halogen (F, Cl, Br, I); -C(CH)NH; and -C(CH)NHBoc. More preferably, R 4 is hydrogen.

[0062] In one embodiment of the present invention, the compounds of general formula (I) contain the following substituents: Y is nitrogen (N); R 1 is H; halogen; -OH; -N; -CHN; -NR (wherein R is independently selected from H or C1-C6 alkyl, which may be branched or straight chain); C6-C 10 Aryl (optionally substituted with -NH, -NO, -COOH and / or -CHCOOH); C to C 10 arylalkyl (optionally substituted with -NH2, -NO2, -COOH, -CH2Cl and / or -CH2COOH); -CF3; -COOR (wherein R is as defined above);

[0063] [ka]

[0064] ;-CH2CH(OMe)2;

[0065] [ka]

[0066] selected from the group consisting of: R 2 teeth

[0067] [ka]

[0068] and; A is H;-(CH2) n COOH (where n is an integer from 1 to 3); -CH(CH3)COOH; -CH((CH2) n -CH3)COOH (wherein n is as defined above); -CH2P(=O)(OR)2 (wherein R is as defined above); -CH2C(=O)(NH2); -CH2C(=O)(NH)-CH2COOH; -CH2P(=O)(OH)(Ar) (wherein Ar is phenyl optionally substituted with C1-C6 alkyl); Z is

[0069] [ka]

[0070] is selected from the group consisting of where: R 3 teeth,

[0071] [ka]

[0072] and; R 4 is H; halogen; piperidinyl; trimethylsilyl; triisopropylsilyl; phenyl; C1-C6 alkyl (which may be branched or linear); C3-C6 cycloalkyl; -CF3; -CH2NHR 6 (where R 6 is selected from H, fluorenylmethyloxycarbonyl and benzyloxycarbonyl; —C(CH3)2(NHR 6 ) (where R 6 is as defined above); adamantyl;

[0073] [ka]

[0074] selected from the group consisting of; R 5 H; -CF3; halogen; -OH; C7~C 10 arylalkyl (optionally substituted with -NH, -NO, -COOH, -CHCl and / or -CHCOOH); or

[0075] [ka]

[0076] (where R 4 is as defined above); and and / or R 2 and R 3 are both expressions

[0077] [ka]

[0078] , or the expression

[0079] [ka]

[0080] a 1,2,3-triazole group (where R 4 is as defined above); However, at most one A may be H.

[0081] In a preferred embodiment, Y is nitrogen, and therefore R 1 and R 2 The compound forms a pendant arm comprising a pyridyl moiety substituted with

[0082] In one embodiment, R 1 is in the meta position from Y, and R 2 It is located in the para position from -CH2-.

[0083] In one embodiment, R 1 is in the meta position from Y, and R 2 It is located in the ortho position relative to -CH2-.

[0084] In another embodiment, R 1 is located para to Y.

[0085] In one preferred embodiment, R 2 teeth

[0086] [ka]

[0087] is.

[0088] Preferably, R 2 teeth

[0089] [ka]

[0090] and R 1 is selected from the group consisting of H, phenyl, carboxyphenyl, halogen (preferably Cl), trifluoromethyl, carboxyl, carboxylic acid ester or amine, more preferably R 1 is H or carboxyphenyl.

[0091] In one embodiment, Z is

[0092] [ka]

[0093] is.

[0094] In one preferred embodiment, Z is

[0095] [ka]

[0096] and R 3 and R 5 is as defined above. Preferably, Z is

[0097] [ka]

[0098] and even more preferably Z is

[0099] [ka]

[0100] is.

[0101] In the compounds of general formula (I), A may be the same or different, and preferably A is the same.

[0102] In one embodiment, Z is

[0103] [ka]

[0104] is.

[0105] In one embodiment, Z is

[0106] [ka]

[0107] is.

[0108] In one embodiment, A is -(CH2)COOH; -(CH2)2COOH; -CH2P(=O)(OEt)2; -CH2P(=O)(OH)2; -CH2P(=O)(OH)(OEt); -CH2P(=O)(Ph)(OH); -(CH2)COONH2; -(CH2)C(=O)NH-CH2COOH; -(CH2)COO t -Bu; -CH(CH2COOH)COOH.

[0109] Preferably, A is selected from the group comprising -(CH2)COOH; -CH2P(=O)(OEt)2; -CH2P(=O)(OH)2; -CH2P(=O)(OH)(OEt); and -CH2P(=O)(Ph)(OH). Most preferably, A is -(CH2)COOH.

[0110] In one embodiment, Y is nitrogen; R 1 is H; Cl; -N(CH3)2; phenyl (optionally substituted with -COOH or -CH2COOH); benzyl (optionally substituted with -COOH or -CH2COOH); -CF3; -COOCH3; -COOCH(CH3)2; -COOtBu;

[0111] [ka]

[0112] -SH; A is H;-(CH2) n independently selected from the group consisting of: -COOH (where n is 1 or 2); -CHP(=O)(OH)2; -CHP(=O)(OH)(OEt); -CHP(=O)(OEt)2; -CHC(=O)(NH2); -CHC(=O)(NH)-CHCOOH; -CHP(=O)(OH)(Ph); Z is

[0113] [ka]

[0114] is selected from the group consisting of where: R 3 teeth

[0115] [ka]

[0116] and; R 4 is selected from the group consisting of H; halogen; piperidinyl; trimethylsilyl; triisopropylsilyl; phenyl; cyclopropyl, terc-butyl; -CF3; -CH2NH2; -CH2N(H)(Fmoc); -C(CH3)2(NH2); -C(CH3)2(NHBoc); adamantyl; R 5 is H, or

[0117] [ka]

[0118] (where R 4 is as defined above); However, compounds in which at most one A is H.

[0119] In one preferred embodiment, Z is

[0120] [ka]

[0121] and;R 3 teeth

[0122] [ka]

[0123] and;R 4 and R 5is as defined above, preferably R 4 and R 5 is H.

[0124] In another embodiment, R 2 and R 3 are both expressions

[0125] [ka]

[0126] , or the expression

[0127] [ka]

[0128] a 1,2,3-triazole group (where R 4 is as defined above, preferably R 4 is hydrogen); in such an embodiment, The bridge is formed between two opposite nitrogen atoms of the cyclic moiety, and the compound of general formula (I) thus forms a cage-type structure. The bridge is formed between two opposite nitrogen atoms of the cyclic moiety, and the compound of general formula (IIa) or general formula (IIb)

[0129] [ka]

[0130] is an entity of; where L is

[0131] [ka]

[0132] is a linker selected from the group consisting of: R 1 , R 4 and R 5is as defined above. The linker L is derived from the Z group as defined above. Preferably, L is

[0133] [ka]

[0134] and more preferably, L is

[0135] [ka]

[0136] is.

[0137] In one preferred embodiment, said compound of general formula (I) is selected from the group comprising compounds having the following combination of substituents: Y is N + -O - and;R 1 ,R 4 and R 5 is H:

[0138] [Table 1]

[0139] Y is N; R 1 , R 4 and R 5 is H:

[0140] [Table 2-1]

[0141] [Table 2-2]

[0142] Y is N; Z is

[0143] [ka]

[0144] and R is

[0145] [ka]

[0146] is:

[0147] [Table 3]

[0148] Y is N; R 2 and R 3 is the expression

[0149] [ka]

[0150] together form a 1,2,3-triazole group of:

[0151] [Table 4]

[0152] Y is N; R 2 and R 3 is the expression

[0153] [ka]

[0154] together form a 1,2,3-triazole group of:

[0155] [Table 5]

[0156] Y is N; R 1 is phenyl; R 2 teeth

[0157] [ka]

[0158] And:R 3 teeth

[0159] [ka]

[0160] is:

[0161] [Table 6]

[0162] Y is N; A is —(CH2)COOH; R 1 is H;R 2 teeth

[0163] [ka]

[0164] is:

[0165] [Table 7-1]

[0166] [Table 7-2]

[0167] Y is N; A is —(CH2)COOH; R 2 teeth

[0168] [ka]

[0169] and Z is

[0170] [ka]

[0171] is:

[0172] [Table 8]

[0173] Y is N; A is -CH(CH3)COOH; R 2 teeth

[0174] [ka]

[0175] and Z is

[0176] [ka]

[0177] is:

[0178] [Table 9]

[0179] In a most preferred embodiment, said compound of general formula (I) is one in which Y is nitrogen and Z is

[0180] [ka]

[0181] and the remaining substituents are present in the following combinations:

[0182] [Table 10-1]

[0183] [Table 10-2]

[0184] [Table 10-3]

[0185] .

[0186] In another aspect, the subject of the present invention is a process for the synthesis of said compounds of general formula (I), comprising the following steps: i) providing an alkyne intermediate of general formula Z-Cl, where Z is as defined above; ii) an azide intermediate of general formula (III)

[0187] [ka]

[0188] provide; where Y, R 1 and R 2 is as defined above; iii) cyclen derivatives of general formula (IV)

[0189] [ka]

[0190] to provide where pA is -(CH2) n COO tBu (where n is an integer from 1 to 3); benzyloxycarbonyl; -CH(CH3)COO t Bu; -CH(CH3)COOR (where R is tert-butyl, methyl, or ethyl); -CH((CH2) n CH3)COOR (where n and R are as defined above; -CH((CH2) n -COOR)COOR (wherein n is as defined above and R is independently selected from tert-butyl, methyl, or ethyl); -CHP(=O)(OR) (wherein R is C1-C6 alkyl which may be branched or straight chain); -CHC(=O)(NH); -CHC(=O)(NH)-CHCOOH; -CHP(=O)(OH)(Ar) (wherein Ar is phenyl which may be optionally substituted with C1-C6 alkyl); preferably, pA is selected from the group comprising -CHCOOtBu; -CHP(O)(OEt); -(CH)COOtBu; iv-A) The cyclene derivative of general formula (IV) is reacted with an alkyne intermediate Z-Cl to give an intermediate of general formula (V):

[0191] [ka]

[0192] get, where Z and pA are as defined above; or iv-B) Reacting the cyclen derivative of general formula (IV) with the azide intermediate of general formula (III) to obtain an intermediate of general formula (VI).

[0193] [ka]

[0194] get, where Y, R 1 , R 2 and pA is as defined above; vA) reacting the intermediate of general formula (V) with the azide intermediate of general formula (III) to obtain an intermediate of general formula (VII)

[0195] [ka]

[0196] get, where Y, R 1 , R 2 , pA and Z are as defined above; or vB) reacting the intermediate of general formula (VI) with the alkyne intermediate Z-Cl to obtain the intermediate of general formula (VII); vi) Optionally, hydrolysis of the protecting groups is carried out to obtain the compound of general formula (I).

[0197] In general, compounds of the general formula Z-Cl are either commercially available or can be synthesized using Pd(0)-catalyzed Sonogashira cross-coupling reactions from the corresponding 2-halopyridines (preferably Br, I) and terminal alkynes or silyl-protected acetylenes.

[0198] Compounds of general formula (III) can be obtained from 2,6-bis(halomethyl)pyridine (preferably Cl) by demethylation using sodium azide as the source of the azide moiety.

[0199] Cyclene derivatives of general formula (IV) are commercially available or can be prepared by reacting a trans-N-bis-protected cyclen (preferably carbamate-protected) with a cyclen or alkyl haloacetate (alkylation), an alkyl acrylate (Michael addition) or a trialkyl phosphite in the presence of (para)formaldehyde (Kabachnik-Fields reaction).

[0200] Steps iv-A), iv-B), vA) and vB) are carried out in a solvent selected from the group comprising MeCN (preferably), DMSO or DMF. The reaction is usually carried out at room temperature for a period of a few hours up to a few days.

[0201] Step vi) of deprotecting the pA group is optional; for example, if pA is -CHP(O)(OEt), without deprotection, A in the resulting general formula (I) is equal to pA. Specific conditions for deprotecting the protecting groups depend on the chemical nature of the protecting groups and are known to those skilled in the art. Typically, terc-butyl ester and Boc protecting groups are hydrolyzed under acidic conditions (TFA) or thermally, methyl or isopropyl protecting groups are hydrolyzed under alkaline conditions, and ethyl phosphonate esters are converted to the corresponding phosphonic acids by transesterification with trimethylsilyl bromide in the presence of a pyridine base.

[0202] Another subject of the present invention is a compound of general formula (I) defined above, which is prepared by combining with a lanthanide(III) cation, Na + , Ba 2+ , Pb 2+ , Sr 2+ , Ca 2+ , Cd 2+ , Zn 2+ , Mn 2+ , Pt 2+ , Cu 2+ , Ni 2+ ,Sc. 3+ , Y 3+ , Bi 3+ , In 3+ , Ru 3+ , Ir 3+ , Ga 3+ , Tl 3+ , Pd 2+ and a coordination compound with a metal cation selected from the group consisting of: 3+ , Ce 3+ , Pr 3+ , Nd 3+ , Sm 3+ ,EU 3+ , Gd 3+ , Tb 3+ , Dy3+ , Ho 3+ , Er 3+ , Tm 3+ , Yb 3+ , Lu 3+ , Y 3+ ,Sc. 3+ , Bi 3+ , In 3+ , Tl 3+ , Pb 2+ , Ca 2+ , Cd 2+ , Zn 2+ , Cu 2+ , Ni 2+ , Mn 2+ , Pd 2+ , Na + is selected from the group consisting of:

[0203] The metal ion is 40 Ca, 42 Ca, 43 Ca, 44 Ca, 46 Ca, 48 Ca, 45 Sc, 89 Y, 138 La, 139 La, 136 Ce, 138 Ce, 140 Ce, 142 Ce, 141 Pr, 142 Nd, 143 Nd, 144 Nd, 145 Nd, 146 Nd, 148 Nd, 150 Nd, 144 Sm, 147 Sm, 148 Sm, 149 Sm, 150 Sm, 152 Sm, 154 Sm, 151 EU, 153 EU, 152 Gd, 154 Gd, 155 Gd, 156 Gd, 157 Gd, 158 Gd, 160 Gd, 159 Tb,156 Dy, 158 Dy, 160 Dy, 161 Dy, 162 Dy, 163 Dy, 164 Dy, 165 Ho, 162 Er, 164 Er, 166 Er, 167 Er, 168 Er, 170 Er, 169 Tm, 168 Yb, 170 Yb, 171 Yb, 172 Yb, 173 Yb, 174 Yb, 176 Y, 175 Lu, 176 stable isotopes such as Lu, and 44 Sc, 47 Sc, 64 Cu, 67 Cu, 86 Y, 90 Y, 140 Nd, 149 Pm, 151 Pm, 153 Sm, 159 Gd, 149 Tb, 161 Tb, 165 Dy, 161 Ho, 166 Ho, 169 Er, 167 Tm, 175 Yb, 177 It should be understood to include all isotopes of a particular metal, including radioactive isotopes such as Lu.

[0204] Lanthanides (Ln) are defined as a group of 15 metallic chemical elements with atomic numbers ranging from 57 to 71 (lanthanum to lutetium).

[0205] Preferably, the coordination compound has the general formula (VIII)

[0206] [ka]

[0207] It has.

[0208] where M is the metal cation and R 1 , R 4 , R 5 , and A is as defined above, and n is selected from 1, 2, 3 and 4; More preferably, the coordination compound has the general formula (VIIIa)

[0209] [ka]

[0210] It has.

[0211] The coordination compounds according to the present invention also include solvates and salts of pharmaceutically acceptable acids. The positive charge of the metal cation is offset by two negative charges arising from the A pendant arm (e.g., acetate). Thus, the overall charge of the coordination compound is -1 (for monovalent metal ions), 0 (for divalent metal ions), or +1 (for trivalent metal ions). The counterions that compensate for the total charge of the coordination compound are selected from the group consisting of anions derived from the following pharmaceutically acceptable acids: 1-Hydroxy-2-naphthoic acid;2,2-Dichloroacetic acid;2-Hydroxyethanesulfonic acid;2-Oxoglutaric acid;4-Acetamidobenzoic acid;4-Aminosalicylic acid;Acetic acid;Adipic acid;Ascorbic acid (L);Aspartic acid (L);Benzenesulfonic acid;Benzoic acid;Camphoric acid (+);Camphor-10-sulfonic acid (+);Capric acid (Decanoic acid);Caproic acid (Hexanoic acid);Caprylic acid (Octanoic acid);Carbohydrate;Cinnamic acid;Citric acid;Cyclamic acid;Dodecylsulfuric acid;Ethan-1,2-disulfonic acid;Ethanesulfonic acid;Formic acid;Fumaric acid;Galactaric acid;Gentisic acid;Glucoheptonic acid (D);Gluconic acid (D); Glucuronic acid (D); Glutamic acid; Glutaric acid; Glycerophosphoric acid; Glycolic acid; Hippuric acid; Hydrobromic acid; Hydrochloric acid; Isobutyric acid; Lactic acid (DL); Lactobionic acid; Lauric acid; Maleic acid; Malic acid (-L); Malonic acid; Mandelic acid (DL); Methanesulfonic acid; Naphthalene-1,5-disulfonic acid; Naphthalene-2-sulfonic acid; Nicotinic acid; Nitric acid; Oleic acid; Oxalic acid; Palmitic acid; Pamoic acid; Phosphoric acid; Propionic acid; Pyroglutamic acid (-L); Salicylic acid; Sebacic acid; Stearic acid; Succinic acid; Sulfuric acid; Tartaric acid (+L); Thiocyanic acid; Toluenesulfonic acid (p); Undecylenic acid; Trifluoroacetic acid.

[0212] The coordination compounds can be prepared from a free ligand of general formula (I) (preferably uncaged, and therefore preferably without a trazole bridge) and a metal salt (typically a water-soluble metal salt, preferably selected from the group including chloride, nitrate, acetate, formate, trifluoroacetate, trifluoromethylsulfonate, more preferably chloride or nitrate) according to Scheme 1 above. By stirring these reactants in an aqueous environment (pH 5-7) at a temperature ranging from 25°C to 100°C, the metal cation is coordinated to the compound of general formula (I), and then the R 2 and R 3 The substituents form the triazole bridge, allowing the metal cation to "click-zip" into the cage (reaction temperature and time are highly dependent on the choice of ligand and metal cation).

[0213] However, suitable ligands are not limited to the non-caged compounds of general formula (I), but are preferred because a wider range of metal cations (such as lanthanides) can form caged coordination compounds. Bridged (caged) compounds of general formula (I) having 1,5-triazole bridges are typically Tl 3+ , Pb 2+ , Bi 3+ , In 3+ , Cd 2+ , Ca 2+ , Cu 2+ , Ni 2+ , Mn 2+ , Pd 2+ , Na + The bridged (cage-type) compound of general formula (I) having a 1,4-triazole bridge is typically capable of forming a coordination compound with a metal cation selected from the group consisting of La 3+ , Ce 3+ , Pr 3+ , Nd 3+ , Sm 3+ ,EU 3+ , Gd 3+ , Tb 3+ , Dy 3+ , Ho 3+ , Er 3+ , Tm 3+ , Yb 3+ , Lu 3+ , Y 3+ , Tl 3+ , Pb 2+ , Bi 3+ , In 3+ , Cd 2+ , Ca 2+ , Cu 2+ , Ni 2+ , Mn 2+ , Pd 2+ , Na + and the like. The metal cations can form coordination compounds with metal cations selected from the group comprising:

[0214] The resulting coordination compounds are highly inert and stable, making them suitable for a variety of applications, including MRI contrast agents, radiodiagnosis or radiotherapy, and drug tracking involving peptides or proteins.

[0215] Coordination to metal cations of said compounds of general formula (I) is via the macrocyclic nitrogen atom of the cyclene moiety, the oxygen atoms present in the A group (pendant arm) and the Y group (when Y is N-oxide), as well as via the nitrogen atoms of the Y group (when Y is nitrogen) and the nitrogen atoms present in the Z group.

[0216] In one embodiment, the coordination compounds according to the present invention are capable of undergoing post-Click-Zip transformation. This means that after the metal cation is coordinated into the cage of the compound of general formula (I), either according to Scheme 1 above or by coordination to a cage compound of general formula (I), it may undergo further reactions if some functional groups are present. Typically, such functional groups are present in the substituents R of the coordination compound. 1 or R 5 The halogen present in R is preferably Cl, NO, or SH, but the post-Click-Zip conversion 1 , R 4 , R 5 and / or reactions of the A group (such as amide coupling reactions of uncoordinated carboxyl or amino groups).

[0217] For example, R 1 is a halogen, preferably Cl; or R 1 C6~C replaced by -CH2Cl 10 If the halogen atom is aryl, the coordination compound of general formula (VIII) can undergo a post-click transformation using a Pd(0) catalyzed Suzuki cross-coupling reaction with phenylboronic acid (3-borono-5-nitrobenzoic acid, or 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetic acid, or 2-(4-boronophenyl)acetic acid), during which the halogen is replaced so that the resulting coordination compound of general formula (VIII) has an R substituted with -COOH or -CHCOOH. 1 =COOH or C6~C 10The resulting aryl group can then be used in amide coupling reactions to synthesize peptide conjugates. The oligoarginine conjugates can cross cell membranes, allowing the metal cage to be internalized in cells.

[0218] Another possible post-click transformation is to convert R to a -N group by reaction with NaN. 1 , R 4 and / or R 5 The resulting azide can be used to couple the metal cage to another alkyne-containing moiety using the copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction.

[0219] Another possible post-click transformation is 1 , R 4 and / or R 5 The TIPS protecting group present in the substituent of R 1 , R 3 and / or R 5 teeth

[0220] [ka]

[0221] The hydrolysis is carried out using aqueous K2CO3, resulting in R 1 , R 3 and / or R 5 teeth

[0222] [ka]

[0223] The resulting deprotected triple bond is suitable for further conjugation via reaction with the alkyne, e.g., a click reaction with an azide, and formation of a triazole bridge (e.g., a CuAAC reaction).

[0224] In another embodiment, the post-click conversion is 1 , R 4 and / or R 5 where R 1 , R 3 and / or R 5 teeth

[0225] [ka]

[0226] Thus, the triple bond is converted to a dimethyl acetal (a protected form of the aldehyde). This represents the conversion of one functional group (an alkyne) to another (an aldehyde). The aldehyde is available for conjugation reactions based on the formation of Schiff bases with amines, hydrazines, and acid hydrazides.

[0227] In yet another embodiment, the post-click conversion can be a deuteration reaction of the CH2 group of the pendant arm in DO in the presence of DBU, converting it to a CD2 group. This conversion does not change the physicochemical properties of the molecule (in terms of stability or reactivity), but it does change the overall mass of the molecule so that it can be completely distinguished from the undeuterated parent molecule using mass spectrometry. Thus, the metal ion cages bearing certain metals can be used as two different mass tags.

[0228] In yet another embodiment, the post-click transformation may be a selective reduction of the pyridyl cycle of the Z substituent, wherein Z is

[0229] [ka]

[0230] and R 3 and R 5is as defined above, and NaBH4 or NaBD4 is used as the reducing agent, thereby converting the Z group to

[0231] [ka]

[0232] ,or,

[0233] [ka]

[0234] Similar to deuteration of the CH2 group, this conversion alters the mass of the molecule for the purpose of metal tagging.

[0235] In yet another embodiment, the post-click conversion is 1 and / or R 4 and / or R 5 and / or the NH group of A (if present) (where R 1 and / or R 4 and / or R 5 -NH2 or -(CH2) n The reaction may be a reaction of a -NH group (containing NH) with FmocCl, resulting in the conversion of the -NH group to an -NHFmoc group. The Fmoc group is the most commonly used amine protecting group in solid phase peptide synthesis (SPPS). Therefore, the introduction of the Fmoc group facilitates the use of the metal cage in SPPS.

[0236] In yet another embodiment, the post-click conversion is 1 and / or R 4 and / or R 5 and / or the reaction of the NH group of A (if present) with a carboxyl group of another molecule of general formula (I) as defined above or a metal complex thereof, resulting in a conjugate in which two or more of said compounds of general formula (I) or metal complexes thereof are linked via peptide (amide) bonds.

[0237] In yet another embodiment, the post-click conversion is 1 and / or R 4 and / or R 5 and / or if present in A, it may be a reaction of the uncoordinated -COOH group with the amino group of an amino acid or peptide, thus forming a peptide bond for conjugation purposes.

[0238] In yet another embodiment, the post-click conversion is 1 , R 4 , and / or R 5 The reaction may be an S-alkylation reaction between a substituent halogen, preferably Cl, and a thiol, resulting in the conversion of the halogen to a sulfide. The thiol may be, for example, NaS, NaSH, N-Boc-cysteine, or a cysteine-containing peptide, and the reaction may be carried out in the presence of DIPEA. Incorporating cysteine ​​(an amino acid) into the structure of the coordination compound according to the present invention further expands the possibilities for amide bonds (e.g., peptide bonds).

[0239] In yet another embodiment, the post-click conversion is 1 , R 3 , and / or R 5 of

[0240] [ka]

[0241] group (where R 1 , R 3 , and / or R 5 teeth

[0242] [ka]

[0243] with an azide, preferably an alkyl- or aryl-azide, whereby the alkyl- or aryl- and R 1 , R 3 , and / or R 5 A triazole bridge is formed connecting the aryl azide and the alkyl azide. The aryl azide is preferably a C6-C10 aryl azide, such as benzyl azide, optionally substituted with -COOH or -CH2COOH. The alkyl azide is preferably a C1-C7 alkyl azide, where the alkyl can be linear or branched.

[0244] In yet another embodiment, the post-click conversion is 1 or R 5 -N3 group (where R 1 or R 5 The reaction may be a reaction of a C7-C10 arylalkynyl group (containing an -N3 group) with a substituent containing a carbon-carbon triple bond, thereby forming a triazole bridge connecting the substituent and the coordination compound. The triple bond-containing substituent may be, for example, a C7-C10 arylalkynyl optionally substituted with -COOH or -CH2COOH (e.g., phenylacetylene), where the phenyl moiety may be optionally substituted with -COOH or -CH2COOH.

[0245] In yet another embodiment, the post-Click-Zip transformation is 1 and / or R 5 with a maleimide-containing substituent, thereby forming a thiosuccinimide linkage connecting the substituent and the coordination compound; or 1 and / or R 5 reaction of the -SH group of R with another substituent containing an -SH group to form a disulfide bridge linking the substituent and the coordination compound; or 1 and / or R 5 Reaction of the -SH group with an alkyl- or aryl halide forms a thioether linkage connecting the alkyl- or aryl to the coordination compound.

[0246] In yet another embodiment, the post-Click-Zip transformation is 1 and / or R 5 The reaction may be of the -NO2 group with a substituent containing an -SH group, thereby forming a thioether bond linking the substituent and the coordination compound.

[0247] In one embodiment, the post-Click conversion can involve reaction with another coordination compound of the present invention, thereby forming a chain or dimer. The coordination compound chain or dimer can contain the same metal cation in both click-zip cages or different metal cations in each chelate. The advantage of the chains and dimers is the possibility of combining the properties, e.g., specific weights, of two cage-type chelates to produce mass tags with a wide range of weights.

[0248] Typically, the coordination compound is R 1 or R 5 group and R of the following coordination compounds 1 or R 5 is attached to the chain by forming a triazole bridge between the R 5 The use of the group Z is

[0249] [ka]

[0250] ,or,

[0251] [ka]

[0252] (This is only possible if

[0253] The general basic structure of the coordination compound chain is, for example, (IXa) or (IXb)

[0254] [ka]

[0255] [ka]

[0256] .

[0257] In one embodiment, the coordination compound comprises an amine group (R 1 and / or R 4 and / or R 5 and / or A) and a carboxyl group (R 1 and / or R 4 and / or R 5 and / or A) is attached to the chain by an amide bond formed between the substituent of another coordination compound containing A).

[0258] The chain typically comprises two coordination compounds according to the invention, but may comprise three, four or more coordination compounds.

[0259] In yet another embodiment, the two types of linkages described above can be in one chain, e.g., two adjacent coordination compounds can be linked by one triazole bridge, and R 5 or R 1 The substituent can be left to link another coordination compound according to the invention via a triazole bridge or, if an amino or carboxyl group is present, via an amide bond.

[0260] In one embodiment, the coordination compound is a coordination compound having a substituent R containing an —SH group. 1 or R 5 and a substituent R of another coordination compound containing an -SH group. 1 or R 5 The chain is linked by a disulfide bond (-SS-) formed between the

[0261] The coordination compound dimer is R 1 group and R of the second coordination compound 5 At the same time, the R 5 group and R of the second coordination compound 1 The present invention comprises two coordination compounds according to the present invention linked together by forming a triazole bridge between the groups.

[0262] A typical structure of the coordination compound dimer is, for example, (X)

[0263] [ka]

[0264] .

[0265] The synthesis of the coordination compound chain or dimer is based on a click reaction between the azide group of the first coordination compound and the triple bond of the second coordination compound, thereby forming a triazole bridge, linking the two coordination compounds into a dimer. The triazole bridge is selected from the group including: R of the first coordination compound 1 or R 4 or R 5 and the -N3 group of the second coordination compound R 1 or R 4 or R 5 The triple bond of the formula

[0266] [ka]

[0267] , or the expression

[0268] [ka]

[0269] 1,2,3-triazole group.

[0270] Preferably, the metal cations are different, for example, the metal cations are Tb 3+ and 176 Yb 3+ is selected from.

[0271] Another object of the present invention is a conjugate suitable for use as a marker for drug tracking. Many drugs are peptide- or protein-based, and their biodistribution can be tracked by attaching fluorescent or radioactive markers to the peptide or protein structure. The coordination compounds and dimers of the present invention are suitable for binding the drug to the peptide or protein of interest. Therefore, isolated cell cultures or tissues can be analyzed for the presence of the metal complex using conventional methods, such as LC-MS, a commonly used and relatively inexpensive instrument. The cells or tissues can be completely hydrolyzed with strong acid without decomposing the coordination compound of the present invention. The coordination compound can then be detected from the hydrolyzed sample. Such methods are inexpensive and reliable due to the extremely high stability of the coordination compounds. Furthermore, LC-MS methods allow the use of multiple coordination compounds with different metal isotopes (multiplexing), thereby allowing all coordination compounds to be analyzed in a single LC-MS run.

[0272] The conjugate according to the present invention comprises the coordination compound according to the present invention as defined above, or a dimer or chain of the coordination compound as defined above, conjugated to a peptide or protein. The conjugation is carried out via an amide bond formed between the amino group of the peptide or protein and the carboxyl group of the coordination compound or the chain or dimer, or vice versa. Therefore, the coordination compound or the chain or dimer must contain either an -NH2 or -COOH group prior to conjugation of the peptide or protein. Preferably, the -NH2 or -COOH group is R1 and / or R 4 and / or R 5 The conjugate is part of the -NH2;-(CH2) group. n NH2;-COOH;-(CH2) n COOH; -NH2, -COOH, -(CH2) n NH2, or -(CH2) n C6-C substituted with COOH 10 R selected from the group including aryl (where n is an integer ranging from 1 to 3) 1 and / or R 4 and / or R 5 The ligand is formed from a coordination compound or a coordination compound dimer having a group.

[0273] The conjugation reaction conditions are known to those skilled in the art, and the reaction is usually carried out in DMSO at 25° C. in the presence of an organic base (triethylamine, ethyldiisopropylamine) using a commonly used peptide coupling agent (preferably HATU, PyAOP). The reaction is usually completed within a few minutes.

[0274] The peptide is preferably selected from the group comprising oligopeptides of 3 to 20 amino acids; Said protein is preferably selected from the group comprising antibodies, preferably monoclonal antibodies.

[0275] Another object of the present invention is a method for tracking a drug, preferably a peptide-based or protein-based drug, comprising the steps of: i) providing a cell culture or tissue to be analyzed, comprising at least one conjugate as defined above, wherein said peptide or said protein is that of said peptide-based drug or said protein-based drug to be tracked; ii) hydrolyzing the cell culture or tissue of step i) using a strong acid, preferably non-oxidizing aq. HCl, to obtain a hydrolysate; iii) qualitatively and / or quantitatively analyzing the hydrolysate of step ii) for the presence of the coordination compound or the presence of the coordination compound dimer according to the present invention, preferably using LC-MS.

[0276] A further object of the present invention is the in vitro use of said coordination compound, or said coordination compound dimer, or said conjugate according to the present invention in pharmacology, preferably for the development and testing of new drugs, more preferably for drug marking and tracking.

[0277] Another object of the present invention is to use the coordination compound or the coordination compound dimer according to the present invention as a medical diagnostic, preferably an MRI contrast agent. Preferably, Gd is used as a contrast agent for medical diagnostics, since Gd has a very high relaxivity. 3+ The coordination compounds or dimers of the formula (I) are used as MRI contrast agents. The extremely high kinetic inertness of the coordination compounds (about 100 times higher than that of GdDOTA, a commonly used MRI contrast agent) allows for the use of higher dosages, and free gadolinium, which is itself toxic to humans or animals, is not released from the coordination compounds according to the present invention. Therefore, the claimed coordination compounds are expected to be safe for use in humans or animals.

[0278] Another object of the present invention is the use of said coordination compound or said coordination compound chain or said coordination compound dimer according to the invention, wherein M is a radionuclide, preferably 44 Sc, 47 Sc, 64 Cu, 67 Cu, 86 Y, 90 Y, 140 Nd, 149 Pm, 151 Pm, 153 Sm, 159 Gd, 149 Tb, 161 Tb, 165 Dy, 161 Ho, 166 Ho, 169 Er, 167Tm, 175 Yb, 177 and R is selected from the group including R and R for use in medicine as a radiodiagnostic agent and / or radiopharmaceutical. Alternatively, the radionuclide may be present in the side chain of said coordination compound, such as a radioisotope of a halogen, e.g., R 1 and / or R 5 teeth, 18 It may be F.

[0279] The present invention is further demonstrated by the following examples.

[0280] [Example] Example 1: Synthesis of an intermediate for an alkyne pendant arm Synthesis of TD701:

[0281] [ka]

[0282] A pear-shaped glass flask (50 mL) was charged with (6-bromopyridin-2-yl)methanol (2.47 g; 13.1 mmol; 1.0 equiv.) and a magnetic stirrer and protected with argon three times. Subsequently, solid CuI (149 mg; 782 μmol; 6.0 mol%) and [Pd(PPh3)2Cl2] (225 mg; 321 μmol; 2.4 mol%) were added and protected with argon three times. Dry THF (25 mL) was then added through a septum under a constant flow of argon, followed by ethynyltrimethylsilane (2.0 mL; 14.5 mmol; 1.1 equiv.) and TEA (5.5 mL; 39.5 mmol; 3.0 equiv.). The mixture then turned dark brown in color. The flask was left stirring at room temperature for 3 hours under a septum (but without external argon). The mixture was then transferred to a separatory funnel containing EtOAc (150 mL) and H2O (100 mL). After shaking, the dark (not completely homogeneous) organic layer was separated, and the aqueous layer was further extracted with EtOAc (5 x 50 mL). The combined organic layers were filtered through a cotton plug, and the filtrate was further dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The resulting dark brown oily residue was purified by flash chromatography (330 g SiO2, 100% DCM to 10% EtOAc in DCM). The combined product-containing fractions were evaporated to dryness and further co-evaporated once with DCM. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a dark yellow oil. Yield: 2.16 g (80%; 1 step; based on (6-bromopyridin-2-yl)methanol). NMR (DMSO-d6): 1 Hδ H 0.24(CH3,s,9H);4.52(CH2,d,2H,J 3 HH =6);5.48(OH,t,1H, 3 J HH =6);7.39(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.46(arom.,dd,1H, 3 J HH =8; 4 J HH=1);7.80(arom.,t,1H, 3 J HH =8). 13 C{ 1 H}δ C -0.3 (CH3, s); 63.9 (CH2, s); 93.4 and 104.5 (C≡C; 2 × s); 120.2 (aromatic, s); 125.4 (aromatic, s); 137.2 (aromatic, s); 140.8 (aromatic, s); 162.7 (aromatic, s). ESI-HRMS: 206.0995 [M+H] + (Theoretical value [C 11 H 16 N1O1Si1] + =206.0996).

[0283] Synthesis of TD557:

[0284] [ka]

[0285] In a round-bottom glass flask (100 mL), TD701 (2.16 g; 10.5 mmol; 1.0 equiv.) was dissolved in DCM (40 mL). A freshly prepared solution of SOCl2 (1.50 mL; 20.7 mmol; 2.0 equiv.) in DCM (10 mL) was then added, and the open flask was stirred at room temperature for 1 h. A dilute aqueous solution of NaHCO3 (50 mL) was then added, and the resulting biphasic mixture was vigorously stirred at room temperature for an additional 1 h, during which time gas bubbles were slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. A brown residue ( 1The resulting crude product (already pure by HNMR) was purified by flash chromatography (330 g SiO2, 60% PE in DCM to 20% PE in DCM) to remove a highly colored impurity. The combined product fractions were evaporated to dryness and co-evaporated once with DCM. The remaining nearly colorless oil solidified on standing in the freezer. The resulting solid was crushed and further dried under high vacuum overnight to give the product in the form of the free base as a fine white powder. Yield: 2.14 g (87%; 1 step; based on TD701). NMR (DMSO-d6): 1 Hδ H 0.25(CH3,s,9H);4.76(CH2,s,2H);7.50(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.55(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.86(arom.,t,1H, 3 J HH =8). 13 C{ 1 H}δ C -0.4 (CH3, s); 46.3 (CH2, s); 94.3 and 103.8 (C≡C; 2 × s); 123.3 (arom., s); 126.7 (arom., s); 138.1 (arom., s); 141.5 (arom., s); 157.0 (arom., s). ESI-HRMS: 224.0657 [M+H] + (Theoretical value [C 11 H 15 N1Cl1Si1] + =224.0657).

[0286] Synthesis of TD558:

[0287] [ka]

[0288] In a pear-shaped glass flask (100 mL), TD557 (1.05 g; 4.69 mmol; 1.0 equiv.) was dissolved in MeCN (40 mL). Next, a freshly prepared solution of KF (406 mg; 7.0 mmol; 1.5 equiv.) in HO (7 mL) was added, and the flask was vigorously stirred at room temperature for 2 h. The mixture was concentrated to ~10 mL and diluted with DCM (50 mL) and dilute aqueous NaHCO (50 mL). The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 × 25 mL). The combined organic layers were dried over anhydrous NaSO, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried briefly under high vacuum to give the product in the form of the free base as a pale yellow oil. Yield: 702 mg (99%; 1 step; based on TD557). NMR (DMSO-d6): 1 Hδ H 4.37(C≡CH,s,1H);4.76(CH2,s,2H);7.53(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.57(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.87(arom.,t,1H, 3 J HH =8). 13 C{ 1 H}46.3(CH2,s);80.6 and 82.6(C≡CH;2×s);123.4(arom.,s);126.8(arom.,s);138.1(arom.,s);141.3(arom.,s);157.0(arom.,s). ESI-HRMS:152.0262[M+H] + (Theoretical value [C8H7N1Cl1] + =152.0262).

[0289] Synthesis of TD712:

[0290] [ka]

[0291] A pear-shaped glass flask (10 mL) was charged with (6-bromopridin-2-yl)methanol (284 mg; 1.51 mmol; 1.0 equiv.) and a magnetic stirrer and protected with argon three times. Subsequently, solid CuI (14.5 mg; 76 μmol; 5.0 mol%) and [Pd(PPh3)2Cl2] (21 mg; 30 μmol; 2.0 mol%) were added and protected with argon three times. Dry THF (4 mL) was then added through a septum under a constant flow of argon, followed by the addition of ethynyltriisopropylsilane (370 μL; 1.65 mmol; 1.1 equiv.) and TEA (630 μL; 4.52 mmol; 3.0 equiv.), after which the mixture turned dark brown. The flask was left stirring under a septum (but without external argon) at room temperature for 16 h. The mixture was then transferred to a separatory funnel containing EtOAc (30 mL) and H2O (30 mL). After shaking, the dark (not completely homogeneous) organic layer was separated, and the aqueous layer was further extracted with EtOAc (5 x 25 mL). The combined organic layers were filtered through a cotton plug, and the filtrate was further dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The resulting dark brown oily residue was purified by flash chromatography (120 g SiO2, 100% DCM to 10% EtOAc in DCM). The combined product-containing fractions were evaporated to dryness and further co-evaporated once with DCM. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a yellow oil. Yield: 387 mg (89%; 1 step; based on (6-bromopyridin-2-yl)methanol). NMR (DMSO-d6): 1 Hδ H 0.92-1.26(i-Pr,m,21H);4.53(CH2,d,2H, 3 J HH =6);5.48(OH,t,1H, 3 J HH =6);7.40(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.47(arom.,dd,1H, 3 J HH =8; 4 JHH =1);7.80(arom.,t,1H, 3 J HH =8). 13 C{ 1 H} δ C 10.7(i-Pr,s);18.5(i-Pr,s);64.0(CH2,s);89.4 and 106.6(C≡C;2×s);120.2(ar om.,s);125.8(arom.,s);137.2(arom.,s);140.9(arom.,s);162.7(arom.,s). ESI-HRMS:290.1932[M+H] + (Theoretical value [C 17 H 28 N1O1Si1] + =290.1935).

[0292] Synthesis of TD723:

[0293] [ka]

[0294] In a round-bottom glass flask (25 mL), TD712 (384 g; 1.33 mmol; 1.0 equiv.) was dissolved in DCM (5 mL). A freshly prepared solution of SOCl2 (200 μL; 2.75 mmol; 2.1 equiv.) in DCM (1 mL) was then added, and the open flask was stirred at room temperature for 1 h. A dilute aqueous solution of NaHCO3 (10 mL) was then added, and the resulting biphasic mixture was vigorously stirred at room temperature for an additional 1 h, during which time gas bubbles were slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 × 10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. A brown residue ( 1The resulting oil (already pure by HNMR) was purified by flash chromatography (120 g SiO2, 20% DCM in PE to 30% DCM in PE) to remove a strongly colored impurity. The combined product fractions were evaporated to dryness and co-evaporated once with DCM. The remaining colorless oil was further dried overnight under high vacuum to give the product in the form of the free base as a colorless solid. Yield: 356 mg (87%; 1 step; based on TD712). NMR (DMSO-d6): 1 Hδ H 0.91-1.28(i-Pr,m,21H);4.78(CH2,s);7.52(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.57(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.86(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 308.1593 [M+H] + (Theoretical value [C 17 H 27 N1Cl1Si1] + =308.1596).

[0295] Synthesis of TD530:

[0296] [ka]

[0297] A pear-shaped glass flask (50 mL) was charged with (6-bromopyridin-2-yl)methanol (1.00 g; 5.35 mmol; 1.0 equiv.), N-Boc-propargylamine (1.00 g; 6.44 mmol; 1.2 equiv.), and a magnetic stirrer and protected with argon three times. Subsequently, solid CuI (102 mg; 536 μmol; 10.0 mol%) and [Pd(PPh3)2Cl2] (187 mg; 266 μmol; 5.0 mol%) were added and protected with argon three times. Dry THF (27 mL) was added through a septum under a constant flow of argon. After the addition of TEA (2.24 mL; 16.1 mmol; 3.0 equiv.), the mixture turned dark brown. The flask was left stirring at room temperature for 18 hours under a septum (but without external argon). The mixture was then transferred to a separatory funnel containing EtOAc (30 mL) and H2O (50 mL). After shaking, the dark (not completely homogeneous) organic layer was separated, and the aqueous layer was further extracted with EtOAc (3 x 30 mL). The combined organic layers were filtered through a cotton plug, and the filtrate was further dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The resulting dark brown oily residue was purified by flash chromatography (220 g SiO2, 100% DCM to 100% EtOAc). The combined product-containing fractions were evaporated to dryness and further co-evaporated once with DCM. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a yellow oil. Yield: 610 mg (43%; 1 step; based on (6-bromopyridin-2-yl)methanol). NMR (DMSO-d6): 1 Hδ H 1.40(CH3,s,9H);3.99(CH2,d,2H, 3 J HH =6);4.51(CH2,d,2H, 3 J HH =6);5.47(OH,t,1H, 3 J HH =6);7.32(arom,dd,1H, 3 J HH =8; 4 J HH =1);7.40(NH,t,1H, 3 J HH=6);7.44(arom.,dd,1H,J 3 HH =8; 4 J HH =1);7.79(arom.,t,1H, 3 J HH =8). 13 C{ 1 H} δ C 28.2(CH3,s);29.9(CH2,s);64.0(CH2,s);78.4(C-CH3,s);81.3 and 86.9(C≡C;2×s);119.8(arom.,s);125. 0(arom.,s);137.2(arom.,s);141.1(arom.,s);155.3(CO,s);162.5(arom.,s).ESI-HRMS:263.1390[M+H] + (Theoretical value [C 14 H 19 N2O3] + =263.1390).

[0298] Synthesis of TD538:

[0299] [ka]

[0300] In a round-bottom glass flask (250 mL), TD530 (568 mg; 2.17 mmol; 1.0 equiv.) was dissolved in DCM (20 mL), followed by the addition of TEA (900 μL; 6.46 mmol; 3.0 equiv.). Next, a freshly prepared solution of MsCl (355 μL; 4.33 mmol; 2.0 equiv.) in DCM (5 mL) was added. The resulting solution was stirred at room temperature for 20 min. The mixture was then diluted with DCM (50 mL), followed by the addition of a dilute aqueous solution of NaHCO3 (50 mL). The resulting biphasic mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. ESI-MS (LC-MS): 341.1 [M+H] + (Theoretical value [C 15 H 21N2O5S1] + =341.1). All of the TD538 obtained in this way was used directly for TD539 without further purification or characterization.

[0301] Synthesis of TD1045:

[0302] [ka]

[0303] A pear-shaped glass flask (25 mL) was charged with (6-bromopyridin-2-yl)methanol (500 mg; 2.67 mmol; 1.0 equiv.), 1,1-dimethylpropargylamine (420 μL; 3.99 mmol; 1.5 equiv.), and a magnetic stirrer, and the flask was then protected with argon three times. Subsequently, solid CuI (21 mg; 110 μmol; 4.0 mol%) and [Pd(PPh3)2Cl2] (19 mg; 27 μmol; 1.0 mol%) were added, and the flask was then protected with argon three times. Under a constant flow of argon, dry THF (10 mL) was added through a septum, followed by TEA (1.10 mL; 7.89 mmol; 3.0 equiv.), after which the mixture turned dark brown. The flask was left stirring at room temperature for 18 hours under a septum (but without external argon). The mixture was filtered through a syringe microfilter (PTFE) and the solid phase was further washed with MeCN. The filtrate was evaporated to dryness and the resulting dark brown solid residue was purified by flash chromatography (120 g SiO2, 100% EtOAc to 40% EtOAc in MeOH). The combined fractions containing the product were evaporated to dryness and further co-evaporated once with DCM. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a yellow solidified oil. Yield: 283 mg (56%; 1 step; based on (6-bromopyridin-2-yl)methanol). NMR (DMSO-d6): 1 Hδ H 1.38(CH3,s,6H);3.17(NH2,s,2H);4.51(CH2,s,2H);5.44(OH,bs,1H);7.26(arom.,dd,1H, 3 J HH =8;4 J HH =1);7.40(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.76(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 191.1180 [M+H] + (Theoretical value [C 11 H 15 N2O1] + =191.1180).

[0304] Synthesis of TD1048:

[0305] [ka]

[0306] A pear-shaped glass flask (25 mL) was charged with TD1045 (280 mg; 1.47 mmol; 1.0 equiv.), followed by the addition of MeCN (10 mL) and a solution of BocO (2.0 m in dry THF; 1.0 mL; 2.00 mmol; 1.4 equiv.). The resulting mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness, and the resulting yellow residue was purified by flash chromatography (120 g SiO, 100% DCM to 100% EtOAc). The combined product-containing fractions were evaporated to dryness and co-evaporated once with DCM. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a colorless solid. Yield: 242 mg (57%; 1 step; based on TD1045). NMR (DMSO-d6): 1 Hδ H 1.40(CH3,s,9H),1.53(CH3,s,6H);4.51(CH2,d,2H, 3 J HH =4);5.44(OH,t,1H, 3 J HH =4);7.12(NH,bs,1H);7.25(arom,dd,1H, 3 J HH =8; 4 J HH=1);7.41(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.77(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 291.1704 [M+H] + (Theoretical value [C 16 H 23 N2O3] + =291.1703).

[0307] Synthesis of TD1050:

[0308] [ka]

[0309] In a glass vial (20 mL), TD1048 (19 mg; 65 μmol; 1.0 equiv.) was dissolved in DCM (3 mL) and TEA (28 μL; 201 μmol; 3.1 equiv.) was added. Next, a freshly prepared solution of MsCl (10 μL; 129 μmol; 2.0 equiv.) in DCM (1 mL) was added. The resulting solution was stirred at room temperature for 20 min. The mixture was then diluted with DCM (15 mL), followed by the addition of a dilute aqueous solution of NaHCO3 (15 mL). The resulting biphasic mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 20 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. ESI-MS (LC-MS): 369.1 [M+H] + (Theoretical value [C 17 H 25 N2O5S1] + =369.1). All TD1050 thus obtained was used directly for TD1054 (and similarly for TD1105) without further purification or characterization.

[0310] Synthesis of TD966:

[0311] [ka]

[0312] A pear-shaped glass flask (100 mL) was charged with (6-bromopyridin-2-yl)methanol (1.00 g; 5.35 mmol; 1.0 equiv.), N-Boc-4-ethynylpiperidine (1.23 g; 5.88 mmol; 1.1 equiv.), and a magnetic stirrer, and the flask was protected with argon three times. Subsequently, solid CuI (41 mg; 215 μmol; 4.0 mol%) and [Pd(PPh3)2Cl2] (75 mg; 107 μmol; 2.0 mol%) were added, and the flask was protected with argon three times. Under a constant flow of argon, dry THF (40 mL) was added through a septum, followed by TEA (2.20 mL; 15.7 mmol; 3.0 equiv.). The mixture turned dark brown. The flask was left stirring at room temperature for 2 days under a septum (but without external argon). The mixture was then transferred to a separatory funnel containing EtOAc (30 mL) and H2O (50 mL). After shaking, the dark (not completely homogeneous) organic layer was separated, and the aqueous layer was further extracted with EtOAc (3 x 30 mL). The combined organic layers were filtered through a cotton plug, and the filtrate was further dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The resulting dark brown oily residue was purified by flash chromatography (220 g SiO2, 100% DCM to 100% EtOAc). The combined product-containing fractions were evaporated to dryness and further co-evaporated once with DCM. The residue was further dried overnight under high vacuum to give the free base form of the product as a yellow oil. Yield: 1.47 g (87%; 1 step; based on (6-bromopyridin-2-yl)methanol). NMR (DMSO-d6): 1 Hδ H 1.40(CH3,s,9H);1.44-1.57(CH2,m,2H);1.75-1.89(CH2,m,2H);2.87(CH-C≡Ctt,1H, 3 J HH =8, 3 J HH =4);3.01-3.17(CH2,m,2H);3.59-3.73(CH2,m,2H);4.51(CH2-O,d,2H, 3 J HH=6);5.44(OH,t,1H, 3 J HH =6);7.32(arom,dd,1H, 3 J HH =8; 4 J HH =1);7.41(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.76(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 317.1860 [M+H] + (Theoretical value [C 18 H 25 N2O3] + =317.1860).

[0313] Synthesis of TD1117:

[0314] [ka]

[0315] In a glass vial (20 mL), TD952 (50.0 mg; 158 μmol; 1.0 equiv) was dissolved in DCM (6 mL), and TEA (66 μL; 473 μmol; 3.0 equiv) and neat MsCl (25 μL; 323 μmol; 2.0 equiv) were added. The resulting solution was stirred at room temperature for 20 min, after which a solution of DCM (10 mL) and diluted aqueous NaHCO3 (10 mL) was added. The resulting biphasic mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 × 10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. ESI-MS (LC-MS): 395.2 [M+H] + (Theoretical value [C 19 H 27 N2O5S1] + =395.2). All TD1117 thus obtained was used directly for TD1118 without further purification or characterization.

[0316] Synthesis of TD936:

[0317] [ka]

[0318] A glass vial (4 mL) containing (6-bromopyridin-2-yl)methanol (135 mg; 722 μmol; 1.0 equiv.), CuI (5.5 mg; 29 μmol; 4.0 mol%), [Pd(PPh3)2Cl2] (10.0 mg; 14 μmol; 2.0 mol%), and a magnetic stirrer was placed in a protective atmosphere with argon three times. Dry THF (3 mL) was then added through a septum under a constant flow of argon, followed by phenylacetylene (95 μL; 865 μmol; 1.2 equiv.) and TEA (300 μL; 2.15 mmol; 3.0 equiv.). The mixture turned dark brown. The vial was left stirring at room temperature for 16 h under a septum (but without external argon). The mixture was filtered through a PTFE syringe microfilter and washed with MeCN. The filtrate was evaporated to dryness, redissolved in MeCN (3 mL), and directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were neutralized with dilute aqueous NaHCO3 and evaporated to dryness. The residue was dissolved in a mixture of DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After brief shaking, the bottom phase was separated, and the aqueous phase was further extracted with DCM (4 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residual solid was briefly dried under high vacuum to give the product in the form of the free base as a yellow oil. Yield: 141 mg (93%; 1 step; based on (6-bromopyridin-2-yl)methanol). NMR (DMSO-d6): 1 Hδ H 4.57(CH2,d,2H, 3 J HH =6);5.50(OH,t,1H, 3 J HH =6);7.42-7.54(arom.,Ph,m,2+3H);7.57-7.64(Ph,m,2H);7.85(arom.,t,1H, 3 JHH = 8). ESI-HRMS: 210.0913 [M+H] + (Theoretical value [C 14 H 12 N1O1] + =210.0913).

[0319] Synthesis of TD939:

[0320] [ka]

[0321] In a glass vial (20 mL), TD936 (22 mg; 105 μmol; 1.0 equiv.) was dissolved in DCM (6 mL), and TEA (44 μL; 316 μmol; 3.0 equiv.) and neat MsCl (16 μL; 207 μmol; 2.0 equiv.) were added. The resulting solution was stirred at room temperature for 20 min, and then a dilute aqueous solution of NaHCO3 (5 mL) was added. The resulting biphasic mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. ESI-MS (LC-MS): 288.1 [M+H] + (Theoretical value [C 15 H 14 N1O3S1] + =288.1). All of the TD939 obtained in this manner was used directly for TD944 without further purification or characterization.

[0322] Synthesis of TD937:

[0323] [ka]

[0324] A glass vial (4 mL) was charged with (6-bromopyridin-2-yl)methanol (135 mg; 722 μmol; 1.0 equiv.), CuI (5.5 mg; 29 μmol; 4.0 mol%), [Pd(PPh3)2Cl2] (10.0 mg; 14 μmol; 2.0 mol%), and a magnetic stirrer, and was protected with argon three times. Then, under a constant flow of argon, dry THF (3 mL) was added through a septum, followed by cyclopropylacetylene (75 μL; 886 μmol; 1.2 equiv.) and TEA (300 μL; 2.15 mmol; 3.0 equiv.), after which the mixture turned dark brown. The vial was left stirring at room temperature under a septum (but without external argon) for 16 h. The mixture was filtered through a PTFE syringe microfilter and washed with MeCN. The filtrate was evaporated to dryness, redissolved in MeCN (3 mL), and directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were neutralized with dilute aqueous NaHCO3 and evaporated to dryness. The residue was dissolved in a mixture of DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After brief shaking, the bottom phase was separated, and the aqueous phase was further extracted with DCM (4 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residual solid was briefly dried under high vacuum to give the product in the form of the free base as a yellow oil. Yield: 98 mg (78%; 1 step; based on (6-bromopyridin-2-yl)methanol). NMR (DMSO-d6): 1 Hδ H 0.69-0.83(CH2-CH,m,2H);0.83-0.99(CH2-CH,m,2H);1.56(CH,tt, 3 J HH =8, 3 J HH =5);4.49(CH2,d,2H, 3 J HH =6);5.42(OH,t,1H, 3 J HH =6);7.27(arom,dd,1H, 3 J HH =8, 4 J HH =1);7.38(arom.,dd,1H,3 J HH =8, 4 J HH =1);7.74(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 174.0913 [M+H] + (Theoretical value [C 11 H 12 N1O1] + =174.0913).

[0325] Synthesis of TD940:

[0326] [ka]

[0327] In a glass vial (20 mL), TD937 (26 mg; 150 μmol; 1.0 equiv.) was dissolved in DCM (9 mL), and TEA (63 μL; 452 μmol; 3.0 equiv.) and neat MsCl (23 μL; 297 μmol; 2.0 equiv.) were added. The resulting solution was stirred at room temperature for 20 min, and then a dilute aqueous solution of NaHCO3 (5 mL) was added. The resulting biphasic mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. ESI-MS (LC-MS): 252.0 [M+H] + (Theoretical value [C 12 H 14 N1O3S1] + =252.1). All of the TD940 obtained in this way was used directly for TD943 without further purification or characterization.

[0328] Synthesis of TD952:

[0329] [ka]

[0330] A glass vial (4 mL) was charged with (6-bromopyridin-2-yl)methanol (150 mg; 802 μmol; 1.0 equiv.), CuI (6.1 mg; 32 μmol; 4.0 mol%), [Pd(PPh3)2Cl2] (11.5 mg; 16 μmol; 2.0 mol%), and a magnetic stirrer, and was protected with argon three times. Then, under a constant flow of argon, dry THF (3 mL) was added through a septum, followed by tert-butylacetylene (200 μL; 1.62 mmol; 2.0 equiv.) and TEA (340 μL; 2.44 mmol; 3.0 equiv.), after which the mixture turned dark brown. The vial was left stirring under a septum (but without external argon) for 24 h at room temperature. The mixture was filtered through a PTFE syringe microfilter and washed with MeCN. The filtrate was evaporated to dryness, redissolved in MeCN (3 mL), and directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were neutralized with dilute aqueous NaHCO3 and evaporated to dryness. The residue was dissolved in a mixture of DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After brief shaking, the bottom phase was separated, and the aqueous phase was further extracted with DCM (4 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residual solid was briefly dried under high vacuum to give the product in the form of the free base as a yellow oil. Yield: 146 mg (96%; 1 step; based on (6-bromopyridin-2-yl)methanol). NMR (DMSO-d6): 1 Hδ H 1.30(CH3,s,9H); 4.51(CH2,d,2H, 3 J HH =6);5.43(OH,t,1H, 3 J HH =6);7.26(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.39(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.74(arom.,t,1H, 3 JHH = 8). ESI-HRMS: 190.1227 [M+H] + (Theoretical value [C 12 H 16 N1O1] + =190.1226).

[0331] Synthesis of TD956:

[0332] [ka]

[0333] In a glass vial (20 mL), TD952 (8 mg; 42 μmol; 1.0 equiv.) was dissolved in DCM (3 mL), and TEA (18 μL; 129 μmol; 3.0 equiv.) and neat MsCl (6.5 μL; 84 μmol; 2.0 equiv.) were added. The resulting solution was stirred at room temperature for 20 min, after which DCM (5 mL) and diluted aqueous NaHCO3 (5 mL) were added. The resulting biphasic mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 × 10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. ESI-MS (LC-MS): 268.1 [M+H] + (Theoretical value [C 13 H 18 N1O3S1] + =268.1). All of the TD956 obtained in this manner was used directly for TD959 without further purification or characterization.

[0334] Synthesis of TD965:

[0335] [ka]

[0336] A glass vial (4 mL) was charged with (6-bromopyridin-2-yl)methanol (53 mg; 283 μmol; 1.0 equiv.), 1-ethynyladamantane (50 mg; 312 μmol; 1.1 equiv.), CuI (2.7 mg; 14 μmol; 5.0 mol%), [Pd(PPh3)2Cl2] (5.0 mg; 7 μmol; 2.5 mol%), and a magnetic stirrer, and was protected with argon three times. Under a constant flow of argon, dry THF (1.5 mL) was added through a septum, followed by TEA (120 μL; 861 μmol; 3.0 equiv.). The vial was left stirring under a septum (but without external argon) for 2 days. The mixture was filtered through a PTFE syringe microfilter and washed with MeCN. The filtrate was evaporated to dryness, redissolved in MeCN (3 mL), and directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were neutralized with dilute aqueous NaHCO3 and evaporated to dryness. The residue was dissolved in a mixture of DCM (25 mL) and HO (25 mL) and transferred to a separatory funnel. After brief shaking, the bottom phase was separated, and the aqueous phase was further extracted with DCM (4 × 10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residual solid was briefly dried under high vacuum to give the product in the form of the free base as a colorless oil. Yield: 67 mg (89%; 1 step; based on (6-bromopyridin-2-yl)methanol). NMR (DMSO-d6): 1 Hδ H 1.61-1.76(Adm.,m,6H);1.82-1.93(Adm.,m,6H);1.94-2.00(Adm.,m,3H);4.50(CH2,d,2H, 3 J HH =6);5.43(OH,t,1H, 3 J HH =6);7.25(arom,dd,1H, 3 J HH =8, 4 J HH =1);7.38(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.74(arom.,t,1H,J3 HH = 8). ESI-HRMS: 268.1694 [M+H] + (Theoretical value [C 18 H 22 N1O1] + =268.1696).

[0337] Synthesis of TD990:

[0338] [ka]

[0339] In a glass vial (20 mL), TD965 (21 mg; 79 μmol; 1.0 equiv.) was dissolved in DCM (3 mL), and TEA (33 μL; 237 μmol; 3.0 equiv.) and neat MsCl (12 μL; 155 μmol; 2.0 equiv.) were added. The resulting solution was stirred at room temperature for 20 min, after which DCM (10 mL) and a dilute aqueous solution of NaHCO3 (10 mL) were added. The resulting biphasic mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 15 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. ESI-MS (LC-MS): 346.1 [M+H] + (Theoretical value [C 19 H 24 N1O3S1] + =346.1). All TD990 thus obtained was used directly for TD992 without further purification or characterization.

[0340] Synthesis of TD1194:

[0341] [ka]

[0342] In a glass vial (4 mL), TD558 (110 mg; 726 μmol; 1.0 equiv.) was dissolved in MeCN (3.3 mL), NIS (180 mg; 800 μmol; 1.1 equiv.) and AcOH (50 μL; 875 μmol; 1.2 equiv.) were added, and the resulting mixture was stirred at 80 °C for 3 h. The mixture was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and diluted with DCM (50 mL). The resulting biphasic mixture was transferred to a separatory funnel, and diluted aqueous NaHCO3 and aqueous NaSO3 solutions were added. After brief shaking, the bottom phase was separated, and the aqueous layer was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous NaSO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was briefly dried under high vacuum to give the product in free base form as a pale red solid. Yield: 61.5 mg (31%; 1 step; based on TD558). NMR (DMSO-d): 1 Hδ H 4.74(CH2,s,2H);7.48(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.54(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.85(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 277.9230 [M+H] + (Theoretical value [C8H6N1I1Cl1] + =277.9228).

[0343] Synthesis of TD1178:

[0344] [ka]

[0345] A glass vial (20 mL) was charged with CuI (60.0 mg; 315 μmol; 40 mol%), Phen (114 mg; 633 μmol; 80.0 mol%), KHCO3 (159 mg; 1.59 mmol; 2.0 equiv.), and a magnetic stirrer, and the vial was protected with argon three times. Next, under a constant flow of argon, a solution of TD558 (120 mg; 792 μmol; 1.0 equiv.) and Togni I (288 mg; 872 μmol; 1.1 equiv.) in dry DCM (8 mL) was added through a septum, and the resulting mixture was stirred at room temperature for 2 days. The mixture was filtered through a syringe microfilter (PTFE), and the solid phase was further washed with DCM. The filtrate was evaporated to dryness, and the residue was resuspended in MeCN (3 mL). The mixture was filtered again through a syringe microfilter (PTFE), and the filtrate was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined and diluted with DCM (50 mL). The resulting biphasic mixture was transferred to a separatory funnel, and a dilute aqueous solution of NaHCO3 was added. After brief shaking, the bottom layer was separated, and the aqueous layer was further extracted with DCM (3 x 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The resulting yellow oil was briefly dried under high vacuum (until the oil crystallized) to give the product in the form of the free base as a pale yellow solid (some of the product sublimed to the top of the flask as colorless crystals). Yield: 48.5 mg (28%; 1 step; TD558 reference). NMR (DMSO-d6): 1 Hδ H 4.83(CH2,s,2H);7.78(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.88(arom.,dd,1H, 3 J HH =8, 4 J HH =1);8.04(arom.,t,1H, 3 J HH =8). 19 F{ 1 H}δ F -49.2(s).ESI-HRMS:220.0140[M+H] +(Theoretical value [C9H6N1F3Cl1] + =220.0135).

[0346] Synthesis of TD797:

[0347] [ka]

[0348] In a round-bottom glass flask (50 mL), methyl 4,6-dibromopicolinate (200 mg; 678 μmol; 1.0 equiv.) was dissolved in a mixture of THF (4 mL) and MeOH (2 mL). To the resulting slightly yellow solution, solid NaBH4 (205 mg; 5.42 mmol; 8.0 equiv.) was added in portions (over 10 min without the stopper) during which time hydrogen gas evolved vigorously and the mixture turned nearly colorless. The mixture was transferred to a separatory funnel and diluted with DCM (100 mL) and H2O (100 mL). After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of a free base as a white solid. Yield: 179 mg (99%; 1 step; based on methyl 4,6-dibromopicolinate). NMR (DMSO-d): 1 Hδ H 4.53(CH2,s,2H);5.68(OH,bs,1H);7.67(arom.,m,1H);7.89(arom.,m,1H). ESI-HRMS:265.8809[M+H] + (Theoretical value [C6H6N1O1Br2] + =265.8811).

[0349] Synthesis of TD804:

[0350] [ka]

[0351] A glass vial (4 mL) containing TD797 (100 mg; 375 μmol; 1.0 equiv.) and a magnetic stirrer was placed in a 4 mL vial and protected with argon three times. Subsequently, solid CuI (4.3 mg; 23 μmol; 6.0 mol%) and [Pd(PPh3)2Cl2] (8.0 mg; 11 μmol; 3.0 mol%) were added and protected with argon three times. Next, dry THF (1.5 mL) was added through a septum under a constant flow of argon, followed by ethynyltrimethylsilane (160 μL; 1.16 mmol; 3.1 equiv.) and TEA (160 μL; 1.15 mmol; 3.1 equiv.), after which the mixture turned dark brown. The flask was left stirring under a septum (but without external argon) at ambient temperature for 16 h. The mixture was then transferred to a separatory funnel containing EtOAc (50 mL) and H2O (50 mL). After shaking, the dark organic layer was separated and the aqueous layer was further extracted with EtOAc (3 x 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The resulting dark brown oily residue was purified by flash chromatography (120 g SiO2, 100% DCM to 20% EtOAc in DCM). The combined product-containing fractions were evaporated to dryness and co-evaporated once with DCM. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a dark brown oil. Yield: 99 mg (88%; 1 step; based on TD797). NMR (DMSO-d6): 1 Hδ H 0.24(CH3,s,9H);0.25(CH3,s,9H);4.51(CH2,d,2H, 3 J HH =6);5.53(OH,t,1H, 3 J HH =6);7.40(arom.,d,1H, 4 J HH =2);7.42(arom.,d,1H, 4 J HH = 2). ESI-HRMS: 302.1389 [M+H] + (Theoretical value [C 16 H 24 N1O1Si2] + =302.1391).

[0352] Synthesis of TD806:

[0353] [ka]

[0354] In a pear-shaped glass flask (50 mL), TD804 (98 mg; 325 μmol; 1.0 equiv.) was dissolved in DCM (3 mL). Next, a freshly prepared solution of SOCl2 (47 μL; 647 μmol; 2.0 equiv.) in DCM (1 mL) was added, and the open flask was stirred at room temperature for 1 h. The reaction mixture was diluted with DCM (16 mL), and dilute aqueous NaHCO3 (20 mL) was added. The resulting biphasic mixture was vigorously stirred at room temperature for an additional 1 h, during which time gas bubbles slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 20 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in the form of the free base as a brown oil. Yield: 102 mg (98%; 1 step; based on TD804). NMR (DMSO-d6): 1 Hδ H 0.25(CH3,s,18H);4.74(CH2,s,2H);7.53(arom.,d,1H, 4 J HH =2);7.60(arom.,d,1H, 4 J HH = 2). ESI-HRMS: 320.1051 [M+H] + (Theoretical value [C 16 H 23 N1O1Cl1Si2] + =320.1052).

[0355] Synthesis of TD807:

[0356] [ka]

[0357] In a round-bottom glass flask (50 mL), TD806 (101 mg; 316 μmol; 1.0 equiv.) was dissolved in MeCN (5 mL). Next, a freshly prepared solution of KF (46.4 mg; 800 μmol; 2.5 equiv.) in HO (800 μL) was added, and the flask was stirred at room temperature for 5 h. The mixture was concentrated to less than 1 mL and diluted with DCM (50 mL) and dilute aqueous NaHCO (50 mL). The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous NaSO, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a brown solid. Yield: 48 mg (87%; 1 step; based on TD806). NMR (DMSO-d): 1 Hδ H 4.48(C≡CH,s,1H);4.75(C≡CH,s,1H);4.76(CH2,s,2H);7.61(arom.,d,1H, 4 J HH =1);7.65(arom.,d,1H, 4 J HH = 1). ESI-HRMS: 176.0267 [M+H] + (Theoretical value [C 10 H7N1Cl1] + =176.0267).

[0358] Synthesis of TD662:

[0359] [ka]

[0360] In a glass vial (4 mL), dipropargylamine (116 μL; 1.12 mmol; 1.0 equiv.) was dissolved in MeCN (2.5 mL), followed by the addition of neat 1,2-dibromoethane (965 μL; 11.2 mmol; 10 equiv.) and NaHCO3 (113 mg; 1.35 mmol; 1.2 equiv.). The resulting mixture was stirred at 80 °C for 2 days. The mixture was then diluted with DCM (50 mL), followed by the addition of a dilute aqueous solution of NaHCO3 (50 mL). The resulting biphasic mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further purified by column chromatography (SiO2; 25 g; DCM). The combined product-containing fractions were evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in free base form as a nearly colorless oil. Yield: 69 mg (31%; 1 step; based on dipropargylamine). NMR (DMSO-d6): 1 Hδ H 2.86(CH2,t,2H, 3 J HH =7);3.21(C≡CH,t,2H, 4 J HH =2);3.21(CH2,t,4H, 4 J HH =2);3.55(CH2,t,2H, 3 J HH =7). 13 C{ 1 H} δ C 30.3 (CH2,s); 41.6 (CH2,s); 53.9 (CH2,s); 76.4 and 78.5 (C≡CH,2×s). ESI-HRMS: 200.0070 [M+H] + (Theoretical value [C8H 11 N1Br1] + =200.0070).

[0361] Example 2: Synthesis of azide pendant arm intermediate Synthesis of TD406:

[0362] [ka]

[0363] In a round-bottom glass flask (250 mL), 2,6-bis(chloromethyl)pyridine (6.17 g; 35.0 mmol; 1.2 equiv.) was dissolved in MeCN (80 mL), followed by the addition of solid NaN (1.90 g; 29.2 mmol; 1.0 equiv.) (90 g; 29.2 mmol; 1.0 equiv.) and anhydrous KCO (4.04 g; 29.2 mmol; 1.0 equiv.), and the resulting suspension was stirred at 50 °C for 5 days. The mixture was filtered through a glass frit S3, and the solid residue was washed with MeCN. The filtrate was evaporated to dryness, and the orange oily residue was purified by column chromatography (SiO, 30% PE in DCM to 100% DCM). The product-containing fractions were combined and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in free base form as a faint yellow oil. Yield: 2.62 g (49%; 1 step; based on NaN). Recovered amount: 2.21 g of 2,6-bis(chloromethyl)pyridine (36% of the initial amount). NMR (DMSO-d): 1 Hδ H 4.53(CH2,s,2H);4.78(CH2,s,2H);7.40(arom.,d,1H, 3 J HH =8; 4 J HH =1);7.52(arom.,d,1H, 3 J HH =8; 4 J HH =1);7.89(arom.,t,1H, 3 J HH =8). 13 C{ 1 H} δ C 46.6(CH2,s);54.2(CH2,s);121.9(arom.,s);122.6(arom.,s);138.5(arom.,s);155.7(arom.,s);156.3(arom.,s). ESI-HRMS:183.0433[M+H] + (Theoretical value [C7H8N4Cl1] + =183.0432).

[0364] Synthesis of TD595:

[0365] [ka]

[0366] In a glass vial (20 mL), TD406 (30 mg; 162 μmol; 1.0 equiv.) was dissolved in DCM (5 mL), followed by the addition of a freshly prepared solution of MCPBA (77%; 72 mg; 320 μmol; 2.0 equiv.) in DCM (1 mL). The resulting solution was stirred at room temperature for 3 h. The mixture was diluted with DCM (20 mL) and dilute aqueous NaHCO3 (25 mL) and transferred to a separatory funnel. After shaking, the bottom layer was separated, and the aqueous layer was further extracted with DCM (3 × 20 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. ESI-MS (LC-MS): 199.0 [M+H] + (Theoretical value [C7H8N4O1Cl1] + =199.0). All TD595 thus obtained was used directly for TD596 without further purification or characterization.

[0367] Synthesis of TD726:

[0368] [ka]

[0369] In a round-bottom glass flask (1000 mL), dimethyl 4-chloropyridine-2,6-dicarboxylate (15.0 g; 65.3 mmol; 1.0 equiv.) was slowly dissolved in a mixture of THF (540 mL) and MeOH (120 mL). The resulting slightly yellow solution was cooled in an ice bath (~5 °C), followed by the addition of solid NaBH4 (12.4 g; 328 mmol; 5.0 equiv.) in portions (over 1 h without the stopper). During this time, hydrogen gas evolved vigorously, and the mixture changed color from red to orange and finally to yellow. After the addition, the flask was allowed to warm to room temperature and stirred at room temperature for 16 h. The resulting slightly yellowish milky solution was filtered through a glass frit (S3), and the filtrate was evaporated to dryness and co-evaporated with DCM (as a suspension). The residue was dissolved in boiling H2O (~600 mL), and the resulting highly alkaline solution was continuously extracted with DCM overnight. The organic layer (containing the partially crystallized product) was evaporated to dryness. The residue was mechanically pulverized and further dried under high vacuum (to constant mass) to give the product in the form of the free base as a nearly colorless microcrystalline powder. Yield: 10.84 g (96%; 1 step; based on dimethyl 4-chloropyridine-2,6-dicarboxylate). NMR (DMSO-d6): 1 Hδ H 4.53(CH2,d,4H, 3 J HH =6);5.54(OH,t,2H, 3 J HH =6);7.36(arom.,s,2H). 13 C{ 1 H} δ C 63.7(CH2,s);118.0(arom.,s);144.0(arom.,s);163.5(arom.,s). ESI-HRMS:174.0317[M+H] + (Theoretical value [C7H9N1O2Cl1] + =174.0316). EA(C7H8N1O2Cl1,M R =173.6):C48.4(48.5);H4.7(4.5);N7.7(8.1);Cl20.4(21.2).

[0370] Synthesis of TD759:

[0371] [ka]

[0372] In a round-bottom glass flask (100 mL), TD726 (626 mg; 3.61 mmol; 1.0 equiv.) was suspended in DCM (20 mL). A freshly prepared solution of SOCl2 (780 μL; 10.7 mmol; 3.0 equiv.) in DCM (10 mL) was then added, and the open flask was stirred at room temperature for 90 min to produce a clear solution. Dilute aqueous NaHCO3 (40 mL) was then added, and the resulting biphasic mixture was vigorously stirred at room temperature for an additional 1 h, during which time gas bubbles slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The resulting crystallized residue was triturated and further dried under high vacuum overnight to give the product in the form of the free base as a nearly colorless microcrystalline powder. Yield: 711 mg (94%; 1 step; based on TD726). NMR (DMSO-d): 1 Hδ H 4.79(CH2,s,4H);7.70(arom.,s,2H). 13 C{ 1 H} δ C 45.7(CH2,s);122.8(arom.,s);144.4(arom.,s);158.2(arom.,s). ESI-HRMS:209.9638[M+H] + (Theoretical value [C7H7N1Cl3] + =209.9639).

[0373] Synthesis of TD760:

[0374] [ka]

[0375] In a glass vial (20 mL), TD759 (708 mg; 3.36 mmol; 1.3 equiv.) was dissolved in MeCN (20 mL). Solid NaN (168 mg; 2.58 mmol; 1.0 equiv.) and dry KCO (360 mg; 2.61 mmol; 1.0 equiv.) were subsequently added, and the resulting suspension was stirred at 80 °C for 24 h. The mixture was filtered through a syringe microfilter (PTFE), and the solid residue was washed with MeCN. The filtrate was evaporated to dryness, and the yellow oily residue was purified by column chromatography (SiO, 40% PE in DCM to 10% PE in DCM). The product-containing fractions were combined and evaporated to dryness. The residue was further dried under high vacuum for 1 h, affording the product in the form of the free base as a faint yellow oil. Yield: 323 mg (58%; 1 step; based on NaN). Recovery: 218 mg of TD759 (31% of the initial amount). NMR (DMSO-d): 1 Hδ H 4.56(CH2,s,2H);4.78(CH2,s,2H);7.58(arom.,d,1H, 4 J HH =2);7.69(arom.,d,1H, 4 J HH =8). 13 C{ 1 H} δ C 45.8(CH2,s);53.6(CH2,s);121.9(arom.,s);122.5(arom.,s);144.4(arom.,s);157.9(arom.,s);158.2(arom.,s). ESI-HRMS:217.0040[M+H] + (Theoretical value [C7H7N4Cl]2 + =217.0042).

[0376] Synthesis of TD1146:

[0377] [ka]

[0378] In a glass vial (20 mL), TD726 (495 mg; 2.85 mmol; 1.0 equiv.) was dissolved in DMF (11.0 mL; 143 mmol; 50 equiv.), followed by the addition of freshly ground KOH (1.6 g; 28.5 mmol; 10 equiv.). The resulting suspension was stirred in the presence of air at 100 °C for 3 days (the vial septum was punctured with two large needles). The mixture was then filtered through a syringe microfilter (PTFE; the solid was washed with DMF). The filtrate was evaporated to dryness and purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined and directly lyophilized to give the product as a white foam in the form of a trifluoroacetate salt. Yield: 187 mg (22%; 1 step; based on TD726). NMR (DMSO-d6): 1 Hδ H 3.18(CH3,s,6H);4.59(CH2,d,4H, 3 J HH =6);5.91(OH,t,2H, 3 J HH =6);6.84(arom.,s,2H).ESI-HRMS:183.1128[M+H] + (Theoretical value [C9H 15 N2O2] + =183.1128). EA(C9H 14 N2O2-1.0TFA,M R =296.2):C44.6(44.1);H5.1(4.9);N9.5(9.1);F19.2(18.2).

[0379] [ka]

[0380] Synthesis of TD1154: In a round-bottom glass flask (100 mL), TD1146 1.0TFA (173.3 mg; 585 μmol; 1.0 equiv.) was suspended in DCM (17 mL) and neat SOCl2 (780 μL; 2.34 mmol; 4.0 equiv.) was added. The resulting mixture was stirred at room temperature for 90 min, resulting in a clear solution. Next, a dilute aqueous solution of NaHCO3 (20 mL) was added, and the resulting biphasic mixture was vigorously stirred at room temperature for an additional 1 h, during which time gas bubbles slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in the form of the free base as a white solid. Yield: 105.6 mg (82%; 1 step; based on TD726). NMR (DMSO-d6): 1 Hδ H 2.98(CH3,s,6H);4.60(CH2,s,4H);6.73(arom.,s,2H).ESI-HRMS:219.0450[M+H] + (Theoretical value [C9H 13 N2Cl2] + =219.0450).

[0381] Synthesis of TD1163:

[0382] [ka]

[0383] In a glass vial (20 mL), TD1154 (103.9 mg; 474 μmol; 1.3 equiv.) was dissolved in MeCN (7 mL). Solid NaN (23.7 mg; 365 μmol; 1.0 equiv.) and dry KCO (50.3 mg; 364 μmol; 1.0 equiv.) were subsequently added, and the resulting suspension was stirred at 70 °C for 24 h. The mixture was filtered through a syringe microfilter (PTFE), and the solid residue was washed with MeCN. The filtrate was evaporated to dryness, and the residue was purified by column chromatography (SiO, 5% EtOAc in DCM to 10% EtOAc in DCM). The product-containing fractions were combined and evaporated to dryness. The residue was further dried under high vacuum for 1 h to give the product in the form of the free base as a colorless oil. Yield: 33.8 mg (41%; 1 step; based on NaN). Recovery: 11.0 mg of TD1154 (11% of the initial amount). NMR (DMSO-d): 1 Hδ H 2.99(CH3,s,6H);4.32(CH2,s,2H);4.60(CH2,s,2H);6.60(arom.,d,1H, 4 J HH =2);6.73(arom.,d,1H, 4 J HH = 2). ESI-HRMS: 226.0853 [M+H] + (Theoretical value [C9H 13 N5Cl1] + =226.0854).

[0384] Synthesis of TD1024:

[0385] [ka]

[0386] A glass vial (100 mL) was charged with CuI (70 mg; 0.37 mmol; 4.0 mol%), [Pd(PPh3)2Cl2] (125 mg; 0.18 mmol; 2.0 mol%), and a magnetic stirrer and protected with argon three times. Subsequently, a solution of dimethyl 4-iodopyridine-2,6-dicarboxylate (2.93 g; 9.13 mmol; 1.0 equiv.) in a mixture of dry THF (40 mL), dry toluene (40 mL), and dry DMF (2 mL) was added through a septum. After the addition of ethynyltriisopropylsilane (2.25 mL; 10.0 mmol; 1.1 equiv.) and TEA (3.80 mL; 27.3 mmol; 3.0 equiv.), the mixture turned pale red. The flask was left stirring at room temperature under a septum (but without external argon) for 20 h. The resulting dark mixture was filtered through a syringe microfilter (PTFE) and the filtrate was evaporated to dryness. The residue was dissolved in DCM (250 mL) and the resulting red solution was washed once with an aqueous solution of NaSO. The organic layer was separated, dried over NaSO, filtered through a glass frit (S3), and evaporated to dryness. The residue was purified by flash chromatography (220 g SiO, 100% DCM to 20% EtOAc in DCM). The combined product-containing fractions were evaporated to dryness and co-evaporated once with DCM. The residue was further dried overnight under high vacuum to give the crude product in the form of the free base as an orange oil. Yield: 3.16 g (92%; 1 step; based on dimethyl 4-iodopyridine-2,6-dicarboxylate). NMR (DMSO-d): 1 Hδ H 0.92-1.27(i-Pr,m,21H);3.92(CH3,s,6H);8.16(arom,s,2H). 13 C{ 1 H} δ C 10.6(i-Pr,s);18.4(i-Pr,s);52.9(CH3,s);99.0(C≡C-Si,s);102.5(C≡C-a rom.;s);129.3(arom.,s);132.7(arom.,s);148.4(arom.,s);164.0(CO,s). ESI-HRMS:376.1939[M+H] + (Theoretical value [C 20H 30 N1O4Si1] + =376.1939).

[0387] Synthesis of TD808:

[0388] [ka]

[0389] In a round-bottom glass flask (250 mL), TD1024 (3.15 g; 8.39 mmol; 1.0 equiv.) was dissolved in a mixture of THF (60 mL) and MeOH (30 mL). Solid NaBH4 (2.55 g; 67.4 mmol; 8 equiv.) was then added portionwise over 1 h (during which time vigorous hydrogen gas evolution occurred), causing the mixture to turn dark red. After the addition, the flask was further stirred at room temperature for 1 h. The mixture was evaporated to dryness. The residue was suspended in DCM (200 mL) and HO (200 mL) and transferred to a separatory funnel. After shaking, the bottom phase (as a suspension) was separated. The aqueous phase was further extracted with DCM (3 × 100 mL). The combined organic layers were diluted with EtOAc (500 mL). The resulting brown solution was dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in free base form as a light brown solid. Yield: 2.61 g (97%; 1 step; TD1024 reference). NMR (DMSO-d): 1 Hδ H 1.04-1.17(i-Pr,m,21H);4.51(CH2,d,4H, 3 J HH =6);5.48(OH,t,2H, 3 J HH =6);7.31(arom,s,2H). 13 C{ 1 H} δ C 10.6(i-Pr,s);18.5(i-Pr,s);63.9(CH2,s);94.6(C≡C-Si,s);105.1(C≡C-arom.;s);119.8(arom.,s);130.8(arom.,s);161.9(arom.,s). ESI-HRMS:320.2036[M+H]+ (Theoretical value [C 18 H 30 N1O2] + =320.2040).

[0390] Synthesis of TD815:

[0391] [ka]

[0392] In a round-bottom glass flask (250 mL), pre-purified TD808 (2.61 g; 8.17 mmol; 1.0 equiv.) was suspended in DCM (100 mL). A freshly prepared solution of SOCl2 (1.80 mL; 24.8 mmol; 3.0 equiv.) in DCM (10 mL) was then added, and the open flask was stirred at room temperature for 1 h. Dilute aqueous NaHCO3 (100 mL) was then added, and the resulting biphasic mixture was vigorously stirred at room temperature for an additional 1 h, during which time gas bubbles slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 75 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in its free base form as a yellow oil. Yield: 2.87g (99%; 1 step; TD808 standard). NMR (DMSO-d6): 1 Hδ H 0.97-1.20(i-Pr,m,21H);4.78(CH2,s,4H);7.59(arom,s,2H). 13 C{ 1 H} δ C 10.6(i-Pr,s);18.4(i-Pr,s);45.9(CH2,s);96.6(C≡C-Si,s);103.5(C≡C-arom.;s);124.6(arom.,s);132.0arom.,s);157.1(arom.,s). ESI-HRMS:356.1368[M+H] + (Theoretical value [C 18 H 28 N1Cl2Si1] + =356.1368).

[0393] Synthesis of TD817:

[0394] [ka]

[0395] In a round-bottom glass flask (250 mL), TD815 (2.86 g; 8.02 mmol; 1.2 equiv.) was dissolved in MeCN (130 mL). Solid NaN (435 mg; 6.69 mmol; 1.0 equiv.) and dry KCO (920 mg; 6.67 mmol; 1.0 equiv.) were subsequently added, and the resulting suspension was stirred at 80 °C for 16 h. The mixture was filtered through a syringe microfilter (PTFE), and the solid residue was washed with MeCN. The filtrate was evaporated to dryness, and the orange oily residue was purified by column chromatography (220 g SiO, 30% PE in DCM to 10% PE in DCM). The product-containing fractions were combined and evaporated to dryness. The residue was further dried under high vacuum for 1 h to give the product in the form of the free base as a slightly yellow oil. Yield: 1.23 g (51%; 1 step; based on NaN). Recovery: 709 mg of TD815 (25% of the initial amount). NMR (DMSO-d6): 1 Hδ H 1.01-1.21(i-Pr,m,21H);4.55(CH2,s,2H);4.78(CH2,s,2H);7.47(arom.,d,1H, 4 J HH =1);7.58(arom,d,1H, 4 J HH =1). 13 C{ 1 H} δ C 10.6(i-Pr,s);18.4(i-Pr,s);46.0(CH2,s);53.7(CH2,s);96.5(C≡C-Si,s);103.7( C≡C-arom,s);124.3(arom.,s);131.8(arom.,s);156.6(arom.,s);157.1(arom.,s). ESI-HRMS:363.1763[M+H] + (Theoretical value [C 18 H 28N4Cl1Si1] + =363.1766).

[0396] Synthesis of TD1052:

[0397] [ka]

[0398] In a glass vial (20 mL), dimethyl 4-iodopyridine-2,6-dicarboxylate (500 mg; 1.56 mmol; 1.0 equiv.), CuI (600 mg; 3.15 mmol; 2.0 equiv.), and [(dppf)PdCl] (7.0 mg, 10 μmol; 0.6 mol%) were dissolved in dry DMF (8 mL), followed by the addition of methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (600 mg; 3.12 mmol; 2.0 equiv.) in dry DMF (2 mL). The resulting dark mixture was stirred at 100 °C for 16 h. After cooling, the mixture was diluted with DCM (10 mL) and filtered through a syringe microfilter (PTFE). The solid was washed with additional DCM. The filtrate was further diluted with DCM (50 mL) and washed with a dilute aqueous solution of NaHCO (5 × 25 mL). The organic layer was dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was purified by column chromatography (120 g SiO2, 100% DCM to 15% EtOAc in DCM). The product-containing fractions were combined and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in the form of the free base as a colorless solid. Yield: 366 mg (89%; 1 step; based on dimethyl 4-iodopyridine-2,6-dicarboxylate). NMR (DMSO-d6): 1 Hδ H 3.97(CH3,s,6H);8.49(arom.,q,2H, 4 J HF =1). 19 F { 1 H} δ F -63.4(s). ESI-HRMS:264.0480[M+H] + (Theoretical value [C 10 H9N1O4F3] +=264.0478).

[0399] Synthesis of TD1053:

[0400] [ka]

[0401] In a round-bottom glass flask (100 mL), TD1052 (365 mg; 1.39 mmol; 1.0 equiv.) was dissolved in a mixture of MeOH (5 mL) and THF (10 mL). The resulting colorless solution was cooled in an ice bath (below 5 °C), and solid NaBH (420 mg, 11.1 mmol, 8.0 equiv.) was added in portions (over 30 min without a stopper) during this time. Vigorous hydrogen gas evolution occurred, and the mixture changed color to red and orange. After the addition, the flask was warmed to room temperature and stirred for an additional 30 min at room temperature. The resulting yellow solution was evaporated to dryness. The residue was purified by column chromatography (120 g SiO, solid loading method, 100% EtOAc to 15% MeOH in EtOAc). The product-containing fractions were combined and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in the form of a free base as a yellow oil. Yield: 171 mg (60%; 1 step; based on TD1052). NMR (DMSO-d): 1 Hδ H 4.62(CH2,s,4H);5.65(OH,s,2H);7.60(arom.,q,2H, 4 J HF =1). 19 F{ 1 H} δ F -63.6(s). ESI-HRMS:208.0580[M+H] + (Theoretical value [C8H9N1O2F3] + =208.0580).

[0402] Synthesis of TD1055:

[0403] [ka]

[0404] In a round-bottom glass flask (100 mL), TD1053 (169 mg; 816 μmol; 1.0 equiv.) was dissolved in DCM (20 mL). Next, a freshly prepared solution of SOCl2 (237 μL; 3.26 mmol; 4.0 equiv.) in DCM (5 mL) was added, and the resulting mixture was stirred at room temperature for 16 h. Dilute aqueous NaHCO3 (25 mL) was then added, and the resulting biphasic mixture was vigorously stirred at room temperature for an additional 10 min, during which time gas bubbles slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a yellow oil. Yield: 168 mg (84%; 1 step; based on TD1053). NMR (DMSO-d6): 1 Hδ H 4.90(CH2,s,4H);7.94(arom.,s,2H). 19 F{ 1 H} δ F -63.4s).APCI-HRMS:243.9902[M+H] + (Theoretical value [C8H7N1F3Cl2] + =243.9902).

[0405] Synthesis of TD1057:

[0406] [ka]

[0407] In a glass vial (4 mL), TD1055 (143 mg; 586 μmol; 1.8 equiv.) was dissolved in MeCN (3.5 mL), followed by the addition of solid NaN (21.5 mg; 331 μmol; 1.0 equiv.) and dry KCO (46 mg; 333 μmol; 1.0 equiv.), and the resulting suspension was stirred at 70 °C for 3 days. The mixture was filtered through a syringe microfilter (PTFE), and the solid residue was washed with MeCN. The filtrate was evaporated to dryness, and the orange oily residue was purified by column chromatography (80 g SiO, 30% PE in DCM to 40% DCM in PE). The product-containing fractions were combined and evaporated to dryness. The residue was further dried under high vacuum for 1 h, affording the product in the form of the free base as a colorless oil. Yield: 43.9 g (53%; 1 step; NaN). Recovery: 32.1 mg of TD1055 (22% of the initial amount). NMR (DMSO-d): 1 Hδ H 4.69(CH2,s,2H);4.90(CH2,s,2H);7.81(arom.,d,1H, 4 J HH =1);7.93(arom.,d,1H, 4 J HH =1). 19 F{ 1 H} δ F -63.4(s). APCI-HRMS:251.0306[M+H] + (Theoretical value [C8H7N4F3Cl1] + =251.0306).

[0408] Synthesis of TD549:

[0409] [ka]

[0410] A pear-shaped glass flask (250 mL) was charged with dimethyl 4-chloropyridine-2,6-dicarboxylate (5.00 g; 21.8 mmol; 1.0 equiv.), phenylboronic acid (3.20 g; 26.2 mmol; 1.2 equiv.), and XPhos Pd G2 (510 mg; 648 μmol; 3.0 mol%) and protected with argon three times. Next, dry DMF (110 mL) was added through a septum under a constant flow of argon, followed by freshly dried (high vacuum using a heat gun) Cs2CO3 (15.6 g; 47.9 mmol; 2.2 equiv.; the flask was briefly opened for the addition). The mixture was then stirred under a septum (but without external argon) at 80 °C for 20 h. The resulting dark mixture was filtered through a glass frit (S3), and the filtrate was poured into a stirred beaker containing HO (500 mL). The precipitate was collected on a glass frit (S2), washed with HO, and dried overnight under high vacuum to give the free base form of the product as an off-white solid. Yield: 3.01 g (51%; 1 step; based on dimethyl 4-chloropyridine-2,6-dicarboxylate). NMR (DMSO-d6): 1 Hδ H 3.95(CH3,s,6H);7.45-7.57(Ph,m,3H);7.82-8.00(Ph,m,2H);8.48(arom.,s,2H); 13 C{ 1 H} δ C 52.7(CH3,s);125.0(arom,s);127.2(Ph.,s);129.5(Ph.,s);130.2(Ph.,s);135.5(Ph.,s);148.6(arom.,s);150.0(arom.,s);164.7(CO.,s). ESI-HRMS:272.0916[M+H] + (Theoretical value [C 15 H 14 N1O4] + =272.0917).

[0411] Synthesis of TD563:

[0412] [ka]

[0413] In a round-bottom glass flask (500 mL), TD549 (2.98 g; 11.0 mmol; 1.0 equiv) was dissolved in a mixture of MeOH (100 mL) and THF (100 mL). The resulting slightly yellow solution was cooled in an ice bath (below 5 °C), followed by the addition of solid NaBH (2.90 g, 76.7 mmol, 9.0 equiv) in portions (over 30 min without a stopper). During this time, hydrogen gas evolved vigorously and the mixture changed color to red and orange. After the addition, the flask was warmed to room temperature and stirred for an additional 30 min at room temperature. The resulting slightly yellow, opaque solution was filtered through a syringe microfilter (PTFE). The filtrate was evaporated to dryness and further co-evaporated with DCM (as a suspension). The residue was dissolved in a mixture of HO (300 mL) and DCM (300 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (8 x 75 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3) and evaporated to dryness. The resulting crystallized residue was crushed and further dried overnight under high vacuum to give the product in the form of the free base as a nearly colorless solid. Yield: 2.29 g (97%; 1 step; TD549 reference). NMR (DMSO-d6): 1 Hδ H 4.59(CH2,s,4H);5.45(OH,s,2H);7.45-7.57(Ph,m,3H);7.60(arom.,s,2H);7.68-7.82(Ph,m,2H). 13 C{ 1 H} δ C 64.2(CH2,s);115.7(arom.,s);126.7(Ph,s);129.1(Ph,s);129.3(Ph,s);138.1(Ph,s);148.2(arom.,s);161.7(arom.,s). ESI-HRMS:216.1022[M+H] + (Theoretical value [C 13 H 14 N1O2] + =216.1019).

[0414] Synthesis of TD564:

[0415] [ka]

[0416] In a round-bottom glass flask (500 mL), TD563 (2.25 g; 10.5 mmol; 1.0 equiv.) was dissolved in DCM (250 mL) (with gentle heating). Next, a freshly prepared solution of SOCl2 (2.27 mL; 31.3 mmol; 3.0 equiv.) in DCM (10 mL) was added. After this, a precipitate began to form. After stirring at room temperature for 1 h, all of the previously formed precipitate had redissolved. Next, a dilute aqueous solution of NaHCO3 (150 mL) was added to the clear yellow solution, and the resulting biphasic mixture was vigorously stirred for an additional 1 h at room temperature, during which time gas bubbles were slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in free base form as a pale yellow solid. Yield: 2.59 g (98%; 1 step; TD563 reference). NMR (DMSO-d6): 1 Hδ H 4.88(CH2,s,4H);7.47-7.58(Ph,m,3H);7.77-7.85(Ph,m,2H);7.86(arom.,s,2H). 13 C{ 1 H} δ C 46.6(CH2,s);120.3(arom.,s);126.9(Ph,s);129.3(Ph,s);129.7(Ph,s);136.5(P h,s);149.5(arom,s);157.1(arom.,s);157.1(arom.,s);ESI-HRMS:252.0342[M+H] + (Theoretical value [C 13 H 12 N1Cl2] + =252.0341).

[0417] Synthesis of TD566:

[0418] [ka]

[0419] In a pear-shaped glass flask (100 mL), TD564 (1.60 g; 6.35 mmol; 1.3 equiv.) was dissolved in MeCN (60 mL) (with gentle heating). Solid NaN (320 mg; 4.92 mmol; 1.0 equiv.) and dry KCO (680 mg; 4.93 mmol; 1.0 equiv.) were then added, and the resulting suspension was stirred at 80 °C for 24 h. The mixture was filtered through a glass frit S3, and the solid residue was washed with MeCN. The filtrate was evaporated to dryness, and the orange oily residue was purified by column chromatography (SiO, 15% PE in DCM to 100% DCM). The product-containing fractions were combined and evaporated to dryness. The residue was further dried under high vacuum for 1 h to give the product in the form of the free base as a white solid. Yield: 691 mg (54%; 1 step; based on NaN). Recovery: 613 mg of TD564 (38% of the initial amount). NMR (DMSO-d): 1 Hδ H 4.59(CH2,s,2H);4.84(CH2,s,2H);7.46-7.60(Ph,m,3H);7.74(arom.,d,1H, 4 J HH =2);7.77-7.85(Ph,m,2H);7.86(arom.,d,1H, 4 J HH =2). 13 C{ 1 H} δ C 46.7(CH2,s);54.2(CH2,s);119.4(arom.,s);120.0(arom.,s);126.9(Ph,s);129.3(P h,s);129.7(Ph,s);136.6(Ph,s);149.3(arom.,s);156.6(arom.,s);157.1(arom.,s). ESI-HRMS:259.0747[M+H] + (Theoretical value [C 13 H 12 N4Cl1] + =259.0745).

[0420] Synthesis of TD1083:

[0421] [ka]

[0422] In a round-bottom glass flask (100 mL), methyl isonicotinate (250 mg; 1.83 mmol; 1.0 equiv) was dissolved in MeOH (10 mL) and concentrated H2SO4 (25 μL) was added. The resulting mixture was stirred at 55 °C for 30 min. After cooling, a solution of (NH4)2SO8 (4.16 g; 18.2 mmol; 10 equiv) in H2O (10 mL) was added dropwise, and the resulting mixture was further stirred at 55 °C for 16 h. The reaction was then carefully quenched with aqueous NaHCO3. The mixture was transferred to a separatory funnel and extracted with EtOAc (5 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was purified by preparative HPLC (C18; H2O-MeCN gradient with FA). Fractions containing the product were combined and directly lyophilized to give the product in free base form as a white fluffy solid. Yield: 99.5 mg (28%; 1 step; based on methyl isonicotinate). NMR (DMSO-d): 1 Hδ H 3.92 (CH3, s, 3H); 4.60 (CH2, d, 4H, 3 J HH =6);5.58(OH,t,2H, 3 J HH =6);7.80(arom.,s,2H).ESI-HRMS:198.0759[M+H] + (Theoretical value [C9H 12 N1O4] + =198.0761). EA(C9H 11 N1O4,M R =197.2):C54.8(54.8);H5.6(5.5);N7.1(7.0).

[0423] Synthesis of TD1087:

[0424] [ka]

[0425] In a round-bottom glass flask (50 mL), TD1083 (96.2 mg; 488 μmol; 1.0 equiv.) was dissolved in DCM (20 mL), followed by the addition of SOCl2 (106 μL; 1.46 mmol; 3.0 equiv.). The mixture was stirred at room temperature for 1 h. The mixture was then quenched by the addition of dilute aqueous NaHCO3 (20 mL). The resulting biphasic mixture was vigorously stirred at room temperature for an additional 1 h, during which time gas bubbles slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight (where the product slowly crystallized) to give the free base form of the product as a bright yellow solid. Yield: 112.8 mg (99%; 1 step; based on TD1083). NMR (DMSO-d6): 1 Hδ H 3.92(CH3,s,3H);4.90(CH2,s,4H);7.98(arom.,s,2H).ESI-HRMS:234.0085[M+H] + (Theoretical value [C9H 10 N1O2Cl2] + =234.0083).

[0426] Synthesis of TD1089:

[0427] [ka]

[0428] In a glass vial (4 mL), TD1087 (112.0 mg; 478 μmol; 1.2 equiv.) was dissolved in MeCN (2 mL). Solid NaN (26.0 mg; 400 μmol; 1.0 equiv.) and dry KCO (55.0 mg; 400 μmol; 1.0 equiv.) were subsequently added, and the resulting suspension was stirred at 70 °C for 2 h. The mixture was filtered through a syringe microfilter (PTFE), and the solid residue was washed with MeCN. The filtrate was evaporated to dryness, and the oily residue was purified by column chromatography (SiO, DCM to 10% EtOAc in DCM). The product-containing fractions were combined and evaporated to dryness. The residue was further dried overnight under high vacuum to give the crude product in the form of the free base (containing up to 20% of the bis-azide by-product) as a yellow oil. Yield: 49.6 mg. ESI-HRMS: 241.0490 [M+H] + (Theoretical value [C9H 10 N4O2Cl1] + =241.0487). A portion of the TD1089 thus obtained was used directly for TD1092 without further purification or characterization.

[0429] Synthesis of TD1101:

[0430] [ka]

[0431] In a round-bottom glass flask (100 mL), isopropyl isonicotinate (600 mg; 3.63 mmol; 1.0 equiv) was dissolved in MeOH (20 mL) and concentrated H2SO4 (50 μL) was added. Next, a solution of (NH4)2S2O8 (8.32 g; 36.5 mmol; 10 equiv) in H2O (20 mL) was added dropwise, and the resulting mixture was further stirred at 70 °C for 16 h. The reaction was then carefully neutralized with aqueous NaHCO3. The mixture was transferred to a separatory funnel and extracted with EtOAc (5 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was purified by preparative HPLC (C18; H2O-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in its free base form as a white fluffy solid. Yield: 284 mg (35%; 1 step; based on isopropyl isonicotinate). NMR (DMSO-d): 1 Hδ H 1.34(CH3,d,6H, 3 J HH =6);4.59(CH2,d,4H, 3 H J HH =6);5.19(CH,hept,1H, 3 J HH =6);5.57(OH,t,2H, 3 J HH =6);7.78(arom.,s,2H).ESI-HRMS:226.1076[M+H] + (Theoretical value [C 11 H 16 N1O4] + =226.1074).

[0432] Synthesis of TD1109:

[0433] [ka]

[0434] In a round-bottom glass flask (100 mL), TD1101 (283 mg; 1.26 mmol; 1.0 equiv.) was dissolved in DCM (40 mL), followed by the addition of SOCl2 (275 μL; 3.79 mmol; 3.0 equiv.). The mixture was stirred at room temperature for 1 h. The mixture was then quenched by the addition of dilute aqueous NaHCO3 (40 mL). The resulting biphasic mixture was vigorously stirred at room temperature for an additional 1 h, during which time gas bubbles slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight (where the product slowly crystallized) to give the product in the form of the free base as a pale yellow solid. Yield: 322 mg (98%; 1 step; based on TD1101). NMR (DMSO-d6): 1 Hδ H 1.35(CH3,d,6H, 3 J HH =6);4.90(CH2,s,4H);5.20(CH,hept,1H, 3 J HH =6);7.96(arom.,s,2H).ESI-HRMS:262.0399[M+H] + (Theoretical value [C 11 H 14 N1O2Cl2] + =262.0396).

[0435] Synthesis of TD1112:

[0436] [ka]

[0437] In a glass vial (20 mL), TD1109 (321 mg; 1.23 mmol; 1.2 equiv.) was dissolved in MeCN (6 mL). Solid NaN (66.5 mg; 1.02 mmol; 1.0 equiv.) and dry KCO (141 mg; 1.02 mmol; 1.0 equiv.) were subsequently added, and the resulting suspension was stirred at 70 °C for 16 h. The mixture was filtered through a syringe microfilter (PTFE), and the solid residue was washed with MeCN. The filtrate was evaporated to dryness, and the oily residue was purified by column chromatography (SiO, DCM to 3% EtOAc in DCM). The product-containing fractions were combined and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a colorless oil. Yield: 116 mg (42%; 1 step; based on NaN). Recovery: 124 mg of TD1109 (39% of the amount originally used). NMR (DMSO-d6): 1 Hδ H 1.35(CH3,d,6H, 3 J HH =6);4.67(CH2,s,2H);4.89(CH2,s,4H);5.19(CH,hept,1H, 3 J HH =6);7.83(arom.,d,1H, 4 J HH =1).7.95(arom.,d,1H, 4 J HH = 1). ESI-HRMS: 269.0800 [M+H] + (Theoretical value [C 11 H 14 N4O2Cl1] + =269.0800).

[0438] Synthesis of TD1119:

[0439] [ka]

[0440] In a glass vial (40 mL), tert-butyl isonicotinate (486 mg; 2.71 mmol; 1.0 equiv.) was dissolved in MeOH (15 mL), followed by the addition of concentrated H2SO4 (40 μL). Next, a solution of (NH4)2SO8 (6.20 g; 27.2 mmol; 10 equiv.) in H2O (15 mL) was added dropwise, and the resulting mixture was further stirred at 80 °C for 30 min. The reaction was then carefully neutralized with NaHCO3. The mixture was transferred to a separatory funnel and extracted with EtOAc (5 × 30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was purified by preparative HPLC (C18; H2O-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in the form of the free base as a white fluffy solid. Yield: 161 mg (25%; 1 step; based on tert-butyl isonicotinate). NMR (DMSO-d): 1 Hδ H 1.57(CH3,s,9H);4.58(CH2,d,4H, 3 J HH =4);5.54(OH,t,2H, 3 J HH =4);7.73(arom.,s,2H).ESI-HRMS:240.1230[M+H] + (Theoretical value [C 12 H 18 N1O4] + =240.1230).

[0441] Synthesis of TD1129:

[0442] [ka]

[0443] In a round-bottom glass flask (100 mL), TD1119 (159 mg; 664 μmol; 1.0 equiv) was dissolved in DCM (20 mL), followed by the addition of SOCl2 (145 μL; 2.00 mmol; 3.0 equiv). The mixture was stirred at room temperature for 1 h. The mixture was then quenched by the addition of dilute aqueous NaHCO3 (20 mL). The resulting biphasic mixture was vigorously stirred at room temperature for an additional 1 h, during which time gas bubbles slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight (where the product slowly crystallized) to give the free base form of the product as a faint yellow solid. Yield: 175 mg (95%; 1 step; based on TD1119). NMR (DMSO-d6): 1 Hδ H 1.57(CH3,s,9H);4.89(CH2,s,4H);7.91(arom.,s,2H).ESI-HRMS:276.0552[M+H] + (Theoretical value [C 12 H 16 N1O2Cl2] + =276.0553).

[0444] Synthesis of TD1130:

[0445] [ka]

[0446] In a glass vial (4 mL), TD1129 (174.0 mg; 630 μmol; 1.2 equiv.) was dissolved in MeCN (3 mL). Solid NaN (34.0 mg; 523 μmol; 1.0 equiv.) and dry KCO (72.0 mg; 522 μmol; 1.0 equiv.) were subsequently added, and the resulting suspension was stirred at 70 °C for 6 h. The mixture was filtered through a syringe microfilter (PTFE), and the solid residue was washed with MeCN. The filtrate was evaporated to dryness, and the oily residue was purified by column chromatography (SiO, DCM to 3% EtOAc in DCM). The product-containing fractions were combined and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a colorless oil. Yield: 58.2 mg (39%; 1 step; based on NaN). Recovery: 90.5 mg of TD1129 (52% of the initial amount). NMR (DMSO-d6): 1 Hδ H 1.57(CH3,s,9H);4.66(CH2,s,2H);4.89(CH2,s,2H);7.79(arom.,d,1H, 4 J HH =2).7.91(arom.,d,1H, 4 J HH = 2). ESI-HRMS: 305.0772 [M+Na] + (Theoretical value [C 12 H 15 N4O2Cl1Na1] + =305.0776).

[0447] Synthesis of TD725:

[0448] [ka]

[0449] A glass vial (40 mL) was charged with TD726 (628 mg; 3.62 mmol; 1.0 equiv.), (4-(tert-butoxycarbonyl)phenyl)boronic acid (880 mg; 3.96 mmol; 1.1 equiv.), and XPhos Pd G2 (57 mg; 7.2 μmol; 2.0 mol%) and protected with argon three times. Next, dry 1,4-dioxane (16 mL) was added through a septum under a constant flow of argon, followed by a freshly prepared solution of K3PO4-HO (920 mg; 4.00 mmol; 1.1 equiv.) in HO (8 mL) (briefly flushed with argon before addition). The mixture was stirred at 80 °C for 16 h under a septum (but without external argon). The resulting dark mixture was transferred to a separatory funnel and diluted with EtOAc (40 mL) and H2O (50 mL). After shaking, the upper layer was separated and the aqueous layer was further extracted with EtOAc (5 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was purified by flash chromatography (120 g SiO2, 100% EtOAc to 30% MeOH in EtOAc). The combined fractions containing the product were evaporated to dryness and further co-evaporated once with DCM. The residue was further dried overnight under high vacuum to give the crude product in the form of the free base as an off-white powder. Yield: 1.04 g, NMR (DMSO-d6): 1 Hδ H 1.57(CH3,s,9H); 4.61(CH2,d,4H, 3 J HH =6);5.48(OH,t,2H, 3 J HH =6);7.64(arom.,s,2H);7.88(arom.,dm,2H, 3 J HH =9);8.04(arom,dm,2H, 3 J HH =9). 13 C{ 1 H} δ C27.8(CH3,s);64.2(CH2,s);81.0(C-CH3,s);115.8(arom.,s);127.0(arom,s);129.9(arom .,s);131.6(arom.,s);142.2(arom.,s);147.0(arom.,s);162.0(arom.,s);164.6(CO,s). ESI-HRMS:316.1539[M+H] + (Theoretical value [C 18 H 22 N1O4] + =316.1543).

[0450] Synthesis of TD730:

[0451] [ka]

[0452] In a round-bottom glass flask (250 mL), prepurified TD725 (1.04 g; ≤3.30 mmol; 1.0 equiv.) was suspended in DCM (35 mL). Next, a freshly prepared solution of SOCl2 (721 μL; 9.93 mmol; ≥3.0 equiv.) in DCM (5 mL) was added. A clear solution immediately formed, and the open flask was stirred at room temperature for 1 h. The mixture was quenched by the addition of dilute aqueous NaHCO3 (60 mL). The resulting biphasic mixture was vigorously stirred at constant temperature for an additional 1 h, during which time gas bubbles slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (5 × 30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was purified by flash chromatography (120 g SiO2, 100% DCM to 20% EtOAc in DCM). The combined fractions containing the product were evaporated to dryness and co-evaporated once with DCM. The remaining faintly yellow oil slowly crystallized. The solid was crushed and further dried under high vacuum overnight to give the product in the form of the free base as an off-white powder. Yield: 839 mg (66%; 2 steps; based on TD726). NMR (DMSO-d6): 1 Hδ H1.58(CH3,s,9H);4.86(CH2,s,4H);7.92(arom.,s,2H);7.94(arom.,dm,2H, 3 J HH =9);8.05(arom.,dm,2H, 3 J HH =9). 13 C{ 1 H} δ C 27.8(CH3,s);46.5(CH2,s);81.2(C-CH3,s);120.6(arom,s);127.2(arom.,s);129.9(arom .,s);132.1(arom.,s);140.6(arom.,s);148.3(arom.,s);157.3(arom.,s);164.5(CO,s). ESI-HRMS:352.0866[M+H] + (Theoretical value[C 18 H 20 [N1O2Cl2] + =352.0866).

[0453] Synthesis of TD733:

[0454]

change

[0455] In a glass vial (20 mL), TD730 (837 mg; 2.38 mmol; 1.3 equiv.) was dissolved in MeCN (18 mL). Solid NaN (119 mg; 1.83 mmol; 1.0 equiv.) and dry KCO (253 mg; 1.83 mmol; 1.0 equiv.) were subsequently added, and the resulting suspension was stirred at 80 °C for 24 h. The mixture was filtered through a syringe microfilter (PTFE), and the solid residue was washed with MeCN. The filtrate was evaporated to dryness, and the oily residue was purified by column chromatography (SiO, 20% pentane in DCM to 1% EtOAc in DCM). The product-containing fractions were combined and evaporated to dryness. The resulting nearly colorless oil was further dried under high vacuum overnight (where the product slowly crystallized) to give the product in the form of the free base as a colorless solid. Yield: 230 mg (35%; 1 step; based on NaN). Recovery: 442 mg of TD730 (53% of the initial amount). NMR (DMSO-d6): 1 Hδ H 1.57(CH3,s,9H);4.62(CH2,s,2H);4.86(CH2,s,2H);7.80(arom.,d,1H, 4 J HH =2);7.91(arom.,d,1H, 4 J HH =2);7.94(arom,dm,2H, 3 J HH =9);8.05(arom.,dm,2H, 3 J HH =9). 13 C{ 1 H}δ C 27.8(CH3,s);46.6(CH2,s);54.2(CH2,s);81.2(C-CH3,s);119.6(arom,s);120.3(arom.,s);127.2(arom.,s);129. 9(arom.,s);132.1(arom.,s);140.7(arom.,s);148.1(arom.,s);156.8(arom.,s);157.3(arom.,s);164.5(CO,s). ESI-HRMS:359.1270[M+H] + (Theoretical value [C 18 H 20 N4O2Cl1]+ =359.1269).

[0456] Synthesis of TD1425:

[0457] [ka]

[0458] A glass vial (40 mL) was charged with methyl 2-bromo-6-methylisonicotinate (1.00 g; 4.35 mmol; 1.0 equiv.), NBS (recrystallized from boiling HO; 770 mg; 4.33 mmol; 1.0 equiv.), (BnO) (53 mg; 219 μmol; 5 mol%), and a magnetic stirrer, and the mixture was protected with argon three times. Under a constant flow of argon, CCl (21 mL) was added through a septum. The vial was left stirring under a septum (but without external argon) at 75 °C for 24 h. After cooling, the mixture was filtered through a syringe microfilter (PTFE), and the solid was washed with DCM. The filtrate was evaporated to dryness, resuspended in c-Hex (25 mL), and filtered through a syringe microfilter (PTFE). The filtrate was evaporated to dryness and the residue was purified by flash chromatography (120 g SiO2, 100% c-Hex to 80% DCM in c-Hex). The combined fractions containing the product were evaporated to dryness. The residual oil was further dried overnight under high vacuum (where the product crystallized) to give the product in the form of the free base as a slightly yellow solid. Yield: 571 mg (43%; 1 step; based on methyl 2-bromo-6-methylisonicotinate). Recovery: 314 mg of methyl 2-bromo-6-methylisonicotinate (31% of the initial amount used). NMR (DMSO-d6): 1 Hδ H 3.91(CH3,s,3H);4.79(CH2,s,2H);7.94(arom.,d,1H, 4 J HH =1);8.05(arom.,d,1H, 4 J HH = 1). ESI-HRMS: 307.8916 [M+H] + (Theoretical value [C8H8N1O2Br2] +=307.8916).

[0459] Synthesis of TD1428:

[0460] [ka]

[0461] A pear-shaped glass flask (25 mL) was charged with TD1425 (523 mg; 1.69 mmol; 1.0 equiv.), N-Boc-1,1-dimethylpyropargylamine (310 mg; 1.69 mmol; 1.0 equiv.), and a magnetic stirrer. The flask was then protected with argon three times. Solid CuI (13.0 mg; 68 μmol; 4.0 mol%) and [Pd(PPh3)2Cl2] (24 mg; 34 μmol; 2.0 mol%) were then added, and the mixture was reprotected with argon (three times). Next, dry THF (7.5 mL) was added through a septum under a constant flow of argon, followed by DIPEA (885 μL; 5.08 mmol; 3.0 equiv.). The flask was left at room temperature under a septum (but without external argon) with stirring for 2 days. The mixture was evaporated to dryness, resuspended in DCM (15 mL), and filtered through a syringe microfilter (PTFE), washing the solid phase with additional DCM. The filtrate was purified by flash chromatography (120 g SiO, 10% EtOAc in c-Hex to 50% EtOAc in c-Hex). The combined product-containing fractions were evaporated to dryness and repurified once by flash chromatography (80 g SiO, 100% EtOAc in DCM to 5% EtOAc in DCM). The combined product-containing fractions were evaporated to dryness and co-evaporated once with DCM. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a slightly yellow solidifying oil. Yield: 175 mg (25%; 1 step; TD1425 reference). NMR (DMSO-d): 1 Hδ H 1.42(CH3-CO,s,9H);1.56(CH3-CN,s,6H);3.91(CH3-O,s,3H);4.79(CH2,s,2H);7.23(NH,bs,1H);7.71(arom.,d,1H, 4 JHH =1);7.97(arom.,d,1H, 4 J HH = 1). ESI-HRMS: 411.0913 [M+H] + (Theoretical value [C 18 H 24 N2O4Br1] + =411.0914).

[0462] Synthesis of TD1386:

[0463] [ka]

[0464] In a round-bottom glass flask (100 mL), pyridine-2,6-diyldimethanol (2.00 g; 14.4 mmol; 1.0 equiv.) and imidazole (0.98 g; 14.4 mmol; 1.0 equiv.) were dissolved in DMF (20 mL), and TBDMSCl (2.17 g; 14.4 mmol; 1.0 equiv.) was added. The resulting solution was stirred at room temperature for 2 h. The mixture was evaporated to dryness, and the residue was dissolved in a mixture of DCM (50 mL) and dilute aqueous NaHCO3 (50 mL) and transferred to a separatory funnel. After brief shaking, the bottom phase was separated, and the aqueous layer was further extracted with DCM (5 × 30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was purified by column chromatography (100 g SiO2, DCM to 50% EtOAc in DCM). The product-containing fractions were combined, evaporated to dryness, and co-evaporated once with DCM. The residue was further dried overnight under high vacuum to give the product in free base form as a nearly colorless oil. Yield: 1.54 g (42%; 1 step; based on pyridine-2,6-diyldimethanol). NMR (DMSO-d6): 1 Hδ H 0.09(CH3,s,6H);0.92(CH3,s,9H);4.51(CH2,d,2H, 3 J HH =5);4.71(CH2,s,2H);5.37(OH,t,1H, 3 J HH=5);7.28(arom,dd,1H, 3 J HH =8; 4 J HH =1);7.34(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.81(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 254.1569 [M+H] + (Theoretical value [C 13 H 24 N1O2Si1] + =254.1571).

[0465] Synthesis of TD1389:

[0466] [ka]

[0467] In a round-bottom glass flask (250 mL), TD1386 (1.54 g; 6.08 mmol; 1.0 equiv.) was dissolved in DCM (60 mL), followed by the addition of SOCl2 (885 μL; 12.0 mmol; 2.0 equiv.). The mixture was stirred at room temperature for 1 h. Afterwards, the mixture was quenched by the addition of a dilute solution of NaHCO3 (60 mL). The resulting biphasic mixture was vigorously stirred at room temperature for an additional 30 min, during which time gas bubbles slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in the form of the free base as a nearly colorless oil. Yield: 1.63 g (99%; 1 step; based on TD1386). NMR (DMSO-d6): 1 Hδ H 0.10(CH3,s,6H);0.92(CH3,s,9H);4.74(CH2,s,2H);4.75(CH2,s,2H);7.40(arom.,dd,1H, 3 J HH =8;4 J HH =1);7.42(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.87(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 272.1231 [M+H] + (Theoretical value [C 13 H 23 N1O1Cl1Si1] + =272.1232).

[0468] Synthesis of TD1390:

[0469] [ka]

[0470] In a glass flask (20 mL), KCN (390 mg; 5.99 mmol; 1.05 equiv.) was dissolved in HO (2 mL), followed by the addition of TD1389 (1.55 g; 5.70 mmol; 1.0 equiv.) in DMF (6 mL). The resulting mixture was vigorously stirred at 100 °C for 70 min. After cooling to room temperature, the mixture was diluted with HO (4 mL) and MeCN (8 mL) and filtered through a syringe microfilter (PTFE). The filtrate was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (100 mL) and HO (100 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The remaining pale yellow oil solidified in the freezer. The resulting solid was crushed and further dried under high vacuum overnight to give the product in free base form as a faint yellow powder. Yield: 1.23 g (82%; 1 step; based on TD1389). NMR (DMSO-d6): 1 Hδ H0.10(CH3,s,6H);0.92(CH3,s,9H);4.17(CH2,s,2H);4.75(CH2,s,2H);7.30(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.40(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.86(arom.,t,1H, 3 J HH =8). ESI-HRMS: 263.1573 [M+H] + (Theoretical value[C 14 H 23 N2O1Si1] + =263.1574).

[0471] Synthesis of TD1393:

[0472]

change

[0473] In a pear-shaped glass flask (100 mL), TD1390 (700 mg; 2.67 mmol; 1.00 equiv.) was dissolved in MeOH (2.70 mL; 66.7 mmol; 25 equiv.), followed by the addition of TMSCl (2.37 mL; 18.7 mmol; 7.0 equiv.). The resulting mixture was vigorously stirred at 60 °C for 2 h. After cooling to room temperature, the mixture was quenched with HO (4 mL) and partially neutralized (to pH 3-4) by the slow addition of saturated aqueous NaHCO (5 mL). The resulting biphasic mixture was carefully evaporated to dryness, and HO (10 mL) was added to the residue. The mixture was filtered through a syringe microfilter (RC), and the filtrate was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO and evaporated to dryness. The residue was dissolved in DCM (100 mL) and H2O (100 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried briefly under high vacuum to give the product in the form of the free base as a slightly yellow oil. Yield: 404 mg (84%; 1 step; based on TD1390). NMR (DMSO-d6): 1 Hδ H 3.61(CH3,s,3H);3.81(CH2,s,2H);4.51(CH2,d,2H, 3 J HH =5);5.39(OH,t,1H, 3 J HH =5);7.20(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.36(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.76(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 182.0811 [M+H] + (Theoretical value [C9H 12 N1O3] + =182.0812).

[0474] Synthesis of TD1395:

[0475] [ka]

[0476] In a round-bottom glass flask (50 mL), TD1393 (340 mg; 1.88 mmol; 1.0 equiv.) and imidazole (190 mg; 2.79 mmol; 1.5 equiv.) were dissolved in DMF (6 mL), followed by the addition of TBDMSCl (425 mg; 2.82 mmol; 1.5 equiv.). The resulting solution was stirred at room temperature for 16 h. The mixture was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in free base form as a pale yellow oil. Yield: 537 mg (97%; 1 step; based on TD1393). NMR (DMSO-d6): 1 Hδ H 0.09(CH3,s,6H);0.91(CH3,s,9H);3.61(CH3,s,3H);3.81(CH2,s,2H);4.70(CH2,s,2H);7.23(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.32(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.79(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 296.1676 [M+H] + (Theoretical value [C 15 H 26 N1O3Si1] +=296.1677).

[0477] Synthesis of TD1396:

[0478] [ka]

[0479] In a round-bottom glass flask (100 mL), TD1395 (537 mg; 1.82 mmol; 1.0 equiv.) was dissolved in a mixture of THF (9 mL) and MeOH (9 mL). To the resulting colorless solution, solid NaBH4 (2.06 g; 54.5 mmol; 30 equiv.) was added in portions (over 10 min without a stopper) during which time hydrogen gas evolved vigorously. The mixture was further stirred at room temperature for 1 h, during which it was diluted twice with MeOH (9 mL each) to ensure stirring. The mixture was diluted with DCM (100 mL) and HO (100 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in free base form as a nearly colorless oil. Yield: 483 mg (99%; 1 step; TD1395 reference). NMR (DMSO-d): 1 Hδ H 0.09(CH3,s,6H);0.92(CH3,s,9H);2.84(CH2,t,2H, 3 J HH =7);3.71(CH2,td,2H, 3 J HH =7,J 3 HH =5);4.62(OH,t,1H, 3 J HH =5);4.71(CH2,s,2H);7.14(arom,dd,1H, 3 J HH =8; 4 J HH =1);7.24(arom.,dd,1H, 3 J HH =8; 4 J HH=1);7.70(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 268.1726 [M+H] + (Theoretical value [C 14 H 26 N1O2Si1] + =268.1727).

[0480] Synthesis of TD1401:

[0481] [ka]

[0482] In a round-bottom glass flask (50 mL), TD1396 (94 mg; 352 μmol; 1.0 equiv.) was dissolved in DCM (3.5 mL), followed by the addition of SOCl2 (51 μL; 702 μmol; 2.0 equiv.). The mixture was stirred at room temperature for 3 h. The mixture was diluted with DCM (20 mL) and quenched by the addition of dilute aqueous NaHCO3 (10 mL). The resulting biphasic mixture was vigorously stirred for an additional 30 min at room temperature, during which time gas bubbles slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 15 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was dissolved in DMF (1.7 mL), followed by the addition of solid NaN3 (46 mg; 708 μmol; 2.0 equiv.). The resulting mixture was stirred at 80 °C overnight. After cooling, the mixture was directly purified by preparative HPLC (C18; TD1390 reference). The product-containing fractions were combined, neutralized with diluted aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (25 mL) and H2O (25 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 x 10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a colorless oil. Yield: 17.6 mg (28%; 2 steps; TD1396 reference). NMR (DMSO-d6): 1 HδH 2.98(CH2,t,2H, 3 J HH =7);3.69(CH2,td,2H, 3 J HH =7, 3 J HH =5);4.53(CH2,d,2H, 3 J HH =6);5.37(OH,t,1H, 3 J HH =6);7.18(arom、dd,1H, 3 J HH =8; 4 J HH =1);7.32(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.73(arom.,t,1H, 3 J HH =8). ESI-HRMS:201.0746[M+Na] + (Theoretical value[C8H 10 N4O1Na1] + =201.0747).

[0483] Synthesis of TD1406:

[0484]

change

[0485] In a round-bottom glass flask (25 mL), TD1401 (16.5 mg; 92.6 μmol; 1.0 equiv) was dissolved in DCM (1.85 mL), followed by the addition of SOCl2 (13.5 μL; 186 μmol; 2.0 equiv). The mixture was stirred at room temperature for 1 h. The mixture was diluted with DCM (10 mL) and quenched by the addition of dilute aqueous NaHCO3 (10 mL). The resulting biphasic mixture was vigorously stirred for an additional 30 min at room temperature, during which time gas bubbles slowly evolved. The mixture was transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in the form of the free base as a colorless solid. Yield: 17.7 mg (97%; 1 step; TD1401 standard). NMR (DMSO-d): 1 Hδ H 3.02(CH2,t,2H, 3 J HH =7);3.71(CH2,td,2H, 3 J HH =7, 3 J HH =5);4.75(CH2,s,2H);7.32(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.41(arom.,dd,1H, 3 J HH =8; 4 J HH =1);7.79(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 197.0589 [M+H] + (Theoretical value [C8H 10 N4Cl1] + =197.0589).

[0486] Synthesis of TD1339:

[0487] [ka]

[0488] A pear-shaped glass flask (250 mL) containing ethyl l-lactate (3.54 g; 30.0 mmol; 1.00 equiv.) and a magnetic stirrer was briefly protected with argon three times. Next, dry DCM (100 mL) was added through the septum under a constant stream of argon. The mixture was cooled in an ice bath (5 °C), followed by the dropwise addition of anhydrous triflic acid (5.3 mL; 31.5 mmol; 1.05 equiv.), followed immediately by the dropwise addition of dry pyridine (2.54 mL; 31.5 mmol; 1.05 equiv.). The resulting mixture was stirred at 5 °C for 30 min. The resulting suspension was directly purified by column chromatography (140 g SiO2, DCM). The combined product-containing fractions were evaporated to dryness and briefly dried under high vacuum to give the product as a slightly pinkish oil. Yield: 5.90 g (79%; 1 step; based on ethyl l-lactate). NMR (DMSO-d6): 1 Hδ H 1.26(CH3-CH2,t,3H, 3 J HH =7);1.51(CH3-CH,d,3H, 3 J HH =7);4.19-4.31(CH2,m,2H);5.27(CH,q,1H, 3 J HH =7). 19 F{ 1 H}δ F -77.7(s). CI-HRMS:251.0193[M+H] + (Theoretical value [C6H 10 O5S1F3] + =251.0196).

[0489] Synthesis of TD1488:

[0490] [ka]

[0491] A pear-shaped glass flask (25 mL) containing dimethyl l-malate (200 mg; 1.23 mmol; 1.00 equiv.) and a magnetic stirrer was briefly guarded with argon three times. Next, dry DCM (4 mL) was added through the septum under a constant flow of argon. The mixture was cooled in an ice bath (5 °C), followed by the dropwise addition of anhydrous triflic acid (220 μL; 1.31 mmol; 1.06 equiv.), immediately followed by the dropwise addition of dry pyridine (105 μL; 1.30 mmol; 1.06 equiv.). The resulting mixture was stirred at room temperature for 30 min. The resulting suspension was directly purified by column chromatography (30 g SiO2, DCM). The combined product fractions were evaporated to dryness and briefly dried under high vacuum to give the product as a colorless oil. Yield: 258 mg (77%; 1 step; based on dimethyl l-malate). NMR (DMSO-d6): 1 Hδ H 3.08(CH2,dd,1H, 2 J HH =17, 3 J HH =5);3.13(CH2,dd,1H, 2 J HH =17, 3 J HH =6);4.65(CH3,s,3H);3.78(CH3,s,3H);5.48(CH,dd,1H, 3 J HH =6, 3 J HH =5). 19 F{ 1 H}δ F -77.7(s). ESI-HRMS:316.9910[M+Na] + (Theoretical value [C7H9O7F3S1Na1] + =316.9913).

[0492] Example 3: Synthesis of protected macrocyclic intermediates Synthesis of TD680:

[0493] [ka]

[0494] In a pear-shaped glass flask (250 mL), Cbz2 cyclen (free base; 1.54 g; 3.50 mmol; 1.6 equiv.) was dissolved in MeCN (100 mL), followed by the addition of dry K2CO3 (300 mg; 2.17 mmol; 1.0 equiv.). To the vigorously stirred reaction mixture, a solution of tert-butyl bromoacetate (427 mg; 2.19 mmol; 1.0 equiv.) in dry MeCN (25 mL) was added dropwise (over 2 h). The solid was filtered off using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (150 mL) and HO (150 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 x 75 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a nearly colorless oil. Yield: 624 mg (51%; 1 step; based on tert-butyl bromoacetate). NMR (CD3CN): 1 Hδ H 1.32-1.49(CH3,m,9H);2.57-2.76(mc,m,4H);2.81-2.98(mc,m,4H);3.23-3.64(mc,CH2-CO,m,8+2H);5.00-5.15 CH2-Ph,m,4H);7.19-7.48(Ph,m,10H).ESI-HRMS:555.3171[M+H] + (Theoretical value [C 30 H 43 N4O6] + =555.3177).

[0495] Synthesis of TD686:

[0496] [ka]

[0497] In a glass vial (4 mL), TD680 (150 mg; 270 μmol; 1.0 equiv) was dissolved in MeCN (3 mL), and BocO (2.0 M solution in THF; 200 μL; 400 μmol; 1.5 equiv) was added. The resulting mixture was stirred at room temperature for 16 h. The reaction mixture was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, immediately neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in free base form as a colorless oil. Yield: 170 mg (96%; 1 step; TD680 standard). NMR (CD3CN): 1 Hδ H 1.17-1.56(CH3,m,18H);2.62-2.99(mc,m,4H);3.10-3.53(mc,CH2-CO,m,12+2H );4.96-5.16(CH2-Ph,m,4H);7.20-7.47(Ph,m,10H).ESI-HRMS:655.3691[M+H] + (Theoretical value [C 35 H 51 N4O8] + =655.3701).

[0498] Synthesis of TD691:

[0499] [ka]

[0500] A pear-shaped glass flask (25 mL) containing TD686 (170 mg; 260 μmol) and a magnetic stirrer was placed in the flask and protected with argon three times. Solid palladium on carbon (17 mg) was then added and the mixture was protected with argon three times. Under a constant flow of argon, MeOH (10 mL) was added through a septum. The argon flow was then removed, and H2 gas (from a balloon) was bubbled into the mixture. The mixture was then left at room temperature for 30 minutes. The catalyst was then filtered using a syringe microfilter (PTFE; the filter was further washed with MeOH). The filtrate was evaporated to dryness and coevaporated once with DCM. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a colorless oil. Yield: 100 mg (>99%; 1 step; based on TD686). ESI-MS (LC-MS): 387.3 [M+H] + (Theoretical value [C 19 H 39 N4O4] + =387.3). The entire amount of TD691 was used directly in TD692 without further characterization.

[0501] Synthesis of TD1106:

[0502] [ka]

[0503] In a pear-shaped glass flask (50 mL), Cbz2-cyclen (free base, 500 mg; 1.14 mmol; 1.0 equiv.) was dissolved in MeCN (10 mL), followed by the addition of dry KCO (870 mg; 5.69 mmol; 5.0 equiv.) and methyl bromoacetate (235 μL; 2.51 mmol; 2.2 equiv.) in MeCN (5 mL). The resulting suspension was stirred at room temperature for 2 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO, and evaporated to dryness. The residue was dissolved in DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in the form of the free base as a colorless oil. Yield: 422 mg (64%; 1 step; based on the Cbz2 cyclen). NMR (CD3CN): 1 Hδ H 2.63-2.88(mc,m,8H);3.19-3.49(mc,CH2-CO,m,8+4H);3.59(CH3,s,6H);5.07(CH2-Ph,s,4H);7.21-7.41(Ph,m,10H).ESI-LCMS:585.3[M+H] + (Theoretical value [C 30 H 41 N4O8] + =585.3).

[0504] Synthesis of TD1116:

[0505] [ka]

[0506] A pear-shaped glass flask (50 mL) was equipped with palladium on carbon (42 mg) and a magnetic stirrer and protected with argon three times. Next, under a constant flow of argon, a solution of TD1106 (418 mg; 715 μmol) in MeOH (25 mL) was added through the septum. The argon flow was then removed, and H2 gas (from a balloon) was bubbled into the mixture, which was left at room temperature for 1 h. The catalyst was then filtered using a syringe microfilter (PTFE; the filter was further washed with MeOH). The filtrate was evaporated to dryness and coevaporated once with DCM. The residue was further dried under high vacuum overnight to give the product in the form of the free base as a colorless oil, which crystallized upon standing. Yield: 223 mg (99%; 1 step; based on TD1106). NMR (CD3CN): 1 Hδ H 2.48-2.57(mc,m,8H);2.63-2.74(mc,m,8H);3.36(CH2-CO,s,4H);3.64(CH3,s,6H).ESI-MS LC-MS):317.2[M+H] + (Theoretical value [C 14 H 29 N4O4] + =317.2).

[0507] Synthesis of TD687:

[0508] [ka]

[0509] In a glass vial (2 mL), TD680 (150 mg; 270 μmol; 1.0 equiv.) and P(OEt) (232 μL; 1.35 mmol; 5.0 equiv.) were mixed, followed by solid (CHO). n(12 mg; 400 μmol; 1.5 equiv.) was added. The resulting suspension was stirred at room temperature for 3 days. The reaction mixture was diluted with MeCN (700 μL) and filtered through a syringe microfilter (PTFE). The filter was further washed with MeCN. The filtrate was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, immediately neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (25 mL) and HO (25 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in free base form as a faint yellow oil. Yield: 167 mg (87%; 1 step; based on TD680). ESI-MS(LC-MS):705.4[M+H] + (Theoretical value [C 35 H 54 N4O9P1] + =705.4). The entire amount of TD687 was used directly in TD695 without further characterization.

[0510] Synthesis of TD695:

[0511] [ka]

[0512] A pear-shaped glass flask (25 mL) was charged with TD687 (167 mg; 237 μmol) and a magnetic stirrer and protected with argon three times. Solid palladium on carbon (17 mg) was then added and protected with argon three times. MeOH (10 mL) was added through a septum under a constant flow of argon. The argon flow was then removed, and H gas (from a balloon) was bubbled into the mixture, which was then left at room temperature for 30 minutes. The reaction mixture was further stirred at room temperature under a hydrogen atmosphere (from a balloon) for 16 hours. The catalyst was then filtered off using a syringe microfilter (PTFE; the filter was further washed with MeOH). The filtrate was evaporated to dryness and coevaporated once with DCM. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a slightly yellow oil. Yield: 100 mg (97%; 1 step; based on TD686). ESI-MS (LC-MS): 437.2 [M+H] + (Theoretical value [C 19 H 41 N4O5P1] + =37.3). The entire amount of TD695 was used directly in TD700 without further characterization.

[0513] Synthesis of TD913:

[0514] [ka]

[0515] In a glass vial (20 mL), Cbz2-cyclen (free base; 242 mg; 549 μmol; 1.0 equiv.) was dissolved in MeCN (12 mL), followed by solid (CHO). nPhP(OMe)2 (50 mg; 1.67 mmol; 3.0 equiv.) and PhP(OMe)2 (350 μL; 2.2 mmol; 4.0 equiv.) were added. The resulting suspension was stirred at 80 °C for 24 h. The reaction mixture was filtered through a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (75 mL) and HO (75 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in free base form as a nearly colorless oil. Yield: 259 mg (61%; 1 step; based on Cbz2 cyclen). NMR (CD3CN): 1 Hδ H 2.30-3.38(mc,CH2-P,m,16+4H);3.47(CH3,d,6H, 3 J HP =11);5.00-5.12(CH2-Ph,m,4H);7.27-7.40(Ph,m,10H);7.40-7.51(Ph,m,4H);7.51-7.60(Ph,m,2H);7.60-7.82(Ph,m,4H). 31 P δ P 42.8(m). ESI-HRMS:777.3178[M+H] + (Theoretical value [C 40 H 51 N4O8P2] + =777.3177).

[0516] Synthesis of TD910:

[0517] [ka]

[0518] A pear-shaped glass flask (50 mL) was charged with TD913 (250 mg; 322 μmol) and a magnetic stirrer and protected with argon three times. Solid palladium on carbon (50 mg) was then added and protected with argon three times. MeOH (25 mL) was added through a septum under a constant flow of argon. The argon flow was then removed, and H2 gas (from a balloon) was bubbled into the mixture, which was then left at room temperature for 2 hours. The catalyst was then filtered using a syringe microfilter (PTFE; the filter was further washed with MeOH). The filtrate was evaporated to dryness and coevaporated once with DCM. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a nearly colorless oil. Yield: 161 mg (98%; 1 step; based on TD913). NMR (CD3CN): 1 Hδ H 2.28-3.14(mc,CH2-P,m,16+4H);3.53(CH3,d,3H, 3 J HP =11);3.54(CH3,d,3H, 3 J HP =11);7.46-7.63(Ph,m,6H);7.71-7.83(Ph,m,4H). 31 P δ P 43.3(m). ESI-HRMS:509.2438[M+H] + (Theoretical value [C 24 H 39 N4O4P2 + ]=509.2441).

[0519] Synthesis of TD571:

[0520] [ka]

[0521] In a glass vial (4 mL), Cbz2cyclene (free base; 1.00 g; 2.27 mmol; 1.0 equiv.) and P(OEt)3 (2.00 mL; 11.7 mmol; 5.1 equiv.) were mixed, followed by solid (CH2) n(164 mg; 5.47 mmol; 2.4 equiv.) was added. The resulting suspension was stirred at room temperature for 24 h. The reaction mixture was evaporated to dryness and the residue was purified by column chromatography (80 g of SiO2; DCM-MeOH-NH3 in water 150:10:1). The fractions containing the product were combined and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a slightly yellow oil. Yield: 1.29 g (77%; 1 step; based on the Cbz2 cycler). ESI-MS (LC-MS): 741.3 [M+H] + Theoretical value [C 34 H 55 N4O 10 P2] + =741.3). The entire amount of TD571 was used directly for TD573 without further characterization.

[0522] Synthesis of TD573

[0523] [ka]

[0524] A pear-shaped glass flask (100 mL) containing TD571 (1.29 g, 1.74 mmol) and a magnetic stirrer was placed in a flask and protected with argon three times. Solid palladium on carbon (129 mg) was then added and protected with argon three times. Under a constant flow of argon, EtOH (96%, 70 mL) was added through a septum. The argon flow was then removed, and H gas (from a balloon) was bubbled into the mixture, which was then left at room temperature for 30 minutes. The reaction mixture was further stirred under a hydrogen atmosphere (from a balloon) for 16 hours at room temperature. The catalyst was then filtered off using a syringe microfilter (PTFE; the filter was further washed with EtOH). The filtrate was evaporated to dryness and coevaporated once with DCM. The residue was further dried under high vacuum overnight to give the product in the form of the free base as a slightly yellow oil. Yield: 776 mg (95%; 1 step; based on TD571). ESI-MS(LC-MS):473.2[M+H] + (Theoretical value [C 18 H 43 N4O4P2]+ =473.3). The majority of TD573 was used directly for TD579 without further characterization.

[0525] Synthesis of TD423:

[0526] [ka]

[0527] In a pear-shaped glass flask (25 mL), tBuDO2A (free base; 410 mg; 1.03 mmol; 1.4 equiv.) was dissolved in dry MeCN (8 mL), followed by the addition of Cs2CO3 (840 mg; 2.58 mmol; 3.4 equiv.) and KI (172 mg; 1.04 mmol; 1.4 equiv.). A solution of 2-chloro-N-(prop-2-yn-1-yl)acetamide (100 mg; 760 μmol; 1.0 equiv.) in dry MeCN (2 mL) was added, and the resulting mixture was stirred at room temperature for 3 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (100 mL) and H2O (100 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness to give the crude product in the form of the free base as a yellow oil. Yield: 180 mg (mixture of ∼75% TD423 and 25% bis(substituted) by-product). ESI-MS (LC-MS): 496.4 [M+H] + (Theoretical value [C 25 H 46 N5O5] + =496.3). All TD423 was used directly in TD425 without further purification or characterization.

[0528] Synthesis of TD635:

[0529] [ka]

[0530] In a pear-shaped glass flask (250 mL), tBuDO2A (free base; 1.21 g; 3.02 mmol; 2.3 equiv.) was dissolved in MeCN (150 mL). To the vigorously stirred reaction mixture, a solution of TD558 (1202 mg; 1.33 mmol; 1.0 equiv.) in MeCN (100 mL) was added dropwise (over 2 h). The resulting mixture was further stirred at room temperature for 2 days and then evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (100 mL) and HO (100 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the free base form of the product as a faint yellow oil, which crystallized on standing. Yield: 453 mg (66%; 1 step; based on TD558). NMR (CD3CN): 1 Hδ H 1.39(CH3,s,18H);2.53(mc,m,4H);2.61(mc,m,4H);2.73mc,m,4H);2.77(mc,m,4H); 3.01(CH2-CO,s,4H);3.44(C≡CH,s,1H);3.66(CH2-arom,s,2H);7.38(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.70(arom.,t,1H, 3 J HH =8);7.76(arom.,dd,1H,J 3 HH =8, 4 J HH =1). 13 C{ 1 H}δ C28.4(CH3,s);48.2(mc,s);51.8(mc,s);52.6(mc,s);54.9(mc,s);57.6(CH2-CO,s);59.3(CH2-arom,s);78.0(C≡CH,s);81.3(C -CH3,s);84.0(C≡CH,s);124.9(arom.,s);126.5(arom.,s);137.8(arom.,s);141.6(arom.,s);162.8(arom.,s);172.0(CO,s). ESI-HRMS:516.3542[M+H] + (Theoretical value [C 28 H 46 N5O4] + =516.3544).

[0531] Synthesis of TD539:

[0532] [ka]

[0533] In a pear-shaped glass flask (100 mL), tBuDO2A (free base; 1.73 g; 4.32 mmol; ≥ 2.0 equiv.) was dissolved in dry MeCN (15 mL), followed by the addition of dry K2CO3 (900 mg; 6.52 mmol; ≥ 3.0 equiv.). To the vigorously stirred reaction mixture, a solution of freshly prepared and isolated TD538 (≤ 2.17 mmol; 1.0 equiv.) in dry MeCN (10 mL) was added dropwise (over 15 min). The resulting mixture was further stirred at room temperature for 20 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (150 mL) and H2O (150 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 x 100 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a yellow solidified oil. Yield: 653 mg (47%; 2 steps; based on TD530). NMR (CD3CN): 1 Hδ H 1.39(CH3,s,18H);1.43(CH3,s,9H);2.51-2.84(mc,m,16H);3.01(CH2-CO,s,4H);3.64(CH2-arom,s,2H);4.05(CH2-C≡C,bd,2H, 3 J HH =5); 5.97(NH-CO, bt, 1H, 3 J HH =5);7.27(arom.,dd,1H, 3 J HH =7, 4 J HH =2);7.66-7.75(arom,m,2H). 13 C{ 1 H} δ C28.4(CH3,s);28.6(CH3,s);31.2(CH2-C≡C,bs);48.1(mc,s);51.8(mc,s);52.8(mc,s);55.1(mc,s);57.6(CH2-CO,s);59.6(CH2-arom,s);8 1.3(C-CH3, 2×s); 82.6 and 87.0(C≡C, 2×s); 124.3(arom, s); 126.0(arom, s); 137.7(arom, s); 142.3(arom, s); 162.7(arom, s); 172.0(CO, s). ESI-HRMS:645.4331[M+H] + (Theoretical value [C 34 H 57 N6O6] + =645.4334).

[0534] Synthesis of TD1118:

[0535] [ka]

[0536] In a pear-shaped glass flask (50 mL), tBuDO2A (free base; 160 mg; 399 μmol; ≥ 2.5 equiv.) was dissolved in MeCN (10 mL), followed by the addition of dry K2CO3 (65 mg; 470 μmol; ≥ 3 equiv.). To the vigorously stirred reaction mixture, a solution of freshly prepared, isolated TD1117 (≤ 470 mmol; 1.0 equiv.) in MeCN (10 mL) was added dropwise (over 5 min). The resulting mixture was further stirred at room temperature for 16 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a colorless solidified oil. Yield: 62.2 mg (57%; 2 steps; TD966 reference). NMR (CD3CN): 1 Hδ H 1.40(CH3,s,18H);1.43(CH3,s,9H);1.53-1.64(CH2,m,2H);1.82-1.91(CH2,m,2H);2.47-2.79(mc,m,16H);2.79-2.88(CH- C≡C,m,1H);3.01(CH2-CO,s,4H);3.06-3.18(CH2,m,2H);3.63(CH2-arom,s,2H);3.71-3.82(CH2,m,2H);7.27(arom.,dd,1H, 3 J HH =7, 4 J HH =2);7.62-7.72(arom.,m,2H).ESI-HRMS:699.4801[M+H] + (Theoretical value [C 38 H 63 N6O6] + =699.4804).

[0537] Synthesis of TD692:

[0538] [ka]

[0539] In a pear-shaped glass flask (250 mL), TD681 (100 mg; 259 μmol; 1.5 equiv.) was dissolved in MeCN (25 mL). To the vigorously stirred reaction mixture, a solution of TD558 (26 mg; 172 μmol; 1.0 equiv.) in MeCN (25 mL) was added dropwise (over 1 h). The resulting mixture was further stirred at room temperature for 2 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, immediately neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (25 mL) and HO (25 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a yellow oil. Yield: 38 mg (44%; 1 step; based on TD558). ESI-MS (LC-MS): 502.4 [M+H] + (Theoretical value [C 27 H 44 N5O4 + ] + =502.3). The entire amount of TD692 was used directly in TD703 without further characterization.

[0540] Synthesis of TD700:

[0541] [ka]

[0542] In a pear-shaped glass flask (100 mL), TD695 (100 mg; 229 μmol; 1.5 equiv.) was dissolved in MeCN (25 mL), followed by the addition of dry K2CO3 (21 mg; 152 μmol; 1.0 equiv.). To the vigorously stirred reaction mixture, a solution of TD558 (23 mg; 152 μmol; 1.0 equiv.) in MeCN (25 mL) was added dropwise (over 1 h). The resulting mixture was further stirred at room temperature for 16 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (75 mL) and HO (75 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 x 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3) and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a yellow oil. Yield: 20 mg (24%; 1 step; based on TD558). ESI-MS (LC-MS): 552.3 [M+H] + (Theoretical value [C 27 H 47 N5O5P1] + =552.3). The entire amount of TD700 was used directly for TD705 without further characterization.

[0543] Synthesis of TD579:

[0544] [ka]

[0545] In a pear-shaped glass flask (25 mL), TD573 (600 mg; 1.27 mmol; 2.0 equiv.) was dissolved in MeCN (10 mL). To the vigorously stirred reaction mixture, a solution of TD558 (30 mg; 488 μmol; 1.0 equiv.) in MeCN (10 mL) was added dropwise (over 30 min). The resulting mixture was further stirred at room temperature for 16 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (125 mL), HO (125 mL), and aqueous NaOH (1%; 20 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a yellow oil. Yield: 137 mg (48%; 1 step; based on TD558). ESI-MS (LC-MS): 588.3 [M+H] + (Theoretical value [C 26 H 48 N5O6P2] + =588.3). TD579 was used directly for TD575 and TD580 without further characterization.

[0546] Synthesis of TD799:

[0547] [ka]

[0548] In a pear-shaped glass flask (100 mL), tBuDO2Prop (free base; 165 mg; 385 μmol; 2.0 equiv.) was dissolved in MeCN (30 mL). To the vigorously stirred reaction mixture, a solution of TD558 (29 mg; 191 μmol; 1.0 equiv.) in MeCN (30 mL) was added dropwise (over 6 h). The resulting mixture was further stirred at room temperature for 10 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in free base form as a slightly yellow oil. Yield: 47 mg (45%; 1 step; based on TD558). NMR (CD3CN): 1 Hδ H 1.38(CH3,s,18H);2.19-2.25(CH2-CO,m,4H);2.44-2.64(mc,CH2-CH2-CO,m,16+4H);3.41(C≡CH,s,1H);3.66(CH2-arom,s,2H);7.36(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.70(arom.,t,1H, 3 J HH =8);7.77(arom.,bd,1H, 3 J HH = 8). ESI-HRMS: 544.3854 [M+H] + (Theoretical value [C 30 H 50 N5O4] + =544.3857).

[0549] Synthesis of TD663:

[0550] [ka]

[0551] In a pear-shaped glass flask (50 mL), tBuDO2A (free base; 210 mg; 524 μmol; 2.7 equiv.) was dissolved in MeCN (15 mL), followed by the addition of dry K2CO3 (27 mg; 195 μmol; 1.0 equiv.). To the vigorously stirred reaction mixture, a solution of TD566 (50 mg; 193 μmol; 1.0 equiv.) in MeCN (15 mL) was added dropwise (over 15 min). The resulting mixture was further stirred at room temperature for 2 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-Me The product was purified by CN gradient (with FA addition). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (50 mL) and H2O (50 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 x 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in the form of the free base as a nearly colorless oil. Yield: 83 mg (69%; 1 step; based on TD566). NMR (CD3CN): 1 Hδ H 1.33(CH3,s,18H);2.62(mc,m,4H);2.66(mc,m,4H);2.74(mc,m,4H);2.80(mc,m,4H);2.82(CH2-CO,s,4H);3.74(CH2-arom,s,2 H);4.46(CH2-N3,s,2H);2.46-7.58(Ph,arom.,m,3+1H);7.78-7.87(Ph,m,2H);8.18(arom.,bs,1H).ESI-HRMS:623.4022[M+H] + (Theoretical value [C 33 H 51 N8O4] + =623.4028).

[0552] Synthesis of TD711:

[0553] [ka]

[0554] In a pear-shaped glass flask (100 mL), tBuDO2A (free base; 564 mg; 1.41 mmol; 2.5 equiv.) was dissolved in MeCN (40 mL). To the vigorously stirred reaction mixture, a solution of TD406 (103 mg; 564 μmol; 1.0 equiv.) in MeCN (40 mL) was added dropwise (over 2 h). The resulting mixture was further stirred at room temperature for 3 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (100 mL) and HO (100 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in free base form as a nearly colorless oil. Yield: 217 mg (70%; 1 step; based on TD406). NMR (CD3CN): 1 Hδ H 1.39(CH3,s,18H);2.55(mc,m,4H);2.60(mc,m,4H);2.76(mc,m,8H);3.03(CH 2-CO,s,4H);3.70(CH2-arom,s,2H);4.39(CH2-N3,s,2H);7.21(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.67(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.73(arom.,t,1H, 3 J HH =8). 13 C{ 1 H} δ C28.4(CH3,s);48.3(mc,s);51.8(mc,s);52.7(mc,s);54.7(mc,s);56.1(CH2-N3,s);57.7(CH2-CO,s);59.0(CH2-arom,s) ;81.2(C-CH3,s);121.2(arom.,s);124.1(arom.,s);138.2(arom.,s);155.6(arom.,s);161.9(arom.,s);172.0(CO,s). ESI-HRMS:547.3716[M+H] + (Theoretical value [C 27 H 47 N8O4] + =547.3715).

[0555] Synthesis of TD596:

[0556] [ka]

[0557] In a pear-shaped glass flask (25 mL), tBuDO2A (free base; 162 mg; 405 μmol; ≥ 2.5 equiv.) was dissolved in MeCN (10 mL). To the vigorously stirred reaction mixture, a solution of crude TD633 (≤ 162 μmol; 1.0 equiv.) in MeCN (10 mL) was added dropwise (over 15 min). The resulting mixture was further stirred at room temperature for 2 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated, and the aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in free base form as a nearly colorless oil. Yield: 44 mg (48%; 2 steps; based on TD406). ESI-MS (LC-MS): 563.4 [M+H] + (Theoretical value [C 27H 47 N8O5] + =563.4). The entire amount of TD596 was used directly in TD604 without further characterization.

[0558] Synthesis of TD1340:

[0559] [ka]

[0560] In a pear-shaped glass flask (50 mL), Cbz2 cyclen (free base; 1.32 g; 3.0 mmol; 1.0 equiv.) was dissolved in dry MeCN (15 mL), followed by the addition of dry K2CO3 (1.65 g; 12.0 mmol; 4.0 equiv.). Next, a solution of TD1339 (1.50 g; 6.0 mmol; 2.0 equiv.) in dry MeCN (5 mL) was added, and the resulting suspension was stirred at room temperature for 8 h. The solid was filtered off using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (150 mL) and HO (150 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in the form of the free base as a colorless oil. Yield: 1.57 g (82%; 1 step; based on the Cbz2 cyclen). NMR (CD3CN): 1 Hδ H 1.09(CH3-CH,d,6H, 3 J HH =7);1.19(CH3-CH2,t,6H, 3 J HH =7);2.58-2.70(mc,m,4H);2.76-2.98(mc,m,4H);3.23-3.52(mc,m,8H);3.52-3.79(CH,m,2H);4.06(CH2-CH3,q,4H, 3 JHH =7);5.00-5.13(CH2-Ph,m,4H);7.24-7.41(Ph,m,10H).ESI-HRMS:641.3539[M+H] + (Theoretical value [C 34 H 49 N4O8] + =641.3545).

[0561] Synthesis of TD1341:

[0562] [ka]

[0563] A pear-shaped glass flask (100 mL) was equipped with palladium on carbon (127 mg) and a magnetic stirrer and protected with argon three times. Next, a solution of TD1340 (1.27 g; 1.98 mmol) in MeOH (50 mL) was added through the septum. The argon was then removed, and H2 (from a balloon) gas was bubbled through the mixture for 1 h at room temperature. The catalyst was then filtered using a syringe microfilter (PTFE; the filter was further washed with MeOH). The filtrate was evaporated to dryness and coevaporated once with DCM. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a nearly colorless oil. Yield: 723 mg (98%; 1 step; based on TD1340). NMR (CD3CN): 1 Hδ H 1.16-1.33(CH3,m,12H);2.35-2.45(mc,m,4H);2.56-2.65(mc,m,4H);2.65-2.76(mc,m,4H);2.76-2.85(mc,m,4H);3.54(CH,q,2H, 3 J HH =7);4.00-4.24(CH2-CH3,q,4H, 3 J HH = 7). ESI-HRMS: 372.2808 [M+H] + (Theoretical value [C 18 H 37 N4O4 + ] + =373.2809).

[0564] Synthesis of TD1343:

[0565] [ka]

[0566] In a pear-shaped glass flask (50 mL), TD1341 (143 mg; 384 μmol; 2.0 equiv.) was dissolved in MeCN (15 mL) and dry K2CO3 (26.3 mg; 191 μmol; 1.0 equiv.) was added. To the vigorously stirred reaction mixture, a solution of TD558 (29.0 mg; 191 μmol; 1.0 equiv.) in MeCN (15 mL) was added dropwise (over 15 min). The resulting mixture was stirred at room temperature for 2 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (25 mL) and HO (25 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 x 10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried briefly under high vacuum to give the product in free base form as a nearly colorless oil. Yield: 46.0 mg (49%; 1 step; based on TD558). NMR (CD3CN): 1 Hδ H 1.14(CH3-CH,d,6H, 3 J HH =7);1.19(CH3-CH2,t,6H, 3 J HH =7);2.28-2.38(mc,m,4H);2.45-2.59(mc,m,4H);2.59-2.94(mc,m,8H);3.23(CH,q,2H, 3 J HH =7);3.41(CH2-arom, d, 1H, 2 J HH =15);3.45(C≡CH,s,1H);3.88(CH2-arom.,d,1H, 2 JHH =15);4.06(CH2-CH3,q,4H, 3 J HH =7);7.35-7.42(arom.,m,1H);7.66-7.79(arom.,m,2H).ESI-HRMS:488.3231[M+H] + (Theoretical value [C 26 H 42 N5O4] + =488.3231).

[0567] Synthesis of TD1345:

[0568] [ka]

[0569] In a pear-shaped glass flask (25 mL), TD1343 (45.0 mg; 92 μmol; 1.0 equiv.) was dissolved in MeCN (2 mL), followed by the addition of dry K2CO3 (51 mg; 196 μmol; 4.0 equiv.). Next, a solution of TD406 (20.0 mg; 110 μmol; 1.2 equiv.) in MeCN (2 mL) was added, and the resulting suspension was stirred at room temperature for 7 days. The solid was filtered through a syringe microfilter (PTFE), and the filtrate was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (25 mL) and HO (25 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 × 10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried briefly under high vacuum to give the product in free base form as a nearly colorless oil. Yield: 52.9 mg (90%; 1 step; based on TD1343). ESI-HRMS: 634.3825 [M+H] + (Theoretical value [C 33 H 48 N9O4] + =634.3824). The entire amount of TD1345 was used directly for TD1346 without further characterization.

[0570] Synthesis of TD1447:

[0571] [ka]

[0572] In a pear-shaped glass flask (100 mL), TD1341 (722 mg; 1.94 mmol; 2.0 equiv) was dissolved in MeCN (40 mL), followed by the addition of dry K2CO3 (133 mg; 964 μmol; 1.0 equiv). To the vigorously stirred reaction mixture, a solution of TD1057 (241 mg; 962 μmol; 1.0 equiv) in MeCN (40 mL) was added dropwise (over 2 h). The resulting mixture was further stirred at room temperature for 16 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (100 mL) and HO (100 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 x 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the formate salt as a faint yellow oil. Yield: 367 mg (60% assuming M-1.0FA; M R = 632.7; 1st step; TD1057 standard). Recovery amount: 352 mg of TD1341 (49% of the initial amount). NMR (CD3CN): 1 Hδ H 1.18(CH3-CH,d,6H, 3 J HH =7);1.21(CH3-CH2,t,6H, 3 J HH =7);2.52-2.62(mc,m,4H);2.64-2.76(mc,m,4H);2.81-2.94(mc,m,4H);2.95-3.06(mc,m,4H);3.39(CH-CH3,q,2H, 3 J HH=7);3.81(CH2-arom、d,1H, 2 J HH =15);3.90(CH2-arom.,d,1H, 2 J HH =15);4.09(CH2-CH3,q,4H, 3 J HH =7);4.62(CH2-N3,s,2H);7.58(arom.,d,1H, 4 J HH =2);7.66(arom.,d,1H, 4 J HH =2). 19 F δ F -65.10(s). ESI-HRMS:587.3266[M+H] + (Theoretical value[C 26 H 42 N8O4F3] + =587.3276).

[0573] Synthesis of TD1449:

[0574]

change

[0575] In a pear-shaped glass flask (25 mL), TD1447 (assuming M-FA; 263.0 mg; 418 μmol; 1.0 equiv.) was dissolved in MeCN (5 mL), followed by the addition of dry K2CO3 (231 mg; 1.67 μmol; 4.0 equiv.). Next, a solution of TD1428 (173.0 mg; 421 μmol; 1.0 equiv.) in MeCN (10 mL) was added, and the resulting suspension was stirred at 40 °C for 1 h. The solid was filtered off using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 x 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried briefly under high vacuum to give the product in the form of the free base as a yellow waxy oil. Yield: 309 mg (81%; 1 step; based on TD1447). NMR (CD3CN): 1 Hδ H 1.10(CH3-CH,d,6H, 3 J HH =7);1.13(CH3-CH2,t,6H, 3 J HH =7);1.43(CH3-CO,s,9H);1.61(CH3-CN,s,6H);2.53-2.80(mc,m,12H);2.89-3.03(mc,m,4H);3.32(CH-CH3,q,2H, 3 J HH =7);3.68(CH2-arom, s,2H);3.71(CH2-arom, d,1H, 2 J HH =15);3.77(CH2-arom.,d,1H, 2 J HH =15);3.89(CH3-O,s,3H);3.99(CH2-CH3,q,4H,J 3 HH =7);4.52(CH2-N3,s,2H);5.56(NH,bs,1H);7.50(arom,d,1H, 4 JHH =2);7.71(arom.,d,1H, 4 J HH =2);8.00(arom.,d,1H, 4 J HH =2);8.16(arom.,d,1H, 4 J HH =2). 19 F δ F -64.98(s). ESI-HRMS:917.4851[M+H] + (Theoretical value[C 44 H 64 N 10 O8F3] + =917.4855).

[0576] Synthesis of rac-TD1489:

[0577]

change

[0578] In a glass vial (20 mL), Cbz2 cyclen (free base; 178 mg; 404 μmol; 1.0 equiv.) was dissolved in MeCN (2 mL), followed by the addition of dry K2CO3 (225 mg; 1.63 mmol; 4.0 equiv.). Next, a solution of TD1488 (250 mg; 850 μmol; 2.1 equiv.) in MeCN (2 mL) was added, and the resulting suspension was stirred at 40 °C for 4 days (racemization occurred during the reaction). The solid was filtered using a syringe microfilter (PTFE) and washed with additional MeCN. The filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and concentrated on a rotary evaporator to remove most of the MeCN. The mixture was diluted with DCM (25 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 x 10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a colorless oil. Yield: 172 mg (73%; 1 step; based on Cbz2 cyclen). ESI-MS (LC-MS): 585.3 [M+H] + (Theoretical value [C 30 H 41 N4O8] + =585.3).

[0579] Synthesis of TD1493:

[0580] [ka]

[0581] In a glass vial (20 mL), rac-TD1489 (free base; 172 mg; 294 μmol; 1.0 equiv.) was dissolved in MeCN (2 mL), followed by the addition of dry K2CO3 (162 mg; 1.17 mmol; 4.0 equiv.). Next, a solution of TD1399 (148 mg; 592 μmol; 2.0 equiv.) in MeCN (2 mL) was added, and the resulting suspension was stirred at 40 °C for 70 min. The solid was filtered using a syringe microfilter (PTFE) and washed with MeCN. The filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the pure product (the more hydrophobic diastereomer, differing in the chirality of the malic acid arm, was successfully separated) were combined, neutralized with dilute aqueous NaHCO3, and concentrated on a rotary evaporator to remove most of the MeCN. The mixture was diluted with DCM (25 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 x 10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried under high vacuum overnight to give the product in the form of the free base as a colorless oil. Yield: 88.1 mg (44%; 1 step; based on rac-TD1489). NMR (CD3CN): 1 Hδ H 1.09(CH3-CH,d,3H, 3 J HH =7);1.19(CH3-CH2,t,3H, 3 J HH =7);2.44(CH2-CH,dd,1H, 2 J HH =17, 3 J HH =6);2.55-2.70(mc,CH2-CH,m,3+2H);2.73-7.43(Ph,m,10H).ESI-HRMS:685.3439[M+H] + (Theoretical value [C 35 H 49 O 10 N4] + =685.3443).

[0582] Synthesis of TD1495:

[0583] [ka]

[0584] A pear-shaped glass flask (25 mL) was equipped with palladium on carbon (9 mg) and a magnetic stirrer and protected with argon three times. Next, a solution of TD1493 (88.0 mg; 129 μmol) in MeOH (6 mL) was added through the septum. The argon flow was then removed, and H2 (from a balloon) gas was bubbled into the mixture, which was then left at room temperature for 1 h. The catalyst was then filtered using a syringe microfilter (PTFE; the filter was further washed with MeOH). The filtrate was evaporated to dryness and coevaporated once with DCM. The residue was further dried under high vacuum overnight to give the product in the form of the free base as a colorless oil. Yield: 52.6 mg (98%; 1 step; based on TD1493). ESI-HRMS: 417.2706 [M+H] + (Theoretical value [C 19 H 37 O6N4] + =417.2708).

[0585] Synthesis of TD1497:

[0586] [ka]

[0587] In a pear-shaped glass flask (25 mL), TD1495 (55 mg; 132 μmol; 1.7 equiv.) was dissolved in MeCN (5 mL), followed by the addition of dry K2CO3 (11.0 mg; 80 μmol; 1.0 equiv.). To the vigorously stirred reaction mixture, a solution of TD558 (12.0 mg; 79 μmol; 1.0 equiv.) in MeCN (5 mL) was added dropwise (over 15 min). The resulting mixture was stirred at room temperature for 4 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (25 mL) and HO (25 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 x 10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried briefly under high vacuum to give the product in free base form as a nearly colorless oil. Yield: 12.1 mg (29%; 1 step; based on TD558). ESI-MS (LC-MS): 532.3 [M+H] + (Theoretical value [C 27 H 42 N5O6] + =532.3).

[0588] Synthesis of TD1500:

[0589] [ka]

[0590] In a glass vial (4 mL), TD1497 (12.1 mg; 23 μmol; 1.0 equiv.) was dissolved in a solution of TD406 (5.8 mg; 32 μmol; 1.4 equiv.) in MeCN (2 mL), followed by the addition of dry K2CO3 (17.5 mg; 127 μmol; 4.0 equiv.). The resulting suspension was stirred at 40 °C for 1 day. The solid was filtered off using a syringe microfilter (PTFE), and the filtrate was diluted with HO (1 mL). The resulting solution was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined, neutralized with diluted aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (25 mL) and HO (25 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (4 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried briefly under high vacuum to give the product in free base form as a faint yellow oil. Yield: 11.9 mg (77%; 1 step; based on TD1497). ESI-MS (LC-MS): 678.3 [M+H] + (Theoretical value [C 34 H 48 N9O6] + =678.4).

[0591] Example 4: Synthesis of uncaged macrocyclic ligands Synthesis of TD425:

[0592] [ka]

[0593] In a pear-shaped glass flask (50 mL), crude TD423 (75% in a mixture with 25% or less of the bis(substituted) by-product; 180 mg; ≥ 1.0 equiv.) was dissolved in dry MeCN (10 mL), followed by the addition of Cs2CO3 (360 mg; 1.11 mmol; 3.8 equiv.). Next, a solution of TD566 (75 mg; 290 μmol; 1.0 equiv.) in MeCN (5 mL) was added, and the resulting suspension was stirred at room temperature for 2 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). Fractions containing the product were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 129 mg (27%; 3 steps; based on 2-chloro-N-(prop-2-yn-1-yl)acetamide). NMR (DO, pD~5): 1 Hδ H 2.62(C≡CH,t,1H, 4 J HH =3);2.89-3.74(mc,CH2-CO,m,16+6H);3.79(CH2-C≡CH,t,2H, 4 J HH =3);4.03(CH2-arom, bs,2H);4.68(CH2-N3,s,2H);7.55-7.67(Ph,arom.,m,3+1H);7.78(arom.,d,1H, 4 J HH =2);7.85-7.92(Ph,m,2H);8.17(arom.,d,1H, 4 J HH = 2). ESI-HRMS: 606.3151 [M+H] + (Theoretical value [C 30 H 40 N9O5] + =606.3147). EA(C30 H 39 N9O5-0.1FA-1.3H2O,M R =633.7):C57.0(56.8);H6.6(6.3);N19.9(19.5). Synthesis of TD669:

[0594] [ka]

[0595] In a glass vial (4 mL), TD663 (30 mg; 48 μmol, 1.0 equiv.) was dissolved in dry MeCN (500 μL), followed by the addition of K2CO3 (20 mg; 145 μmol; 3 equiv.). Next, a solution of TD662 (10 mg; 50 μmol; 1.0 equiv.) in MeCN (500 μL) was added, and the resulting suspension was stirred at room temperature for 2 days. Another portion of TD662 (6 mg; 30 μmol; 0.6 equiv.) in MeCN (200 μL) was then added, and the mixture was stirred at room temperature for another 2 days. The solid was filtered through a syringe microfilter (PTFE), and the filtrate was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the product were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 27 mg (≦88% M-0.1FA-×HO; M R = 634.4; 2-phase; assumed to be TD663 standard). NMR (DO, pD ~ 5): 1 Hδ H2.59-3.82(mc,CH2-CO,CH2-CH2-N(CH2-C≡CH)2,m,16+4+10H);4.00(CH2-arom,bs,2H);4.64(CH2-N3,s,2H);7.55-7.65(Ph,m,3H);7.74(arom.,d,1H, 4 J HH =2);7.83-7.91(Ph,m,2H);8.17(arom.,d,1H, 4 J HH =2). ESI-HRMS: 630.3510 [M+H] + (Theoretical value[C 33 H 44 N9O4] + =630.3511).

[0596] Synthesis of TD604:

[0597]

change

[0598] In a glass vial (4 mL), TD596 (44 mg; 78 μmol; 1.0 equiv.) was dissolved in MeCN (3 mL) and dry K2CO3 (33 mg; 239 μmol; 3.0 equiv.) was added. Next, a solution of TD558 (13 mg; 86 μmol; 1.1 equiv.) in MeCN (600 μL) was added, and the resulting suspension was stirred at room temperature for 16 days. Next, an additional portion of TD558 (2 mg; 13 μmol; 0.2 equiv.) in MeCN (400 μL) was added. The reaction mixture was further stirred at room temperature for 2 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the product were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 38 mg (81%; 2 steps; TD596 reference). NMR (DO, pD~4): 1 Hδ H 2.80-3.68(mc,CH2-CO,m,16+4H);3.72(C≡CH,s,1H);3.93(CH2-arom.,bs,2H);4.17(CH2 -arom.,s,2H);4.74(CH2-N3,s,2H);7.48-7.67(arom.,m,3H);7.74-7.95(arom.,m,3H). 13 C{ 1 H}δ C48.3(mc,s);48.9(mc,s);50.6(CH2-N3,s);51.4(CH2-arom.,s);51.5(mc,s);51.7 (mc,s);56.7(CH2-CO,s);58.8(CH2-arom.,s);80.7 and 82.2(C≡CH;2×s);126.0(aro m, s);126.5(arom, s);127.9(arom, s);128.5(arom, s);131.5(arom, s);140.3(arom s);141.2(arom, s);148.4(arom, s);148.7(arom, s);157.3(arom, s);169.5(CO, s). ESI-HRMS: 566.2836 [M+H] + (Theoretical value [C 27 H 36 N9O5] + =566.2834). EA(C 27 H 35 N9O5-0.1FA-1.6H2O,M R =599.1):C54.3(54.4);H6.5(6.1);N21.0(20.7).

[0599] Synthesis of TD556:

[0600] [ka]

[0601] In a pear-shaped glass flask (25 mL), TD635 (216 mg; 419 μmol; 1.0 equiv.) was dissolved in MeCN (12 mL), followed by the addition of dry K2CO3 (174 mg; 1.26 mmol; 3.0 equiv.). Next, a solution of TD406 (92 mg; 504 μmol; 1.2 equiv.) in MeCN (3 mL) was added, and the resulting suspension was stirred at room temperature for 3 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 161 mg (66%; 2 steps; TD635 reference). NMR (DO, pD~4): 1 Hδ H 2.80-3.60(mc,CH2-CO,m,16+4H);3.74(C≡CH,s,1H);3.86(CH2-arom.,bs,2H);3.94(CH2-arom.,bs,2H);4.72(CH2-N3,s,2H);7.65(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.67(arom,dd,1H, 3 J HH =8, 4 J HH =1);7.95-8.03(arom.,m,3H);8.12(arom.,t,1H, 3 J HH =8); 13 C{ 1 H} δ C48.3(mc,s);48.4(mc,s);51.4(mc,s);51.5(mc,s);54.3(CH2-N3,s);56.6(CH2-CO, s);58.0(CH2-arom, s);58.9(CH2-arom.,s);80.9 and 82.0(C≡CH;2×s);124.5(arom., s);126.2(arom.,s);126.3(arom.,s);128.5(arom, s);140.5(arom.s);141.2(arom .s);142.7(arom.s);155.0(arom.s);155.5(arom.s);157.2(arom.s);169.2(CO, s). ESI-HRMS: 550.2880 [M+H] + (Theoretical value [C 27 H 36 N9O4] + =550.2885). EA(C 27 H 35 N9O4-0.1FA-1.6H2O,M R =583.1):C55.8(55.5);H6.6(6.1);N21.6(21.2).

[0602] Synthesis of TD810:

[0603] [ka]

[0604] In a glass vial (20 mL), TD711 (103 mg; 188 μmol; 1.0 equiv.) was dissolved in MeCN (5 mL) and dry K2CO3 (130 mg; 942 μmol; 5.0 equiv.) was added. Next, a solution of TD807 (33 mg; 188 μmol; 1.0 equiv.) in MeCN (2 mL) was added, and the resulting suspension was stirred at room temperature for 3 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined and directly lyophilized to give the product as a mixed trifluoroacetate / formate salt as an off-white fluffy solid. Yield: 55 mg (43%; 2 steps; TD711 reference). NMR (DO, pD~4): 1 Hδ H 2.79-3.63(mc,CH2-CO,m,16+4H);3.74(C≡CH,s,1H);3.82(CH2-arom.,bs,2H);3. 87(CH2-arom.,bs,2H);3.92(C≡CH,s,1H);4.63(CH2-N3,s,2H);7.55(arom, dd,1H, 3 J HH =8, 4 J HH =1);7.72(arom.,d,1H, 4 J HH =1);8.10(arom.,dd,1H, 3 J HH =8, 4 J HH =1);8.16(arom.,t,1H, 3 J HH =8);8.26(arom.,d,1H, 4 J HH = 1). ESI-HRMS: 574.2882 [M+H] +(Theoretical value [C 29 H 36 N9O4] + =574.2885). EA(C 29 H 35 N9O4-0.5TFA-0.1FA-2.1H2O,M R =673.1):C53.7(53.8);H6.0(6.2);N18.7(18.7).

[0605] Synthesis of TD827:

[0606] [ka]

[0607] In a glass vial (20 mL), TD635 (157 mg; 304 μmol; 1.0 equiv.) was dissolved in dry MeCN (7 mL) and dry K2CO3 (210 mg; 1.52 mmol; 5.0 equiv.) was added. Next, a solution of TD817 (110 mg; 303 μmol; 1.0 equiv.) in dry MeCN (3 mL) was added, and the resulting suspension was stirred at room temperature for 2 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl-protected product were combined, concentrated to approximately half the volume (to remove most of the MeCN), and diluted with DCM (125 mL). The resulting biphasic mixture was transferred to a separatory funnel. The aqueous phase was neutralized with dilute aqueous NaHCO3. After shaking, the bottom layer was separated, and the aqueous layer was further extracted with DCM (3 x 50 mL). The combined organic layers were filtered through a glass frit (S3) with anhydrous Na2SO4 and evaporated to dryness. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated once with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; H2O-MeCN gradient, FA added). The product-containing fractions were combined and directly lyophilized to give the product as an off-white fluffy solid in the form of the formate salt. Yield: 149 mg (62%; 2 steps; TD711 reference). NMR (DO+FA, pD~3):1 H δ H 0.86 - 1.33 (i-Pr, m, 21H); 2.58 - 3.78 (mc, CH2-CO, C≡CH, bm, 16 + 4 + 1H); 3.93 (CH2-arom., bs, 2H); 4.06 (CH2-arom, bs, 2H); 4.56 (CH2-N3, s, 2H); 7.37 (arom., s, 1H); 7.53 (arom., d, 1H, 3 J HH = 8); 7.74 (arom., s, 1H); 7.80 (arom., d, 1H, 3 J HH = 8); 7.96 (arom., t, 1H, 3 J HH = 8). ESI-HRMS: 730.4216 [M + H] + (Theoretical value [C 38 H 56 N9O4Si1] + = 730.4219). EA (C 38 H 55 N9O4Si1-0.8FA-1.6H2O, M R = 795.6): C 58.6 (58.7); H 7.6 (7.7); N 15.8 (15.7); Si 3.5 (3.2).

[0608] Synthesis of TD718:

[0609]

Chem.

[0610] In a pear-shaped glass flask (10 mL), TD711 (31 mg; 57 μmol; 1.0 equiv.) was dissolved in dry MeCN (2 mL), followed by the addition of dry K2CO3 (31 mg; 225 μmol; 4.0 equiv.). Next, a solution of TD706 (13 mg; 58 μmol; 1.0 equiv.) in dry MeCN (2 mL) was added, and the resulting suspension was stirred at room temperature for 24 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). The fractions containing the tert-butyl-protected product were combined, diluted with DCM (100 mL), and transferred to a separatory funnel. The aqueous phase was neutralized with dilute aqueous NaHCO3. After shaking, the bottom phase was separated, dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated once with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the product were combined and directly lyophilized to give the zwitterionic form of the product as a white fluffy solid. Yield: 23 mg (61%; 2 steps; TD706 reference). NMR (DO, pD~5): 1 Hδ H 0.28(CH3,s,9H);2.28-3.67(mc,CH2-CO,m,16+4H);3.89(CH2-arom.,bs,4H);4.57(CH2-N3,s,2H);7.49(arom.,dd,1H, 3 J HH =7, 4 J HH =2);7.58-7.64(arom.,m,1H);7.78-7.98(arom.,m,4H).ESI-HRMS:622.6277[M+H] + (Theoretical value [C 30 H 44 N9O4Si1] + =622.3280). EA(C 30 H 43 N9O4Si1-2.6H2O,M R=668.7):C53.9(53.9);H7.3(7.0);N18.9(18.7). Synthesis of TD728:

[0611] [ka]

[0612] In a pear-shaped glass flask (10 mL), TD711 (66 mg; 121 μmol; 1.0 equiv.) was dissolved in MeCN (2 mL), followed by the addition of dry K2CO3 (67 mg; 486 μmol; 4.0 equiv.). Next, a solution of TD723 (37 mg; 120 μmol; 1.0 equiv.) in MeCN (2 mL) was added, and the resulting suspension was stirred at room temperature for 24 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). The fractions containing the tert-butyl-protected product were combined and diluted with DCM (150 mL), and the resulting biphasic mixture was transferred to a separatory funnel. The aqueous phase was neutralized with dilute aqueous NaHCO3. After shaking, the bottom phase was separated, and the aqueous phase was further extracted with DCM (2 × 50 mL). The combined organic layers were filtered through a glass frit (S3) with anhydrous Na2SO4 and evaporated to dryness. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated once with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined and directly lyophilized to give the zwitterionic form of the product as a white fluffy solid. Yield: 64 mg (71%; 2 steps; TD723 reference). NMR (DO, pD~5): 1 Hδ H 0.84-1.34(i-Pr,m,21H);2.41-3.78(mc,CH2-CO,m,16+4H);3.89(CH2-arom.,bs,2H);3.95(CH2-arom.,bs,2H);4.52(CH2-N3,s,2H);7.42(arom,d,1H, 3 J HH=8);7.46(arom.,d,1H, 3 J HH =8);7.74(arom.,d,1H, 3 J HH =8);7.78(arom.,d,1H, 3 J HH =8);7.85(arom.,t,1H, 3 J HH =8);7.87(arom.,t,1H, 3 J HH =8). ESI-HRMS: 706.4221 [M+H] + (Theoretical value[C 36 H 56 N9O4Si1] + =706.4219). EA(C 36 H 55 N9O4Si1-2.6H2O,M R =752.8):C57.4(57.7);H8.1(8.0);N16.7(16.5);Si3.7(3.8).

[0613] Synthesis of TD1204:

[0614]

change

[0615] In a glass vial (20 mL), TD711 (35.0 mg; 64.0 μmol; 1.0 equiv.) was dissolved in MeCN (1 mL) and dry K2CO3 (44.2 mg; 320 μmol; 5.0 equiv.) was added. Next, a solution of TD1194 (21.4 mg; 77.1 μmol; 1.2 equiv.) in MeCN (2 mL) was added, and the resulting suspension was stirred at room temperature for 2 h. The solid was filtered off using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in the formate form as a white fluffy solid. Yield: 23.3 mg (49%; 2 steps; based on TD711). NMR (DO, pD ~ 7): 1 Hδ H 2.83-3.67(mc,CH2-CO,m,16+4H);3.88(CH2-arom.,bs,2H);3.92(CH2-arom.,bs,2H);4.56(CH2-N3,s,2H);7.49(arom.,d,1H, 3 J HH =8);7.54(arom.,d,1H, 3 J HH =8);7.87-8.00(arom.,m,4H).ESI-HRMS:676.1855[M+H] + (Theoretical value [C 27 H 35 N9O4I1 + ]=676.1851). EA(C 27 H 34 N9O4I1-0.3FA-2.7H2O,M R =738.0):C44.4(44.6);H5.5(5.1);N17.1(16.7).

[0616] Synthesis of TD944:

[0617] [ka]

[0618] In a pear-shaped glass flask (10 mL), TD711 (58 mg; 106 μmol; 1.0 equiv.) was dissolved in MeCN (2 mL), followed by the addition of dry K2CO3 (87 mg; 629 μmol; 6.0 equiv.). Next, a solution of freshly prepared and isolated TD939 (≦105 μmol; ≦1.0 equiv.) in MeCN (2 mL) was added, and the resulting suspension was stirred at room temperature for 2 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in zwitterionic form as a slightly yellow solid. Yield: 38 mg (55%; 3 steps; TD936 reference). NMR (DO, pD ~ 7): 1 Hδ H 2.82-3.67(mc,CH2-CO,m,16+4H);3.91(CH2-arom.,bs,2H);3.95(CH2-arom.,bs,2 H);4.58(CH2-N3,s,2H);7.40-8.07(Ph,arom.,m,5+6H).ESI-HRMS:626.3197[M+H] + (Theoretical value [C 33 H 40 N9O4] + =626.3198). EA(C 30 H 39 N9O4-1.5H2O,M R =652.8):C60.7(61.0);H6.5(6.3);N19.3(18.8).

[0619] Synthesis of TD943:

[0620] [ka]

[0621] In a pear-shaped glass flask (10 mL), TD711 (82 mg; 150 μmol; 1.0 equiv.) was dissolved in MeCN (2 mL), followed by the addition of dry K2CO3 (125 mg; 904 μmol; 6.0 equiv.). Next, a solution of freshly prepared and isolated TD940 (≦150 μmol; ≦1.0 equiv.) in MeCN (2 mL) was added, and the resulting suspension was stirred at room temperature for 2 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in zwitterionic form as a slightly yellow solid. Yield: 66 mg (71%; 3 steps; TD937 reference). NMR (DO, pD ~ 7): 1 Hδ H 0.74-1.06(CH2-CH,m,4H);1.35-1.66(CH2-CH,m,1H);2.69-3.65(mc,CH2-CO,m,16+4H);3.86(CH2-arom,bs,2H);3.92( CH2-arom.,bs,2H);4.57(CH2-N3,s,2H);7.42-7.53(arom.,m,2H);7.83-8.01(arom.,m,4H).ESI-HRMS:590.3196[M+H] + (Theoretical value [C 30 H 40 N9O4] + =590.3198). EA(C 30H 39 N9O4-1.5H2O,M R =616.7):C58.4(58.6);H6.9(6.9);N20.4(20.3).

[0622] Synthesis of TD959:

[0623] [ka]

[0624] In a pear-shaped glass flask (10 mL), TD711 (23 mg; 42 μmol; 1.0 equiv.) was dissolved in MeCN (1 mL), followed by the addition of dry K2CO3 (35 mg; 253 μmol; 6.0 equiv.). Next, a solution of freshly prepared and isolated TD956 (≦42 μmol; ≦1.0 equiv.) in MeCN (1 mL) was added, and the resulting suspension was stirred at room temperature for 2 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in zwitterionic form as a nearly colorless solid. Yield: 18 mg (67%; 3 steps; based on TD952). NMR (DO, pD ~ 6): 1 Hδ H 1.34(CH3,s,9H);2.72-3.69(mc,CH2-CO,m,16+4H);3.88(CH2-arom.,bs,2H);3.92(CH2-arom.,bs,2H );4.57(CH2-N3,s,2H);7.46-7.55(arom.,m,2H);7.84-8.03(arom.,m,4H).ESI-HRMS:606.3513[M+H] + (Theoretical value [C31 H 44 N9O4] + =606.3511). EA(C 31 H 43 N9O4-1.8H2O,M R =638.2):C58.3(58.7);H7.4(7.0);N19.8(19.4) Synthesis of TD992:

[0625] [ka]

[0626] In a glass vial (4 mL), TD711 (43 mg; 79 μmol; 1.0 equiv.) was dissolved in MeCN (1 mL), followed by the addition of dry K2CO3 (44 mg; 318 μmol; 4.1 equiv.). Next, a solution of freshly prepared and isolated TD990 (≦79 μmol; ≦1.0 equiv.) in MeCN (1 mL) was added, and the resulting suspension was stirred at 40 °C for 16 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined and directly lyophilized to give the product in the formate form as an off-white fluffy solid. Yield: 42.0 mg (73%; 3 steps; TD965 reference). NMR (DO, pD~5): 1 Hδ H1.62 - 1.83 (Adm., m, 6H); 1.86 - 2.04 (Adm., m, 9H); 2.77 - 3.70 (mc, CH2-CO, m, 16 + 4H); 3.90 (CH2-arom., bs, 2H); 3.94 (CH2-arom., bs, 2H); 4.57 (CH2-N3, s, 2H); 7.44 - 7.56 (arom., m, 2H); 7.82 - 8.00 (arom., m, 4H). ESI-HRMS: 684.3982 [M+H] + (Theoretical value [C 37 H 50 N9O4] + = 684.3980). EA (C 37 H 49 N9O4-0.3FA-1.9H2O, M R = 731.9): C 61.2 (61.2); H 7.1 (7.1); N 17.2 (17.0). Synthesis of TD703:

[0627]

Chemical formula

[0628] In a glass vial (20 mL), TD692 (38 mg; 76 μmol; 1.0 equiv.) was dissolved in MeCN (3 mL), followed by the addition of dry K2CO3 (31 mg; 217 μmol; 3.0 equiv.). Next, a solution of TD406 (14 mg; 77 μmol; 1.0 equiv.) in MeCN (2 mL) was added, and the resulting suspension was stirred at room temperature for 16 h. Then, another portion of dry K2CO3 (31 mg; 217 μmol; 3.0 equiv.) and TD406 (10 mg; 55 μmol; 0.7 equiv.) in MeCN (1 mL) was added, and the resulting suspension was further stirred at room temperature for 3 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). Fractions containing the protected product were combined, evaporated to dryness, and coevaporated twice with MeOH. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated three times with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the product were combined and directly lyophilized to give the product as a slightly yellow viscous oil in the form of a mixed trifluoroacetate / formate salt. Yield: 16 mg (34%; 2 steps; TD692 reference). NMR (DO, pD ~ 5): 1 Hδ H 2.59-3.73(mc,CH2-CO,m,16+2H);3.75(C≡CH,s,1H);3.84(CH2-arom.,bs,2H);4.29-4.62(CH2-arom.,CH2-N3,m,2+2H);7.37(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.45(arom,dd,1H, 3 J HH =8, 4 J HH =1);7.52(arom.,t,1H, 3 J HH =8);7.56(arom.,dd,1H, 3 J HH =8, 4 J HH=1);7.59(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.89(arom.,t,1H, 3 J HH =8).ESI-HRMS: 492.2829 [M+H] + (Theoretical value [C 25 H 34 N9O2] + =492.2830). EA(C 25 H 33 N9O2-0.3TFA-0.7FA-3.5H2O,M R =621.1):C50.9(51.1);H6.8(6.8);N20.3(20.0).

[0629] Synthesis of TD705:

[0630] [ka]

[0631] In a glass vial (20 mL), TD700 (20 mg; 36 μmol; 1.0 equiv.) was dissolved in MeCN (3 mL), followed by the addition of dry K2CO3 (20 mg; 145 μmol; 4.0 equiv.). Next, a solution of TD406 (9 mg; 49 μmol; 1.4 equiv.) in MeCN (1 mL) was added, and the resulting suspension was stirred at room temperature for 2 days. The solid was filtered off using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in free base form as a nearly colorless oil. Yield: 22 mg (87%; 1 step; TD406 reference). ESI-MS (LC-MS): 698.4 [M+H]+ (Theoretical value [C 34 H 53 N9O5P1] + =698.4). The entire amount of TD705 was used directly in TD714 without further characterization.

[0632] Synthesis of TD714:

[0633] [ka]

[0634] In a glass vial (4 mL), TD705 (22 mg; 32 μmol; 1.0 equiv.) was dissolved in dry pyridine (1 mL), and neat TMSBr (50 μL; 379 μmol; 12.0 equiv.) was added. The resulting mixture was stirred at room temperature for 16 h. The reaction was quenched with 50% aqueous MeOH (1 mL), and the mixture was further stirred at room temperature for 1 h. The mixture was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and further coevaporated twice with MeOH. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 12 mg (≦64% M-0.1FA-xH2O; M R = 590.2; 2-phase; assumed to be TD705 standard). NMR (DO, pD ~ 5): 1 Hδ H 2.73-3.71(mc,CH2-CO,CH2-P,m,16+2+1H);3.74(C≡CH,s,1H);3.84(CH2-arom,bs,1H);3.99(CH2-arom,bs,1H);4.17(CH2-arom.,bs,2H);4.47(CH2-P;bd, 2 J H=13);4.67(CH2-N3,s,1H);4.68(CH2-N3,s,1H);7.54(arom,d,1H, 3 J HH =8);7.61(arom.,d,1H, 3 J HH =8);7.62(arom.,d,1H, 3 J HH =8);7.64(arom.,d,1H, 3 J HH =8);7.90(arom.,t,1H, 3 J HH =8);7.93(arom.,t,1H, 3 J HH =8). ESI-HRMS: 586.2649 [M+H] + (Theoretical value[C 26 H 37 N9O5P1] + =586.2650).

[0635] Synthesis of TD921:

[0636]

change

[0637] In a pear-shaped glass flask (25 mL), TD910 (160 mg; 315 μmol; 2.0 equiv.) was dissolved in MeCN (10 mL), followed by the addition of a solution of dry K2CO3 (22 mg; 160 μmol; 1.0 equiv.). Next, a solution of TD558 (24 mg; 158 μmol; 1.0 equiv.) in MeCN (5 mL) was added dropwise over 15 min, and the resulting suspension was stirred at room temperature for 24 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The resulting alkyne-bearing intermediate (along with a partially N-methylated / demethylated by-product resulting from the ongoing automethylation; 49 mg; ≤78 μmol; ≤1.0 equiv.) was dissolved in MeCN (7 mL). TD406 (18 mg; 119 μmol; ≥1.5 equiv.) and K2CO3 (54 mg; 391 μmol; ≥5.0 equiv.) were subsequently added, and the resulting suspension was stirred at room temperature for 16 h. The solid was filtered through a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous NaSO, filtered through a glass frit (S3), and evaporated to dryness. The resulting methyl ester of the product (together with a partially N-methylated / demethylated by-product as a result of ongoing automethylation; 37 mg; ≤48 μmol; ≤1.0 equiv.) was dissolved in dry pyridine (2 mL), followed by the addition of neat TMSBr (65 μL; 492 μmol; ≥12.0 equiv.), and the resulting mixture was stirred at room temperature for 16 h.The reaction was quenched with 50% aqueous MeOH (1 mL), and the mixture was further stirred at room temperature for 1 h. The mixture was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the product were combined and directly lyophilized to give the product in zwitterionic form as a white fluffy solid. Yield: 25.6 mg (20%; 3 steps; TD558 reference). NMR (DO, pD ~ 4): 1 Hδ H 2.56-3.75(mc,CH2-P,C≡CH,m,16+4+1H);4.20(CH2-arom.,bs,2H);4.29(CH2-arom. , bs,2H);4.45(CH2-N3,s,2H);7.23-7.77(Ph,arom.,m,10+4H);7.96(arom.,t,1H, 3 J HH =8);7.97(arom.,t,1H, 3 J HH =8). 31 P{ 1 H} δ P 25.1(bs).ESI-HRMS:796.2334[M+H] + (Theoretical value [C 44 H 34 N 10 O2P2] + =796.2336). EA(C 37 H 45 N9O4P2-4.6H2O,M R =824.6):C53.9(54.5);H6.6(6.3);N15.3(14.7).

[0638] Synthesis of TD580:

[0639] [ka]

[0640] In a glass vial (20 mL), TD579 (137 mg; 233 μmol; 1.0 equiv.) was dissolved in MeCN (7 mL) and dry K2CO3 (141 mg; 1.02 mmol; 4.4 equiv.) was added. Next, a solution of TD406 (50 mg; 274 μmol; 1.2 equiv.) in MeCN (3 mL) was added, and the resulting suspension was stirred at room temperature for 3 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in the form of the free base as a colorless oil. Yield: 125 mg (73%; 1 step; based on TD579). NMR (CD3CN): 1 Hδ H 1.21(CH3,t,12H, 3 J HH =7);2.90(mc,m,8H);2.97(CH2-P,d,4H, 2 J HP =10);2.99(mc,m,4H);3.24(mc,m,8H);3.55(C≡CH,s,1H);4.01(CH2-OP,m,8H);4.21 (CH2-arom, s,2H);4.48(CH2-N3,s,2H);4.55(CH2-arom,s,2H);7.37(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.49(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.51(arom,dd,1H,J 3 HH =8, 4 J HH =1);7.54(arom.,dd,1H, 3 J HH=8, 4 J HH =1);7.80(arom.,t,1H, 3 J HH =8);7.84(arom.,t,1H, 3 J HH =8). 31 P{ 1 H} δ P 24.8(s). ESI-MS(LC-MS):734.5[M+H] + (Theoretical value [C 33 H 54 N9O6P2] + =734.4).

[0641] Synthesis of TD582:

[0642] [ka]

[0643] In a glass vial (4 mL), TD580 (25 mg; 34 μmol; 1.0 equiv.) was dissolved in a mixture of aqueous NaOH (10%; 1 mL) and EtOH (300 μL), and the resulting solution was stirred at room temperature for 2 days. The reaction was quenched with AcOH (200 μL), and the mixture was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). Fractions containing the product were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 9 mg (≦39% M-0.1FA-×HO; M R =682.3; 1 stage; assuming TD580 standard). ESI-MS(LC-MS):678.4[M+H] + (Theoretical value [C 29 H 46 N9O6P2] + =678.3).

[0644] Synthesis of TD581:

[0645] [ka]

[0646] In a glass vial (4 mL), TD580 (25 mg; 34 μmol; 1.0 equiv.) was dissolved in dry pyridine (1 mL), neat TMSBr (110 μL; 833 μmol; ∼25 equiv.) was added, and the resulting mixture was stirred at room temperature for 16 h. The reaction was quenched with 50% aqueous MeOH (1 mL), and the mixture was further stirred at room temperature for 1 h. The mixture was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the product were combined and directly lyophilized to give the product in mostly zwitterionic form as a slightly yellow fluffy solid. Yield: 6 mg (≤28% M-0.1FA-xHO; M R =626.2; 1 stage; assuming TD580 standard). ESI-MS(LC-MS):622.3[M+H] + (Theoretical value [C 25 H 38 N9O6P2] + =622.2).

[0647] Synthesis of TD575:

[0648] [ka]

[0649] In a glass vial (20 mL), TD579 (51 mg; 87 μmol; 1.2 equiv.) was dissolved in MeCN (3 mL), followed by the addition of dry K2CO3 (30 mg; 217 μmol; 3.0 equiv.). Next, a solution of TD566 (20 mg; 73 μmol; 1.0 equiv.) in MeCN (2 mL) was added, and the resulting suspension was stirred at room temperature for 16 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined, neutralized with dilute aqueous NaHCO3, and evaporated to dryness. The residue was dissolved in DCM (50 mL) and HO (50 mL) and transferred to a separatory funnel. After shaking, the bottom phase was separated. The aqueous phase was further extracted with DCM (3 × 25 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered through a glass frit (S3), and evaporated to dryness. The residue was further dried overnight under high vacuum to give the product in free base form as a nearly colorless oil. Yield: 46 mg (90%; 1 step; based on TD566). ESI-MS (LC-MS): 810.4 [M+H] + (Theoretical value [C 39 H 58 N9O6P2] + =810.4). TD575 was used directly in TD576 without further characterization.

[0650] Synthesis of TD576:

[0651] [ka]

[0652] In a glass vial (4 mL), TD575 (23 mg; 28 μmol; 1.0 equiv.) was dissolved in dry pyridine (1 mL), neat TMSBr (220 μL; 1.67 mmol; ∼60 equiv.) was added, and the resulting mixture was stirred at room temperature for 16 h. The reaction was quenched with 50% aqueous MeOH (1 mL), and the mixture was further stirred at room temperature for 1 h. The mixture was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the product were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 15 mg (≤75% M-0.1FA-xHO; M R = 702.3; 1 stage; assumed to be TD575 standard). NMR (DO + NaOD, pD ~ 8): 1 Hδ H 2.54-3.76(mc,CH2-P,m,16+4H);3.93(CH2-arom.,bs,2H);4.10(CH2-arom.,bs,2H);4.60(CH2-N3,s,2H);7.44(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.46-7.51(Ph,m,3H);7.53(arom, dd,1H, 3 J HH =8, 4 J HH =1);7.66-7.70(Ph,m,2H);7.71arom.,d,1H, 4 J HH =2);7.77(arom.,d,1H, 4 J HH =2);7.81(arom.,t,1H, 3 J HH =8). 31 P{ 1 H} δ P 8.2(bs).ESI-HRMS:696.2575[MH] - (Theoretical value [C 31 H 40 N9O6P2] - =696.2582).

[0653] Synthesis of TD801:

[0654] [ka]

[0655] In a glass vial (20 mL), TD799 (46 mg; 85 μmol; 1.0 equiv.) was dissolved in MeCN (5 mL) and dry K2CO3 (48 mg; 348 μmol; 4.1 equiv.) was added. Next, a solution of TD406 (20 mg; 110 μmol; 1.3 equiv.) in MeCN (1 mL) was added, and the resulting suspension was stirred at room temperature for 2 days. The solid was filtered off using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 33 mg (48%; 2 steps; TD799 reference). NMR (DO, pD~3): 1 Hδ H 2.41-3.78(mc,CH2-CO,CH2-P,m,16+2+2H);3.89(CH2-arom.,bs,2H);3.95(CH2-arom.,bs,2H);4.52(CH2-N3,s,2H);7.42(arom,d,1H, 3 J HH =8);7.46(arom.,d,1H, 3 J HH =8);7.74(arom.,d,1H, 3 J HH =8);7.78(arom.,d,1H, 3 J HH =8);7.85(arom.,t,1H, 3 J HH =8);7.87(arom.,t,1H,3 J HH = 8). ESI-HRMS: 578.3208 [M+H] + (Theoretical value [C 29 H 40 N9O4] + =578.3203). EA(C 29 H 39 N9O4-2.0TFA,M R =805.7):C49.2(49.3);H5.1(5.4);N15.6(15.7). Synthesis of TD764:

[0656] [ka]

[0657] In a glass vial (20 mL), TD635 (304 mg; 589 μmol; 1.1 equiv.) was dissolved in MeCN (7 mL) and dry K2CO3 (305 mg; 2.21 mmol; 4.0 equiv.) was added. Next, a solution of TD760 (120 mg; 553 μmol; 1.0 equiv.) in MeCN (3 mL) was added, and the resulting suspension was stirred at room temperature for 2 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in the form of the trifluoroacetate salt as a white fluffy solid. Yield: 196 mg (51%; 2 steps; TD635 reference). NMR (DO, pD~3): 1 Hδ H2.86-3.60(mc,CH2-CO,m,16+4H);3.77(C≡CH,s,1H);3.79-4.06(CH2-arom.,bm,4H);4.62(CH2-N3,s,2H);7.62(arom,d,1H, 4 J HH =2);7.67-7.73(arom.,m,1H);8.07-8.14(arom.,m,2H);8.17(arom.,d,1H, 4 J HH =2). ESI-HRMS: 582.2344 [MH] - (Theoretical value[C 27 H 33 [N9O4Cl1]=582.2350). EA(C 27 H 34 N9O4Cl1-0.7TFA-1.7H2O,M R =694.5):C49.1(49.0);H5.5(5.4);N18.2(18.4);Cl5.1(5.3).

[0658] Synthesis of TD1063:

[0659]

change

[0660] In a glass vial (4 mL), TD635 (50 mg; 97 μmol; 1.0 equiv.) was dissolved in MeCN (1 mL) and dry K2CO3 (67 mg; 485 μmol; 5.0 equiv.) was added. Next, a solution of TD1057 (24 mg; 96 μmol; 1.0 equiv.) in MeCN (1 mL) was added, and the resulting suspension was stirred at room temperature for 2 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 33.4 mg (55%; 2 steps; TD635 standard). NMR (DO, pD ~ 7): 1 Hδ H 2.81-3.68(mc,CH2-CO,m,16+4H);3.72(C≡CH,s,1H);3.82(CH2-arom.,bs,2H);4.05(CH2-arom,bs,2H);4.70 (CH2-N3,s,2H);7.62-7.69(arom.,m,1H);7.82(arom.,s,1H);7.92-7.99(arom.,m,2H);8.38(arom.,s,1H). 19 F{ 1 H} δ F -64.2(s).ESI-HRMS:618.2759[M+H] + (Theoretical value [C 28 H 35 N9O4F3] + =618.2759). EA(C 28 H 34 N9O4F3-0.1FA-1.2H2O,M R =643.8):C52.4(52.8);H5.7(5.4);N19.6(19.4);F8.8(7.8).

[0661] Synthesis of TD1092:

[0662] [ka]

[0663] In a glass vial (4 mL), TD635 (35 mg; 68 μmol; 1.0 equiv.) was dissolved in MeCN (2 mL), followed by the addition of dry K2CO3 (44 mg; 318 μmol; 4.7 equiv.). A solution of pre-purified TD1089 (containing less than 20% of the bis-azide by-product; 20 mg; <83 μmol; <1.2 equiv.) in MeCN (1 mL) was then added, and the resulting suspension was stirred at room temperature for 24 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). Fractions containing the product were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 31.9 mg (74%; 2 steps; TD635 reference). NMR (DO, pD ~ 6): 1 Hδ H 2.76-3.62(mc,CH2-CO,m,16+4H);3.73(C≡CH,s,1H);3.83(CH2-arom.,bs,2H);3. 99(CH3,s,3H);4.05(CH2-arom.,bs,2H);4.69(CH2-N3,s,2H);7.70(arom,dd,1H, 3 J HH =8, 4 J HH =1);8.02(arom.,d,1H, 4 J HH =1);8.05(arom.,dd,1H,3 J HH =8, 4 J HH =1);8.15(arom.,t,1H, 3 J HH =8);8.58(arom.,d,1H, 4 J HH =1). ESI-HRMS: 608.2939 [M+H] + (Theoretical value[C 29 H 38 N9O6] + =608.2940). EA(C 29 H 37 N9O6-0.1FA-1.1H2O,M R =632.1):C55.3(55.1);H6.3(6.0);N19.9(19.6).

[0664] Synthesis of TD1148:

[0665]

change

[0666] In a glass vial (4 mL), TD635 (44 mg; 85 μmol; 1.0 equiv.) was dissolved in MeCN (2.5 mL), followed by the addition of dry K2CO3 (47 mg; 340 μmol; 4.0 equiv.). Next, a solution of TD1112 (23 mg; 86 μmol; 1.0 equiv.) in MeCN (1 mL) was added, and the resulting suspension was stirred at room temperature for 3 days. The solid was filtered off using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 30.7 mg (55%; 2 steps; TD635 standard). NMR (DO, pD ~ 6): 1 Hδ H 1.40(CH3,d,6H, 3 J HH =6);2.77-3.63(mc,CH2-CO,m,16+4H);3.70(C≡CH,s,H);3.80(CH2-arom.,bs,2H);4.06(CH2-arom,bs,2H);4.65(CH2-N3,s,2H);5.26(CH,hept,1H, 3 J HH =6);7.65(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.93-8.00(arom.,m,2H);8.12(arom.,t,1H, 3 J HH =8);8.45(arom.,d,1H, 4 J HH =2).ESI-HRMS: 636.3247 [M+H] + (Theoretical value [C 31 H 42 N9O6]+ =636.3253). EA(C 31 H 41 N9O6-0.1FA-1.0H2O,M R =658.3):C56.7(56.4);H6.6(6.4);N19.1(18.8). Synthesis of TD1160:

[0667] [ka]

[0668] In a pear-shaped glass flask (100 mL), TD1116 (50 mg; 158 μmol; 2.0 equiv.) was dissolved in MeCN (20 mL), followed by the addition of dry K2CO3 (11 mg; 80 μmol; 1.0 equiv.). Next, a solution of TD558 (12 mg; 79 μmol; 1.0 equiv.) in MeCN (20 mL) was added dropwise over 2 h, and the resulting suspension was stirred at room temperature for 7 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). Fractions containing the mono-alkylated intermediate were combined and directly lyophilized. The resulting solid (15.4 mg; ≦36 μmol; 1.0 equiv.) was dissolved in MeCN (2 mL), and dry K2CO3 (25 mg; 181 μmol; ≧5.0 equiv.) was added. Next, a solution of TD1130 (15 mg; 53 μmol; ≥1.5 equiv.) in MeCN (1 mL) was added, and the resulting suspension was stirred at room temperature for 7 days. The solid was filtered off using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the methyl-protected product were combined and directly lyophilized. The residue was redissolved in a mixture of MeOH (2 mL) and HO (1 mL), followed by the addition of solid LiOH-HO (15 mg; 360 μmol; >10 equiv.). The resulting mixture was stirred at room temperature for 15 min, quenched with FA (15 μL), and directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the product were combined and directly lyophilized to give the product as a white solid. Yield: 1.7 mg (<5%; 3 stages; TD558 standard). ESI-HRMS:650.3407[M+H] + (Theoretical value [C 32 H 44 N9O6] + =650.3409).

[0669] Synthesis of TD1176:

[0670] [ka]

[0671] In a glass vial (4 mL), TD635 (43.5 mg; 84.4 μmol; 1.1 equiv.) was dissolved in MeCN (2 mL) and dry K2CO3 (42.5 mg; 308 μmol; 4.0 equiv.) was added. Next, a solution of TD1163 (17.3 mg; 76.7 μmol; 1.0 equiv.) in MeCN (1 mL) was added, and the resulting suspension was stirred at room temperature for 2 days. The solid was filtered off using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined and directly lyophilized to give the product in the formate form as a white fluffy solid. Yield: 43.6 mg (80%; 2 steps; TD1163 reference). NMR (DO, pD ~ 7): 1 Hδ H 2.74-3.62 (mc, CH2-CO, CH3, m, 16+4+6H); 3.70 (C≡CH, s, 1H); 3.76 (CH2-arom., bs, 2H); 3.84 (CH2-arom., bs, 2H); ~4.7-4.8 (CH2-N3, obscured by HOD signal); 6.85 (arom., bs, 1H); 7.02 (arom., bs, 1H); 7.64 (arom., dd, 1H, 3 J HH =8, 4 J HH =1);7.88(arom.,bt,1H, 3 J HH =8);7.94(arom.,bd,1H, 3 J HH = 8). ESI-HRMS: 593.3310 [M+H] + (Theoretical value [C 29 H 41 N 10 O4] + =593.3307). EA(C29 H 40 N 10 O4-0.8FA-4.4H2O,M R =708.8):C50.5(50.8);H7.2(7.1);N19.8(19.5).

[0672] Synthesis of TD647

[0673] [ka]

[0674] In a pear-shaped glass flask (50 mL), TD635 (375 mg; 727 μmol; 1.0 equiv.) was dissolved in dry MeCN (20 mL), followed by the addition of dry K2CO3 (300 mg; 2.17 mmol; 3.0 equiv.). A solution of TD566 (189 mg; 731 μmol; 1.0 equiv.) in dry MeCN (5 mL) was then added, and the resulting suspension was stirred at room temperature for 24 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 342 mg (72%; 2 steps; TD635 reference). NMR (DO, pD~4): 1 Hδ H 2.70-3.59(mc,CH2-CO,m,16+4H);3.66(CH2-arom.,bs,2H);3.69(C≡CH,s,1H);3.90(CH2 -arom.,bs,2H);4.64(CH2-N3,s,2H);7.46-7.60(Ph,arom,m,3+1H);7.64(arom.,dd,1H, 3J HH = 8, 4 J HH = 2); 7.68 (arom., t, 1H, 3 J HH = 8); 7.69 (arom., d, 1H, 4 J HH = 2); 7.79 - 7.88 (Ph, m, 2H); 8.28 (arom., d, 1H, 4 J HH = 2). 13 C{ 1 H} δ C 48.2 (mc, s); 48.3 (mc, s); 51.2 (mc, s); 51.3 (mc, s); 54.7 (CH2 - N3, s); 56.4 (CH2 - CO, s); 57.8 (CH2 - arom., s); 59.1 (CH2 - arom., s); 80.3 and 82.4 (C≡CH; 2×s); 121.9 (arom., s); 124.4 (arom, s); 125.6 (arom, s); 128.2 (Ph, s); 128.5 (Ph, s); 130.0 (Ph, s); 130.8 (arom, s); 137.7 (Ph, s); 140.3 (arom, s); 141.2 (arom, s); 153.6 (arom, s); 155.4 (arom, bs); 156.2 (arom, s); 157.6 (arom, s); 168.7 (CO, s). ESI - HRMS: 626.3195 [M + H] + (Calculated value for [C 33 H 40 N9O4] + = 626.3198). EA (C 33 H 39 N9O4 - 0.2FA - 0.9H2O, M R = 651.2): C 61.2 (61.6); H 6.4 (6.1); N 19.4 (19.1).

[0675] Synthesis of TD722:

[0676]

Chem.

[0677] In a glass vial (20 mL), TD539 (350 mg; 543 μmol; 1.0 equiv.) was dissolved in MeCN (15 mL), followed by the addition of dry K2CO3 (375 mg; 2.72 mmol; 5.0 equiv.). Next, a solution of TD566 (140 mg; 541 μmol; 1.0 equiv.) in MeCN (3 mL) was added, and the resulting suspension was stirred at room temperature for 24 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl / Boc-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (5 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined and directly lyophilized to give the product in the form of the trifluoroacetate salt as a slightly yellow solid. Yield: 295 mg (69%; 2 steps; TD566 reference). NMR (DO, pD ~ 7): 1 Hδ H 2.74-3.64(mc,CH2-CO,m,16+4H);3.72(CH2-arom.,bs,2H);3.98(CH2-arom.,bs,2H);4.1 0(CH2-C≡C,s,2H);4.65(CH2-N3,s,2H);7.47-7.69(Ph,arom,m,3+3H);7.72(arom.,d,1H, 4 J HH =2);7.85-7.96(Ph,m,2H);8.41(arom.,d,1H, 4 J HH = 2). ESI-HRMS: 655.3460 [M+H] + (Theoretical value C 34 H 43 N 10 O4] + =655.3463). EA(C 34 H 42 N 10 O4-0.7TFA-2.0H2O,M R=770.6):C55.2(54.9);H6.1(5.8);N18.2(18.3);F5.2(5.3).

[0678] Synthesis of TD1054:

[0679] [ka]

[0680] In a glass vial (4 mL), TD663 (30.0 mg; 45.3 μmol; 1.0 equiv.) was dissolved in MeCN (1 mL) and dry K2CO3 (37.5 mg; 271 μmol; 5.0 equiv.) was added. Next, a solution of freshly prepared and isolated TD1050 (≦65.3 μmol; ≦1.5 equiv.) in MeCN (1 mL) was added, and the resulting suspension was stirred at room temperature for 3 days. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl / Boc-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined and directly lyophilized to give the product as a colorless solid in the form of a trifluoroacetate / formate mixed salt. Yield: 23.0 mg (65%; 2 steps; TD663 reference). NMR (DO, pD~8): 1 Hδ H 1.72(CH3,s,6H);2.67-3.61(mc,CH2-CO,m,16+4H);3.70(CH2-arom.,bs,2H);3.97(CH2 -arom, bs,2H);4.64(CH2-N3,s,2H);7.45-7.67(Ph,arom.,m,3+3H);7.72(arom.,d,1H, 4 J HH=2);7.86-7.93(Ph,m,2H);8.40(arom.,d,1H, 4 J HH =2). ESI-HRMS: 683.3775 [M+H] + (Theoretical value[C 36 H 47 N 10 O4] + =683.3776). EA(C 36 H 46 N 10 O4-0.3TFA-0.8FA-2.3H2O,M R =777.3):C57.8(58.0);H6.5(6.5);N18.0(17.7).

[0681] Synthesis of TD1105:

[0682]

change

[0683] In a glass vial (4 mL), TD711 (101 mg; 185 μmol; 1.0 equiv.) was dissolved in MeCN (2 mL), followed by the addition of dry K2CO3 (102 mg; 738 μmol; 4.0 equiv.). Next, a solution of freshly prepared and isolated TD1050 (≦222 μmol; ≦1.2 equiv.) in MeCN (1 mL) was added, and the resulting suspension was stirred at 40 °C for 24 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl / Boc-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the fully deprotected intermediate were combined and directly lyophilized. The resulting solid was dissolved in a mixture of HO (6 mL), MeCN (6 mL), and THF (2 mL), followed by the addition of BocO (2.0 M in dry THF; 315 μL; 630 μmol; ≥ 3.4 equiv.) and NaHCO (210 mg; 2.50 mmol; ≥ 13.5 equiv.). The resulting mixture was stirred at room temperature for 2 days, after which another portion of BocO (2.0 M in dry THF; 315 μL; 630 μmol; ≥ 3.4 equiv.) was added. The mixture was further stirred at room temperature for 16 h. The mixture was evaporated to dryness, and the residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 36.8 mg (27%; 3 steps; TD711 standard). NMR (DO, pD ~ 7): 1 Hδ H 1.47(CH3,s,9H);1.65(CH3,s,6H);2.85-3.65(mc,CH2-CO,m,16+4H);3.91(CH2- arom.,bs,2H);3.95(CH2-arom,bs,2H);4.59(CH2-N3,s,2H);7.51(arom.,dd,1H, 3 JHH =7, 4 J HH =2);7.59(arom.,dd,1H,J 3 HH =7,J 4 HH =2);7.89-8.05(arom.,m,4H).ESI-HRMS:707.3988[M+H] + (Theoretical value[C 35 H 51 N 10 O6] + =707.3988). EA(C 35 H 50 N 10 O6-0.1FA-2.0H2O,M R =747.5):C56.4(56.7);H7.3(7.0);N18.7(18.5).

[0684] Synthesis of TD1127:

[0685]

change

[0686] In a glass vial (20 mL), TD1118 (56.9 mg; 81.4 μmol; 1.0 equiv.) was dissolved in MeCN (2 mL), followed by the addition of dry K2CO3 (45.0 mg; 326 μmol; 4.0 equiv.) and TD406 (16.0 mg; 87.6 μmol; 1.1 equiv.) in MeCN (1 mL). The resulting suspension was stirred at room temperature for 16 h. The solid was filtered through a syringe microfilter (PTFE), and the filtrate was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). Fractions containing the tert-butyl / Boc-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined and directly lyophilized to give the product as a colorless solid in the form of a mixed trifluoroacetate / formate salt. Yield: 49.0 mg (71%; 2 steps; based on TD1118). NMR (DO, pD ~ 7): 1 Hδ H 1.96-2.11(CH2,m,2H);2.14-2.29(CH2,m,2H);2.86-3.63(mc,CH2-CO,CH2,CH,m,16+4+4+1H);3.90(CH2-a rom.,bs,2H);3.94(CH2-arom,bs,2H);4.59(CH2-N3,s,2H);7.49-7.55(arom.,m,1H);7.60(arom.,dd,1H, 3 J HH =8, 4 J HH =2);7.92-8.04(arom.,m,4H).ESI-HRMS:633.3612[M+H] + (Theoretical value [C 32 H 45 N 10 O4] + =633.3620). EA(C 32 H 44 N 10 O4-1.8TFA-0.1FA,M R=878.6):C50.9(50.7);H5.5(5.9);N16.6(16.9).

[0687] Synthesis of TD742:

[0688] [ka]

[0689] In a glass vial (20 mL), TD635 (90 mg; 163 μmol; 1.1 equiv.) was dissolved in MeCN (6 mL), followed by the addition of dry K2CO3 (85 mg; 616 μmol; 4.0 equiv.). Next, a solution of TD733 (55 mg; 153 μmol; 1.0 equiv.) in MeCN (2 mL) was added, and the resulting suspension was stirred at room temperature for 18 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 87 mg (80%; 2 steps; TD733 reference). NMR (DO, pD~4): 1 Hδ H 2.76-3.64(mc,CH2-CO,m,16+4H);3.69(C≡CH,s,1H);3.72(CH2-arom.,bs,2H);3.99( CH2-arom.,bs,2H);4.77(CH2-N3,s,2H);7.54-7.70(arom,m,3H);7.85(arom.,d,1H, 4 J HH =2);7.96(arom.,d,2H, 3 J HH =8);8.11(arom.,d,2H,3 J HH =8);8.38(arom.,d,1H, 4 J HH = 2). ESI-HRMS: 670.3094 [M+H] + (Theoretical value [C 34 H 40 N9O6] + =670.3096). EA(C 34 H 40 N9O6-0.2FA-1.6H2O,M R =707.8):C58.0(58.4);H6.1(5.9);N17.8(17.5).

[0690] Synthesis of TD744:

[0691] [ka]

[0692] In a glass vial (20 mL), TD539 (80 mg; 124 μmol; 1.1 equiv.) was dissolved in MeCN (3 mL), followed by the addition of dry K2CO3 (65 mg; 471 μmol; 4.0 equiv.). Next, a solution of TD733 (42 mg; 117 μmol; 1.0 equiv.) in MeCN (3 mL) was added, and the resulting suspension was stirred at room temperature for 24 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient, FA added). Fractions containing the protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined and directly lyophilized to give the product in the form of a trifluoroacetate / formate mixed salt as an off-white fluffy solid. Yield: 44 mg (45%; 2 steps; TD733 reference). NMR (DO, pD~4): 1 Hδ H2.71-3.64(mc,CH2-CO,m,16+4H);3.70(CH2-arom.,bs,2H);3.98(CH2-arom.,bs,2H);4 .12(CH2-C≡C,s,2H);4.70(CH2-N3,s,2H);7.50-7.59(arom, m,2H);7.63(arom.,dd,1H,J 3 HH =7,J 4 HH =2);7.78(arom.,d,1H, 4 J HH =2);7.95(arom.,d,2H, 3 J HH =9);8.07(arom.,d,2H, 3 J HH =9);8.42(arom.,d,1H, 4 J HH = 2). ESI-HRMS: 699.3358 [M+H] + (Theoretical value [C 35 H 43 N 10 O6] + =699.3362). EA(C 35 H 42 N 10 O6-1.0TFA-1.3H2O,M R =836.2):C53.1(52.8);H5.5(5.8);N16.8(16.9).

[0693] Synthesis of TD750: In a glass vial (2 mL), TD744-1.0TFA-1.3HO (1.5 mg; ~1.8 μmol; 1.0 equiv.) was dissolved in aqueous boric acid / NaOH buffer (200 mM; pH 9.0; 900 μL; 180 mmol; ~100 equiv.). A freshly prepared solution of FmocCl (0.6 mg; ~2.3 μmol; 1.3 equiv.) in MeCN (900 μL) was then added. The resulting clear solution was stirred at room temperature for 2 h. The mixture was directly purified by preparative HPLC (C18; HO-MeCN gradient, FA addition). Fractions containing the product were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 0.8 mg (<48% M-0.2FA-xHO; M R= 930.2; 1 stage; TD744-1.0TFA-1.3H2O (assuming standard). ESI-HRMS: 921.4044 [M+H] + (Theoretical value [C 50 H 53 N 10 O8] + =921.4042).

[0694] Synthesis of TD779:

[0695] [ka]

[0696] In a glass vial (4 mL), TD647-0.2FA-0.9HO (10 mg; 15 μmol; 1.0 equiv.) and ammonium chloride (2 mg; 38 μmol; 2.5 equiv.) were dissolved in DMSO (2 mL), and DIPEA (22 μL; 126 μmol; 8.2 equiv.) and solid HATU (18 mg; 47 μmol; 3.1 equiv.) were added. The resulting yellow solution was stirred at room temperature for 15 min. The mixture was quenched with FA (6 μL; 169 μmol; 10 equiv.) and directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). Fractions containing the product were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 7 mg (≦73% M-0.1FA-xHO; M R = 628.4; 1 stage; TD647 - 0.2FA - 0.9HO (assuming standard). NMR (DO + FA, pD ~ 3): 1 Hδ H 2.83-3.74(mc,CH2-CO,bm,16+4H);3.65(C≡CH,s,1H);4.13-4.68(CH2-arom., bm4H);4.69(CH2-N3,s,2H);7.52-7.66(Ph,arom,m,3+1H);7.71(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.80-7.88(Ph,m,2H);7.90(arom.,d,1H,J 4 HH=2);7.91(arom.,t,1H, + J HH =8);8.00(arom.,bs,1H).ESI-HRMS:624.3503[M+H] + (Theoretical value [C 33 H 42 N 11 O2] + =624.3504).

[0697] [ka]

[0698] Synthesis of TD778: In a glass vial (4 mL), TD647-0.2FA-0.9HO (12 mg; 18 μmol; 1.0 equiv.) and glycine tert-butyl ester hydrochloride (8 mg; 48 μmol; 2.6 equiv.) were dissolved in DMSO (2 mL), followed by the addition of DIPEA (26 μL; 149 μmol; 8.1 equiv.) and solid HATU (21 mg; 55 μmol; 3.0 equiv.). The resulting yellow solution was stirred at room temperature for 20 min. The mixture was then quenched with FA (7 μL; 186 μmol; 10 equiv.) and directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 9 mg (62%; 2 steps; based on TD647-0.2FA-0.9HO). NMR (DO, pD~5): 1 Hδ H 2.83-3.93-(mc,CH2-CO,C≡CH,bm,16+8+1H);4.35-4.69(CH2-arom.,CH2-N3.,bm,4+2H);7.49(arom.,d,1H,3 J HH =8);7.56-7.73(Ph,arom.,m,3+1H);7.75-7.92(Ph,arom.,m,2+2H);8.06(arom.,s,1H). ESI-HRMS:738.3471[MH] - (theor.[C 37 H 44 N 11 O6] - =738.3482). EA(C 37 H 45 N 11 O6-0.1FA-2.4H2O,M R =787.7):C56.6(55.9);H6.4(5.8);N19.6(20.3).

[0699] [ka]

[0700] Synthesis of TD1408: In a glass vial (25 mL), TD635 (51 mg; 99 μmol; 1.1 equiv) was dissolved in MeCN (1 mL), followed by the addition of dry K2CO3 (54 mg; 391 μmol; 4.4 equiv). Next, a solution of TD1406 (17.5 mg; 89 μmol; 1.0 equiv) in MeCN (1 mL) was added, and the resulting suspension was stirred at 40 °C for 16 h. The solid was filtered using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 41.7 mg (77%; 2 steps; based on TD1406). NMR (DO, pD ~ 7):1 Hδ H 2.74 - 4.16 (mc, CH2 - CO, CH2 - arom., CH2 - CH2 - N3, C≡CH, m, 16 + 4 + 4 + 4 + 1H); 7.37 - 7.45 (arom., m, 1H); 7.65 (arom., dd, 1H, 3 J HH = 8, 4 J HH = 1); 7.85 - 7.93 (arom., m, 2H); 7.96 (arom., t, 1H, 3 J HH = 8); 8.06 (arom., dd, 1H, 3 J HH = 8, 4 J HH = 1). ESI - HRMS: 564.3044 [M + H] + (theor. [C 28 H 38 N9O4] + = 564.3041). EA (C 28 H 37 N9O4 - 0.1FA - 2.4H2O, M R = 611.5): C 55.2 (56.0); H 6.9 (6.7); N 20.6 (19.8).

[0701]

Chem.

[0702] Synthesis of TD1346: In a pear-shaped glass flask (25 mL), TD1345 (52.7 mg; 83 μmol; 1.0 equiv.) was dissolved in 60% aq. MeCN (2.7 mL), followed by the addition of freshly prepared aq. LiOH (1.0 M; 830 μL; 830 μmol; 10 equiv.). The resulting solution was stirred at room temperature for 5 h. The mixture was quenched by the addition of FA (31 μL; 822 μmol; 10 equiv.). The resulting solution was briefly concentrated to remove most of the MeCN and then directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in the form of the free base as a white fluffy solid. Yield: 35.4 mg (71%; 1 step; based on TD1345). NMR (DO, pD ~ 7): 1 Hδ H 1.24-1.42(CH3,m,6H);2.44-3.81(mc,CH2-arom.,C≡CH,m,16+4+1H);4.41-4.53(CH-CH3,m,2H);4.56(CH2-N3,s,2H);7.49(arom.,d,1H, 3 J HH =8);7.63(arom.,d,1H, 3 J HH =8);7.93(arom.,t,1H, 3 J HH =8);7.96(arom.,t,1H, 3 J HH =8);8.05(arom.,d,1H, 3 J HH =8);8.10(arom.,d,1H, 3 J HH = 8). ESI-HRMS: 578.3197 [M+H] + (theor.[C 29 H 40 N9O4] + =578.3198). EA(C 29 H 39 N9O4-1.2H2O,M R =599.3):C58.1(58.0);H7.0(6.5);N21.0(20.6).

[0703] [ka]

[0704] Synthesis of TD1451: In a pear-shaped glass flask (25 mL), TD1449 (302 mg; 330 μmol; 1.0 equiv.) was dissolved in 60% aq. MeCN (17 mL), followed by the addition of freshly prepared aq. LiOH (1.0 M; 3.3 mL; 3.3 mmol; 10 equiv.). The resulting solution was stirred at room temperature for 2 h. The mixture was briefly concentrated to remove most of the MeCN and directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in mostly zwitterionic form as a white fluffy solid. Yield: 257 mg (87%; 1 step; based on TD1449). NMR (DO, pD ~3): 1 Hδ H 1.37-1.60(CH3-CO,CH3-CH,m,9+6H);1.67(CH3-CN,s,6H);2.66-4.30(mc,CH2-arom.,CH-CH3,m,16+4+2 H);4.68(CH2-N3,s,2H);7.80(arom.,s,1H);7.92(arom.,s,1H);7.98(arom.,s,1H);8.02(arom.,s,1H). 19 Fδ F -64.44(s). ESI-HRMS:847.4070[M+H] + (theor.[C 39 H 54 N 10 O8F3] + =634.3824). EA(C 39 H 53 N 10 O8F3-0.1FA-2.7H2O,M R =900.1):C52.2(53.1);H6.6(6.2);N15.6(15.5);6.3(5.4).

[0705] [ka]

[0706] Synthesis of TD1504: In a glass vial (25 mL), TD1500 (11.9 mg; 18 μmol; 1.0 equiv) was dissolved in MeCN (800 μL) and HO (550 μL), followed by the addition of freshly prepared aq. LiOH (1.0 M; 263 μL; 263 μmol; 15 equiv). The resulting solution was stirred at room temperature for 3 h. The mixture was quenched by the addition of FA (8 μL; 212 μmol; 12 equiv). The resulting solution was diluted with HO (2 mL) and then directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product in the form of the free base as a white fluffy solid. Yield: 8.9 mg (~77% M-2HO; M R = 657.7; 2 steps; assumed based on TD1500). NMR (DO, pD ~ 3): 1 Hδ H 1.28-1.45(CH3,m,3H);2.49-3.70(mc,CH-CH3,CH-CH2,CH2-arom.,m,16+1+3+4H);3.76(C≡CH,s,1H);4.35-4.51(CH2-N3,m,2H);7.69(arom.,dd,2H, 3 J HH =8, 4 J HH =1);7.98(arom.,t,1H, 3 J HH =8);8.05-8.21(arom.,m,3H). ESI-MS(LC-MS):622.3[M+H] + (theor.[C 30 H 40 N9O6] + =622.3).

[0707] Example 5: Synthesis of a (triazole-bridged) cage macrocyclic ligand

[0708] [ka]

[0709] Synthesis of cz-TD425: Obtained as a white fluffy solid in a mostly zwitterionic form as a by-product during the synthesis of TD425. Yield: 3 mg (≤1% M-0.1FA-xHO; M R = 609.9; 3 steps; assumed to be based on 2-chloro-N-(prop-2-yn-1-yl)acetamide). NMR (DO, pD ~ 3): 1 Hδ H 2.52-5.20(mc,CH2-CO,CH2-NH-CO-CH2,CH2-arom.,bm,16+4+4+2H);6.24(CH2-N3,bs,2 H);7.56-7.72(Ph,m,3H);7.88(CH-N3,s,1H);7.88-7.94(Ph,m,2H);8.33(arom.,d,1H, 4 J HH =2);8.73(arom.,d,1H, 4 J HH = 2). ESI-HRMS: 606.3143 [M+H] + (theor.[C 30 H 40 N9O5] + =606.3147).

[0710] [ka]

[0711] Synthesis of cz-TD556: Obtained as a white fluffy solid in the form of the trifluoroacetate salt as a by-product during the synthesis of TD556. Yield: 9 mg (~2%; 2 steps; based on TD635). NMR (DO, pD~3): 1 Hδ H 2.54-4.06(mc,CH2-CO,bm,16+4H);4.59(CH2-arom.,bs,2H);5.02(CH2-arom.,bs,2H);5.86(CH2-N3,bs,2H);7.67(arom.,bd,1H, 3 J HH =8);7.70(arom.,bd,1H, 3 J HH =8);7.83(arom.,bd,1H, 3 JHH =8);7.87(arom.,bd,1H, 3 J HH =8);8.14(arom.,bt,1H, 3 J HH =8);8.16(CH-N3,bs,1H);8.17(arom.,bt,1H, 3 J HH = 8). ESI-HRMS: 550.2889 [M+H] + (theor.[C 27 H 36 N9O4] + =550.2885). EA(C 27 H 35 N9O4-2.7TFA-0.5H2O,M R =866.4):C44.9(44.8);H4.5(4.8);N14.5(14.8).

[0712] [ka]

[0713] Synthesis of cz-TD764: Obtained as a white fluffy solid in the form of the trifluoroacetate salt as a by-product during the synthesis of TD764. Yield: 9 mg (~2%; 2 steps; based on TD635). NMR (DO, pD~3): 1 Hδ H 2.48-3.94(mc,CH2-CO,bm,16+4H);4.57(CH2-arom.,bs,2H);5.00(CH2-arom.,bs,2H);5.94(CH2-N3,bs,2H);7.64(arom.,d,1H, 3 J HH =8);7.79(arom.,s,1H);7.84(arom.,d,1H, 3 J HH =8);7.92(arom.,s,1H);8.14(arom.,t,1H, 3 J HH =8);8.15(CH-N3,s,1H). ESI-HRMS:582.2344[MH] - (theor.[C 27 H 33N9O4Cl1] - =582.2350). EA(C 27 H 34 N9O4Cl1-1.8TFA-3.3H2O,M R =848.7):C43.3(43.3);H5.0(4.5);N14.9(14.5);Cl4.2(4.2);F12.1(12.0).

[0714] [ka]

[0715] Synthesis of cz-TD1063: Obtained as a white fluffy solid in the form of the trifluoroacetate salt as a by-product during the synthesis of TD1113. Yield: 1.7 mg. NMR (DO, pD~3): 1 Hδ H 2.57-3.84(mc,CH2-CO,m,16+4H);4.70(CH2-arom.,s,2H);4.99(CH2-arom.,bs,2H);6.03(CH2-N3,bs,2H);7.65(arom.,d,1H, 3 J HH =8);7.86(arom.,d,1H, 3 J HH =8);8.05(arom.,s,1H);8.12-8.18(arom.,CH-N3,m,1+1H);8.19(arom.,s,1H). 19 Fδ F -65.82(s). NMR (~0.5mM cz-TD1063 in ~50mM MOPS / NaOH buffer in 10% HO, pH 7.0): 19 Fδ F -65.66(s;ΔH 1 / 2 =4.6Hz). ESI-HRMS:618.2754[M+H] + (theor.[C 29 H 35 N9O4F3] + =618.2759).

[0716] [ka]

[0717] Synthesis of TD1188: In a glass vial (4 mL), TD711 (70.0 mg; 128 μmol; 1.0 equiv) was dissolved in MeCN (1.5 mL), followed by the addition of dry K2CO3 (71 mg; 514 μmol; 4.0 equiv). A solution of TD1178 (31 mg; 141 μmol; 1.1 equiv) in MeCN (1.5 mL) was then added, and the resulting suspension was stirred at room temperature for 7 days. The solid was filtered off using a syringe microfilter (PTFE), and the filtrate was evaporated to dryness. The residue was purified by preparative HPLC (C18; HO-MeCN gradient with FA). Fractions containing the tert-butyl-protected product were combined, evaporated to dryness, and coevaporated twice with MeOH to remove most of the MeCN. The residue was dissolved in TFA (3 mL), and the resulting clear mixture was stirred at room temperature for 16 h. The mixture was evaporated to dryness and coevaporated twice with MeOH to remove most of the TFA. The residue was directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the zwitterionic form of the product as a white fluffy solid. Yield: 9.5 mg (10%; 2 steps; based on TD711). NMR (DO, pD~5): 1 Hδ H 2.11-4.00(mc,CH2-CO,bm,16+4H);4.52(CH2-arom.,bs,2H);4.88-6.58(CH2-arom.,CH2-N3,bm,4H);7.66(arom.,bd,1H, 3 J HH =8);7.68(arom.,d,1H, 3 J HH =8);7.75(arom.,bd,1H, 3 J HH =8);7.85(arom.,d,1H, 3 J HH =8);8.08(arom.,bt,1H, 3 J HH =8);8.16(arom.,t,1H, 3 J HH =8). 19 FδF -59.35(s). ESI-HRMS:618.2756[M+H] + (theor.[C 28 H 35 N9O4F3] + =618.2759). EA(C 28 H 34 N9O4F3-0.2TFA-0.4FA-4.3H2O,M R =736.3):C47.0(46.6);H6.0(5.6);N17.1(16.8);F9.3(10.1).

[0718] [ka]

[0719] Synthesis of TD650: In a glass vial (40 mL), TD647-0.2FA-0.9HO (130 mg; 200 μmol; 1.0 equiv.) was dissolved in HO (20 mL), followed by the addition of aq. citric acid (100 mM; 20 mL; 2.0 mmol; 10 equiv.) to pH 2.4. The resulting mixture was then stirred at 80 °C for 2 days. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to give the product as a white fluffy solid in the form of the formate salt. Yield: 109 mg (77%; 1 step; based on TD647-0.2FA-0.9HO). NMR (DO, pD ~ 5, 95 °C): 1 Hδ H 2.69-3.63(mc,CH2-CO,m,16+4H);4.22(CH2-arom.,s,2H);4.58(CH2-arom.,s,2H);6.22(CH2-N3,s,2H);7.61(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.60-7.67(Ph,m,3H);7.83(arom.,s,1H);7.85(arom.,dd,1H, 3 JHH =8, 4 J HH =1); 7.86 - 7.92 (Ph, m, 2H); 8.00 (arom., s, 1H); 8.11 (arom., t, 1H, 3 J HH =8); 8.21 (CH - N3, s, 1H). 13 C{ 1 H}δ C 48.5 (mc, s); 49.9 (mc, s); 50.2 (mc, s); 50.4 (mc, s); 53.4 (CH2 - N3, s); 55.6 (CH2 - CO, s); 57.8 (CH2 - arom., s); 59.2 (CH2 - arom., s); 122.6 (arom., s); 124.2 (arom., s); 125.4 (arom., s); 125.9 (arom., s); 127.8 (Ph, s); 129.9 (Ph, s); 130.7 (Ph, s); 134.5 (CH - N3, s); 137.0 (Ph, s); 138.0 (arom., s); 139.9 (arom., s); 147.1 (arom., s); 152.3 (arom., bs); 153.6 (arom., bs); 153.8 (arom., bs); 156.8 (arom., s); 172.6 (CO, s). ESI - HRMS: 626.3200 [M + H] + (theor. [C 33 H 40 N9O4] + =626.3198). EA (C 33 H 39 N9O4 - 0.9FA - 2.0H2O, M R =703.2): C 57.9 (58.3); H 6.4 (5.9); N 17.9 (17.5).

[0720]

Chem.

[0721] Synthesis of TD871: In a glass vial (40 mL), TD647-1.3TFA (78.0 mg; 103 μmol; 1.0 equiv.) was dissolved in HO (34 mL), followed by the addition of aq. borate / CsOH buffer (200 mM; pH 9.0; 6.0 mL; 1.2 mmol; ∼12 equiv.). The resulting mixture was then stirred at 80 °C for 6 days. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to obtain the product as a white fluffy solid in the form of the formate salt. Yield: 49.3 mg (70%; 1 step; based on TD647-1.3TFA). NMR (DO, pD ∼5, 95 °C): 1 Hδ H 2.93-3.56(mc,CH2-CO,m,16+4H);4.41(CH2-arom.,s,2H);4.49(CH2-arom.,s,2H);6.00(CH2-N3,s,2H);7.54(arom.,dd,1H, 3 J HH =7, 4 J HH =2);7.56-7.66(Ph,m,3H);7.77(arom.,d,1H, 4 J HH =1);7.79-7.87(Ph,m,2H);7.94(arom.,d,1H, 4 J HH =1);8.05(arom.,dd,1H, 3 J HH =8, 4 J HH =2);8.07(arom.,dd,1H, 3 J HH =8, 3 J HH =7);8.93(CH-N3,s,1H). 13 C{ 1 H}δ C50.0(mc,s);50.7(mc,bs);52.0(mc,s);52.3(mc,s);54.8(CH2-N3,s);56.6(CH2-CO,s);59.9(CH2-arom .,s);60.2(CH2-arom.,bs);121.2(arom.,s);121.7(arom.,s);123.3(arom.,s);124.5(arom.,s);127. 0(CH-N3,s);127.7(Ph,s);129.9(Ph,s);130.5(Ph,s);137.1(Ph,s);140.2(arom.,s);148.0(arom.,s) ;150.3(arom.,s);151.7(arom.,s);152.5(arom.,bs);154.9(arom.,s);155.0(arom.,s);174.4(CO,s). ESI-HRMS:626.3199[M+H] + (theor.[C 33 H 40 N9O4] + =626.3198). EA(C 33 H 39 N9O4-0.6FA-2.3H2O,M R =696.8):C58.1(57.9);H6.5(6.2);N18.1(18.2).

[0722] [Example 6: Synthesis of coordination compounds]

[0723]

change

[0724] Synthesis of TD734: In a glass vial (20 mL), TD714-0.1FA-xHO (6 mg; ~10 μmol; 1.0 equiv.) was dissolved in HO (4 mL). Subsequently, aq. MOPS / NaOH buffer (500 mM; pH 7.0; 100 mL; 500 μmol; ~50 equiv.) and aq. LuCl (100 mM; 110 μL; 11 μmol; ~1.1 equiv.) were added, and the mixture was stirred at 80 °C for 16 h. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient, FA added). The product-containing fractions were combined and directly lyophilized to give the product as a white fluffy solid in the form of the formate salt as a mixture of two diastereoisomers (~11:1). Yield: 5mg (≦61% [M] + [FA] - -xH2O;M R = 803.6; 1 step; assumed based on TD714-0.1FA-xH2O). NMR (DO, pD ~ 4, signals of major isomer): 1 Hδ H 2.50-3.82(mc,CH2-CO,CH2-P,m,16+2+1H);3.85-4.00(CH2-arom.,CH2-P,m,1+1H);4.10(CH2-arom.,d,1H, 2 J HH =16);4.23(CH2-arom.,d,1H, 2 J HH =15);4.99(CH2-arom.,d,1H, 2 J HH =15); 5.53(CH2-N3, d, 1H, 2 J HH =15);7.56(CH2-N3,d,1H, 2 J HH =15);7.67(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.73(arom.,dd,1H, 3 J HH =8, 4 J HH= 1); 7.81 (arom., dd, 1H, 3 J HH = 8, 4 J HH = 2); 7.83 (CH - N3, s, 1H); 7.98 (arom., dd, 1H, 3 J HH = 8, 4 J HH = 2); 7.18 (arom., t, 1H, 3 J HH = 8); 8.20 (arom., t, 1H, 3 J HH = 8); 8.35 (FA, s, 1H). 13 C{ 1 H}δ C 48.9 (mc, s); 51.3 (mc, s); 51.6 (CH2 - P, d, 1 J CP = 73); 52.0 (mc, s); 52.3 (mc, s); 53.4 (mc, s); 53.5 (CH2 - N3, s); 53.6 (mc, s); 55.0 (mc, s); 56.3 (mc, s); 59.7 (CH2 - arom., s); 60.0 (CH2 - arom., s); 61.0 (CH2 - CO, s); 125.9 (arom., s); 127.6 (arom., s); 128.9 (arom., s); 130.4 (arom., s); 136.4 (CH - N3, s); 138.7 (arom., s); 142.0 (arom., s); 142.9 (arom., s); 146.2 (arom., s); 156.8 (arom., s); 157.6 (arom., s); 158.6 (arom., s); 169.5 (FA, s); 178.2 (CO, s). 31 P{ 1 H}δ P 17.8 (s). ESI - HRMS: 758.1822 [M] + (theor. [C 26 H 34 N9O5P1Lu1] + = 758.1823).

[0725]

Chem.

[0726] Synthesis of TD925: In a glass vial (20 mL), TD921-4.6HO (6.1 mg; 7.4 μmol; 1.0 equiv.) was dissolved in HO (18 mL) and i-PrOH (1 mL). Subsequently, aq. MOPS / NaOH buffer (500 mM; pH 7.0; 400 μL; 200 μmol; 27 equiv.) and aq. GdCl (100 mM; 95 μL; 9.5 μmol; 1.3 equiv.) were added, and the mixture was stirred at 80 °C for 7 days. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined and directly lyophilized to obtain the product as a white fluffy solid in the form of a trifluoroacetate salt. Yield: 2.5 mg (≦33% [M] + [TFA] - -xH2O;M R = 1010.0; 1 step; assumed based on TD921-4.6H2O). ESI-HRMS: 897.2158 [M] + (theor.[C 37 H 43 N9O4P2Gd1] + =897.2149).

[0727] [ka]

[0728] Synthesis of TD560: In a glass vial (20 mL), TD556-0.1FA-1.6HO (39 mg; 69 μmol; 1.0 equiv.) was dissolved in HO (15 mL). Then, aq. MOPS / NaOH buffer (500 mM; pH 7.0; 3.40 mL; 1.70 mmol; 25 equiv.) and aq. EuCl (100 mM; 745 μL; 75 μmol; 1.1 equiv.) were added, and the mixture was stirred at 80 °C for 4 h. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to obtain the product as a white fluffy solid in the form of the formate salt. Yield: 24 mg (43%; 1 step; based on TD556-0.1FA-1.6H2O). ESI-HRMS: 700.1867 [M] + (theor.[C 27 H 33 N9O4Eu1] + =700.1862). EA([C 27 H 33 N9O4Eu1] + [FA] - -4.5H2O,M R =825.7):C40.7(40.7);H5.2(4.8);N15.3(15.0);Eu18.4(17.7).

[0729] [ka]

[0730] Synthesis of TD561: In a glass vial (20 mL), TD556-0.1FA-1.6HO (40 mg; 69 μmol; 1.0 equiv.) was dissolved in HO (15 mL). Subsequently, aq. MOPS / NaOH buffer (500 mM; pH 7.0; 3.40 mL; 1.70 mmol; 25 equiv.) and aq. GdCl (100 mM; 745 μL; 75 μmol; 1.1 equiv.) were added, and the mixture was stirred at 80 °C for 4 h. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to obtain the product as a white fluffy solid in the form of the formate salt. Yield: 24 mg (42%; 1 step; based on TD556-0.1FA-1.6H2O). ESI-HRMS: 705.1893 [M] + (theor.[C 27 H 33 N9O4Gd1] + =705.1891). EA([C 27 H 33 N9O4Gd1] + [FA] - -4.6H2O,M R =832.8):C40.4(40.8);H5.2(4.9);N15.1(15.5);Gd18.9(18.1).

[0731] [ka]

[0732] Synthesis of TD562: In a glass vial (20 mL), TD556-0.1FA-1.6HO (40 mg; 69 μmol; 1.0 equiv.) was dissolved in HO (15 mL). Subsequently, aq. MOPS / NaOH buffer (500 mM; pH 7.0; 3.40 mL; 1.70 mmol; 25 equiv.) and aq. TbCl (100 mM; 745 μL; 75 μmol; 1.1 equiv.) were added, and the mixture was stirred at 80 °C for 4 h. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to obtain the product as a white fluffy solid in the form of the formate salt. Yield: 27 mg (47%; 1 step; based on TD556-0.1FA-1.6H2O). ESI-HRMS: 706.1906 [M] + (theor.[C 27 H 33 N9O4Tb1] + =706.1903). EA([C 27 H 33 N9O4Tb1] + [FA] - -4.7H2O,M R =836.2):C40.2(40.5);H5.2(4.9);N15.1(14.7);Tb19.0(18.7).

[0733] [ka]

[0734] Synthesis of TD1069: In a glass vial (20 mL), TD556-0.1FA-1.6HO (20.8 mg; 35.7 μmol; 1.0 equiv.) was dissolved in HO (17 mL). Subsequently, aq. MOPS / NaOH buffer (500 mM; pH 7.0; 1.80 mL; 900 μmol; 25 equiv.) and aq. LuCl (100 mM; 400 μL; 40.0 μmol; 1.1 equiv.) were added, and the mixture was stirred at 80 °C for 16 h. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined and directly lyophilized to obtain the product as a white fluffy solid in the form of a trifluoroacetate salt. Yield: 26.6 mg (82%; 1 step; based on TD556-0.1FA-1.6H2O). NMR (DO, pD~3): 1 Hδ H 2.64-3.82(mc,CH2-CO,m,16+4H);3.95(CH2-arom.,d,1H, 2 J HH =15);4.15(CH2-arom.,d,1H, 2 J HH =16);4.29(CH2-arom.,d,1H, 2 J HH =15);4.85(CH2-arom.,d,1H, 2 J HH =16);5.60(CH2-N3,d,1H, 2 J HH =15);7.10(CH2-N3,d,1H, 2 J HH =15);7.73(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.80(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.85(arom.,dd,1H, 3 J HH =8, 4 J HH=1);7.88(CH-N3,s,1H);8.02(arom.,dd,1H, 3 J HH =8, 4 J HH =1);8.25(arom.,t,1+1H, 3 J HH = 8). ESI-HRMS: 722.2060 [M] + (theor.[C 27 H 33 N9O4Lu1] + =722.2058). EA([C 27 H 33 N9O4Lu1] + [TFA] - -0.3TFA-2.4H2O,M R =913.0):C38.9(38.7);H4.2(3.8);N13.8(13.7);Lu19.2(16.5).

[0735] [ka]

[0736] Synthesis of TD751: In a glass vial (20 mL), TD718-2.6HO (14 mg; 21 μmol; 1.0 equiv.) was dissolved in HO (18 mL). Subsequently, aq. MES / NaOH buffer (500 mM; pH 5.2; 1.30 mL; 650 μmol; 31 equiv.) and aq. LuCl (100 mM; 236 μL; 24 μmol; 1.1 equiv.) were added, and the mixture was stirred at 80 °C for 3 h. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to obtain the product as a white fluffy solid in the form of the formate salt. Yield: 13 mg (65%; 1 step; based on TD718-2.6H2O). NMR (DO, pD ~ 6): 1 Hδ H0.10(CH3,s,9H);2.61-3.83(mc,CH2-CO,m,16+4H);3.94(CH2-arom.,d,1H, 2 J HH =15);4.24(CH2-arom.,d,1H, 2 J HH =16);4.24(CH2-arom.,d,1H, 2 J HH =15);4.84(CH2-arom.,d,1H, 2 J HH =16);5.54(CH2-N3,d,1H, 2 J HH =15);6.92(CH2-N3,d,1H, 2 J HH =15);7.70(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.75(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.85(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.98(arom.,dd,1H, 3 J HH =8, 4 J HH =1);8.22(arom.,t,1H, 3 J HH =8);8.26(arom.,t,1H, 3 J HH =8);8.45(FA,s,1H)。 13 C{ 1 H}δ C-1.85 (CH3, s); 48.4 (mc, s); 48.7 (mc, s); 50.7 (mc, s); 51.9 (mc, s); 52.0 (CH2-N3, s); 53.2 (mc, s); 53.8 (mc, s); 54.4 (mc, s); 55.5 (mc, s); 58.4 (CH2-CO, s); 59.0 (CH2-arom., s); 59.1 (CH2-arom., s); 60.9 (CH2-CO, s); 125.9 (arom., s); 126.7 (arom., s); 128.3 (arom., s); 130.4 (arom., s); 141.4 (arom., s); 142.3 (arom., s); 142.6 (arom., s); 146.3 (arom., s); 147.5 (arom., s); 155.1 (arom., s); 157.7 (arom., s); 158.7 (arom., s); 171.0 (FA, s); 176.8 (CO, s); 178.6 (CO, s). ESI-HRMS: 794.2445 [M] + (theor. [C 30 H 41 N9O4Si1Lu1] + = 794.2453). EA ([C 30 H 41 N9O4Si1Lu1] + [FA] - -6.6H2O, M R = 958.7): C 38.8 (39.0); H 5.8 (5.3); N 13.1 (13.0); Si 2.9 (3.0); Lu 18.2 (17.4).

[0737]

Chem.

[0738] Synthesis of TD737: In a glass vial (20 mL), TD728-2.6HO (15 mg; 20 μmol; 1.0 equiv.) was dissolved in HO (17 mL). Subsequently, aq. MOPS / NaOH buffer (500 mM; pH 7.0; 2.00 mL; 1.00 mmol; 50 equiv.) and aq. LuCl (100 mM; 220 μL; 22 μmol; 1.1 equiv.) were added, and the mixture was stirred at 80 °C for 5 days. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with FA). The product-containing fractions were combined and directly lyophilized to obtain the product as a white fluffy solid in the form of the formate salt. Yield: 12 mg (58% in one step based on TD728-2.6H2O). NMR (DO, pD~5): δ H 0.96(i-Pr,d,9H, 3 J HH =8);0.99(i-Pr,d,9H, 3 J HH =8);1.26(i-Pr,hept,3H, 3 J HH =8);2.62-3.84(mc,CH2-CO,m,16+4H);4.05(CH2-arom.,s,2H);4.29(CH2-arom.,d,1H, 2 J HH =17);4.88(CH2-arom.,d,1H, 2 J HH =17); 5.43(CH2-N3, d, 1H, 2 J HH =15);6.74(CH2-N3,d,1H, 2 J HH =15);7.67(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.82(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.83(arom.,dd,1H, 3 J HH =8, 4 JHH = 1); 7.94 (arom., dd, 1H, 3 J HH = 8, 4 J HH = 1); 8.20 (arom., t, 1H, 3 J HH = 8); 8.22 (arom., t, 1H, 3 J HH = 8); 8.43 (FA, s, 1H). 13 C{ 1 H}δ C 12.1 (i - Pr, s); 18.8 (i - Pr, 2×s); 47.9 (mc, bs); 50.0 (mc, bs); 50.4 (mc, bs); 52.2 (mc, bs); 53.0 (CH2 - N3, s); 54.0 (mc, s); 55.2 (mc, s); 57.2 (mc, s); 57.6 (mc, s); 59.3 (CH2 - CO, s); 60.0 (CH2 - arom., s); 62.3 (CH2 - arom., bs); 64.0 (CH2 - CO, s); 126.1 (arom., s); 127.4 (arom., s); 128.9 (arom., s); 130.8 (arom., s); 141.6 (arom., s); 143.3 (arom., s); 143.8 (arom., s); 144.6 (arom., s); 147.1 (arom., s); 156.1 (arom., s); 158.4 (arom., s); 159.6 (arom., s); 171.5 (FA, s); 178.0 (CO, s); 179.1 (CO, s). ESI - HRMS: 878.3396 [M] + (theor. [C 36 H 53 N9O4Si1Lu1] + = 878.3392). EA ([C 36 H 53 N9O4Si1Lu1] + [FA] - - 5.9H2O, M R = 1030.2): C 43.1 (43.4); H 12.2 (11.8); N 12.2 (11.8); Si 2.7 (2.9); Lu 17.0 (16.9).

[0739]

Chem.

[0740] Synthesis of TD946: In a glass vial (20 mL), TD943-1.5HO (28.1 mg; 45.6 μmol; 1.0 equiv.) was dissolved in HO (16 mL). Subsequently, aq. MOPS / NaOH buffer (500 mM; pH 7.0; 3.00 mL; 1.50 mmol; 33 equiv.) and aq. LuCl (100 mM; 540 μL; 54 μmol; 1.2 equiv.) were added, and the mixture was stirred at 80 °C for 3 days. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined and directly lyophilized to give the product as a white fluffy solid in the form of a trifluoroacetate salt. Yield: 32.6 mg (77%; 1 step; based on TD943-1.5H2O). NMR (DO, pD~5): δ H 0.69-0.94(CH2-CH,m,2H);0.94-1.07(CH2-CH,m,2H);1.65(CH2-CH,tt,1H, 3 J HH =8, 3 J HH =5);2.58-3.86(mc,CH2-CO,m,16+4H);3.97(CH2-arom.,d,1H, 2 J HH =15);4.19(CH2-arom.,d,1H, 2 J HH =16);4.29(CH2-arom.,d,1H, 2 J HH =15);4.87(CH2-arom.,d,1H, 2 J HH =16);5.53(CH2-N3,d,1H, 2 J HH =15);6.99(CH2-N3,d,1H, 2 J HH =15);7.73(arom.,dd,1H, 3 J HH =8, 4 J HH= 1); 7.86 (arom., dd, 1H, 3 J HH = 8, 4 J HH = 1); 7.93 (arom., dd, 1H, 3 J HH = 8, 4 J HH = 1); 7.99 (arom., dd, 1H, 3 J HH = 8, 4 J HH = 1); 8.24 (arom., t, 1H, 3 J HH = 8); 8.29 (arom., t, 1H, 3 J HH = 8). ESI-HRMS: 762.2375 [M] + (theor. [C 30 H 37 N9O4Lu1] + = 762.2371). EA ([C 30 H 37 N9O4Lu1] + [TFA] - -0.2TFA - 1.8H2O, M R = 930.8): C 41.8 (42.2); H 4.4 (4.2); N 13.5 (13.1); F 7.3 (7.0).

[0741]

Chem.

[0742] Synthesis of TD961: In a glass vial (20 mL), TD959-1.5HO (14.7 mg; 23.0 μmol; 1.0 equiv.) was dissolved in HO (16 mL). Subsequently, aq. MOPS / NaOH buffer (500 mM; pH 7.0; 1.50 mL; 0.75 mmol; 33 equiv.) and aq. LuCl (100 mM; 280 μL; 28 μmol; 1.2 equiv.) were added, and the mixture was stirred at 80 °C for 3 days. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined and directly lyophilized to obtain the product as a white fluffy solid in the form of the trifluoroacetate salt. Yield: 20.1 mg (90% in one step based on TD959-1.8H2O). NMR (DO, pD~4): δ H 1.18(CH3,s,9H);2.64-3.86(mc,CH2-CO,m,16+4H);4.01(CH2-arom.,d,1H, 2 J HH =15);4.16(CH2-arom.,d,1H, 2 J HH =15);4.35(CH2-arom.,d,1H, 2 J HH =17);4.87(CH2-arom.,d,1H, 2 J HH =17); 5.48(CH2-N3, d, 1H, 2 J HH =15);6.66(CH2-N3,d,1H, 2 J HH =15);7.70(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.84(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.87(arom.,dd,1H, 3 J HH =8, 4 J HH=1);7.93(arom.,dd,1H, 3 J HH =8, 4 J HH =1);8.22(arom.,t,1H, 3 J HH =8);8.26(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 778.2685 [M] + (theor.[C 31 H 41 N9O4Lu1] + =778.2684). EA([C 31 H 41 N9O4Lu1] + [TFA] - -0.2TFA-3.0H2O,M R =968.5):C41.4(41.5);H4.9(4.6);N13.0(12.7);F7.1(7.4).

[0743] [ka]

[0744] Synthesis of TD951: In a glass vial (20 mL), TD944-1.5HO (14.4 mg; 22.1 μmol; 1.0 equiv.) was dissolved in HO (18 mL). Subsequently, aq. MOPS / NaOH buffer (500 mM; pH 7.0; 1.20 mL; 600 μmol; 27 equiv.) and aq. LuCl (100 mM; 265 μL; 26.5 μmol; 1.2 equiv.) were added, and the mixture was stirred at 80 °C for 4 days. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined and directly lyophilized to obtain the product as a white fluffy solid in the form of the trifluoroacetate salt. Yield: 17.4 mg (80% in one step based on TD944-1.5H2O). NMR (D2O, pD~2): δ H2.69-3.88(mc,CH2-CO,m,16+4H);3.97(CH2-arom.,d,1H, 2 J HH =15);4.16-4.36(CH2-arom.,m,2H);4.88(CH2-arom.,d,1H, 2 J HH =17);5.51(CH2-N3,d,1H, 2 J HH =15);6.97(CH2-N3,d,1H, 2 J HH =15);7.16-7.25(Ph,m,2H);7.28-7.41(Ph,m,3H);7.49(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.73(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.83(arom.,dd,1H, 3 J HH =8, 4 J HH =1);8.03(arom.,dd,1H, 3 J HH =8, 4 J HH =1);8.06(arom.,t,1H, 3 J HH =8);8.25(arom.,t,1H, 3 J HH =8)。ESI-HRMS:798.2379[M] + (theor.[C 33 H 37 N9O4Lu1] + =798.2371)。EA([C 33 H 37 N9O4Lu1] + [TFA] - -0.2TFA-3.0H2O,M R =988.5):C43.0(43.0);H4.4(4.3);N12.8(12.6);F6.9(6.9)。

[0745] [ka]

[0746] Synthesis of TD994: In a glass vial (20 mL), TD992-0.3FA-1.9HO (41.6 mg; 56.8 μmol; 1.0 equiv.) was dissolved in HO (16 mL). Subsequently, aq. MOPS / NaOH buffer (500 mM; pH 7.0; 2.85 mL; 1.43 mmol; 25 equiv.) and aq. LuCl (100 mM; 625 μL; 62.5 μmol; 1.1 equiv.) were added, and the mixture was stirred at 80 °C for 8 days. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined and directly lyophilized to obtain the product as a white fluffy solid in the form of a trifluoroacetate salt. Yield: 52.3 mg (86%; 1 step; based on TD992-0.3FA-1.9H2O). NMR (DO, pD~4): δ H 1.52-1.78(Adm.,m,9H);1.80-1.95(Adm.,m,6H);2.60-3.83(mc,CH2-CO,m,16+4H);3.99(CH2-arom.,d,1H, 2 J HH =15);4.11(CH2-arom.,d,1H, 2 J HH =15);4.33(CH2-arom.,d,1H, 2 J HH =17);4.84(CH2-arom.,d,1H, 2 J HH =17); 5.43(CH2-N3, d, 1H, 2 J HH =15);6.58(CH2-N3,d,1H, 2 J HH =15);7.66(arom.,d,1H, 3 J HH =8);7.80(arom.,d,1H, 3 J HH=8);7.88-7.92(arom.,m,2H);8.18(arom.,t,1H, 3 J HH =8);8.22(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 856.3154 [M] + (theor.[C 37 H 47 N9O4Lu1] + =856.3153). EA([C 37 H 47 N9O4Lu1] + [TFA] - -0.4TFA-3.0H2O,M R =1069.4):C44.7(44.3);H5.0(4.6);N11.8(11.5);F7.5(7.4);Lu16.4(14.0).

[0747] [ka]

[0748] Synthesis of TD1221: In a glass vial (20 mL), TD1204-0.3FA-2.7HO (22.5 mg; 30.5 μmol; 1.0 equiv.) was dissolved in HO (18 mL). Subsequently, aq. MOPS / NaOH buffer (500 mM; pH 7.0; 1.57 mL; 785 μmol; 26 equiv.) and aq. LuCl (100 mM; 375 μL; 37.5 μmol; 1.2 equiv.) were added, and the mixture was stirred at 80 °C for 24 h. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined and directly lyophilized to obtain the product as a white fluffy solid in the form of the trifluoroacetate salt. Yield: 24.5 mg (74%; 1 step; based on TD1204-0.3FA-2.7H2O). NMR (DO, pD~): 1 Hδ H2.59-3.83(mc,CH2-CO,m,16+4H);3.95(CH2-arom.,d,1H, 2 J HH =15);4.23(CH2-arom.,d,1H, 2 J HH =17);4.27(CH2-arom.,d,1H, 2 J HH =15);4.85(CH2-arom.,d,1H, 2 J HH =17);5.62(CH2-N3,d,1H, 2 J HH =15);7.01(CH2-N3,d,1H, 2 J HH =15);7.72(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.87(arom.,d,1H, 4 J HH =2);7.90(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.97(arom.,dd,1H, 3 J HH =8, 4 J HH =1);8.23(arom.,t,1H, 3 J HH =8);8.29(arom.,t,1H, 3 J HH =8)。ESI-HRMS:848.1020[M] + (theor.[C 27 H 32 N9O4I1Lu1] + =848.1024)。EA([C 27 H 32 N9O4I1Lu1] + [TFA] - -0.5TFA-4.1H2O,M R =1092.3):C33.0(33.8);H3.8(3.6);N11.5(11.8);I11.6(10.9);F7.8(7.0)。

[0749] [ka]

[0750] Synthesis of TD782: In a glass vial (40 mL), TD764-0.7TFA-1.7HO (190 mg; 274 μmol; 1.0 equiv.) was dissolved in HO (21 mL). Subsequently, aq. MOPS / NaOH buffer (500 mM; pH 7.0; 15.4 mL; 7.70 mmol; 28 equiv.) and aq. LuCl (100 mM; 3.30 mL; 330 μmol; 1.2 equiv.) were added, and the mixture was stirred at 80 °C for 16 h. The mixture was then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined and directly lyophilized to give the product as a white fluffy solid in the form of a trifluoroacetate salt. Yield: 224 mg (85%; 1 step; based on TD764-0.7TFA-1.7H2O). NMR (DO, pD~3): 1 Hδ H 2.61-3.84(mc,CH2-CO,m,16+4H);3.94(CH2-arom.,d,1H, 2 J HH =15);4.15(CH2-arom.,d,1H, 2 J HH =16);4.29(CH2-arom.,d,1H, 2 J HH =15);4.84(CH2-arom.,d,1H, 2 J HH =16);5.58(CH2-N3,d,1H, 2 J HH =15);7.10(CH2-N3,d,1H, 2 J HH =15);7.80(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.83(arom.,d,1H, 4 JHH =2);7.85(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.88(CH-N3,s,1H);8.13(arom.,d,1H, 4 J HH =2);8.24(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 756.1664 [M] + (theor.[C 27 H 32 N9O4Cl1Lu1] + =756.1668). EA([C 27 H 32 N9O4Cl1Lu1] + [TFA] - -5.1H2O,M R =961.9):C36.2(36.4);H4.4(4.0);N13.1(12.9);Cl3.7(3.7);F5.9(6.4);Lu18.2(16.8).

[0751] [ka]

[0752] Synthesis of TD819: In a glass vial (4 mL), [TD782] + [TFA] -A mixture of 5.1HO (10.0 mg; 10.4 μmol; 1.0 equiv.), 3-borono-5-nitrobenzoic acid (4.4 mg; 21 μmol; 2.0 equiv.), and XPhosPdG (0.5 mg; ~0.6 μmol; 6 mol%) was charged and then flushed with argon three times. Next, under a constant flow of argon, dry DMF (420 μL) was added through a septum, followed by a freshly prepared (and briefly flushed with argon) aq. solution of KPO-HO (326 mM; 160 μL; 52 μmol; 5.0 equiv.). The mixture was then stirred at 80 °C for 16 h under a septum (but without external argon). The resulting heterogeneous mixture was diluted with HO (2 mL), and the resulting slightly opaque, pale yellow solution was filtered through a syringe microfilter (RC). The filtrate was then directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined and directly lyophilized to give the product as a white fluffy solid. Yield: 10 mg (≦96% [M] + [TFA] - -xH2O;M R =1000.7;1 stage;[TD782] + [TFA] - -5.1H2O). NMR (D2O, pD~3): 1 Hδ H 2.66-3.84(mc,CH2-CO,m,16+4H);4.07(CH2-arom.,d,1H, 2 J HH =15);4.17(CH2-arom.,d,1H, 2 J HH =16);4.39(CH2-arom.,d,1H, 2 J HH =15);4.87(CH2-arom.,d,1H, 2 J HH =15); 5.72(CH2-N3, d, 1H, 2 J HH =15);7.19(CH2-N3,d,1H, 2 J HH =15);7.83(arom.,dd,1H, 3 J HH =8,4 J HH =1);7.87(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.90(CH-N3,s,1H);8.14(arom.,d,1H, 4 J HH =2);8.26(arom.,t,1H, 3 J HH =8);8.45(arom.,d,1H, 4 J HH =2);8.82(arom.,dm,1H, 4 J HH =2);8.92(arom.,dd,1H, 4 J HH =2, 4 J HH =1);8.97(arom.,dm,1H, 4 J HH = 2). ESI-HRMS: 887.2121 [M] + (theor.[C 34 H 36 N 10 O8Lu1] + =887.2120).

[0753] [ka]

[0754] Synthesis of TD891: In a glass vial (4 mL), add [TD782] + [TFA] -After adding 5.1H2O (15.0 mg; 15.6 μmol; 1.0 equiv.), 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetic acid (8.0 mg; 30.5 μmol; 2.0 equiv.), and XPhosPdG2 (0.7 mg; ~0.9 μmol; 6 mol%), the mixture was flushed with argon three times. Next, dry DMF (600 μL) was added through a septum under a constant flow of argon, followed by a freshly prepared (briefly flushed with argon) aq. solution of K3PO4-H2O (391 mM; 200 μL; 78 μmol; 5.0 equiv.). The mixture was stirred at 80 °C for 16 h under a septum (but without external argon). The resulting heterogeneous mixture was diluted with HO (2 mL), and the resulting slightly opaque, pale yellow solution was filtered through a syringe microfilter (RC). The filtrate was then directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined and directly lyophilized to give the product as a white fluffy solid. Yield: 7.2 mg (41%; 1 step; [TD782] + [TFA] - -5.1H2O). NMR (D2O, pD~3): 1 Hδ H 2.60-3.82(mc,CH2-CO,m,16+4H);3.86(OC-CH2-arom.,s,2H);3.99(CH2-arom.,d,1H, 2 J HH =15);4.15(CH2-arom.,d,1H, 2 J HH =16);4.32(CH2-arom.,d,1H, 2 J HH =15);4.84(CH2-arom.,d,1H, 2 J HH =16);5.65(CH2-N3,d,1H, 2 J HH =15);7.13(CH2-N3,d,1H, 2 J HH =15);7.54(arom.,dm,2H, 3 J HH =8);7.81(arom.,dd,1H,3 J HH =8, 4 J HH =1);7.85(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.90(CH-N3,s,1H);7.92(arom.,dm,2H, 3 J HH =8);7.99(arom.,d,1H, 4 J HH =1);8.25(arom.,t,1H, 3 J HH =8);8.30(arom.,d,1H, 4 J HH = 1). ESI-HRMS: 856.2418 [M] + (theor.[C 35 H 39 N9O6Lu1] + =856.2426). EA([C 35 H 39 N9O6Cl1Lu1] + [TFA] - -0.7TFA-4.6H2O,M R =1132.4):C40.7(40.6);H4.4(4.3);N11.1(11.3).

[0755] [ka]

[0756] Synthesis of TD891-OH: TD891-OH was obtained as a by-product in the synthesis of TD891 (as a white fluffy solid in the form of the trifluoroacetate salt). Yield: 5.7 mg. NMR (DO, pD ~ 3): 1 Hδ H 2.60-3.83(mc,CH2-CO,CH2-arom.,m,16+4+1H);4.13(CH2-arom.,d,1H, 2 J HH =16);4.20(CH2-arom.,d,1H, 2 J HH=15);4.82(CH2-arom.,d,1H, 2 J HH =16); 5.43(CH2-N3, d, 1H, 2 J HH =15);6.97(CH2-N3,d,1H, 2 J HH =15);7.09(arom.,d,1H, 4 J HH =2);7.40(arom.,d,1H, 4 J HH =2);7.79(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.84(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.89(CH-N3,s,1H);8.23(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 738.2003 [M] + (theor.[C 27 H 33 N9O5Lu1] + =738.2007).

[0757] [ka]

[0758] Synthesis of TD786: In a glass vial (4 mL), [TD782] + [TFA] --5.1HO (37 mg; 38 μmol; 1.0 equiv) was dissolved in DMSO (3 mL), followed by the addition of solid NaN (54 mg; 831 μmol; 20 equiv), and the mixture was stirred at 80 °C for 2 h. The mixture was then concentrated to remove most of the DMSO. The residue was dissolved in HO and lyophilized. The crude product was purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined and directly lyophilized to give the product in the form of the trifluoroacetate salt as a white fluffy solid. Yield: 21 mg (55%; 1 step; [TD782] + [TFA] - -5.1H2O). NMR (D2O, pD ~ 4): 1 Hδ H 2.62-3.81(mc,CH2-CO,m,16+4H);3.87(CH2-arom.,d,1H, 2 J HH =15);4.14(CH2-arom.,d,1H, 2 J HH =16);4.25(CH2-arom.,d,1H, 2 J HH =15);4.83(CH2-arom.,d,1H, 2 J HH =16);5.52(CH2-N3,d,1H, 2 J HH =15);7.05(CH2-N3,d,1H, 2 J HH =15);7.38(arom.,d,1H, 4 J HH =2);7.71(arom.,d,1H, 4 J HH =2);7.80(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.84(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.88(CH-N3,s,1H);8.24(arom.,t,1H, 3 J HH= 8). ESI-HRMS: 763.2075 [M] + (theor.[C 27 H 32 N 12 O4Lu1] + =763.2072). EA([C 27 H 32 N 12 O4Lu1] + [TFA] - -0.2TFA-5.2H2O,M R =993.0):C35.6(35.6);H4.3(4.0);N16.9(16.6);F6.9(6.8).

[0759] [ka]

[0760] Synthesis of TD787: Obtained as a white fluffy solid in the form of the trifluoroacetate salt as a by-product during the synthesis of TD786. Yield: 5 mg (~10%; 1 step; based on TD782-5.1H2O). NMR (DO, pD~2): 1 Hδ H 2.59-3.79(mc,CH2-CO,CH2-arom.,m,16+4+1H);4.02-4.19(CH2-arom.,m,2H);4.25(CH2-arom.,d,1H, 2 J HH =15);4.79(CH2-arom.,d,1H, 2 J HH =15);5.27(CH2-N3,d,1H, 2 J HH =15);6.74(arom.,d,1H, 4 J HH =2);6.83(CH2-N3,d,1H, 2 J HH =15);7.03(arom.,d,1H, 4 J HH =2);7.78(arom.,dd,1H, 3 J HH =8, 4 J HH=1);7.82(arom.,dd,1H, 3 J HH =8, 4 J HH =1);7.89(CH-N3,s,1H);8.22(arom.,t,1H, 3 J HH = 8). ESI-HRMS: 737.2165 [M] + (theor.[C 27 H 34 N 10 O4Lu1] + =737.2167).

[0761] [ka]

[0762] Synthesis of TD830: In a glass vial (20 mL), TD764-0.7TFA-1.7HO (60 mg; 86 μmol; 1.0 equiv) was dissolved in HO (5 mL), followed by aq. MOPS / NaOH buffer (500 mM; pH 7.0; 5.0 mL; 2.50 mmol; 29 equiv.), and freshly prepared aq. 176 YbCl3 (~90 μmol; ~1.0 eq.; 18.1 mg 176 YbO was dissolved in 0.5 mL of 6 M HCl at 80 °C for several hours, after which the acid was evaporated and 10 mL of HO was added to the residue. The mixture was then stirred at 80 °C for 6 hours and then filtered through a syringe microfilter (RC). The filtrate was concentrated on a rotary evaporator and then directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined and directly lyophilized to give the product as a white fluffy solid in the form of the trifluoroacetate salt. Yield: 71 mg (≦94% [M] + [TFA] - -xH2O;M R = 871.0; 1 step; assumed based on TD764-0.7TFA-1.7H2O). ESI-HRMS: 757.1680 [M] + (theor.[C 27 H 32N9O4Cl1 176 Yb1] + =757.1686).

[0763] [ka]

[0764] Synthesis of TD836: In a glass vial (4 mL), [TD830] + [TFA] - -xHO (12.1 mg; ∼14 μmol; 1.0 equiv) was dissolved in DMSO (500 μL), followed by the addition of solid NaN (9.0 mg; 138 μmol; ∼10 equiv), and the mixture was stirred at 80 °C for 30 min. The mixture was then diluted with HO (3 mL) and directly purified by preparative HPLC (C18; HO-MeCN gradient with TFA). The product-containing fractions were combined and directly lyophilized to give the product as a white fluffy solid. Yield: 9.9 mg (≤81% [M] + [TFA] - -xH2O;M R =877.6;1 stage;[TD830] + [TFA] - -xH2O). ESI-HRMS: 764.2085 [M] + (theor.[C 27 H 32 N 12 O4 176 Yb1] + =764.2090).

[0765] [ka]

[0766] Synthesis of TD888: In a glass vial (20 mL), TD764-0.6TFA-1.0HO (56.5 mg; 84.3 μmol; 1.0 equiv.) was dissolved in HO (15 mL). Subsequently, aq. MOPS / NaOH buffer (500 mM; pH 7.0; 4.0 mL; 2.00 mmol; 24 e...

Claims

1. Compounds of general formula (I) 【Chemistry 1】 Here, Y is selected from the group consisting of nitrogen; N-oxide; Ris H; halogen; -OH; -N ３ ; -(CH ２ ) ｎ N ３ (where n is an integer in the range of 1 to 3); -NR ２ (where R is H, or C １ to C ６ alkyl which can be branched or linear and is independently selected); -(CH ２ ) ｎ NR ２ (where n and R are as defined above); C ６ to C １０ aryl (optionally, -NH ２ , -NO ２ , -N ３ ,​ 【Chemistry 2】 , -COOH, -CH ２ Cl and / or -CH ２ (Can be substituted with COOH); C ７ ~C １０ Arylalkyl (optionally, -NH ２ , -NO ２ , -N ３ , 【Transformation 3】 , -COOH, -CH ２ Cl, and / or -CH ２ (May be substituted with COOH); -CF ３ ;-COOR(where R is as defined above);-(CH ２ ) ｎ COOR (where n and R are as defined above); 【Chemistry 4】 ;-CH ２ CH(OMe) ２ ; 【Transformation 5】 ;-SH;-SO ３ H; -SO ２ Ar (where Ar is phenyl); NO ２ Selected from the group consisting of; R ２ teeth, 【Transformation 6】 ； or 【Transformation 7】 (where n is an integer in the range of 1 to 3); and; A is H; -(CH ２ ) ｎ COOH (where n is an integer from 1 to 3); -CH (CH ３ )COOH;-CH((CH ２ ) ｎ CH ３ )COOH (where n is as defined above); -CH ２ P (= O) (OR) ２ (Here, R is as defined above); -CH((CH ２ ) ｎ COOH)COOH(where n is an integer from 1 to 3); -CH((CH ２ ) ｎ NH ２ )COOH (where n is an integer from 1 to 3); -CH ２ C(=O)(NH ２ ); -CH ２ C(=O)(NH)-CH ２ COOH; -CH ２ P(=O)(OH)(Ar) (where Ar is arbitrarily C) １ ~C ６ Independently selected from the group consisting of; (phenyl which can be substituted with alkyl); Z is, 【Transformation 8】 Selected from the group consisting of, Here, R ３ teeth, 【Chemistry 9】 And, R ４ H; halogen; piperidinyl; trimethylsilyl; triisopropylsilyl; phenyl; C １ ~C ６ Alkyl (can be branched or linear); C ３ ~C ６ Cycloalkyl; -CF ３ ;-(CH ２ ) ｎ NHR ６ (Here, n is an integer in the range of 1 to 3, and R ６ (is selected from H, fluorenylmethyloxycarbonyl, tert-butyloxycarbonyl and benzyloxycarbonyl); C (CH ３ ) ２ (NHR ６ ) (Here, R ６ (as defined above); adamantyl; 【Chemistry 10】 ;-(CH ２ ) ｎ COOH (where n is as defined above); C ６ ~C １０ Aryl (optional, -NH) ２ , -NO ２ , -N ３ 【Chemistry 11】 , -COOH, -CH ２ Cl, and / or -CH ２ Selected from the group consisting of (which can be substituted with COOH); R ５ R １ ,or, 【Chemistry 12】 (Here, R ４ (is as defined above); and / or, R ２ and R ３ Both are, formula 【Chemistry 13】 , or formula 【Chemistry 14】 The 1,2,3-triazole group (where R ４ (as defined above); form; However, at most one A is H.

2. A compound of general formula (I) as described in claim 1: Y is nitrogen; R １ is H; Cl; -N(CH ３ ) ２ ; phenyl (optionally substituted with -COOH or -CH ２ COOH); benzyl (optionally substituted with -COOH or -CH ２ COOH); -CF ３ ; -COOCH ３ ; -COOCH(CH ３ ) ２ ; -COOtBu; 【Chemistry 15】 Selected from the group consisting of; A is H; -(CH ２ ), ｎ COOH, (where n is 1 or 2); -CH ２ P(=O)(OH) ２ ; -CH ２ P(=O)(OH)(OEt); -CH ２ P(=O)(OEt) ２ ; -CH ２ C(=O)(NH ２ ); -CH ２ C(=O)(NH)-CH ２ COOH; -CH ２ P(=O)(OH)(Ph); and is independently selected from the group consisting of; Z is, 【Chemistry 16】 Selected from the group consisting of, Here, R ３ teeth 【Chemistry 17】 And; R ４ H; halogen; piperidinyl; trimethylsilyl; triisopropylsilyl; phenyl; cyclopropyl; tert-butyl; -CF ３ ;-CH ２ NH ２ ;-CH ２ N(H)(Fmoc);-C(CH ３ ) ２ (NH ２ ); -C(CH ３ ) ２ Selected from the group consisting of (NHBoc); adamantyl; R ５ H, or, [Chemistry 18] (Here, R ４ (as defined above); However, it is a compound in which at most one A is H.

3. The compound according to claim 1, Z is, 【Chemistry 19】 And; R ３ teeth 【Chemistry 20】 And; R ４ and R ５ A compound as defined in claim 1.

4. The compound according to claim 1, R ２ and R ３ Both are such that the bridge between two opposing nitrogen atoms in the cyclone portion is an entity of general formula (IIa) or (IIb), 【Chemistry 21】 ,formula 【Chemistry 22】 , or formula 【Chemistry 23】 Forms a 1,2,3-triazole group; Here, L is 【Chemistry 24】 Selected from the group consisting of; R １ , R ４ and R ５ A compound as defined in claim 1.

5. Y is nitrogen, Z 【Chemistry 25】 And, A compound of general formula (I) according to claim 1, selected from the group consisting of compounds in which the remaining substituents are present in the following combinations: Table 1-1 Table 1-2 Table 1-3 Table 1-4 。

6. A method for preparing a compound of general formula (I) according to any one of claims 1 to 5, comprising the following steps: i) Provide an alkyne intermediate of general formula Z-Cl, Here, Z is as defined in claim 1; ii) Azide intermediate of general formula (III) 【Chemistry 26】 To provide Here, Y, R １ and R ２ This is as defined in claim 1; iii) Cyclene derivatives of general formula (IV) 【Chemistry 27】 To provide Here, pA is -(CH ２ ) ｎ COO ｔ Bu (where n is an integer between 1 and 3); benzyloxycarbonyl; -CH(CH ３ )COOR(where R is tert-butyl, methyl, or ethyl);-CH((CH ２ ) ｎ CH ３ )COOR(where n is an integer from 1 to 3 and R is tert-butyl, methyl, or ethyl);-CH((CH ２ ) ｎ COOR)COOR(where n is an integer from 1 to 3, and R is independently selected from tert-butyl, methyl, or ethyl);-CH ２ P (= O) (OR) ２ (Here, R can be branched or linear C) １ ~C ６ (It is alkyl); -CH ２ C(=O)(NH ２ ); -CH ２ C(=O)(NH)-CH ２ COOH; -CH ２ P(=O)(OH)(Ar) (where Ar is arbitrarily C) １ ~C ６ Selected from the group consisting of: (phenyl that can be substituted with alkyl); iv-A) The cyclone derivative of general formula (IV) is reacted with the alkyne intermediate Z-Cl to obtain the intermediate of general formula (V). 【Chemistry 28】 To obtain Here, Z is as defined in claim 1, and pA is as defined above; or iv-B) The cyclene derivative of general formula (IV) is reacted with the azide intermediate of general formula (III) to obtain the intermediate of general formula (VI). 【Chemistry 29】 To obtain Here, Y, R １ , R ２ And pA are as defined above; v-A) The intermediate of general formula (V) is reacted with the azide intermediate of general formula (III) to obtain the intermediate of general formula (VII). 【Transformation 30】 To obtain Here, Y, R １ , R ２ , pA and Z are as defined above; or v-B) The intermediate of general formula (VI) is reacted with the alkyne intermediate Z-Cl to obtain the intermediate of general formula (VII); vi) Optionally, hydrolysis of the protecting group is performed to obtain the compound of general formula (I).

7. A compound of general formula (I) as described in claim 1, a lanthanide (III) cation, and Na ＋ Ba ２＋ Pb ２＋ , Sr ２＋ Ca ２＋ , Cd ２＋ , Zn ２＋ Mn ２＋ , Pt ２＋ ,Cd ２＋ Ni ２＋ , Sc ３＋ , Y ３＋ , Bi ３＋ In ３＋ Ru ３＋ , Ir ３＋ Ga ３＋ , Tl ３＋ , Pd ２＋ A coordination compound with a metal cation selected from the group consisting of the following.

8. A method for preparing a coordination compound according to claim 7, characterized by comprising the following steps: i) Synthesize the compound of general formula (I) described in claim 1; ii) Inorganic acids, lanthanide (III) cations, Na ＋ Ba ２＋ Pb ２＋ , Sr ２＋ Ca ２＋ , Cd ２＋ , Zn ２＋ Mn ２＋ , Pt ２＋ ,Cd ２＋ Ni ２＋ , Sc ３＋ , Y ３＋ , Bi ３＋ In ３＋ Ru ３＋ , Ir ３＋ Ga ３＋ , Tl ３＋ , Pd ２＋ To provide a salt with a metal cation selected from the group consisting of; iii) The compound of general formula (I) derived from step i) and the metal salt derived from step ii) are mixed in an aqueous solution, resulting in the chelation of the metal cation by the compound of general formula (I), and the coordination compound described in claim 7 is formed; iv) Optionally, convert the coordination compound derived from step iii), the conversion being selected from at least one of the following reactions: -R １ and / or R ４ and / or R ５ A Pd(0)-catalyzed Suzuki cross-coupling reaction between a halogen group present and phenylboronic acid, thereby introducing -COOH into -R1 and / or R4 and / or R5; -NaN ３ R １ and / or R ４ and / or R ５ The halogen group present and -N ３ Substitution reaction with a group; -R １ and / or R ４ and / or R ５ Hydrolysis of the TIPS protecting group present in the substituent, where R １ and / or R ３ and / or R ５ teeth, 【Chemistry 31】 And as a result, R １ and / or R ３ and / or R ５ teeth, 【Chemistry 32】 This will result in; - Methanol and R １ and / or R ４ and / or R ５ (Here, R １ and / or R ３ and / or R ５ teeth, 【Transformation 33】 The addition reaction with (which converts the triple bond into a dimethyl acetal; - D in which DBU exists ２ CH of group A in O ２ Deuteration of the base, CH ２ The base is CD ２ Convert to base; - Selective reduction of the pyridyl ring of the Z substituent, where Z is 【Transformation 34】 And R ３ and R ５ This is as defined in claim 1; -Z substituent (where Z is 【Chemistry 35】 And R ３ and R ５ The pyridyl ring of (as defined in claim 1), with NaBH as the reducing agent. ４ or NaBD ４ Selective reduction using the following method, thereby removing the Z groups, 【Transformation 36】 or 【Chemistry 37】 Convert to; -R １ and / or R ４ and / or R ５ and / or NH of A ２ Base (R here) １ and / or R ４ and / or R ５ and / or A is -NH ２ or - (CH ２ ) ｎ NH ２ (including) and the reaction with FmocCl, resulting in -NH ２ Convert the group to the -NHFmoc group; -R １ and / or R ４ and / or R ５ and / or the reaction of the -COOH group present in A with the amino group of an amino acid or peptide, thereby forming a peptide bond; -R １ , R ４ , R ５ S-alkylation reaction of substituted halogens with thiols, resulting in the conversion of halogens to sulfides; -R １ , R ３ , and / or R ５ of 【Transformation 38】 Base (R here) １ , R ３ , and / or R ５ teeth, 【Chemistry 39】 The reaction of (including) with azide, thereby the alkyl or aryl and R １ , R ３ , and / or R ５ Forms triazole crosslinks that bridge the groups; -R １ and / or R ５ -N ３ Base (here, R １ and / or R ５ is, -N ３ A reaction between a group (including a carbon-carbon triple bond) and a substituent containing a carbon-carbon triple bond, thereby forming a triazole crosslink that bridges the substituent and the coordination compound; -R １ and / or R ５ The reaction of the -SH group with a substituent containing maleimide, thereby forming a thiosuccinimide bond that bridges the substituent and the coordination compound; -R １ and / or R ５ The reaction of the -SH group with another substituent containing a -SH group, thereby forming a disulfide bridge that bridges the substituent with the coordination compound; -R １ and / or R ５ The reaction of the -SH group with an alkylhalogenide or arylhalogenide, thereby forming a thioether bond between the alkyl or aryl and the coordination compound; -R １ and / or R ５ NO ２ The reaction between the group and the substituent containing the -SH group forms a thioether bond that connects the substituent to the coordination compound.

9. A coordination compound chain comprising at least two coordination compounds according to claim 7, R of the first coordination compound １ or R ５ The group and the R of the following coordination compound １ or R ５ Binding via triazole crosslinks formed between the group and / or, A coordination compound substituent R containing a certain amine group １ or R ４ or R ５ Alternatively, A and substituent R of another coordination compound containing a carboxyl group. １ or R ４ or R ５ or bonded via an amide bond formed between A and, and / or, A coordination compound substituent R containing a certain -SH group １ or R ５ And substituent R of another coordination compound containing an -SH group １ or R ５ A coordination compound chain, linked via a disulfide bond formed between the two.

10. Coordination compound dimer of general formula (X) 【Chemistry 40】 Here, M １ and M ２ This is a metal cation as defined in claim 7, R １ , R ４ and R ５ This is as defined in claim 1.

11. A conjugate for drug tracking comprising the coordination compound described in claim 7 and conjugated to a peptide or protein; Here, the coordination compound described in claim 7 is -NH ２ or R containing a -COOH group １ and / or R ４ and / or R ５ Includes the base; R １ and / or R ４ and / or R ５ The peptide or protein is bound to the -COOH group present in the peptide or protein via an amide bond formed between the -COOH group present in the peptide and the amino group of the peptide or protein, or R １ and / or R ４ and / or R ５ -NH ２ The peptide or protein is bound via an amide bond formed between the group and the carboxyl group of the peptide or protein; further, The peptide is selected from the group comprising oligopeptides of 3 to 20 amino acids; further, The aforementioned protein is a conjugate selected from the group including antibodies.

12. A conjugate for drug tracking comprising the coordination compound chain described in claim 9 and conjugated to a peptide or protein; Here, the coordination compound chain described in claim 9 is -NH ２ or R containing a -COOH group １ and / or R ４ and / or R ５ Includes the base; R １ and / or R ４ and / or R ５ The peptide or protein is bound to the -COOH group present in the peptide or protein via an amide bond formed between the -COOH group present in the peptide and the amino group of the peptide or protein, or R １ and / or R ４ and / or R ５ -NH ２ The peptide or protein is bound via an amide bond formed between the group and the carboxyl group of the peptide or protein; further, The peptide is selected from the group comprising oligopeptides of 3 to 20 amino acids; further, The aforementioned protein is a conjugate selected from the group including antibodies.

13. A method for tracking drugs, characterized by comprising the following steps: i) supplying a cell culture or tissue to be analyzed, comprising at least one conjugate according to claim 11, wherein the peptide or protein is a peptide-based or protein-based drug to be tracked; ii) Hydrolyze the cell culture or tissue from step i) using a strong acid to obtain a hydrolysate; iii) The presence of the coordination compound of claim 7 is qualitatively and / or quantitatively analyzed by the hydrolysate of step ii).

14. A method for tracking drugs, characterized by comprising the following steps: i) supplying a cell culture or tissue to be analyzed, comprising at least one conjugate according to claim 12, wherein the peptide or protein is a peptide-based or protein-based drug to be tracked; ii) Hydrolyze the cell culture or tissue from step i) using a strong acid to obtain a hydrolysate; iii) The presence of the coordination compound chain of claim 9 is qualitatively and / or quantitatively analyzed by the hydrolysate of step ii).

15. In vitro use of the coordination compound according to claim 7, or the coordination compound chain according to claim 9, or the conjugate according to claim 11 or 12, in pharmaceuticals, for the development and testing of new drugs.

16. In vitro use of the coordination compound dimer according to claim 10 for the development and testing of new drugs in pharmaceuticals.

17. A coordination compound according to claim 7, or a coordination compound chain according to claim 9, for use in medical diagnosis.

18. A coordination compound dimer according to claim 10 for use in medical diagnosis.

19. A coordination compound according to claim 7, or a coordination compound chain according to claim 9, Here, the metal is ４４ Sc, ４７ Sc, ６４ Cd, ６７ Cd, ８６ Y, ９０ Y, １４０ Nd, １４９ Pm, １５１ Pm, １５３ Sm, １５９ Gd, １４９ Tb, １６１ Tb, １６５ Dy, １６１ Ho, １６６ Ho, １６９ Er, １６７ Tm, １７５ Yb, １７７ Selected from the group including Lu, and / or R １ , R ４ , and / or R ５ It contains radioactive halogens, A coordination compound according to claim 7, or a coordination compound chain according to claim 9, for use in medicine as a radiodiagnostic agent and / or radiopharmaceutical.

20. A coordination compound dimer according to claim 10, Here, M １ and M ２ teeth, ４４ Sc, ４７ Sc, ６４ Cd, ６７ Cd, ８６ Y, ９０ Y, １４０ Nd, １４９ Pm, １５１ Pm, １５３ Sm, １５９ Gd, １４９ Tb, １６１ Tb, １６５ Dy, １６１ Ho, １６６ Ho, １６９ Er, １６７ Tm, １７５ Yb, １７７ Selected from the group including Lu, and / or R １ , R ４ , and / or R ５ It contains radioactive halogens, The coordination compound dimer according to claim 10, for use in medicine as a radiodiagnostic agent and / or radiopharmaceutical.

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