Chelating agents and related methods

Abstract

The present disclosure generally relates to the field of chelating agents, including chelating agents useful for radiopharmaceuticals and their use in nuclear medicine as tracers, imaging agents and for the treatment of various diseases. The chelating agents include compounds of Formula (I), Formula (II) and Formula (III) disclosed herein.

Description

[0001] CHELATING AGENTS AND RELATED METHODS

[0002] TECHNICAL FIELD

[0003] This disclosure provides chelating compounds, including the use of chelating compounds in the field of radiopharmaceuticals.

[0004] BACKGROUND

[0005] Radiopharmaceuticals are utilized both for diagnostics and therapy and play an important role in modern medicine. Typical radiopharmaceutical compounds include three parts: a targeting moiety (vector) which ensures delivery to the desired location within body; a chelator that retains a radionuclide; and a linker that covalently couples chelator to the targeting moiety. Depending on the application, different properties of a chelator are desired such as strong affinity to the radionuclide, stability of the formed complex in the in vivo conditions, rapid complexation kinetics under mild conditions, high versality of linker incorporation without sacrificing coordination integrity, selectivity of binding of the desired radionuclide, lipophilicity and physical chemical properties of a chelator, and the like. Most chelators include a bidentate substructure such as ethylenediamine

[0006]

[0007] -AO - OO

[0008] N-A 0- / 0-1

[0009] (s), aminoethanol ( ), or ethylene glycol ( * ). Modulation of properties of these moieties may substantially improve properties of chelators. For example, multidentate aza ligands able to complex metal ions and useful in diagnostics and therapy are disclosed in W003 / 008390. The compound 6-amino-6-methylperhydro-l,4-diazepinetetraacetic acid (AAZTA) is a chelating agent that forms stable complexes with metal ions.

[0010] Targeted radionuclide therapy include FDA approved radiopharmaceuticals with chelating moieties such as l,4,7,10-tetraazacyclododecane-N, N', N", N"'-tetraacetic acid (DOTA), a macrocyclic chelator for binding radioactive metal isotopes, conjugated to a tumor targeting moiety. Other macrocyclic chelators are disclosed in WO2020 / 229974. The compound -amino-6-((16-((6-carboxypyridin-2-yl)methyl)- 1,4, 10, 13-tetraoxa-7, 16-diazacyclooctadecan-7-yl)methyl)picolinic acid (Macropa-NH2) is a stable and selective chelator for large lanthanide ions such as225Ac.

[0011] Ongoing development of targeted radiopharmaceuticals along with insufficient supply of currently used radionuclides requires the development of novel chelators for alternative radionuclides. There remains an unmet medical need for novel chelating agents useful for chelating metals such as radionuclides, such as chelators with improved stability of complexation of radionuclides, or greater affinity for metal ions.SUMMARY

[0012] In some embodiments, novel chelating agents containing one or more pyrrolidine bidentate moieties. For example, novel chelating agents can have chemical structures that include one or more pyrrolidine bidentate moieties in place of ethylenediamine, aminoethanol or ethylene glycol moieties in various known chelating agents. The nitrogen in the incorporated pyrrolidine ring may serve a variety of functions. For example, the nitrogen in the incorporated pyrrolidine moiety may serve as a coordination site itself, provide an opportunity for an additional point of attachment of a pendant to improve coordination properties, or serve as a point of attachment of a conjugated linker in a larger molecule. In some embodiments, the incorporation of pyrrolidine in the chemical structure of a chelator, as described herein, may favorably change the charge and lipophilicity of a chelator (e.g., the nitrogen in the pyrrolidine ring incorporated in a chelator may improve pK properties of a radiopharmaceutical molecule). In some embodiments, the introduction of the pyrrolidine ring may change rigidity of the structure resulting in better affinity to a radionuclide. Utilization of more than one pyrrolidine moiety allows for the incorporation of a second vector for improved uptake at the desired location in the body.

[0013] The configuration of each of the two chiral centers in the incorporated pyrrolidine may be oriented to obtain desired properties in a chelator, such as better coordination and / or better specificity to some of radionuclides. In some embodiments, a chelating agent as described herein has a chemical structure characterized by incorporating a pyrrolidine substructure selected from

[0014] R.>RV-N

[0015] R I >-R

[0016]

[0017] orN / ' in place of an ethylenediamine (

[0018]

[0019] * ) substructure in a chelating agent. In some embodiments, a chelating agent as described herein has a chemical structure

[0020] R- V R r >— R characterized by incorporating a pyrrolidine substructure selected from

[0021]

[0022] or o / -

[0023]

[0024] in place of an aminoethanol ( ° ' ) substructure in a chelating agent. In some embodiments, a chelating agent as described herein has a chemical structure characterized by incorporating a

[0025] R R..

[0026] R R

[0027] pyrrolidine substructure selected from

[0028]

[0029] or in place of an ethylene glycol (

[0030]

[0031] ) substructure in a chelating agent. One or more pyrrolidine type bidentate moieties as described above can be introduced in a chelator, as described herein.

[0032] In some embodiments, a chelating agent comprising a polydentate chelating ligand comprises one or more bidentate moieties having the chemical structure -X-P-Y- where X and Y are each independently oxygen or nitrogen and P is a pyrrolidine covalently bonded to X and Y, and wherein the pyrrolidine containing chelating agent chelates one or more metal or nonmetal ions. In some embodiments, the one or more bidentate donor moieties are independently selected from the group

[0033]

[0034] some embodiments, the pyrrolidine containing chelating agent comprises a single pyrrolidine bidentate donor moiety. In some embodiments, the chelator is selected from the group consisting of: a chelating agent comprising at least one modified ethylene glycol moiety wherein the ethylene of the modified ethylene glycol moiety is replaced by a pyrrolidine; or an ethylenediamine chelating agent comprising at least one ethylenediamine moiety is replaced by at least one modified ethylenediamine moiety wherein the ethylene of the modified ethylenediamine moiety is replaced by a pyrrolidine; or an aminoethanol chelating agent comprising at least one aminoethanol replaced by at least one modified aminoethanol moiety wherein the ethylene moiety of the modified aminoethanol moiety is replaced by a pyrrolidine.

[0035] In some embodiments, the chelator is a modified AAZTA chelator where the ethylenediamine chelating moiety is replaced by a modified ethylenediamine chelating moiety wherein the ethylene of the modified ethylenediamine chelating moiety is replaced by pyrrolidine. In some embodiments, the chelator is a compound of Formula (I)

[0036] R6 R25a

[0037] a ^2 -4— R25b

[0038] Zi

[0039] X1"f~R25d

[0040] R2R25e

[0041]

[0042] (I),

[0043] wherein

[0044] X1 isN;

[0045] X2isN;

[0046] Zi is C(R24)(R25C);

[0047] RI, R2, Rfi is a linker to a vector, a chelating pendant moiety, or hydrogen;R24 and Rise are independently NHRi, CO2R1, a linker to a vector, a chelating pendant moiety or hydrogen;

[0048] R25a, R25b, R25d, and R25C are each independently hydrogen or C1-C4 alkyl; and the one or more chelating pendant moieties are each independently carboxy, C1-C20 alkyl optionally substituted with one or more carboxy, amino or sulfur, or aryl or heteroaryl each optionally substituted with one or more carboxy, sulfur or amino or optionally substituted with C1-C4 alkyl optionally substituted with one or more carboxy, sulfur or amino.

[0049] In some embodiments, each chelating pendant moiety is independently selected from the group consisting of: -COOH, -(C1-C4 alkyl)COOH, -(C1-C4 alkyl)COOH, -(C1-C4 alkyl)-S-CH3, -(C1-C4 alkyl)-Pyr-COOH (wherein Py is pyridinyl), and-(Ci-C4 alkyl)-Ph-COOH (wherein Ph in phenyl). In some embodiments, each chelating pendant moiety is independently selected from the group consisting of: -COOH, -(CH2)COOH, -(CH2)2COOH, -(CH2)2-S-CH3, -CH2-Pyr-COOH (wherein Py is pyridinyl), and -CH2-PI1-COOH (wherein Ph in phenyl).

[0050] In some embodiments, the linker to the vector in Formula (I) comprises C1-C20alkyl, amide, carboxy, oxo, ether, phenyl or heteroaryl. In some embodiments, the linker to the vector in Formula (I) comprises one or more of C1-C20 alkyl, -CONH-, -NHCO-, phenyl, and C3-C6 cycloalkyl optionally substituted with one or more C1-C4 alkyl, carboxy, and amino. In some embodiments, the vector comprises one or more amino acids or amino acid derivatives.

[0051] In some embodiments, the chelator is a compound of Formula (I-A)

[0052] R2with HO group

[0053]

[0054] (IA),

[0055] wherein R1, R2, and R7 are each independently a linker to a vector, a chelating pendant moiety, or hydrogen. In some embodiments, the chelator is a compound of Formula (I-B)

[0056] Formula (I-B) structure labels

[0057]

[0058] *7 (IB),

[0059] wherein R1, R2, R3, and R7 are each independently a linker to a vector, an additional chelating pendant moiety, or hydrogen.In some embodiments, the compound is a compound of Formula (IA) or Formula (IB), wherein R2, R3 and R7 are each independently a chelating pendant moiety selected from the group o 0

[0060] consisting of but not limited to: -

[0061]

[0062] COOH, -(CH2)COOH, -(CH2)2COOH, X, and \ In some embodiments, the chelator is a modified DOTA chelator comprising one or more pyrrolidine moieties. In some embodiments, the chelator is a compound of Formula (II)

[0063] / 1

[0064] Z N—R1

[0065] Y2*2

[0066]

[0067] (II),

[0068] wherein:

[0069] Xi is NR2 or 0;

[0070] X2 is NR7 or 0;

[0071] Yi is NR3 or 0;

[0072] Y2 is NRe or 0;

[0073] Ri, R2, R3, Re, and R7 are independently a hydrogen, a linker to a vector, an additional chelating pendant moiety as defined above with respect to Formula (I) (e.g., C1-C4 alkyl optionally substituted with carboxy, amino or C1-C4 alkylamino);

[0074] Z is –(CH2)2-X3-[(CH2)2-Y3]n-(CH2)2-, wherein

[0075] X3 is NR4, 0, methylene or a direct bond;

[0076] Y3 is NR5, or 0;

[0077] n is 0 or 1; and

[0078] R4, and R5 are each independently a linker to a vector, a hydrogen, or a chelating pendant moiety as defined above with respect to Formula (I).

[0079] In some embodiments, a chelating compound is a compound of Formula (II) wherein Ri is hydrogen, a linker to a vector, an additional chelating pendant moiety as defined above with respect to Formula (I) (e.g., C1-C4 alkyl optionally substituted with carboxy, amino or C1-C4 alkylamino); and R2, R3, R, Rs, Re and R7 are independently a hydrogen, a linker to a vector or chelating pendant moiety (e.g., C1-C4 alkyl optionally substituted with carboxy, amino or C1-C4 alkylamino).In some embodiments, the chelator is a compound of formula (IIA):

[0080]

[0081] (IIA),

[0082] wherein: Xi, Yi, X2, Y2, Ri and Z are each as defined with respect to Formula (II). In some embodiments, the chelator is a compound of formula (IIB):

[0083]

[0084] wherein: Xi, Yi, X2, Y2, Ri and Z are each as defined with respect to Formula (II).

[0085] In some embodiments, the chelator is a compound of Formula (II), (IIA) or (IIB), wherein Xi and X2 are each oxygen and Yi is NR3 and Y2 is NRe, wherein R3 and R6 are as defined with respect to Formula (II);and Z is -(CH2)2-, -(CH2)3-, -(CH2)2-O-(CH2)2-, -(CH2)2-NH-(CH2)2-, or -(CH2)2-O-(CH2)2-O-(CH2)2-.

[0086] In some embodiments, the chelating agent is a chelator of Formula (III):

[0087] R1

[0088]

[0089] (III)

[0090] wherein

[0091] each ring A and ring B are independently a 6-10 membered aryl or a 5-10 membered heteroaryl, wherein each of ring A and ring B is optionally substituted with one or more substituents independently selected from the group consisting of halo, alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, -OR13, -SR13, -(CH2)pCOOR13, -OC(O)R13, -N(R13)2, -CON(R13)2, -NO2, -CN, -OC(O)N(R13)2, and X;

[0092] Zi and Z2 are independently -(C(Ri2)2)m-, or-(CH2)n-C(Ri2)(X)-(CH2)n-;

[0093] each X is independently -L1-R11;

[0094] each n is independently 0, 1, 2, 3, 4, or 5;each m is independently 1, 2, 3, 4 or 5;

[0095] each p is independently 0 or 1;

[0096] Li is absent or a linker;

[0097] R11 is a nucleophilic moiety or an electrophilic moiety, or R11 comprises a targeting ligand;

[0098] each R12 is independently hydrogen, alkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl;

[0099] each R13 is independently hydrogen or alkyl;

[0100] provided the chelator comprises at least one X and when X is present on ring A or ring B, then Li is a linker or R12 is not hydrogen;

[0101] Ri is a linker to a vector, a chelating pendant moiety, or a group modulating physical chemical properties.

[0102] In some embodiments, the compound is a radiometal complex of Formula (III) further comprising a radiometal ion, such as an alpha-emitting radiometal ion such as actinium-225.

[0103] Embodiments of the present disclosure provide a pyrrolidine containing chelating compound -X— p-y- - comprising one or more bidentate donor moieties having a chemical structure of5 5, wherein X and Y are each independently oxygen or nitrogen and P is a pyrrolidine covalently bonded to X and Y, and wherein the pyrrolidine containing chelating compound chelates to one or more metal or nonmetal ions.

[0104] Embodiments of the present disclosure further provide a chelating ligand, comprising:

[0105] Ri2r' / O E R 01 one or more bidentate donor moieties having

[0106]

[0107] 13, *,12*

[0108]

[0109] , provided that at least one of the

[0110] electron donor unit i

[0111]

[0112] s wherein:the one or more bidentate donor moieties are covalently bonded by C1-C4 alkyl; R10is, each independently, H, a nitrogen protecting group, a linker to a vector, a chelating pendant moiety, or a vector; R12and R13are, each independently, H, Ci-Ce alkyl optionally substituted with aryl or heteroaryl, which are optionally substituted with -COOR16, -P(O)(OR16)2, or Ci-Ce heteroalkyl; R14a, R14b, R15a, R15b, and R16are, each independently, H, Ci-Ce alkyl, or Ci-Ce heteroalkyl; and the chelating ligand has from 4 to 12 coordination sites for one or more metals or nonmetal ions to bind to the one or more bidentate donor moieties.

[0113] BRIEF DESCRIPTION OF THE DRAWINGS

[0114] In the figures, identical reference numbers identify similar elements. The sizes and relative positions of elements in the figures are not necessarily drawn to scale and some of these elements are arbitrarily enlarged and positioned to improve figure legibility. Further, the particular shapes of the elements as drawn are not intended to convey any information regarding the actual shape of the particular elements, and have been solely selected for ease of recognition in the figures.

[0115] FIG. 1 illustrates i) an HPLC trace of compound 8 and Macropa, ii) an HPLC trace of compound 8 and Macropa with excess chelators with regards to metal (La), and iii) an HPLC trace of compound 8 and Macropa with excess metal (La).

[0116] FIG. 2A illustrates a comparison of minimal concentration of Compound 8 required for complexation with La at room temperature (25°C) and minimal concentration of DOTA required for complexation with La at room temperature (25°C) and elevated temperature (95°C).

[0117] FIG. 2B illustrates a comparison of minimal concentration of Compound 8 required for complexation with Lu at room temperature (25°C) and minimal concentration of DOTA required for complexation with Lu at room temperature (25°C) and elevated temperature (95°C).

[0118] FIG. 2C illustrates a comparison of minimal concentration of Compound 8 required for complexation with Tb at room temperature (25°C) and minimal concentration of DOTA required for complexation with Tb at room temperature (25°C) and elevated temperature (95°C).

[0119] FIG. 3 illustrates HPLC / ICP / MS quantification of La bound to compound 8 compared to free La at different concentrations of compound 8..

[0120] DETAILED DESCRIPTION

[0121] In some embodiments, the chelator is any compound comprising the chelating compound of Formula (I)R25a

[0122] R25b

[0123] z1

[0124] X1—R25d

[0125] R2R25e (I),

[0126]

[0127] wherein

[0128] Xi isN;

[0129] X2isN;

[0130] Zi is C(R24)(R25C);

[0131] R1, R2, R6 is a linker to a vector, a chelating pendant moiety, or hydrogen;

[0132] R24, and R25care each independently NHR1, CO2R1, a linker to a vector, a chelating pendant moiety or hydrogen;

[0133] R25a, R25b, R25d, and R25e are each independently hydrogen or C1-C4 alkyl; and the one or more chelating pendant moieties are each independently carboxy, C1-C20 alkyl optionally substituted with one or more carboxy, amino or sulfur, or aryl or heteroaryl each optionally substituted with one or more carboxy, sulfur or amino or optionally substituted with C1-C4 alkyl optionally substituted with one or more carboxy, sulfur or amino.

[0134] In some embodiments, R1, R2, R6, R24 and R25c are each independently chelating pendant moieties. In some embodiments, R1, R2, R6, and R25c are each independently chelating pendant moieties and R24 is hydrogen. For example, in some embodiments Ri, R2, Rs and R25c are each the same or different, and are each independently carboxy, C1-C20 alkyl optionally substituted with one or more carboxy, amino or sulfur, or aryl or heteroaryl each optionally substituted with one or more carboxy, sulfur or amino or optionally substituted with C1-C4 alkyl optionally substituted with one or more carboxy, sulfur or amino. In some embodiments R1, R2 and R6 are each independently a pendant metal chelating moiety comprising (Ci-Cs)alkyl substituted with a (Ci-Cs)alkyl-carboxy, such as -(C1-C4 alkyl)C(O)O-(Ci-C4 alkyl). In some embodiments R24 is hydrogen and R25C is a metal chelating moiety comprising a (Ci-Cs)alkyl, an amide and a carboxy. In some embodiments R24 is hydrogen and R25c is -NR10aC(O)-(C1-C4 alkyl)C(O)OR10b or -C(O) NR10a-(C1-C4 alkyl)C(O)OR10b wherein R10a and R10b are each independently H or C1-C4 alkyl. In some embodiments Ri, R2 and Rs are each independently -(CH2)C(O)O-C(CH3)3, R 4 is hydrogen and R25c is -NH(CO)-(CH2)2-C(O)O-CH3. In some embodiments Ri, R2, and Rs are each independently -(CH2)C(O)O-C(CH3)3, R24 is hydrogen and R2scis -COOH.

[0135] In some embodiments, the linker to the vector in Formula (I) comprises (C1-C20alkyl) optionally substituted with one or more oxygen, nitrogen or sulfur heteroatoms, one or more amide,Ce-Cio aryl or 5-10 member heteroaryl, C3-C8 cycloalkyl, 3-10 member heterocycloalkyl, carboxy, amino, or sulfonyl.

[0136] In some embodiments, the chelator is a modified DOTA chelator comprising one or more pyrrolidine moieties. In some embodiments, the chelator is a compound of Formula (II)

[0137] Formula (II) structure labels with X1, Z, N-R1, Y2, X2

[0138]

[0139] (II),

[0140] wherein:

[0141] Xi is NR2 or 0;

[0142] X2 is NR7 or O;

[0143] Yi is NR3 or 0

[0144] Y2 is NRe or 0;

[0145] Ri;R2, R3, Re, and R? are each independently a linker to a vector, a protecting group, hydrogen or C1-4 alkyl, or a chelating pendant moiety;

[0146] Z is –(CH2)2-X3-[(CH2)2-Y3]n-(CH2)2-, wherein

[0147] X3 is NR4, 0, methylene or a direct bond;

[0148] Y3 is NR5, or 0;

[0149] n is 0 or 1;

[0150] R4, and R5 are each independently a linker to a vector, a hydrogen, or a chelating pendant moiety as defined above with respect to Formula (I).

[0151] In some embodiments, the compound is a compound of Formula (II), wherein Ri, R2, R3, and R are each independently hydrogen, a protecting group (e.g., Boc), a linker to a vector, an additional chelating pendant moiety as defined above with respect to Formula (I), (e.g., C1-C4 alkyl optionally substituted with carboxy, amino or C1-C4 alkylamino). In some embodiments, a chelating compound is a compound of Formula (II) wherein Ri, R2, R3, R4, R5, Rs, and R7 is hydrogen, a protecting group (e.g., Boc), a linker to a vector, an additional chelating pendant moiety as defined above with respect to Formula (I), (e.g., C1-C4 alkyl optionally substituted with carboxy, amino or C1-C4 alkylamino).

[0152] In some embodiments, the chelator is a compound of formula (HA):

[0153]

[0154] wherein: Xi, Yi, X2, Y2, Ri, and Z are each as defined with respect to Formula (II). In some embodiments, the chelator is a compound of formula (IIB):

[0155]

[0156] wherein: Xi, Yi, X2, Y2, Ri, and Z are each as defined with respect to Formula (II).

[0157] In some embodiments, the compound is a compound of Formula (II), Formula (II- A) or Formula (II-B), wherein Xi and X2 are each 0 and Y 1 and Y2 are each NH. In some embodiments, the compound is a compound of Formula (II), Formula (II- A) or Formula (II-B), wherein Z is -(CH2)2- or-(CH2)3-. In some embodiments, the compound is a compound of Formula (II), Formula (II- A) or Formula (II-B), wherein Xi and X2 are each 0 and Y 1 and Y2 are each NH and Z is -(CH2)2- or - (CH2)S-. In some embodiments, the compound is a compound of Formula (II), Formula (II-A) or Formula (II-B), wherein Z is -(CH2)2-O-(CH2)2- or -(CH2)2-NH-(CH2)2-. In some embodiments, the compound is a compound of Formula (II), Formula (II-A) or Formula (II-B), wherein Xi and X2 are each 0 and Yi and Y2 are eachNH and Z is -(CH2)2-O-(CH2)2- or -(CH2)2-NH-(CH2)2-. In some embodiments, the compound is a compound of Formula (II), Formula (II-A) or Formula (II-B), wherein Z is -(CH2)2-O-(CH2)2- O-(CH2)2-. In some embodiments, the compound is a compound of Formula (II), Formula (II-A) or Formula (II-B), wherein Xi and X2 are each 0 and Yi and Y2 are each NH, and Z is -(CH2)2-O-(CH2)2-O-(CH2)2-.

[0158] In some embodiments, the chelator is a compound of Formula (II), (IIA) or (IIB), wherein Xi and X2 are each nitrogen and Yi and Y2 are each a direct bond and Z is N. In some embodiments, the chelator is a compound of formula (1):

[0159] / >

[0160]

[0161] R< 7(1),

[0162] wherein Ri, R2, R4, and R7 is as defined in Formula (II). In some embodiments, the compound is a compound of formula (1), wherein chelating pendant in R2, R4 and R7 is -(CH2)COOH.

[0163] In some embodiments, the chelator is a compound of Formula (II), (IIA), or (IIB), wherein Xi and X2 are each oxygen and Yi and Y2 are NR3 and NRe, respectively, and Z is -(CH2)2- or - (CH2)3-, and Ri, R3, and Re are as defined in Formula (II). In some embodiments, the chelator is a compound selected from the group consisting of:

[0164]

[0165] R6X—7(2c), and (2d),

[0166] wherein Ri, R3 and Re are as defined with respect to Formula (II).

[0167] In some embodiments, the chelator is a compound of Formula (II), (IIA) or (IIB), wherein Xi and X2 are each NR2 and NR7, respectively, and Y 1 and Y2 are each NR3 and NRe, respectively, and Z is - (CH2)2- or - (CH2)3-. In some embodiments, the compound is a compound of formula (2e):

[0168] p y \ n

[0169]

[0170] (2e),

[0171] wherein Ri, R2, R3, Re, and R7 are as defined in Formula (II).

[0172] In some embodiments, the chelator is a compound of Formula (II), (IIA) or (IIB), wherein Xi and X2 are each oxygen and Y 1 and Y2 are NR3 and NRe, respectively, and Z is -(CH2)2-NR4-(CH2)2-, wherein R3, R4, and Re are each as defined with respect to Formula (II) above. In some embodiments, the chelator is a compound selected from the group consisting of:

[0173]

[0174] wherein Ri, R3, R, and Re are as defined in Formula (II).

[0175] In some embodiments, the chelator is a compound of Formula (II), (IIA) or (IIB), wherein Xi and X2 are NR2 and NR7, respectively, and Y 1 and Y2 are each NR3 and NRe, respectively, and Z is - (CH2)2-NR5- (CH2)2-, wherein R2, R3, R4, Re and R7 are as defined in Formula (II) above. In some embodiments, the compound is a compound of formula (4a), formula (4b) or formula (4c):

[0176]

[0177] wherein Ri, R2, R3, R4, Re, and R7 are as defined in Formula (II). In some embodiments, the chelator is a compound is a compound of formula (5a) or formula (5b):

[0178]

[0179] wherein R3 and Rg are as defined in Formula (II).

[0180] In some embodiments, the chelating agent is a chelator of Formula (III):

[0181] R!

[0182]

[0183] (Ill)

[0184] wherein:

[0185] each ring A and ring B is independently a 6- 10 membered aryl or a 5- 10 membered heteroaryl, wherein each of ring A and ring B is optionally substituted with one or more substituents independently selected from the group consisting of halo, alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, -OR13, -SR13, -(CH2)pCOOR13, -OC(O)R13, -N(R13)2, -CON(R13)2, -NO2, -CN, -OC(O)N(R13)2, and X;

[0186] Zi and Z2 are independently -(C(Ri2)2)m-, or -(CH2)n-C(Ri2)(X)-(CH2)n-; each X is independently -L1-R11;

[0187] each n is independently 0, 1, 2, 3, 4, or 5;

[0188] each m is independently 1, 2, 3, 4 or 5;

[0189] each p is independently 0 or 1;Li is absent or a linker;

[0190] Rn is a nucleophilic moiety or an electrophilic moiety, or Rn comprises a targeting ligand;

[0191] each R12 is independently hydrogen, alkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl;

[0192] each R13 is independently hydrogen or alkyl;

[0193] provided the chelator comprises at least one X and when X is present on ring A or ring B, then Li is a linker or R12 is not hydrogen;

[0194] Ri is a linker to a vector, a chelating pendant moiety, or a group modulating physical chemical properties.

[0195] In some embodiments, the compound is a compound of Formula (III), wherein Ri is C1-C20 alkyl optionally substituted with one or more amino, ether, sulfoether, carboxy, or oxo, such as -(CH2)-S-(CI-C6alkyl)-NH2.

[0196] In some embodiments, the compound is a compound of Formula (III), wherein Zi is -CH2- or -CHR12-, Z2 is -L1-R11 or -CR12-, ring A and ring B are each independently phenyl, or a 5-6 member heteroaryl (e.g., a phenyl, diazole, pyridine, or pyrimidine) optionally substituted with one or more carboxy, and Li is absent or a linker and R11 is a nucleophilic moiety or an electrophilic moiety, or R11 comprises a targeting ligand, and each R12 is independently hydrogen, -methyl, or ethyl, provided at least one R12 is methyl or ethyl.

[0197] In some embodiments, the compound is a compound of Formula (III), wherein ring A and ring B are each independently, Zi is -CH2- or -CHR12- and Z2 is -CHL1-R11, wherein Li and R11 are as defined with respect to Formula (III).

[0198] In some embodiments, Li in Formula (II) is selected from the group consisting of:

[0199]

[0200] wherein n is an integer of 0 to 10, preferably an integer of 1 to 4, and m is an integer of 0 to 12, preferably an integer of 0 to 6.

[0201] In some embodiments, the compound is a compound of Formula (III) wherein Rn is

[0202] or In some embodiments, the compound is a compound of Formula (III)

[0203] mAh

[0204]

[0205] wherein the mAh is a monoclonal antibody or antibody binding fragment such as PSMB127, pertuzumab, cetuximab, panitumumab, Herceptin or Hl 1B6. In some embodiments, a compound is a compound of Formula (III) wherein R11 is cyclooctynyl, or a cyclooctynyl derivative selected from the group consisting of bicyclononynyl (BCN), difluoroniated cyclooctynyl (DIFO), dibenzocyclooctynyl (DIBO), keto-DIBO, biarylazacyclooctynonyl (BARAC), dibenzoazacyclooctynyl (DIBAC, DBCO, ADIBO), dimethoxyazacyclooctynyl (DIMAC), difluorobenzocyclooctynyl (DIFBO), momobenzocyclooctynyl (MOBO) or tetramethoxy dibenzocyclooctynyl (TMDIBO).

[0206] In one embodiment, the chelating ligand has a structure of Formula (III):

[0207]

[0208] wherein:

[0209] each ring A and ring B is independently a 6-10 membered aryl or a 5-10 membered heteroaryl, wherein each of ring A and ring B is optionally substituted with one or more substituents independently selected from the group consisting of halo, alkyl, alkenyl, cycloalkyl, cycloalkenyl,aryl, heterocyclyl, heteroaryl, -OR13, -SR13, -(CH2)pCOOR13, -0C(0)Ri3, -N(R13)2, -CON(R13)2, -NO2, -CN, and -OC(O)N(Ri3)2; Zi and Z2 are independently -(C(Ri2)2)m-, or -(CH2)n-C(R12)(X)-(CH2)n-; each n is independently 0, 1, 2, 3, 4, or 5; each m is independently 1, 2, 3, 4 or 5; each p is independently 0 or 1; each R12 is independently hydrogen, alkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl; each R13 is independently hydrogen or alkyl; and Ri is H, a linker to a vector, a chelating pendant moiety, a group modulating physical chemical properties, or a nitrogen protecting group.

[0210] In some embodiments, Zi is -CH2-; Z2 is -CH2-; ring A and ring B are each a 6 member heteroaryl substituted with -COOH or with -COOH and -N(CH3)2; and Ri is a nitrogen protecting group for the compound of Formula (III).

[0211] In some embodiments, the nitrogen protecting group of R1is t-butyl carbamate (BOC), 9-fluorenylmethyl carbamate (Fmoc), benzyloxycarbonyl (Cbz), acyl (Ac), trifluoroacetamide, benzyl

[0212]

[0213] In some embodiments, the nitrogen

[0214] protecting group of R1is t-butyl carbamate (

[0215]

[0216] BOC), " T",, or

[0217]

[0218] In some specific embodiments, the compound is a compound selected from Table 1A.

[0219]

[0220]

[0221]

[0222]

[0223] Compound ID Structure

[0224] III-9

[0225] III- 10

[0226]

[0227] In one embodiment, a pyrrolidine containing chelating compound is disclosed. The pyrrolidine containing chelating compound comprises one or more bidentate donor moieties having a -X— p— y-j>-chemical structure of5 5. X and Y are each independently oxygen or nitrogen and P is a pyrrolidine covalently bonded to X and Y, and wherein the pyrrolidine containing chelating compound chelates to one or more metal or nonmetal ions.

[0228] In some embodiments, a chelating ligand is disclosed. The chelating ligand comprises one or

[0229] ”12R * 0 / - R Elm

[0230]

[0231] ore bidentate donor moieties having13,,12■

[0232]

[0233] provided that at least one of the electron donor unit is

[0234]

[0235] wherein: the one or more bidentate donor moieties are covalently bonded by C1-C4 alkyl; R10is, each independently, H, a nitrogen protecting group, a linker to a vector, a chelating pendant moiety, or a vector; R12and R13are, each independently, H, Ci-Ce alkyl optionally substituted with aryl or heteroaryl, which are optionally substituted with -COOR16, -P(O)(OR16)2, or Ci-Ce heteroalkyl; R14a, R14b, R15a, R15b, and R16are, each independently, H, Ci-Ce alkyl, or Ci-Ce heteroalkyl; and the chelating ligand has from 4 to 12 coordination sites for one or more metals or nonmetal ions to bind to the one or more bidentate donor moieties.

[0236] In some embodiments, one or more bidentate donor moieties have

[0237]

[0238]

[0239] In some embodiments, the bidentate donor moiety

[0240] comprises In some embodiments, the bidentate donor moiety comprises

[0241]

[0242] In some embodiments, the bidentate donor moiety comprises In some embodiments, R14a, R14b, R15a, R15b, and R16are H. In some embodiments, one or

[0243] more bidentate donor moieties are independently selected from the group consisting of:

[0244]

[0245]

[0246] In some embodiments, R10is a nitrogen protecting group. In some embodiments, the nitrogen protecting group of R10is t-butyl carbamate (BOC), 9-fluorenylmethyl carbamate (Fmoc),

[0247] benzyloxycarbonyl (Cbz), acyl (Ac), trifluoroacetamide, benzyl (

[0248]

[0249] Bn),

[0250]

[0251] In some embodiments, the nitrogen protecting group of R10

[0252]

[0253] In some embodiments, R12and R13are H or Bn.

[0254] In some embodiments, Ci-Cg heteroalkyl of R12and R13is -CH2C(=O)OH,

[0255]

[0256]

[0257] In some embodiments, n1and n2are each an integer of 1.

[0258] In some specific embodiments, the compound is a compound selected from Table IB.

[0259] Table 1B. Exemplary Compounds

[0260] Compound ID Structure

[0261] 7 ox / ^\ / ~A / / °

[0262] V* j vjH

[0263] HONA Z~NOH

[0264] 8 0 fA P

[0265] J HON~\ / ~NOH

[0266] 0^0^

[0267] 9 ox / ===\ ^A,p

[0268] Y^ / / v W

[0269] HONA 7~nOH

[0270] H°X0^5 o^OH

[0271] 0

[0272] .

[0273] 10 Ox / ^\ / " A / P

[0274] HONA J ANOH HN NH

[0275]

[0276] Compound ID Structure

[0277] 11

[0278] yn m

[0279] HO OH HN NH

[0280] V

[0281] o=s=o

[0282] ox

[0283] 12

[0284] HO-^0C )-N*H O°H

[0285] 1 CH 0°H0

[0286] 13

[0287] W CH HONA / -~NOH

[0288] H V <r0H

[0289] 14

[0290] yn CH Ho'NA OH

[0291] HVO U °koH

[0292]

[0293]

[0294] Structure

[0295]

[0296] Structure

[0297]

[0298]

[0299] Preparation of Chelating Agents

[0300] In some embodiments, methods of making chelating agents are provided, such as the methods described below. For example, chelating agents can be obtained by the following scheme:

[0301] H 11

[0302] __.. N NsPG Z PG

[0303]

[0304] X = N(R)orO Z = chain of atoms (carbon N(R)orO

[0305] R = any substituent oxygen, nitrogen)

[0306]

[0307] chain of atoms (carbon LG = leaving group, PG = protecting group oxygen, nitrogen) as example (MsO-, TsO-, CI-, Br-) as example (Bn-, Bz-, Boc-, Fmoc-) R, R2, R3= any substituent Using 3-4-disubstituted BOC-pyrrolidine derivatives as a starting material (according to the scheme above) allows obtaining a wide range of macrocycles containing a pyrrolidine fragment, and may provide one or more potential advantages, including (1) chelators having an additional point for selective functionalization of target macrocyclic compounds; (2) chelators hsaving a more rigid and controllable geometric structure in comparison with the common analogs; and / or (3) flexibility to design a variety of chelator configurations using a diverse set of “chain amines” as building blocks. A variety of “chain amines” can be used for macrocyclization, to obtain cyclic chelating agents. These “chain amines” suitable for macrocyclization include:

[0308] Where PG is a generic protecting group such as, but not limited to, Fmoc, Bn, Bz, and Boc.In some embodiments, the synthesis of the 3-4-disubstituted BOC-pyrrolidine derivatives, can be performed according to the following general scheme:

[0309] HCI (aq), MeOH

[0310] o~. / ~OMsMsCl, Et3N, DCMBOC"NCAO / X^OMS

[0311]

[0312] In some embodiments, the macrocyclization and deprotection to form cyclic chelating agents, can be performed according to the following general scheme:

[0313] OMsH H

[0314] BOCYY Bn Z Bn Pd / c, H2(g), \U Na2CO3, MeCN Boc-N MeOH

[0315]

[0316] I Z = chain of atoms (carbon OMs oxygen, nitrogen)

[0317] For example, in some embodiments, the following cyclic chelating agents were prepared using the general methods described herein:

[0318] NH

[0319] NH

[0320]

[0321]

[0322] In some embodiments, the synthesis of the 3-4-disubstituted BOC-pyrrolidine derivatives, can be performed according to the following general scheme:

[0323] 1. Benzaldehyde 2. NaBH4

[0324] - Na2CO3) MeCN

[0325] o^-OMs MsCl, Et3N, DCM

[0326]

[0327] In some embodiments, the macrocycle chelating agents, can be prepared according to general scheme:

[0328] XNH HN,,

[0329] JT N-BocXNH

[0330]

[0331] Additional Embodiments

[0332] In some embodiments, additional chelating agents can be obtained by structurally modifying a benchmark chelator with a pyrrolidine, as described below.

[0333] Benchmark Chelator Modified Structure

[0334] 0^ ^0

[0335] RX<N

[0336] HO N OH r >-R HO N rr QH Replace one -N-(CH2)2-N- moiety with * 0 0 VN

[0337] 1 5~~R

[0338] DOTA

[0339] N £

[0340] or *

[0341] CRYPT V

[0342] [ 'HR ' / N / \

[0343] 0- / - Replace one -N-(CH2)2-O- moiety with * fX° °i o °)

[0344] i i *RV-N

[0345] R I y~R

[0346] or °; or

[0347] RV-N

[0348] I y~R o-A Replace one -O-(CH2)2-O- moiety with *RV. N

[0349] | >-R

[0350] or ° / s~

[0351] HO^O

[0352] RV'N

[0353] I )“RN- / - Replace one -N-(CH2)2-N- moiety with * XN

[0354] Hl £

[0355] CB-TE2A ° °H

[0356] N / '

[0357]

[0358] orsBenchmark Chelator Modified Structure

[0359] HO^OR^N | >~~R N

[0360] / \ 0 ' / ? Y“

[0361] N-A VN-V Replace one -N-(CH2)2-N- moiety with * °YJT )~R

[0362] NOTA0H

[0363] or

[0364] CROWNRX^N OH I XR

[0365] 'Ar \ rNi Replace one -N-(CH2)2-O- moiety with

[0366] Y> <kRv-N

[0367] ° C J? I y~R

[0368] k, N. J 0 / .

[0369] ors; or

[0370] RK^N r y_ROH NZ

[0371] Replace one -N-(CH2)2-N- moiety withsRV-N

[0372] r >“R

[0373] orNfs

[0374] AAZTARV-N _ z0Hr y~R HO-^Z^N \\

[0375] V0N- / . o / A Replace one -N-(CH2)2-N- moiety with

[0376] 1 N-\

[0377] VoHRV-N

[0378] | }-R

[0379] 1 °

[0380] HO^O

[0381] or

[0382] MACROPA

[0383] 0 I XRHO-Z

[0384] oz Replace one -N-(CH2)2-O- moiety with

[0385] ®

[0386] N NVN

[0387] X

[0388] 1 }~R

[0389] or °; or

[0390] XoH

[0391]

[0392] 0Benchmark Chelator Modified Structure

[0393] 1 )— R O' / - Replace one -O-(CH2)2-O- moiety with * V

[0394] r >-R

[0395] or ° / '

[0396] DTPARVN

[0397] 0 I \_RHQA] 0 AT A

[0398] N-AHOY-N-^NXAOH Replace one -N-(CH2)2-N- moiety with *

[0399] 00 0

[0400] JL JL HO ^ ^ OH

[0401] N- / .

[0402]

[0403] ors

[0404] Radionuclides

[0405] In some embodiments, the chelating agents are provided comprising the pyrrolidine chelators herein and their chelates with bi-trivalent ions of the metal elements having atomic number ranging between 20 and 31, 39, 42, 43, 44, 49, and between 57 and 83, and radioisotopes selected from:

[0406] 203Pb,67Ga,68Ga,72As,111In,113In,90Y,97Ru,62Cu,64Cu,52Fe,52mMn,140La,175Yb,153Sm,166Ho,149Pm,177LU,142Pr,159Gd,212Bi,47Sc,149Pm,67Cu,111Ag,199Au,161Tb and51Cr as well as the salts thereof with physiologically compatible bases or acids.

[0407] In some embodiments, the chelates further comprise Gd(3+), Eu(3+), Dy(3+), La(3+), Yb(3+) or Mn(2+) metal ions, including radionuclides thereof.

[0408] In some embodiments, chelating agents provided herein are to be used as radio-imaging agents or radio-pharmaceuticals different radionuclides are complexed to the chelator. Illustrative radionuclides include, for example,89Zr,44Sc,111In,90Y,66Ga,67Ga,68Ga,177Lu,99mTc,61Cu,62Cu,64Cu,67Cu,149Tb,152Tb,155Tb,161Tb,153Gd,155Gd,157Gd,213Bi,225Ac,230U,223Ra,165Er and52Fe. According to one aspect of this invention, the radionuclide is111In,90Y,68Ga,64Cu,153Gd,155Gd,213Bi,225Ac,52Fe, or177Lu.

[0409] In some embodiments, a radiopharmaceutical compound comprises a chelating agent disclosed herein and one or more of177Lu,225Ac,211At,67Cu,161Tb,67Ga,203Pb,223Ra, and212Pb.Pharmaceutical Compositions

[0410] The invention also relates to pharmaceutical preparations which contain a therapeutically effective amount of the active ingredients (compound according to the invention of Formula (I), Formula (II) or Formula (III) together with organic or inorganic solid or liquid, pharmaceutically acceptable carriers which are suited for the intended administration. The phrase “pharmaceutically acceptable” is employed herein to refer to those compounds, material, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of a patient without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. The compounds according to the invention can be formulated, where appropriate, together with further active substances and with excipients and carriers common in pharmaceutical compositions, e.g. — depending on the preparation to be produced — talcum, gum arabic, lactose, starch, magnesium stearate, cocoa butter, aqueous and non-aqueous carriers, fatty bodies of animal or vegetable origin, paraffin derivatives, glycols (in particular polyethylene glycol), various plasticizers, dispersants or emulsifiers, pharmaceutically compatible gases (e.g., air, oxygen, carbon dioxide, etc.), preservatives.

[0411] When solutions for infusion or injection are used, they are preferably aqueous solutions or suspensions, it being possible to produce them prior to use, e.g., from lyophilized preparations which contain the active substance as such or together with a carrier, such as mannitol, lactose, glucose, albumin and the like. The ready made solutions are sterilized and, where appropriate, mixed with excipients, e.g., with preservatives, stabilizers, emulsifiers, solubilizers, buffers and / or salts for regulating the osmotic pressure. The sterilization can be obtained by sterile filtration using filters having a small pore size according to which the composition can be lyophilized, where appropriate. Small amounts of antibiotics can also be added to ensure the maintenance of sterility.

[0412] In some embodiments, radiopharmaceutial compositions comprise a radionuclide (e.g., 100-1,000 MBq / mL). The radiopharmaceutical can be provided as an injectable formulation supplied as a sterile solution for intravenous use. The radiopharmaceutical can be provided in a single-dose vial comprising acetic acid (0.30 mg / mL), sodium acetate (0.41 mg / mL), gentisic acid (0.39 mg / mL), sodium ascorbate (50.0 mg / mL), pentetic acid (0.10 mg / mL), and water for injection (q.s. to 1 mL). The pH range of the solution is 4.5 to 7.0.

[0413] In some embodiments, the radiopharmaceutical can be provided in a single-dose vial comprising acetic acid (0.48 mg / mL), sodium acetate (0.66 mg / mL), gentisic acid (0.63 mg / mL), sodium hydroxide (0.64 mg / mL), ascorbic acid (2.8 mg / mL), diethylene triamine pentaacetic acid(0.05 mg / mL), sodium chloride (6.85 mg / mL), and water for injection (q.s. to ImL). The pH range of the solution is 4.5 to 6.

[0414] Methods of Imaging and Treatment

[0415] The compounds of the invention are also therapeutically useful. For example, compounds of the invention labeled with the appropriate therapeutic radionuclide are useful in radioisotope therapy. Radioisotope therapy (radiotherapy) involves the administration of a radiolabeled compound in sufficient quantity to damage or destroy the targeted tissue. After administration of the compound (by e.g., intravenous, subcutaneous, or intraperitonal injection), the radiolabeled pharmaceutical localizes preferentially at the disease site (e.g„ tumor tissue, etc,,) Once localized, the radiolabeled compound then damages or destroys the diseased tissue with the energy that is released during the radioactive decay of the isotope that is administered. As discussed herein, the compounds of the invention may be used in radiotherapy in combination with adjuvant chemotherapy (or in combination with any other appropriate therapeutic agent).

[0416] Imaging may be carried out in the normal manner, for example by injecting a sufficient amount of the imaging composition to provide adequate imaging and then scanning with a suitable imaging or scanning machine, such as a tomograph or gamma camera. In certain embodiments, a method of imaging a region in a patient includes the steps of: (i) administering to a patient a diagnostically effective amount of a compound complexed with a radionuclide; exposing a region of the patient to the scanning device; and (ii) obtaining an image of the region of the patient.

[0417] The design of a successful radiotherapeutic involves several critical factors:

[0418] • selection of an appropriate targeting group to deliver the radioactivity to the disease site;

[0419] • selection of an appropriate radionuclide that releases sufficient energy to damage that disease site, without substantially damaging adjacent normal tissues; and

[0420] • selection of an appropriate combination of the targeting group and the radionuclide without adversely affecting the ability of this conjugate to localize at the disease site. For radiometals, this often involves a chelating group that coordinates tightly to the radionuclide, combined with a linker that couples said chelate to the targeting group, and that affects the overall biodistribution of the compound to maximize uptake in target tissues and minimize uptake in normal, non-target organs.

[0421] The chelators of the present invention when complexed with an appropriate radionuclide and, preferably targeting group and linker are useful for radiotherapy. The selective targeting of cancercells with radiopharmaceuticals, either for imaging or therapeutic purposes is challenging. A variety of radionuclides are known to be useful for radio-imaging or cancer radiotherapy, including111In,90Y,68Ga,177Lu,99mTc,123I and131I. Depending on whether the compounds herein including compounds of Formula (I), Formula (II) or Formula (III) are to be used as radio-imaging agents or radio-pharmaceuticals different radionuclides are complexed to the chelator.

[0422] The amount of the compound of the present invention, or a formulation which may comprise a complex of a metal and a compound according to Formula (I), Formula (II) or Formula (III) or its salt, solvate, stereoisomer, or tautomer that is administered to a patient depends on several physiological factors that are routinely used by the physician, including the nature of imaging to be carried out, tissue to be targeted for imaging or therapy and the body weight and medical history of the patient to be imaged or treated using a radiopharmaceutical.

[0423] Illustrative radionuclides include, for example:89Zr,44Sc,111In,90Y,66Ga,67Ga,68Ga,177Lu,99mTc,61Cu,62Cu,64Cu,67Cu,149Tb,152Tb,155Tb,161Tb,153Gd,155Gd,157Gd,213Bi,225Ac,230U,223Ra,165Er and52Fe. According to one aspect of this invention, the radionuclide isniIn,90Y,68Ga,64Cu,153Gd,155Gd,213Bi,225Ac,52Fe, or177Lu.

[0424] According to another aspect, a pharmaceutical composition is provided, which is suitable for in vivo imaging and radiotherapy. Suitable pharmaceutical compositions may contain a radio imaging agent, or a radiotherapeutic agent that has a radionuclide either as an element, i.e. radioactive iodine, or a radioactive metal chelate complex of the compound of Formula (la) and / or (lb) in an amount sufficient for imaging, together with a pharmaceutically acceptable radiological vehicle. The radiological vehicle should be suitable for injection or aspiration, such as human serum albumin; aqueous buffer solutions, e.g., tris(hydromethyl) aminomethane (and its salts), phosphate, citrate, bicarbonate, etc; sterile water physiological saline; and balanced ionic solutions containing chloride and or dicarbonate salts or normal blood plasma cautions such as calcium potassium, sodium and magnesium.

[0425] Accordingly in another aspect, the invention provides a method for treating a patient by administering to a patient a therapeutically effective amount of a Formula (I), Formula (II) and / or (III) compound complexed to a radionuclide, or a pharmaceutically acceptable salt or solvate of the complex to treat a patient suffering from a cell proliferative disease or disorder. Specifically, the cell proliferative disease or disorder to be treated or imaged using a compound, pharmaceutical composition or radiopharmaceutical in accordance with this invention is a cancer, for example, prostate cancer and / or prostate cancer metastasis in e.g., lung, liver, kidney, bones, brain, spinal cord, bladder, etc.The complexes of compounds can be administered as MRI contrast agents or radiopharmaceuticals parenterally, preferably formulated as a sterile aqueous solution or suspension, whose pH can range for example from 6.0 to 8.5. Said aqueous solutions or suspensions can be administered in concentrations ranging from 0.002 to 1.0 molar.

[0426] Said formulations can be freeze-dried and supplied as such, to be reconstituted prior to use. For the gastrointestinal use or for injection to body cavities, these agents can be formulated as a solution or suspension containing suitable additives in order to, for example, control viscosity.

[0427] For the oral administration they can be formulated according to preparation methods routinely used in pharmaceutical technique, optionally also as coated formulations to gain extra protection from the acid pH of the stomach, inhibiting the release of the chelated metal ion, which usually occurs at typical pH values of gastric juices.

[0428] Other excipients, such as sweetening agents and / or flavoring agents, can also be added according to known techniques of pharmaceutical formulation. The compounds according to the invention can be formulated, where appropriate, together with further active substances and with excipients and carriers common in pharmaceutical compositions, e.g. — depending on the preparation to be produced — talcum, gum arabic, lactose, starch, magnesium stearate, cocoa butter, aqueous and non-aqueous carriers, faty bodies of animal or vegetable origin, paraffin derivatives, glycols (in particular polyethylene glycol), various plasticizers, dispersants or emulsifiers, pharmaceutically compatible gases (e.g., air, oxygen, carbon dioxide, etc.), preservatives.

[0429] Compounds of the invention useful for radiotherapy are advantageously complexed with a 3+ metal ion from the class of elements known as the lanthanides (elements of atomic number 57-71) and their analogs (i.e. M3+ metals such as ytrium and indium). Typical radioactive metals in this class include the isotopes 90- Ytrium, 111-Indium, 149-Promethium, 153-Samarium, 166-Dysprosium, 166-Holmium, 175- Yterbium, and 177-Lutetium. All of these metals (and others in the lanthanide series) have very similar chemistries, in that they remain in the +3 oxidation state, and prefer to complex to ligands that bear hard (oxygen / nitrogen) donor atoms, such as the chelators of the invention.

[0430] The chealtors of the invention complex with and prevent the release of free (unbound) radiometal into the body. This is important, as in vivo dissociation of 3+ radiometals from their chelate can result in uptake of the radiometal in the liver, bone and spleen [Brechbiel M W, Gansow O A, “Backbone-substituted DTPA ligands for90Y radioimmunotherapy”, Bioconj. Chem. 1991; 2: 187-194; Li, W P, Ma D S, Higginbotham C, Hoffman T, Ketring A R, Cutler C S, Jurisson, S S, “Development of an in vitro model for assessing the in vivo stability of lanthanide chelates.” Nucl.Med. Biol. 2001; 28(2): 145-154; Kasokat T, Urich, K. Arzneim.-Forsch, “Quantification of dechelation of gadopentetate dimeglumine in rats”. 1992; 42(6): 869-76]. Unless one is specifically targeting these organs, such non-specific uptake is highly undesirable, as it leads to non-specific irradiation of non-target tissues, which can lead to such problems as hematopoietic suppression due to irradiation of bone marrow.

[0431] The selection of a proper radionuclide for use in a particular radiotherapeutic application depends on many factors, including:

[0432] • Physical half-life — This should be long enough to allow synthesis and purification of the radiotherapeutic construct from radiometal and conjugate, and delivery of said construct to the site of injection, without significant radioactive decay prior to injection. Preferably, the radionuclide should have a physical half-life between about 0.5 and 8 days.

[0433] • Energy of the emission(s) from the radionuclide — Radionuclides that are particle emitters (such as alpha emitters, beta emitters and Auger electron emitters) are particularly useful, as they emit highly energetic particles that deposit their energy over short distances, thereby producing highly localized damage. Beta emitting radionuclides are particularly preferred, as the energy from beta particle emissions from these isotopes is deposited within 5 to about 150 cell diameters. Radiotherapeutic agents prepared from these nuclides are capable of killing diseased cells that are relatively close to their site of localization, but cannot travel long distances to damage adjacent normal tissue such as bone marrow.

[0434] • Specific activity (i.e. radioactivity per mass of the radionuclide)-Radionuclides that have high specific activity (e.g., generator produced 90- Y, 111-In, 177-Lu) are particularly preferred. The specific activity of a radionuclide is determined by its method of production, the particular target that is used to produce it, and the properties of the isotope in question.

[0435] Many of the lanthanides and lanthanoids include radioisotopes that have nuclear properties that make them suitable for use as radiotherapeutic agents, as they emit beta particles.

[0436] Methods for the preparation of radiometals such as beta-emitting lanthanide radioisotopes are known to those skilled in the art, and have been described elsewhere [e.g., Cutler C S, Smith C J, Ehrhardt G J.; Tyler T T, Jurisson S S, Deutsch E. “Cunent and potential therapeutic uses of lanthanide radioisotopes.” Cancer Biother, Radiopharm. 2000; 15(6): 531-545]. Many of these isotopes can be produced in high yield for relatively low cost, and many (e.g.,90-Y,149-Pm,177-Lu) can be produced at close to carrier-free specific activities (i.e. the vast majority of atoms are radioactive). Since non-radioactive atoms can compete with their radioactive analogs for binding toreceptors on the target tissue, the use of high specific activity radioisotope is important, to allow delivery of as high a dose of radioactivity to the target tissue as possible.

[0437] Therapeutic application of compounds of the invention complexed with therapeutic radionuclides can be defined either as radiopharmaceutical that will be used as a first line therapy in the treatment of a disease such as cancer, as a combination therapy where the radiotherapeutic agents of the invention could be utilized in conjunction with adjuvant chemotherapy (e.g, with one of the other therapeutic agents disclosed herein), or as the therapeutic part of a matched pair therapeutic agent. The matched pair concept refers to a single unmetallated compound which can serve as both a diagnostic and a therapeutic agent depending on the radiometal that has been selected for binding to the appropriate chelate. If the chelator cannot accommodate the desired metals appropriate substitutions can be made to accommodate the different metal while maintaining the pharmacology such that the behaviour of the diagnostic compound in vivo can be used to predict the behaviour of the radiotherapeutic compound.

[0438] The concentration of the imaging agent or the therapeutic agent in the radiological vehicle should be sufficient to provide satisfactory imaging. For example, when using an aqueous solution, the dosage is about 1.0 to 100 millicuries. The actual dose administered to a patient for imaging or therapeutic purposes, however, is determined by the physician administering treatment. The imaging agent or therapeutic agent should be administered so as to remain in the patient for about 1 hour to 10 days, although both longer and shorter time periods are acceptable. Therefore, convenient ampoules containing 1 to 10 mL of aqueous solution may be prepared.

[0439] EXAMPLES EXAMPLE 1:

[0440] SYNTHESIS OF 4-OXO-4-(((5,8-TRANS)-1,5,7-TRIS(2-(TERT-BUTOXY)-2- OXOETHYL)DECAHYDROPYRROLO[3,4-B] [1,4]DIAZEPIN-3-YL)AMINO)BUTANOIC ACID (COMPOUND#!).

[0441] This example describes the synthesis of 4-oxo-4-(((5,8-trans)-l,5,7-tris(2-(tert-butoxy)-2-oxoethy l)decahydropyrrolo [3,4-b] [1,4] diazepin-3 -yl)amino)butanoic acid (Compound# 1 )

[0442] For the LCMS analytical characterization of all intermediates and products in Example 1, the following system and method were used:

[0443] • Mass spectrometer API 150EX, ELSD - Sedex 75, Shimadzu LC system: SCL-lOAvp UV detector, SCL-lOAvp system controller, LC-lOADvp liquid chromatograph.Column: Chromolith ® Speed ROD, RP-18e, 50-4.5mm, MP A: 0.05% TFA in H2O, MP B: 0.05% TFA in Acetonitrile, 5 to 100% B over 12 min, Flow rate: 1.2 mL / min, Runtime: 12 min, UV254 / ELSD detectors.

[0444] • Proton NMR spectra were recorded on the Varian Unity Inova spectrometer operating at 500 MHz utilizing Agilent VnmrJ4software. For all samples D6DMSO was used as a solvent.

[0445]

[0446] Compound #1

[0447] In this case (for trans-dibenzyl pyrrolidine), condensation with nitro-ethanol requires more time compared to the model N, N-dibenzylethane-l,2-diamine. The resulting tert-butyl (trans-5,8)-1,5-dibenzyl-3-(hydroxymethyl)-3-nitrooctahydropyrrolo[3,4-b] [1,4]diazepine-7(lH)-carboxyIate (Int 6) was obtained as seen in Scheme A-1.

[0448] 1. Benzaldehyde °2NXX^OH 2. NaBH4Paraformaldehyde EtOH / PhMe

[0449]

[0450]

[0451] Scheme A-l

[0452] Scheme A-2 shows the synthetic route that we used to create a pyrrolidine-AAZTA analog based on the diazepine (Int 6). Thus, initially we carried out the mild elimination of the hydroxymethyl fragment with potassium tert-butoxide in anhydrous tetrahydrofuran. After isolation we carried out the reduction of the nitro group to an amine with hydrogen on Raney nickel. The obtained stable tert-butyl (trans-5,8)-3-amino-l,5-dibenzyloctahydropyrrolo[3,4-b][l,4]diazepine-7(lH)-carboxyIate (Int 8) was treated with the succinic anhydride and purified. Then, we carried out a mild esterification of the free acid group and sequential deprotection of secondary amino groups included in the cyclic core, thus obtaining methyl 4-(((trans-5,8)-decahydropyrrolo[3,4-b][l,4]diazepine- 3-yl)amino)-4-oxobutanoate trifluoroacetate (Int 12). The target intermediate 13 was purified by chromatography on silica gel.

[0453] Raney Ni H2MeOH Bn Int 6 Int 7 Int 8

[0454]

[0455] Scheme A-2.

[0456] The saponification of intermediate 13 with an equivalent amount of lithium hydroxide in a mixture of water and tetrahydrofuran gave us 4-oxo-4-(((trans-5,8)-l,5,7-tris(2-(tert-butoxy)-2-oxoethyl) decahydropyrrolo[3,4-b][l,4]diazepin-3-yl)amino)butanoic acid (Compound #1) which was purified by HPLC to afford 53 mg and 67% yield.

[0457] Compound la

[0458] Synthesis of tert-butyl trans-3, 4-bis(benzylamino)pyrrolidine-l-carboxylate.

[0459] Powdered molecular sieves (4A, 6 g) and benzaldehyde (6.73 ml, 66 mmol, 3 eq) were added to a solution of tert-butyl trans-3,4-diaminopyrrolidine-l -carboxylate (4.44 g, 22 mmol, 1 eq) inanhydrous MeOH (100 ml). The suspension was stirred for 1 hour at ambient temperature.

[0460] Subsequently, sodium borohydride (3.3 g, 88 mmol, 4 eq) was added by small portions at 0°C in a period of 2 hours. After that the mixture was stirred for 1 hour at 0°C.

[0461] Then, ethyl acetate (150 ml) and a saturated sodium bicarbonate solution (150 ml) were carefully added. The suspension was filtered and extracted with ethyl acetate (3x200 ml). The combined organic extracts were dried over magnesium sulfate (20 g), filtered, and concentrated under vacuum to dryness at 35°C. The residue was diluted with DCM (10ml) and purified by silica gel column chromatography. The fractions containing product were combined and evaporated under vacuum to give tert-butyl trans-3,4-bis(benzylamino)pyrrolidine-l -carboxylate as light-yellow solid (4.1g, 47% yield). Calculated m / z [M+H] = 382.24 Found m / z [M+H] = 492.4

[0462] Compound lb

[0463] Synthesis of tert-butyl (5,8-trans)-l,5-dibenzyl-3-(hydroxymethyl)-3-nitrooctahydropyrrolo[ 3,4-b ][1, 4 [diazepine-7 (1 H) -carboxylate.

[0464] Tert-butyl trans-3,4-bis(benzylamino)pyrrolidine-l -carboxylate (1.6 g, 4.19 mmol, 1 eq) and nitroethanol (0.6 ml, 8.4 mmol, 2 eq) were dissolved in toluene / EtOH (1:1 v / v) (40 ml). Then, paraformaldehyde (450 mg, 14.8 mmol, 3.5 eq) was added and the suspension was vigorously stirred under reflux for 6 h. The volatiles were removed under reduced pressure. The residue was dissolved in dichloromethane (30 ml) and the solution was washed with water (3x10 ml). The organic phase was dried over magnesium sulfate (1 g), filtered, and concentrated under vacuum to dryness at 35°C. The residue was purified by silica gel column chromatography. The fractions containing product were combined and evaporated under vacuum to give tert-butyl (5,8-trans)-l,5-dibenzyl-3-(hydroxymethyl)-3-nitrooctahydropyrrolo [3,4-b] [ 1,4]diazepine-7( 1 H)-carboxylate as off-white glassy solid (1.6g, 77% yield). Calculated m / z [M+H] = 497.27 Found m / z [M+H] = 497.3

[0465] Compound lc

[0466] Synthesis of tert-butyl (5,8-trans)-l,5-dibenzyl-3-nitrooctahydropyrrolo[3,4-b ][1, 4 ]diazepine-7(lH)-carboxylate.

[0467] Tert-butyl (5, 8-trans)- 1,5 -dibenzyl-3 -(hydroxymethyl)-3 -nitrooctahy dropyrrolo[3,4-b][l,4]diazepine-7(lH)-carboxylate (660 mg, 1.33 mmol, 1 eq) was dissolved in anhydrous THF (3.5 ml). Then, to the solution potassium tert-butoxide (224 mg, 2 mmol, 1.5 eq) was added and the suspension was vigorously stirred for 1.5 hours at ambient temperature. Then the solvent evaporated under vacuum at 25°C. The residue was dissolved in saturated aqueous ammonium chloride solution(10 ml) and extracted with dichloromethane (3x30 ml). The organic phase was dried over magnesium sulfate (1 g) and the volatiles were evaporated under vacuum at 25°C to give crude tertbutyl (5,8-trans)-l,5-dibenzyl-3-nitrooctahydropyrrolo[3,4-b][l,4]diazepine-7(lH)-carboxylate as a yellow solid (630 mg, 100% yield). Calculated m / z [M+H] = 467.26 Found m / z [M+H] = 467.5

[0468] Compound Id

[0469] Synthesis of tert-butyl (5,8-trans)-3-amino-l,5-dibenzyloctahydropyrrolo[3,4-b] [1,4 J diazepine- 7 ( lH)-carboxylate.

[0470] To a flask flushed with argon a solution of tert-butyl (5,8-trans)-l,5-dibenzyl-3-nitrooctahydropyrrolo[3,4-b][l,4]diazepine-7(lH)-carboxylate ( crude from previous step, 630mg, 1.33 mmol, 1 eq) in methanol (15 ml), was added Raney nickel (0.2g, wet). The argon atmosphere was replaced with 2ATM hydrogen, and the reaction mixture was stirred for 1 hour at ambient temperature. The suspension was filtered through a Celite pad. The filtrate was evaporated and dried at ambient temperature under high vacuum to give crude tert-butyl (5,8-trans)-3-amino-l,5-dibenzyloctahydropyrrolo[3,4-b][l,4]diazepine-7(lH)-carboxylate as a yellow solid (600 mg, 100% yield). Calculated m / z [M+H] = 437.28 Found m / z [M+H] = 437.5

[0471] Compound le

[0472] Synthesis of 4-(((5,8-trans)-l,5-dibenzyl-7-(tert-butoxycarbonyl) decahydropyrrolo[3,4-b ][1, 4 ]diazepin-3-yl)amino)-4-oxobutanoic acid.

[0473] A mixture of tert-butyl (5,8-trans)-3-amino-l,5-dibenzyloctahydropyrrolo[3,4-b][l,4]diazepine-7(lH)-carboxylate (600mg crude from previous step, ~1.33mmol, 1 eq), triethylamine (0.4 ml, 5.4 mmol, 4 eq) and dihydrofuran-2, 5-dione (200mg, 2 mmol, 1.5 eq) in dry acetonitrile (5ml) was stirred for 6 hours at 50°C. Then, the mixture was evaporated under vacuum to dryness, diluted with water (15ml) and extracted with dichloromethane (4x10 ml). The organic extract was dried with MgSO4 (lg), filtered, and concentrated under vacuum to dryness to give crude 4-(((5,8-trans)-l,5-dibenzyl-7-(tert-butoxycarbonyl) decahydropyrrolo[3,4-b][l,4]diazepin-3-yl)amino)-4-oxobutanoic acid as yellow solid (600mg, 80% yield). Calculated m / z [M+H] = 537.30 Found m / z [M+H] = 537.4

[0474] Compound If

[0475] Synthesis of tert-butyl (5,8-trans)-l,5-dibenzyl-3-(4-methoxy-4-oxobutanamido) octahydropyrrolo[ 3, 4-b ][ 1, 4 ]diazepine-7(l H) -carboxylate.To a mixture of 4-(((5,8-trans)-l,5-dibenzyl-7-(tert-butoxycarbonyl)decahydropyrrolo[3,4-b][l,4]diazepin-3-yl)amino)-4-oxobutanoic acid (crude from previous step, 600 mg, ~1.33mmol, 1 eq) and 4-dimethylaminopyridine (16 mg, 0.13 mmol, 0.1 eq) in anhydrous dichloromethane (15 ml) anhydrous methanol (0.54 ml, 13.3 mmol, 10 eq) was added. This mixture was cooled to 5 °C and 1-ethyl-3 -(3 -dimethylamino propyl) carbodiimide (250mg, 1.6 mmol, 1.2 eq) was added in three portions (3x83mg) in a period of 1.5 hours. Then, the mixture was allowed to warm to ambient temperature and stirred for 3 hours. Then, the mixture was evaporated under vacuum to dryness, diluted with water (15ml) and extracted with dichloromethane (4x10 ml).

[0476] The organic extract was concentrated under vacuum to dryness. The residue was diluted with dichloromethane (2ml) and purified by silica gel column chromatography. The fractions containing product were combined and evaporated under vacuum to give tert-butyl (5,8-trans)-l,5-dibenzyl-3-(4-methoxy-4-oxobutanamido)octahydropyrrolo[3,4-b][l,4]diazepine-7(lH)-carboxylate as lightyellow viscous oil (340mg, 57% yield). Calculated m / z [M+H] = 551.32 Found m / z [M+H] = 557.5

[0477] Compound 1g

[0478] Synthesis of methyl 4-(((5,8-trans)-decahydropyrrolo[3,4-b][l,4]diazepin-3-yl)amino)-4-oxobutanoate trifluoroacetate.

[0479] To a flask flushed with argon a solution of tert-butyl (5,8-trans)-l,5-dibenzyl-3-(4-methoxy-4-oxobutanamido)octahydropyrrolo[3,4-b][l,4]diazepine-7(lH)-carboxylate (340 mg, 0.62 mmol, 1 eq) in methanol (20 ml), was added palladium on activated carbon (0.2g, 5% mass, wet). The argon atmosphere was replaced with 2 atm hydrogen, and the reaction mixture was stirred for 20 hours at ambient temperature.

[0480] The suspension was filtered through a Celite pad. The filtrate was evaporated and dried at ambient temperature under a high vacuum to give de-benzylated intermediate (216 mg, 100% yield) as a light-yellow oily residue.

[0481] The residue was diluted with dichloromethane (10ml) and re-evaporated in a high vacuum. Then, the residue was diluted with dichloromethane (2 ml) and trifluoroacetic acid (1 ml) was added. The mixture was stirred for 4 hours at ambient temperature.

[0482] The mixture was evaporated to dryness, re-evaporated with dichloromethane (20 ml) and dried under a high vacuum for 12 hours at ambient temperature to give methyl 4-(((5,8-trans)-decahydropyrrolo[3,4-b][l,4]diazepin-3-yl)amino)-4-oxobutanoate trifluoroacetate (430 mg, >100% yield) as a yellow viscous oil. Calculated m / z [M+H] = 271.17 Found m / z [M+H] = 271.3Compound Ih

[0483] Synthesis of tri-tert-butyl 2,2',2"-((5,8-trans)-3-(4-methoxy-4-oxobutanamido) hexahydropyrrolo[3, 4-b ][1, 4 ]diazepine-l, 5, 7 (2H)-triyl)triacetate.

[0484] To a mixture of methyl 4-(((5,8-trans)-decahydropyrrolo[3,4-b][l,4]diazepin-3-yl)amino)-4-oxobutanoate trifluoroacetate (430mg, ~0.62mmol, 1 eq, crude from previous step) and sodium carbonate (1.64g, 15.5mmol, 25eq) in anhydrous acetonitrile (10ml) tert-butyl bromoacetate (0.46ml, 3.1 mmol, 5 eq) was added. This mixture was stirred 18hrs at ambient temperature, then 24 hours at 40°C. Then, the mixture was cooled to room temperature, filtered and the solid was washed with acetonitrile (10ml). The combined filtrate was evaporated to dryness to give yellow oily residue. The residue was diluted with dichloromethane (2ml) and purified by silica gel column chromatography. The fractions containing product were combined and evaporated under vacuum to give tri-tert-butyl 2,2',2"-((5,8-trans)-3-(4-methoxy-4-oxobutanamido)hexahydropyrrolo[3,4-b][l,4]diazepine-l,5,7(2H)-triyl)triacetate as light-yellow viscous oil (80mg, 22% yield). Calculated m / z [M+H] = 613.37 Found m / z [M+H] = 613.2

[0485] Compound li

[0486] Synthesis of 4-oxo-4-(((5,8-trans)-l,5, 7-tris(2-(tert-butoxy)-2-oxoethyl) decahydropyrrolo[3,4-b ][1, 4 ]diazepin-3-yl)amino)butanoic acid.

[0487] To a solution of tri-tert-butyl 2,2',2"-((5,8-trans)-3-(4-methoxy-4-oxobutanamido)hexahydropyrrolo[3,4-b][l,4]diazepine-l,5,7(2H)-triyl)triacetate (80mg, 0.13mmol, 1 eq) in tetrahydrofuran (5ml) previously made 2M aqueous LiOH solution (0.1ml, 0.2mmol, 1.5 eq) was added. This mixture was stirred for 8 hours at ambient temperature. Then, the mixture was acidified with glacial acetic acid (0.022ml, 0.39mmol, 3 eq) and evaporated to dryness under vacuum. The residue was purified by preparative HPLC chromatography (53mg, 67% yield).

[0488] Calculated m / z [M+H] = 599.36 Found m / z [M+H] = 599.5

[0489] EXAMPLE 2:

[0490] SYNTHESIS OF (TRA S-5,8)-1,5,7-TRIS(2-(TERT-BLTOXY)-2-OXOETHYL)DECAH YDROPYRROLO [3,4- B][1,4]DIAZEPINE-3-CARBOXYLIC ACID (COMPOUND #2)

[0491] Example 2 describes the synthesis of (trans-5,8)-l,5,7-tris(2-(tert-butoxy)-2-oxoethyl)decahydropyrrolo[3,4-b][l,4]diazepine-3-carboxylic acid (Compound #2)

[0492] For the LCMS analytical characterization of all intermediates and products in Example 2, the following system and method were used:• Mass spectrometer API 150EX, ELSD - Sedex 75, Shimadzu LC system: SCL-lOAvp UV detector, SCL-lOAvp system controller, LC-lOADvp liquid chromatograph.

[0493] Column: Chromolith ® Speed ROD, RP-18e, 50-4.5mm, MP A: 0.05% TFA in H2O, MP B: 0.05% TFA in Acetonitrile, 5 to 100% B over 12 min, Flow rate: 1.2 mL / min, Runtime: 12 min, UV254 / ELSD detectors.

[0494] A synthetic route was developed for creating substance (trans-5,8)-l,5,7-tris(2-(tert-butoxy)-2-oxoethyl)decahydropyrrolo[3,4-b][l,4]diazepine-3-carboxylic acid (Compound #2) is presented.

[0495]

[0496] (2)

[0497] The optimized conditions of Scheme B-l were able to achieve a 50% yield of (trans-5,8)-l,5-dibenzyl-7-(tert-butoxycarbonyl) decahydropyrrolo — [3,4-b] [1,4] diazepine-3 -carboxylic acid (Int 1), which served as the starting material for the synthetic route for creating target compounds applying the route presented in Scheme B-2.

[0498] Bn 0 0 Bn

[0499] Paraformaldehyde

[0500] MeCN

[0501]

[0502] Bn

[0503] Scheme B-l.

[0504] We obtained Intermediate 2 by sequential benzyl- and BOC- deprotections, which was used in the alkylation reaction with tert-butyl bromoacetate without additional purification. Compound #2thus obtained was purified by silica gel chromatography.

[0505] Bn 1. Pd / C, H2— 2. T —FA —, - DCM Na2CO3, MeCN - ► Bn

[0506]

[0507] Scheme B-2.

[0508] Compound 2a

[0509] Synthesis of (trans-5,8)-l,5-dibenzyl-7-(tert-butoxycarbonyl)decahydropyrrolo[3,4-b ][1, 4 ]diazepine-3 -carboxylic acid.

[0510] Malonic acid (523mg, 5.03mmol, 1.2eq) was added to a solution of tert-butyl trans-3,4-bis(benzylamino) pyrrolidine- 1 -carboxylate (1.6g, 4.19mmol, leq) in acetonitrile (20 ml) at ambient temperature. This mixture was stirred for 30 min, and then formaldehyde (37% solution in water, 0.78ml, 10.48mmol, 2.5eq) was added. The mixture was stirred at room temperature for 20 hours and then heated at 60°C for another 20 hours. Then the mixture was evaporated to dryness. The residue was dissolved in ethyl acetate (100 ml) and the solution was washed with water (2x20 ml). The organic phase was concentrated under vacuum to dryness. The residue was diluted with DCM (5ml) and purified by silica gel column chromatography. The fractions containing product were combined and evaporated under vacuum to give (trans-5,8)-l,5-dibenzyl-7-(tert-butoxycarbonyl)decahydropyrrolo[3,4-b][l,4]diazepine-3-carboxylic acid as yellowish glassy solid (1.2g, 50% yield). Calculated m / z [M+H] = 466.26 Found m / z [M+H] = 466.3

[0511] Compound 2b

[0512] Synthesis of (trans-5,8)-decahydropyrrolo[3,4-b] [1,4]diazepine-3-carboxylic acid trifluoroacetate.

[0513] The method used was identical to the synthesis of Compound 1g to afford methyl (trans-5,8)-decahydropyrrolo[3,4-b][l,4]diazepine-3-carboxylate trifluoroacetate (0.49g, >100% yield) as a yellow viscous oil. Calculated m / z [M+H] = 186.23 Found / z [M+H] = 186.4Compound 2c

[0514] Synthesis of (trans-5,8)-l,5, 7-tris(2-(tert-butoxy)-2-oxoethyl)decahydropyrrolo[3,4-b ][1, 4 ]diazepine-3 -carboxylic acid.

[0515] The method used was identical to the synthesis of Compound Ih to provide 94mg (20% yield) of tri-tert-butyl 2,2',2"-((trans-5,8)-3-(methoxycarbonyl)hexahydropyrrolo[3,4-b][l,4]diazepine-l,5,7(2H)-triyl)triacetate as a light-yellow solid. Calculated m / z [M+H] = 527.55 Found m / z [M+H] = 528.5

[0516] EXAMPLE 3:

[0517] SYNTHESIS OF TERT-BUTYL (CIS)-3,4-BIS(2-((METHYLSULFONYL)OXY)ETHOXY)PYRROLIDINE-1-CARBOXYLATE, TERT-BUTYL(TRA S)-3,4-BIS(2-((METHYLSULFONYL)OXY)ETHOXY)PYRROLIDINE- 1-CARBOXYLATE.

[0518] This example describes the synthesis of tert-butyl (cis)-3,4-bis(2-((methylsulfonyl)oxy)ethoxy)pyrrolidine- 1 -carboxylate, tert-butyl (trans)-3,4-bis(2-((methylsulfonyl)oxy)ethoxy)pyrrolidine- 1 -carboxylate.

[0519] For the LCMS analytical characterization of all the materials the following system and method were used:

[0520] • Mass spectrometer API 150EX, ELSD - Sedex 75, Shimadzu LC system: SCL-lOAvp UV detector, SCL-lOAvp system controller, LC-lOADvp liquid chromatograph.

[0521] Column: Chromolith ® Speed ROD, RP-18e, 50-4.5mm, MP A: 0.05% TFA in FhO, MP B: 0.05% TFA in Acetonitrile, 5% to 100% B over 12 min, Flow rate: 1.2 mL / min, UV254 / ELSD detectors.

[0522] This example provides a synthesis of two stereoisomers of tert-butyl-3,4-bis(2-((methylsulfonyl)oxy)ethoxy)pyrrolidine- 1 -carboxylate.

[0523] Scheme C-l shows the general synthetic route for stereoisomers of tert-butyl-3,4-bis(2-((methylsulfonyl)oxy)ethoxy)pyrrolidine- 1 -carboxylate.

[0524] HCI (aq), MeOH,

[0525] o^OMs

[0526] MsCI, Et3N, DCI^fi

[0527]

[0528] Scheme C-l.

[0529] The first step was the treatment of tert-butyl-3,4-dihydroxypyrrolidine-l -carboxylate with sodium hydride and 2-(2-bromoethoxy)tetrahydro-2H-pyran and the subsequent thorough purification on silica gel. The following step was the removal of the tetrahydropyran protection group with hydrochloric acid in methanol. The subsequent treatment of the aliphatic alcohols with mesyl chloride at ambient temperature for 20-30 hours gave us the desired tert-butyl -3,4-dihydroxypyrrolidine-1 -carboxylates, which were purified on silica gel.

[0530] Scheme C-2 shows the general method for the preparation of pyrrolidine-containing macrocycles.

[0531] ?MSH HBnR0 J „ ^N,, NSrf V „ Bn Z Bn O N Pd / C, Hz, Boe— N I \

[0532] _ Na2CO3, MeCN Boc-N Z MeOH o O N

[0533]

[0534] I Z = chain of atoms (carbon \ — / 'SnOMs oxygen, nitrogen)

[0535] Scheme C-2.

[0536] We used a series of benzylated aliphatic amines that were commercially available. These amines were used for macrocyclic alkylation with previously prepared tert-butyl -3,4-dihydroxypyrrolidine-1 -carboxylates. The macrocyclic derivatives obtained in this way were purified on silica gel and subjected to the catalytic debenzylation with a palladium catalyst.

[0537] Several macrocyclic derivatives containing a pyrrolidine fragment were obtained. The corresponding structures are presented in Scheme C-3.

[0538]

[0539]

[0540] Scheme C-3.

[0541] Compound 3a

[0542] Synthesis oftert-butyl-trans-3,4-bis(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)pyrrolidine-l-carboxylate.

[0543] A suspension of sodium hydride (11.7 g, 292 mmol, 4 eq, 55% dispersion in mineral oil) in anhydrous DMF (220 ml) under argon atmosphere was cooled to 0°C and treated with a solution of tert-butyl trans-3,4-diazidopyrrolidine-l -carboxylate (14.9 g, 73 mmol, 1 eq) in anhydrous DMF (50 ml). Then, the mixture warmed to ambient temperature and stirred for 90 min. Then, the mixture was cooled to 0°C and a solution of 2-(2-bromoethoxy) tetrahydro-2H-pyran (61g, 292 mmol, 4 eq) in anhydrous DMF (50 ml) was added slowly at 0°C. The resulting light-yellow solution was warmed to ambient temperature and stirred for 18 hrs. (LCMS analysis showed conversion of the starting material to the target product). Then, the reaction mixture was diluted with EtOAc (IL) and washed with a saturated sodium chloride solution (3x200 ml). The organic layer was dried with magnesium sulfate (30 g), filtered and concentrated under vacuum. This residue was dissolved in DCM (30 ml) and purified by silica gel column chromatography (330 g of silica gel, 0% to 80% ethyl acetate in heptane). The fractions containing product (as determined by TLC, eluent - EA / Hex 1 / 1 by volume, plate - SiliaPlate TLA-AUT0337-323N, detection- ninhydrin stain (1% in acetone)) were combined and evaporated. The residue was dried under high vacuum at 35 °C for 6 hours to provide tert-butyl-trans-3,4-bis(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)pyrrolidine-l-carboxylate as a yellowish viscous oil (29g, 85% yield). Calculated m / z [M+H] = 460.28 Found m / z [M+H] = 460.4

[0544] Compound 3b

[0545] Synthesis of tert-butyl (trans)-3,4-bis(2-hydroxyethoxy)pyrrolidine-l -carboxylate.To a solution of tert-butyl-trans-3,4-bis(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)pyrrolidine-l -carboxylate (29 g, 63 mmol, 1 eq) in methanol (250 ml) hydrochloric acid (37% aqueous, 20 ml, 252 mmol, 4 eq) was added dropwise at ambient temperature for 2 hours at vigorous stirring (LCMS analysis shows full conversion of the starting material to the product). Then, the mixture was carefully evaporated at 35 °C to dryness to provide a very viscous yellow residue. The residue was diluted with methylene chloride (30 ml) and purified by silica gel chromatography to provide tert-butyl (trans)-3,4-bis(2-hydroxyethoxy)pyrrolidine-l -carboxylate (17.4 g, 95% yield) as a light-yellow viscous oil. Calculated mJz [M+H] =291.17 Found m / z [M+H] = 292.2

[0546] Compound 3c

[0547] Synthesis of tert-butyl (trans)-3,4-bis(2-((methylsulfonyl)oxy)ethoxy)pyrrolidine-l-carboxylate.

[0548] To a solution of tert-butyl (trans)-3,4-bis(2-hydroxyethoxy)pyrrolidine-l -carboxylate (17.4 g, 59.7 mmol, 1 eq) and triethylamine (24.8 ml, 149 mmol, 3 eq) in an anhydrous dichloromethane (600 ml) at 0°C mesyl chloride (11.5 ml, 149 mmol, 2.5 eq) was added in an one hour. The reaction mixture was kept at 0°C for one hour, followed by warming to an ambient temperature and stirring for 3 hours. After the reaction completion (LCMS analysis) the reaction mixture was diluted with water (500 ml) and stirred for 15 min. The layers were separated, and the aqueous layer was extracted with dichloromethane (2x100 ml). The combined organic extracts were washed with a 5% aqueous hydrochloric acid solution (100 ml), brine (2x100 ml), dried over sodium sulfate (10 g) and evaporated under vacuum to give yellow oily residue. The residue was diluted with dichloromethane (30 ml) and purified by silica gel chromatography to provide tert-butyl (trans)-3,4-bis(2-((methylsulfonyl)oxy)ethoxy)pyrrolidine-l -carboxylate (16.1 g, 55% yield) as a light-yellow viscous oil. Calculated m / z [M+H] = 448.12 Found m / z [M+H] = 448.2

[0549] Compound 3e

[0550] Synthesis of tert-butyl-cis-3, 4-bis(2-( (tetrahydro-2H-pyran-2-yl)oxy)ethoxy)pyrrolidine-l-carboxylate.

[0551] The method used was identical to the synthesis of Compound 3a to provide tert-butyl-cis-3,4-bis(2-((tetrahydro-2H-pyran-2-yl)oxy)ethoxy)pyrrolidine-l -carboxylate as a yellowish viscous oil (31g, 89% yield). Calculated m / z [M+H] = 460.28 Found m / z [M+H] = 460.3Compound 3f

[0552] Synthesis of tert-butyl (cis)-3,4-bis(2-hydroxyethoxy)pyrrolidine-l -carboxylate.

[0553] The method used was identical to the synthesis of Compound 3b to provide tert-butyl (cis)-3,4-bis(2-hydroxyethoxy)pyrrolidine-l -carboxylate (19.1 g, 97% yield) as a light-yellow viscous oil. Calculated m / z [M+H] = 291.17 Found m / z [M+H] = 292.3

[0554] Compound 3g

[0555] Synthesis of tert-butyl (cis)-3, 4-bis(2-( (methylsulfonyl)oxy)ethoxy)pyrrolidine-l-carboxylate. The method used was identical to the synthesis of Compound 3b to provide tert-butyl (cis)-3,4-bis(2-((methylsulfonyl)oxy)ethoxy)pyrrolidine-l -carboxylate (18 g, 60% yield) as a light-yellow viscous oil. Calculated m / z [M+H] = 448.12 Found m / z [M+H] = 448.6

[0556] Compound 3h

[0557] Synthesis of tert-butyl (trans)-4,13-dibenzyltetradecahydro-2H, HH,18H-pyrrolo[3,4-b] [1,4, 10, 13 ]tetraoxa[7, 16 ]diazacyclooctadecine-l 8-carboxylate.

[0558] To a solution of tert-butyl (trans)-3,4-bis(2-((methylsulfonyl)oxy)ethoxy)pyrrolidine-l -carboxylate (2 g, 4.5 mmol, 1 eq) and 2,2'-(ethane-l,2-diylbis(oxy))bis(N-benzylethan-l-amine) (1.7 g, 5.2 mmol, 1.14 eq) in acetonitrile (100 ml) sodium carbonate (4.8 g, 45 mmol, 10 eq) was added. The suspension was refluxed for 40 hours. Then, the reaction mixture was cooled to ambient temperature and filtered. The filtrate was evaporated to dryness and dissolved with ethyl acetate (100 ml). The organic solution was extracted with water (2x30ml), brine (30 ml), dried with MgSCh (2 g), filtered, and evaporated to give a yellow glassy residue. The residue was dissolved in DCM (10 ml) and purified by silica gel column chromatography. The fractions containing product were combined and evaporated to give tert-butyl (trans)-4,13-dibenzyltetradecahydro-2H,llH,18H-pyrrolo[3,4-b][l,4,10,13]tetraoxa[7,16] diazacyclooctadecine-18-carboxylate as a colorless glassy solid (1.5 g, 55% yield). Calculated m / z [M+H] = 584.77 Found m / z [M+H] = 584.4

[0559] Compound 3i

[0560] Synthesis of tert-butyl (trans)-4, 7-dibenzyldodecahydro-12H-pyrrolo[3,4-b] [1,4 ] 'dioxa [7, 10 ]diazacyclododecine-l 2-carboxylate.

[0561] The method used was identical to the synthesis of Compound 3h to give tert-butyl (Irans)-4,7-dibenzyldodecahydro- 12H-pyrrolo [3,4-b] [ 1,4]dioxa[7, 10]diazacyclododecine- 12-carboxylate (1.3 g, 55% yield). Calculated m / z [M+H] = 496.29 Found m / z [M+H] = 496.3Compound 3j

[0562] Synthesis of tert-butyl (trans)-dodecahydro-12H-pyrrolo[3,4-b ][1, 4 ]dioxa[7, 10]diazacyclododecine-12-carboxylate.

[0563] To a 250 ml stainless steel vessel flushed with argon a solution of tert-butyl (trans)-4,7-dibenzyldodecahydro- 12H-pyrrolo [3,4-b] [ 1,4]dioxa[7, 10]diazacyclododecine- 12-carboxylate (1.3 g, 2.67 mmol, 1 eq) in methanol (100 ml) and palladium on activated carbon (1.3 g, 5% mass, wet) was added. The argon atmosphere was replaced with 3 ATM hydrogen and the reaction was stirred for 40 hours at an ambient temperature. The suspension was filtered through a celite pad. The filtrate was evaporated and dried at ambient temperature under high vacuum to give crude tert-butyl (trans)-dodecahydro-12H-pyrrolo[3,4-b][l,4]dioxa[7,10]diazacyclododecine-12-carboxylate as a yellowish oil (0.94 g, >100% yield). Calculated m / z [M+H] = 315.22 Found m / z [M+H] = 316.1

[0564] Compound 3k

[0565] Synthesis of tert-butyl (trans)-tetradecahydro-2H, 11H, 18H-pyrrolo[3, 4-b] [1,4, 10, 13 ]tetraoxa[7, 16 ]diazacyclooctadecine-l 8-carboxylate.

[0566] The method used was identical to the synthesis of Compound 3j to give crude tert-butyl (trans)-tetradecahydro-2H, 11 H, 18H-pyrrolo [3,4-b] [ 1,4, 10, 13]tetraoxa[7, 16]diazacyclooctadecine-18-carboxylate as a yellowish oil (1.2 g, >100% yield). Calculated m / z [M+H] = 403.27 Found m / z [M+H] = 404.2

[0567] Compound 31

[0568] Synthesis of tert-butyl (trans)-4, 8-dibenzyldodecahydro-2H, 13H-pyrrolo[3, 4-b ][1, 4 ] dioxa [7, 11 ]diazacyclotridecine-l 3 -carboxylate.

[0569] The method used was identical to the synthesis of Compound 3h to give tert-butyl (trans)-4,8-dibenzyldodecahydro-2H,13H-pyrrolo[3,4-b][l,4]dioxa[7,ll]diazacyclotridecine-13-carboxylate (2.3 g, 58% yield). Calculated m / z [M+H] = 510.33 Found m / z [M+H] = 510.2

[0570] Compound 3m

[0571] Synthesis of tert-butyl (trans)-4, 10-dibenzyldodecahydro-2H, 8H, 15H-pyrrolo[3, 4-b] [1,4, 10 ]trioxa[7, 13 ]diazacyclopentadecine-15-carboxylate.

[0572] The method used was identical to the synthesis of Compound 3h to give tert-butyl (trans)-4, 10-dibenzyldodecahydro-2H,8H, 15H-pyrrolo [3,4-b] [1,4,10]trioxa[7, 13]diazacyclopentadecine- 15 -carboxylate (2.4 g, 57% yield). Calculated m / z [M+H] = 540.34 Found m / z [M+H] = 540.3Compound 3n

[0573] Synthesis of tert-butyl (trans)-4, 7, 10-tribenzyltetradecahydro-2H, 15H-pyrrolo[3, 4-b ][1, 4 ]dioxa[7, 10, 13]triazacyclopentadecine-15-carboxylate.

[0574] The method used was identical to the synthesis of Compound 3h to give tert-butyl (trans)-4,7,10-tribenzyltetradecahydro-2H, 15H-pyrrolo [3,4-b] [1,4]dioxa[7, 10,13] triazacyclopentadecine-15-carboxylate (0.7 g, 14% yield). Calculated m / z [M+H] = 629.40 Found m / z [M+H] = 629.6

[0575] Compound 3o

[0576] Synthesis of tert-butyl (trans)-dodecahydro-2H,8H,15H-pyrrolo[3,4-b] [1,4, 10 ]trioxa[7, 13 ]diazacyclopentadecine-15-carboxylate.

[0577] The method used was identical to the synthesis of Compound 3j to give crude tert-butyl (trans)-dodecahydro-2H, 8H, 15H-pyrrolo [3,4-b] [1,4,10]trioxa[7, 13 ] diazacyclopentadecine- 15-carboxylate as a yellowish oil (1.1 g, 66% yield). Calculated m / z [M+H] = 360.24 Found m / z [M+H] = 360.4

[0578] Compound 3p

[0579] Synthesis of tert-butyl (trans) -dodecahydro-2H, 13H-pyrrolo[3, 4-b ][ 1, 4 ]dioxa[7, 11] diazacyclotridecine-13-carboxylate.

[0580] The method used was identical to the synthesis of Compound 3j to give crude tert-butyl (trans)-dodecahydro-2H,13H-pyrrolo[3,4-b][l,4]dioxa[7,l l]diazacyclotridecine- 13 -carboxylate as a yellowish oil (0.99 g 67% yield).

[0581] Compound 3q

[0582] Synthesis of tert-butyl (trans)-tetradecahydro-2H,15H-pyrrolo[3,4-b ][1, 4 ]dioxa[7, 10,13 ]triazacyclopentadecine-15-carboxylate.

[0583] The method used was identical to the synthesis of Compound 3j to give tert-butyl (trans)-tetradecahydro-2H, 15H-pyrrolo[3,4-b] [1,4]dioxa[7, 10, 13 ]triazacyclopentadecine-l 5 -carboxylate as a yellowish oil (0.4 g, 100% yield). Calculated m / z [M+H] = 359.26 Found m / z [M+H] = 359.2SYNTHESIS OF MACROCYCLIC DERIVATIVES BASED OF TERT-BUTYL (CIS)-3,4-BIS(2- ((METHYLSULFONYL)OXY)ETHOXY)PYRROLIDINE-1- ARBOXYLATE:

[0584] Compound 3r

[0585] Synthesis of tert-butyl (cis)-4, 7-dibenzyldodecahydro-12H-pyrrolo[3,4-b] [1,4 ] dioxa, [7, 10 ]diazacyclododecine-l 2-carboxylate.

[0586] The method used was identical to the synthesis of Compound 3h to give tert-butyl (cis)-4,7-dibenzyldodecahydro- 12H-pyrrolo [3,4-b] [ 1,4]dioxa[7, 10]diazacyclododecine- 12-carboxylate (1.5 g, 60% yield). Calculated m / z [M+H] = 496.31 Found m / z [M+H] = 496.3

[0587] Compound 3s

[0588] Synthesis of tert-butyl (cis)-4, 13-dibenzyltetradecahydro-2H, HH, 18H-pyrrolo[3,4-b] [1,4, 10, 13 ]tetraoxa[7,16]diazacyclooctadecine-18-carboxylate.

[0589] The method used was identical to the synthesis of Compound 3h to give tert-butyl (cis)-4, 13-dibenzyltetradecahydro-2H, 11 H, 18H-pyrrolo [3,4-b] [ 1,4, 10, 13 ]tetraoxa[7, 16] diazacyclooctadecine-18-carboxylate as a colorless glassy solid (1.5 g, 55% yield). Calculated m / z [M+H] - 584.77 Found m / z [M+H] - 584.4

[0590] Compound 3t

[0591] Synthesis of tert-butyl (cis)-dodecahydro-12H-pyrrolo[3,4-b ][1, 4 ] dioxa [7, 10]diazacyclododecine-12-carboxylate.

[0592] The method used was identical to the synthesis of Compound 3j to give crude tert-butyl (cis)-dodecahydro- 12H-pyrrolo [3,4-b] [ 1,4]dioxa[7, 10]diazacyclododecine- 12-carboxylate as a yellowish oil (0.98 g, >100% yield). Calculated m / z [M+H] = 316.22 Found m / z [M+H] = 316.2

[0593] Compound 3u

[0594] Synthesis of tert-butyl (cis)-tetradecahydro-2H, HH,18H-pyrrolo[3,4-b] [1,4, 10,13 ]tetraoxa[7, 16 ]diazacyclooctadecine-l 8-carboxylate.

[0595] The method used was identical to the synthesis of Compound 3j to give crude tert-butyl (cis)-tetradecahydro-2H, 11H, 18H-pyrrolo[3,4-b] [1,4, 10,13]tetraoxa[7,l 6]diazacyclooctadecine- 18-carboxylate as a yellowish oil (1.2 g, >100% yield). Calculated m / z [M+H] = 403.27 Found m / z [M+H] = 404.3Compound 3v

[0596] Synthesis of tert-butyl (cis)-4,8-dibenzyldodecahydro-2H,13H-pyrrolo[3,4-b ][1, 4 ] 'dioxa [7, 11 ]diazacyclotridecine-l 3 -carboxylate. =

[0597] The method used was identical to the synthesis of Compound 3h to give tert-butyl (cis)-4,8-dibenzyldodecahy dro-2H, 13 H-pyrrolo[3,4-b] [ 1,4] dioxa[7, 11 ]diazacyclotridecine- 13 -carboxylate (1.9 g, 70% yield). Calculated m / z [M+H] = 510.33 Found m / z [M+H] = 510.3

[0598] Compound 3w

[0599] Synthesis of tert-butyl (cis)-4,10-dibenzyldodecahydro-2H,8H,15H-pyrrolo[3,4-b] [ 1,4, 10]trioxa[7, 13 Jdiazacy clopentadecine- 15 -carboxylate.

[0600] The method used was identical to the synthesis of Compound 3h to give tert-butyl (cis)- 4.10-dibenzyldodecahydro-2H,8H, 15H-pyrrolo [3,4-b] [1,4,10]trioxa[7, 13]diazacy clopentadecine- 15-carboxylate (2.9 g, 53% yield). Calculated m / z [M+H] = 540.34 Found m / z [M+H] = 540.3

[0601] Compound 3x

[0602] Synthesis of tert-butyl (cis)-4, 7, 10-tribenzyltetradecahydro-2H, 15H-pyrrolo[3, 4-b ][1, 4 ]dioxa[7, 10, 13]triazacyclopentadecine-15-carboxylate.

[0603] The method used was identical to the synthesis of Compound 3h to give tert-butyl (cis)- 4.7.10-tribenzyltetradecahydro-2H, 15H-pyrrolo [3,4-b] [1,4]dioxa[7, 10,13]triazacyclopentadecine-15-carboxylate (2.5 g, 41% yield). Calculated m / z [M+H] = 629.40 Found m / z [M+H] = 629.3

[0604] Compound 3y

[0605] Synthesis of tert-butyl (cis)-dodecahydro-2H,13H-pyrrolo[3,4-b] [ 1,4]dioxa[7, 11 ]diazacyclotridecine- 13 -carboxylate.

[0606] The method used was identical to the synthesis of Compound 3j to give crude tert-butyl (cis)-dodecahydro-2H,13H-pyrrolo[3,4-b][l,4]dioxa[7,l l]diazacyclotridecine-13-carboxylate as a yellowish oil (1.2 g, 97% yield). Calculated m / z [M+H] = 330.23 Found m / z [M+H] = 330.3

[0607] Compound 3z

[0608] Synthesis of tert-butyl (cis)-dodecahydro-2H,8H,15H-pyrrolo[3,4-b] [1,4, 10 ]trioxa[7, 13 ]diazacyclopentadecine-15-carboxylate.

[0609] The method used was identical to the synthesis of Compound 3j to give crude tert-butyl (cis)-dodecahydro-2H,8H, 15H-pyrrolo[3,4-b] [1,4,10]trioxa[7, 13 ]diazacy clopentadecine- 15-carboxylate as a yellowish oil (1.94 g, >100% yield). Calculated m / z [M+H] = 360.24 Found m / z [M+H] = 360.4

[0610] Compound 3aa

[0611] Synthesis of tert-butyl (cis)-tetradecahydro-2H,15H-pyrrolo[3,4-b] [ 1,4]dioxa[7, 10, 13]triazacyclopentadecine- 15-carboxylate.

[0612] The method used was identical to the synthesis of Compound 3j to give tert-butyl (cis)-tetradecahydro-2H, 15H-pyrrolo [3,4-b] [ 1,4]dioxa[7, 10, 13 ]triazacyclopentadecme- 15 -carboxylate as a yellowish oil (1.49 g, >100% yield). Calculated m / z [M+H] = 359.26 Found m / z [M+H] = 359.3

[0613] EXAMPLE 5:

[0614] PREPARATION AND USE OF TETRAKIS(T-BU)-DOTA-TTX-ACOH RADIOLIGAND CHELATING AGENT

[0615] In some embodiments, pyrrolidine-derivatives of DOTA chelating agents are provided, including chelating gents such as tetrakis(t-bu)-DOTA-TTX-AcOH (Compound 4).

[0616] 0 O

[0617] O OH

[0618]

[0619] (4)

[0620] Compound 4 is useful as a starting material for the preparation of compound (5):

[0621]

[0622] wherein the LINKER is selected from the group consisting of:

[0623]

[0624] (MB2),

[0625]

[0626] shown above.

[0627]

[0628] (5) Compound 5 is modified form of lutetium Lu 177 vipivotide tetraxetan, a radioligand agent combination with lutetium Lu 177. In some embodiments, Compound 5 is useful in a mannerindicated for lutetium Lu 177 vipivotide tetraxetan. Lutetium Lu 177 vipivotide tetraxetan is a radioligand therapeutic agent. The active moiety of lutetium Lu 177 vipivotide tetraxetan is the radionuclide lutetium- 177 which is linked to a moiety that binds to PSMA, a transmembrane protein that is expressed in prostate cancer, including mCRPC. Upon binding of lutetium Lu 177 vipivotide tetraxetan to PSMA- expressing cells, the beta-minus emission from lutetium- 177 delivers radiation to PSMA-expressing cells, as well as to surrounding cells, and induces DNA damage which can lead to cell death. The compound lutetium Lu 177 vipivotide tetraxetan is a radioligand therapeutic agent indicated for the treatment of adult patients with prostate-specific membrane antigen (PSMA)-positive metastatic castration-resistant prostate cancer (mCRPC) who have been treated with androgen receptor (AR) pathway inhibition and taxane-based chemotherapy. The compound lutetium Lu 177 vipivotide tetraxetan is useful for treating select patients for treatment using LOCAMETZ® or an approved PSMA-11 imaging agent based on PSMA expression in tumors. The recommended dosage of Lu 177 vipivotide tetraxetan is 7.4 GBq (200 mCi) every 6 weeks for up to 6 doses, with dose interruption, reduction, or permanent discontinuation may be required due to adverse reactions. The compound lutetium Lu 177 vipivotide tetraxetan 1,000 MBq / mL (27 mCi / mL) Injection is supplied as a sterile, clear, colorless to slightly yellow solution for intravenous use. Each single-dose vial contains acetic acid (0.30 mg / mL), sodium acetate (0.41 mg / mL), gentisic acid (0.39 mg / mL), sodium ascorbate (50.0 mg / mL), pentetic acid (0.10 mg / mL), and water for injection (q.s. to 1 mL). The pH range of the solution is 4.5 to 7.0. In some embodiments, a linker of

[0629] R10has the following structure

[0630]

[0631] ,as shown in compound 5.EXAMPLE 6:

[0632] PREPARATION AND USE OF TETRAKIS(T-BU)-DOTA-TTX-ACOH RADIOLIGAND CHELATING AGENT

[0633] Compound 4 is useful as a starting material for the preparation of compound (6):

[0634] 0 O

[0635] HO OH HO NT pH

[0636] o^J

[0637]

[0638] D-Phe Cy$Tyr~D-Trp LysThrCys-Thr

[0639] 1 - - I

[0640] (6).

[0641] Compound 6 is modified form of lutetium Lu 177 dotatate, a radioligand agent combination with lutetium Lu 177. In some embodiments, Compound 6 is useful in a manner indicated for lutetium Lu 177 dotatate. The compound lutetium Lu 177 dotatate is a radiolabeled somatostatin analog indicated for the treatment of adult and pediatric patients 12 years and older with somatostatin receptor-positive gastroenteropancreatic neuroendocrine tumors (GEP-NETs), including foregut, midgut, and hindgut neuroendocrine tumors. The compound lutetium Lu 177 dotatate is administered as 7.4 GBq (200 mCi) every 8 weeks (± 1 week) for a total of 4 doses. In some embodiments, a linker of R10is D-Phe-cys-tyr-D-trp-lys-cys-thr-NH2 (disulfide bridge: 2-6) acetate as shown in compound 6.

[0642] EXAMPLE 7

[0643] PREPARATION OF 2,2 ',2 '',2'"-(((TR NS)-CYCLOUEXANE-1,2-DIYL)BIS(AZ ETRIYL))TETRAACETIC ACID

[0644] Synthesis of tetramethyl 2,2',2",2'"-(((trans)-l-(tert-butoxycarbonyl)pyrrolidine-3,4-diyl)bis(azanetriyl))tetraacetate

[0645] To a solution of tert-butyl trans-3,4-diaminopyrrolidine-l -carboxylate (200 mg, 1 mmol, 1 eq) in acetonitrile (20 ml) sodium carbonate (2.12 g, 20 mmol, 20 eq) and methyl 2- bromoacetate (1.86 g, 20 mmol, 20 eq) were added. This suspension was stirred for 12 hours at 40oC. LCMS analysis showed full conversion of starting material. The suspension was filtered, and the filtrate was evaporated to dryness. The residue was diluted with water (50 ml) and extracted with ethyl acetate(3x30 ml). The organic extract was evaporated to dryness. The residue was dissolved in DCM (5 ml) and purified by silica gel column chromatography (40 g of silica gel, 0% to 100% ethyl acetate in heptane). The fractions containing the product (TLC control, eluent - ethyl acetate / heptane 1 / 1 by volume, plate - SiliaPlate TLA-AUT0337-323N, detection - ninhydrin stain) were combined and evaporated. The residue was dried under high vacuum at 35oC for 6 hours to give crude tetramethyl 2,2',2",2'"-(((trans)-l -(tert-butoxycarbonyl)pyrrolidine-3,4- diyl)bis(azanetriyl))tetraacetate (370 mg, 75%) as a colorless glassy solid.

[0646] Synthesis of tetramethyl 2,2',2",2'"-(((trans)-l-(4-methoxybenzoyl)pyrrolidine-3,4-diyl)bis(azanetriyl))tetraacetate

[0647] To a solution of tetramethyl 2,2',2",2"'-(((trans)-l-(tert-butoxycarbonyl)pyrrolidine-3,4-diyl)bis(azanetriyl))tetraacetate (370 mg, 0.76 mmol, 1 eq) in dichloromethane (5 ml) trifluoracetic acid (2 ml) was added. The solution was then stirred for 6 hours at ambient temperature. LCMS showed the full conversion of the starting material to the target product. The solution was evaporated under vacuum at 35oC. The residue was dried to constant weight to give tetramethyl 2, 2', 2", 2"'-(((trans)-pyrrolidine-3,4- diyl)bis(azanetriyl))tetraacetate trifluoroacetate (550 mg) as a yellowish gum.

[0648] The intermediate was dissolved in acetonitrile (20 ml). To the mixture sodium carbonate (810 mg, 7.6 mmol, 10 eq) and 4-methoxybenzoyl chloride (195 mg, 1.14 mmol, 1.5 eq) was added. The suspension was stirred for 4 hours at ambient temperature (LCMS control of full conversion of intermediate to the product). The reaction mixture was filtered. The filtrate was evaporated to dryness. The residue was dissolved in DCM (5 ml) and purified by silica gel column chromatography (40 g of silica gel, 0% to 100% ethyl acetate in heptane). The fractions containing the product (TLC control, eluent - ethyl acetate / heptane 1 / 1 by volume, plate - SiliaPlate TLA-AUT0337-323N, detection - ninhydrin stain) were combined and evaporated. The residue was dried under high vacuum at 35oC for 6 hours to give crude tetramethyl 2,2',2",2"'-(((trans)-l-(4-methoxybenzoyl)pyrrolidine-3,4- diyl)bis(azanetriyl))tetraacetate (390 mg, 98%) as a colorless glassy solid.

[0649] Synthesis of 2,2 2 ", 2 '"-(((trans)-l-(4-methoxybenzoyl)pyrrolidine-3, 4-diyl)bis(azanetriyl))tetraacetic acid trifluoroacetate

[0650] To a solution of tetramethyl 2,2',2",2"'-(((trans)-l-(4-methoxybenzoyl)pyrrolidine-3,4-diyl)bis(azanetriyl))tetraacetate (390 mg, 0.74 mmol, 1 eq) in tetrahydrofuran (20 ml) a previously prepared IM aqueous lithium hydroxide solution (5.9 ml, 5.9 mmol, 8 eq) was added. The reaction mixture was stirred for 4 hours at ambient temperature (LCMS control of full saponification of the ester groups). Acetic acid (2 ml) was then added, and the mixture was evaporated to dryness. The residue was diluted with DMSO (5 ml) and purified by preparative HPLC: Column: Silicycle Cl 8, 100A, lOum in 50 x 300mm load and lock column. Buffer A: 0.1% TFA in Water I -Buffer B: 0.1% TFA in Acetonitrile. Method: 5 to 80% B, 20ml / min, 40 minutes. Column Temperature: Ambient Temperature. Detection: LCMS control (UV254).

[0651] After HPLC purification all the fractions containing the product were combined, then concentrated on a rotavapor. The temperature for the rotavapor bath was held at 30oC. Then, the resultant residue was poured into a lyophilization jar and dried to constant weight to give 2, 2', 2", 2"'-(((trans)-l-(4- methoxybenzoyl)pyrrolidine-3,4-diyl)bis(azanetriyl))tetraacetic acid trifluoroacetate as a colorless glassy solid (304 mg).

[0652] EXAMPLE 8

[0653] PREPARATION OF TETRAMETHYL 2,2',2”,2’"-(((CIS)-PYRROLIDINE-3,4- DIYL)BIS(AZANETRIYL))TETRAACETATE TRIFLUOROACETATE

[0654] Synthesis of tetramethyl 2,2’,2",2”’-(((cis)-l-(tert-butoxycarbonyl)pyrrolidine-3,4-diyl)bis(azanetriyl))tetraacetate

[0655] To a solution of tert-butyl cis-3,4-diaminopyrrolidine-l -carboxylate (150 mg, 0.75 mmol, 1 eq) in acetonitrile (20 ml) sodium carbonate (1.4 g, 15 mmol, 20 eq) and methyl 2-bromoacetate (1.4 g, 7.5 mmol, 10 eq) were added. This suspension was stirred for 12 hours at 40oC. LCMS analysis showed full conversion of starting material. The suspension was filtered, and the filtrate was evaporated to dryness. The residue was diluted with water (50 ml) and extracted with ethyl acetate (3x30 ml). The organic extract was evaporated to dryness. The residue was dissolved in DCM (5 ml) and purified by silica gel column chromatography (40 g of silica gel, 0% to 100% ethyl acetate in heptane). The fractions containing the product (TLC control, eluent - ethyl acetate / heptane 1 / 1 by volume, plate - SiliaPlate TLA-AUT0337-323N, detection - ninhydrin stain) were combined and evaporated. The residue was dried under high vacuum at 35oC for 6 hours to give crude tetramethyl 2,2',2",2"'-(((cis)- 1 -(tert-butoxycarbonyl)pyrrolidine-3,4- diyl)bis(azanetriyl))tetraacetate (230 mg, 78%) as a colorless glassy solid.To a solution of the intermediate in dichloromethane (5 ml) trifluoracetic acid (2 ml) was added. The solution was then stirred for 6 hours at ambient temperature. LCMS showed the full conversion of the intermediate to the target product. The solution was evaporated under vacuum at 35oC. The residue was dried to constant weight to give tetramethyl 2,2',2",2"'-(((cis)-pyrrolidine-3,4-diyl)bis(azanetriyl))tetraacetate trifluoroacetate (330 mg) as a yellowish gum.

[0656] Synthesis of 2,2’,2”,2’”-(((cis)-l-(4-methoxybenzoyl)pyrrolidme-3,4-diyl)bis(azanetriyl))tetraacetic acid trifluoroacetate

[0657] To a solution of tetramethyl 2,2',2",2"'-(((cis)-pyrrolidine-3,4-diyl)bis(azanetriyl))tetraacetate trifluoroacetate (330 mg, -0.47 mmol, 1 eq) in acetonitrile (20 ml) sodium carbonate (1 g, 9.4 mmol, 20 eq) and 4-methoxybenzoyl chloride (121 mg, 0.71 mmol, 1.5 eq) were added. The suspension was stirred for 4 hours at ambient temperature (LCMS control of full conversion of intermediate to the product). The reaction mixture was filtered. The filtrate was evaporated to dryness. The residue was dissolved in DCM (5 ml) and purified by silica gel column chromatography (40 g of silica gel, 0% to 100% ethyl acetate in heptane). The fractions containing the product (TLC control, eluent - ethyl acetate / heptane 1 / 1 by volume, plate - SiliaPlate TLA-AUT0337-323N, detection - ninhydrin stain) were combined and evaporated. The residue was dried under high vacuum at 35oC for 6 hours to give crude tetramethyl 2,2',2",2"'-(((cis)-l-(4-methoxybenzoyl)pyrrolidine-3,4- diyl)bis(azanetriyl))tetraacetate (240 mg, 98%) as a colorless glassy solid.

[0658] The intermediate was dissolved in tetrahydrofuran (20 ml) and a previously prepared IM aqueous lithium hydroxide solution (3.76 ml, 3.76 mmol, 8 eq) was added. The reaction mixture was stirred for 4 hours at ambient temperature (LCMS control of full saponification of the ester groups). Then, acetic acid (2 ml) was added, and the mixture was evaporated to dryness. The residue was diluted with DMSO (5 ml) and purified by preparative HPLC: Column: Silicycle C18, 100A, lOum in 50 x 300mm load and lock column. Buffer A: 0.1% TFA in Water I -Buffer B: 0.1% TFA in Acetonitrile. Method: 5 to 80% B, 20ml / min, 40 minutes. Column Temperature: Ambient Temperature. Detection: LCMS control (UV254). After HPLC purification all the fractions containing the product were combined, then concentrated on a rotavapor. The temperature for the rotavapor bath was held at 30oC. Then, the resultant residue was poured into a lyophilization jar and dried to constant weight to give 2,2',2'',2'"-(((cis)-l-(4-methoxybenzoyl)pyrrolidine-3,4-diyl)bis(azanetriyl))tetraacetic acid trifluoroacetate as a colorless glassy solid (65 mg).EXAMPLE 9

[0659] PREPARATION OF 6,6’-((((TRANS)-1-(TERT-BUTOXYCARBONYL)PYRROLIDINE-3,4- IYL)BIS((CA BOXYMETHYL)AZA EDIYL)) BIS(METHYLENE))DIPICOLINIC ACID

[0660] Synthesis of dimethyl 6,6'-((((trans)-l-(tert-butoxycarbonyl)pyrrolidine-3,4-diyl)bis(azancdiyl))bis(mcthylcne))dipicolinate

[0661] To a solution of tert-butyl (trans)-3,4-diaminopyrrolidine-l -carboxylate (300 mg, 1.5 mmol, 1 eq) in methanol (30 ml) methyl 6-formylpicolinate (500 mg, 3 mmol, 2 eq) was added. The mixture was refluxed for 4 hours and cooled to lOoC. To the cooled solution sodium triacetoxyborohydride (1.28 g, 6 mmol, 4 eq) was added and the mixture was stirred for 4 hours keeping temperature at 10°C. The mixture then was allowed to warm to ambient temperature and stirred overnight. LCMS analysis showed full conversion of the starting material to the target intermediate. The mixture was evaporated to dryness. The residue was diluted with water (100 ml) and extracted with dichloromethane (3x50 ml). The organic extract was evaporated to dryness. The residue was diluted with DCM (10 ml) and purified by silica gel column chromatography (40 g of silica gel, 0% to 5% methanol in dichloromethane). The fractions containing product (TLC control, eluent - EA / Hex 1 / 1 by volume, plate - SiliaPlate TLA- UT0337-323N, detection - UV254) were combined, evaporated and dried under high vacuum to give dimethyl 6,6'-((((trans)-l-(tertbutoxy carbonyl) pyrrolidine-3,4- diyl)bis(azanediyl))bis(methylene))dipicolinate as yellowish solid (430 mg, 57%).

[0662] Synthesis of dimethyl 6,6'-((((trans)-l-(tert-butoxycarbonyl)pyrrolidine-3,4-diyl)bis((2-methoxy-2- oxoethyl)azanediyl))bis(methylene))dipicolinate

[0663] To a solution of dimethyl 6,6'-((((trans)-l-(tert-butoxycarbonyl)pyrrolidine-3,4-diyl)bis(azanediyl))bis(methylene))dipicolinate (430 mg, 0.86 mmol, 1 eq) in dimethylformamide (20 ml) sodium carbonate (910 mg, 8.6 mmol, 10 eq) and methyl 2-bromoacetate (640 mg, 3.44 mmol, 4 eq) were added. This suspension was stirred for 12 hours at 40oC. LCMS analysis showed full conversion of starting material. The suspension was filtered, and the filtrate was evaporated to dryness. The residue was diluted with water (100 ml) and extracted with ethyl acetate (3x50 ml). The organic extract was evaporated to dryness.

[0664] The residue was dissolved in DCM (5 ml) and purified by silica gel column chromatography (40 g of silica gel, 0% to 100% ethyl acetate in heptane). The fractions containing the product (TLC control, eluent - ethyl acetate / heptane 1 / 1 by volume, plate - SiliaPlate TLA-AUT0337-323N,detection - UV254) were combined and evaporated. The residue was dried under high vacuum at 35oC for 6 hours to give crude dimethyl 6,6'-((((trans)-l-(tert-butoxycarbonyl)pyrrolidine-3,4-diyl)bis((2-methoxy-2- oxoethyl)azanediyl))bis(methylene))dipicolinate (230 mg, 41%) as yellowish glassy solid.

[0665] Synthesis of 6,6’-((((trans)-l-(tert-butoxycarbonyl)pyrrolidine-3,4-diyl)bis((carboxymethyl)azanediyl)) bis(methylene))dipicolinic acid

[0666] To a solution of crude dimethyl 6,6'-((((trans)-l-(tert-butoxycarbonyl)pynolidine-3,4-diyl)bis((2- methoxy-2-oxoethyl)azanediyl))bis(methylene))dipicolinate (230 mg, 0.36 mmol, 1 eq) in tetrahydrofuran (10 ml) a previously made IM solution of lithium hydroxide (2.88 ml, 2.88 mmol, 8 eq) was added. The mixture was stirred for 2 hours (LCMS control of full conversion of the intermediate to the target product). To the mixture glacial acetic acid (1 ml) was added and the mixture was evaporated to dryness. The residue was diluted with DMSO (2 ml) and purified by preparative HPLC: Column: Silicycle Cl 8, 100A, lOum in 50 x 300mm load and lock column. Buffer A: 0.1% formic acid in Water / -Buffer B: 0.1% formic acid in acetonitrile. Method: 5 to 80% B, 20ml / min, 40 minutes. Column Temperature: Ambient Temperature. Detection: LCMS control (UV254). After HPLC purification all the fractions containing the product were combined then concentrated on a rotavapor. The resultant residue was poured into a lyophilization jar and dried to constant weight to give 6,6'-((((trans)-l-(tert-butoxycarbonyl)pyrrolidine-3,4-diyl)bis((carboxymethyl)azanediyl))bis(methylene)) dipicolinic acid (140 mg, 66% yield) as a white solid.

[0667] EXAMPLE 10

[0668] PREPARATION OF 6,6’-((((TRANS)-1-(4-METHOXYBENZOYL)PYRROLIDINE-3,4- DIYL)BIS(AZANEDIYL))BIS(METHYLENE))DIPICOLINIC ACID TRIFLUOROACETATE

[0669] Synthesis of dimethyl 6,6'-((((cis)-pyrrolidine-3,4-iyl)bis(azanediyl))bis(methylene))dipicolinate To a solution of dimethyl 6,6'-((((trans)-l-(tert-butoxycarbonyl)pyrrolidine-3,4-diyl)bis(azanediyl))bis(methylene))dipicolinate (0.65 g, 1.3 mmol, 1 eq) in dichloromethane (10 ml) trifluoracetic acid (3 ml) was added. The solution was then stirred for 6 hours at an ambient temperature. LCMS showed the full conversion of the starting material to the target product. The solution was evaporated under vacuum at 35oC. To the residue water (50 ml) and sodium carbonate (1.38 g, 13 mmol, 10 eq) were added and the mixture was stirred for 10 minutes. The mixture wasextracted with dichloromethane (3x30 ml). The organic extract was dried with magnesium sulfate, filtered and evaporated. The resultant residue was poured into a lyophilization jar and dried to constant weight to give dimethyl 6,6'-((((cis)-pyrrolidine-3,4-diyl)bis(azanediyl))bis(methylene))dipicolinate (520 mg, 98%) as a yellowish solid.

[0670] Synthesis of 6,6’-((((trans)-l-(4-methoxybenzoyl)pyrrolidme-3,4-diyl)bis(azanediyl))bis(methylene))dipicolinic acid trifluoroacetate

[0671] To a stirred suspension of dimethyl 6,6'-((((trans)-pyrrolidine-3,4-diyl)bis(azanediyl))bis(methylene))dipicolinate (174 mg, 0.43 mmol, 1 eq) and sodium carbonate (230 mg, 2.15 mmol, 5 eq) in acetonitrile (10 ml) 4-methoxybenzoyl chloride (109 mg, 0.64 mmol, 1.5 eq) was added. The suspension was stirred for 4 hours at ambient temperature (LCMS control of full conversion of starting material). The reaction mixture was filtered. The filtrate was evaporated to dryness. The residue was diluted in tetrahydrofuran (5 ml) and a previously prepared IM aqueous lithium hydroxide solution (1.72 ml, 1.72 mmol, 4 eq) was added. The reaction mixture was stirred for 4 hours at ambient temperature (LCMS control of full saponification of the ester groups). Then, acetic acid (1 ml) was added, and the mixture was evaporated to dryness. The residue was diluted with DMSO (2 ml) and purified by preparative HPLC: Column: Silicycle C18, 100A, lOum in 50 x 300mm load and lock column. Buffer A: 0.1% TFA in Water I -Buffer B: 0.1% TFA in Acetonitrile. Method: 5 to 80% B, 20ml / min, 40 minutes. Column Temperature: Ambient Temperature.

[0672] Detection: LCMS control (UV254).

[0673] After HPLC purification all the fractions containing the product were combined, then concentrated on a rotavapor. The temperature for the rotavapor bath was held at 30oC. Then, the resultant residue was poured into a lyophilization jar and dried to constant weight to give 6,6'-((((trans)-l -(4- methoxybenzoyl)pyrrolidine-3,4-diyl)bis(azanediyl))bis(methylene))dipicolinic acid trifluoroacetate as a colorless viscous gum (163 mg).

[0674] EXAMPLE 11

[0675] PREPARATION OF 6,6’-((((TRANS)-1-((4-METHOXYPHENYL)SULFONYL)PYRROLIDINE-3,4- DIYL)BIS(AZANEDIYL))BIS(METHYLENE))DIPICOLINIC ACID TRIFLUOROACETATE

[0676] To a stirred suspension of dimethyl 6,6'-((((trans)-pyrrolidine-3,4-diyl)bis(azanediyl))bis(methylene))dipicolinate (174 mg, 0.43 mmol, 1 eq) and sodium carbonate (230 mg, 2.15 mmol, 5 eq) in acetonitrile (10 ml) 4-methoxybenzenesulfonyl chloride (132 mg, 0.64 mmol, 1.5 eq) was added. The suspension was stirred for 4 hours at ambient temperature (LCMScontrol of full conversion of starting material). The reaction mixture was filtered. The filtrate was evaporated to dryness. The residue was diluted in tetrahydrofiiran (5 ml) and a previously prepared IM aqueous lithium hydroxide solution (1.72 ml, 1 - 72 mmol, 4 eq) was added. The reaction mixture was stirred for 4 hours at ambient temperature (LCMS control of full saponification of the ester groups). Then, acetic acid (1 ml) was added, and the mixture was evaporated to dryness. The residue was diluted with DMSO (2 ml) and purified by preparative HPLC: Column: Silicycle C18, 100A, lOum in 50 x 300mm load and lock column. Buffer A: 0.1% TFA in Water / -Buffer B: 0.1% TFA in Acetonitrile. Method: 5 to 80% B, 20ml / min, 40 minutes. Column Temperature: Ambient Temperature. Detection: LCMS control (UV254).

[0677] After HPLC purification all the fractions containing the product were combined, then concentrated on a rotavapor. The temperature for the rotavapor bath was held at 30oC. Then, the resultant residue was poured into a lyophilization jar and dried to constant weight to give 6,6'-((((trans)-l -((4- methoxyphenyl)sulfonyl)pyrrolidine-3,4-diyl)bis(azanediyl))bis(methylene))dipicolinic acid trifluoroacetate as a colorless viscous gum (159 mg).

[0678] EXAMPLE 12

[0679] PREPARATION OF 6,6'-((((TRANS)-1-(4-METHOXYBENZYL)PYRROLIDINE-3,4- DIYL)BIS(AZANEDIYL))BIS(METHYLENE))DIPICOLINIC ACID TRIFLUOROACETATE To a stirred suspension of dimethyl 6,6'-((((trans)-pyrrolidine-3,4-diyl)bis(azanediyl))bis(methylene))dipicolinate (174 mg, 0.43 mmol, 1 eq) and sodium carbonate (230 mg, 2.15 mmol, 5 eq) in acetonitrile (10 ml) l-(chloromethyl)-4-methoxybenzene (73 mg, 0.47 mmol, 1.1 eq) was added. The suspension was stirred for 40 hours at ambient temperature (LCMS control of full conversion of starting material). The reaction mixture was filtered. The filtrate was evaporated to dryness. The residue was diluted in tetrahydrofuran (5 ml) and a previously prepared IM aqueous lithium hydroxide solution (1.72 ml, 1.72 mmol, 4 eq) was added. The reaction mixture was stirred for 4 hours at ambient temperature (LCMS control of full saponification of the ester groups). Then, acetic acid (1 ml) was added, and the mixture was evaporated to dryness. The residue was diluted with DMSO (2 ml) and purified by preparative HPLC: Column: Silicycle C18, 100A, lOum in 50 x 300mm load and lock column. Buffer A: 0.1% TFA in Water / -Buffer B: 0.1% TFA in Acetonitrile. Method: 5 to 80% B, 20ml / min, 40 minutes. Column Temperature: Ambient Temperature. Detection: LCMS control (UV254).After HPLC purification all the fractions containing the product were combined, then concentrated on a rotavapor. The temperature for the rotavapor bath was held at 30oC. Then, the resultant residue was poured into a lyophilization jar and dried to constant weight to give 6,6'-((((trans)-l -(4- methoxybenzyl)pyrrolidine-3,4-diyl)bis(azanediyl))bis(methylene))dipicolinic acid trifluoroacetate as a colorless viscous gum (179 mg).

[0680] EXAMPLE 13

[0681] PREPARATION OF 6,6’-((((TRANS)-1-(TERT-BUTOXYCARBONYL)PYRROLIDINE-3,4- DIYL)BIS(AZA EDIYL))BIS(METHYLENE)) DIPICOLINIC ACID

[0682] To a solution of tert-butyl (trans)-3,4-diaminopyrrolidine-l -carboxylate (60 mg, 0.3 mmol, 1 eq) in methanol (6 ml) methyl 6-formylpicolinate (100 mg, 0.6 mmol, 2 eq) was added. The mixture was refluxed for 4 hours and cooled to lOoC. To the cooled solution sodium triacetoxyborohydride (255 g, 1.2 mmol, 4 eq) was added and the mixture was stirred for 4 hours keeping temperature lOoC. The mixture then was allowed to warm to ambient temperature and stirred overnight. LCMS analysis showed full conversion of the starting material to the target intermediate. The mixture was evaporated to dryness. The residue was diluted with water (20 ml) and extracted with dichloromethane (3x10 ml). The organic extract was evaporated to dryness.

[0683] The residue was diluted with DCM (3 ml) and purified by silica gel column chromatography (24 g of silica gel, 0% to 5% methanol in dichloromethane). The fractions containing product (TLC control, eluent - EA / Hex 1 / 1 by volume, plate - SiliaPlate TLA-AUT0337-323N, detection -UV254) were combined, evaporated and dried under high vacuum to give crude dimethyl 6,6'-((((trans)- 1 -(tertbutoxy carbonyl) pyrrolidine-3,4-diy l)bis(azanediy l))bis(methylene))dipicolinate as yellowish solid (120 mg, 80% yield). The intermediate was dissolved in tetrahydrofuran (5 ml) and a previously made IM solution of lithium hydroxide (0.6 ml, 0.6 mmol, 2 eq) was added. The mixture was stirred for 2 hours (LCMS control of full conversion of the intermediate to the target product). To the mixture glacial acetic acid (0.5 ml) was added and the mixture was evaporated to dryness. The residue was diluted with DMSO (2 ml) and purified by preparative HPLC: Column: Silicycle C18, 100A, lOum in 50 x 300mm load and lock column. Buffer A: 0.1% formic acid in Water / -Buffer B: 0.1% formic acid in acetonitrile. Method: 5 to 80% B, 20ml / min, 40 minutes. Column Temperature: Ambient Temperature. Detection: LCMS control (UV254). After HPLC purification all the fractions containing the product were combined then concentrated on a rotavapor. Then, the resultant residue was poured into a lyophilization jar and dried to constant weight to give 6,6'-((((trans)- 1 -(tert- butoxy carbonyl)pyrrolidine-3,4-diyl)bis(azanediyl))bis(methylene))dipicolinic acid, 83 mg, 59% for two steps) as a white solid.

[0684] EXAMPLE 14

[0685] PREPARATION OF TERT-BUTYL (TRANS)-3,4-BIS(((8-HYDROXYQUINOLIN-2- YL)METHYL)AMINO)PY OLIDINE-1-CARBOXYLATE

[0686] To a solution of tert-butyl (trans)-3,4-diaminopyrrolidine-l -carboxylate (60 mg, 0.3 mmol, 1 eq) in ethanol (6 ml) 8-hydroxyquinoline-2-carbaldehyde (104 mg, 0.6 mmol, 2 eq) was added. The mixture was refluxed for 4 hours and cooled to lOoC. To the cooled solution sodium borohydride (25 mg, 0.6 mmol, 4 eq) was added and the mixture was stirred for 4 hours keeping temperature lOoC. The mixture then was allowed to warm to ambient temperature and stirred overnight. LCMS analysis showed full conversion of the starting material. The mixture was evaporated to dryness. The residue was diluted with water (20 ml) and extracted with dichloromethane (3x10 ml). The organic extract was evaporated to dryness. The residue was diluted with DMSO (2 ml) and purified by preparative HPLC: Column: Silicycle Cl 8, 100A, lOum in 50 x 300mm load and lock column. Buffer A: 0.1% formic acid in Water / -Buffer B: 0.1% formic acid in acetonitrile. Method: 5 to 80% B, 20ml / min, 40 minutes. Column

[0687] Temperature: Ambient Temperature. Detection: LCMS control (UV254). After HPLC purification all the fractions containing the product were combined then concentrated on a rotavapor. Then, the resultant residue was poured into a lyophilization jar and dried to constant weight to give tert-butyl (trans)-3,4-bis(((8-hydroxyquinolin-2-yl)methyl)amino)pyrrolidine-l-carboxylate (43 mg, 27%) as a white solid.

[0688] EXAMPLE 15

[0689] PREPARATION OF TERT-BUTYL (CIS)-3,4-BIS(((8-HYDROXYQUINOLIN-2- YL)METHYL)AMINO)PYRROLIDINE-1-CARBOXYLATE

[0690] To a solution of tert-butyl (cis)-3,4-diaminopyrrolidine-l -carboxylate (150 mg, 0.75 mmol, 1 eq) in ethanol (15 ml) 8-hydroxyquinoline-2-carbaldehyde (260 mg, 1.5 mmol, 2 eq) was added. The mixture was refluxed for 4 hours and cooled to lOoC. To the cooled solution sodium borohydride (200 mg, 5.2 mmol, 7 eq) was added and the mixture was stirred for 4 hours keeping temperature at lOoC. LCMS analysis showed the transformation of the starting material into a stable cyclic fivemembered intermediate. The reaction mixture was heated to 40°C and over the next 8 hours more sodium borohydride (1 g) was added in portions keeping temperature at 40-50oC. During thisprocess, decomposition of the intermediate into the target product and several by-products was observed (according to LCMS). At the end of the reaction, the content of the target product in the mixture did not exceed 30%.

[0691] The mixture was evaporated to dryness. The residue was diluted with water (30 ml) and extracted with dichloromethane (3x20 ml). The organic extract was evaporated to dryness.

[0692] The residue was diluted with DMSO (2 ml) and purified by preparative HPLC: Column: Silicycle C18, 100A, lOum in 50 x 300mm load and lock column. Buffer A: 0.1% formic acid in Water / -Buffer B: 0.1% formic acid in acetonitrile. Method: 5 to 80% B, 20ml / min, 40 minutes. Column Temperature: Ambient Temperature. Detection: LCMS control (UV254). After HPLC purification all the fractions containing the product were combined then concentrated on a rotavapor. Then, the resultant residue was poured into a lyophilization jar and dried to constant weight to give tert-butyl (cis)-3,4-bis(((8-hydroxyquinolin-2-yl)methyl)amino)pyrrolidine-l-carboxylate (14 mg, 3.6%) as a white solid.

[0693] EXAMPLE 16

[0694] COMPLEXATION RATE OF COMPOUND 8-LA(III) COMPLEX FORMATION IN COMPARISON WITH MACROPA

[0695] The complexation rates were assessed utilizing HPLC / UV analysis employing chelators competition for a metal ion as described below.

[0696] Retention times of compound 8, Macropa

[0697] 5ul of a solution of compound 8 (approximate nominal concentration 100|JM) in an aqueous NaOAc buffer (pH = 4.6) was injected into Agilent 1260 Infinity Series HPLC System for UV analysis using the following method:

[0698] Mobile Phase A: lOmM NaOAc pH 4.6

[0699] Mobile phase B: MeOH

[0700] Gradient: 5% to 85%B over 10 min, 0.5 min at 85%B, equilibrate to 1%B over 1%B for 3.5 min

[0701] Flowrate: 0.55mL / min

[0702] Column: Restek Raptor Inert ARC- 182.7 |Jm, 50x2.1mm

[0703] Retention time of compound 8 was determined (2.83 min)

[0704] Retention time of Macropa was determined (4.39 min)Retention times of complexes of compound 8-La(III) and Macropa-La(III)

[0705] Excess of lanthanum(III) chloride was added to a solution of compound 8 described and was let stand for 15 min at 25°C maintaining a pH between 4.5 and 5.5. 5 pl of the solution was injected into Agilent 1260 Infinity Series HPLC System for UV analysis using the same HPLC method as described above.

[0706] Retention time of complex of compound 8-La(III) was determined (3.55 min) Retention time of complex Macropa-La(III) was determined (4.95 min)

[0707] Competition study of compound 8 and Macropa for La (III)

[0708] An approximately equimolar mixture of compound 8 and Macropa in an aqueous NaOAc buffer (pH = 4.6) was prepared. The mixture was injected into the HPLC with the conditions described above. Retention times of compound 8 and Macropa were confirmed (2.83 min and 4.39 min, respectively). Lanthanum(III) chloride was added to the equimolar mixture of compound 8 and Macropa. The quantity of Lanthanum(III) chloride may vary; however, it is essential to add an amount sufficient to ensure chelator excess within the mixture. The solution was allowed to stand for 15 minutes at 25°C, with the pH maintained between 4.5 and 5.5. 5 pl of the solution was injected into Agilent 1260 Infinity Series HPLC System for UV analysis using the same HPLC method as described above. The area under the curve (AUC) of compound 8, Macropa, compound 8-La(III) complex, and Macropa-La(III) complex were determined using retention times as reference for identification.

[0709] A qualitative evaluation of the complexation rate between compound 8 and La(III) relative to Macropa

[0710] HPLC / UV traces utilized in the complexation study are included in FIGs. 1.

[0711] A qualitative evaluation of the complexation rate between compound 8 and La(III) relative to Macropa was conducted by analyzing the AUC values of HPLC peaks corresponding to compound 8, compound 8-La(III), as well as Macropa and Macropa-La(III) as illustrated in FIGs. 1.

[0712] Further, additional amount of Lanthanum(III) chloride was added to the equimolar mixture of compound 8 and Macropa. The quantity of Lanthanum(III) chloride may vary; however, it is essential to add an amount sufficient to ensure Lanthanum (III) excess compared to chelators within the mixture. The solution was allowed to stand for 15 minutes at 25°C, with the pH maintained between 4.5 and 5.5. 5pl of that solution was injected into Agilent 1260 Infinity Series HPLCSystem for UV analysis using the same HPLC method as described above. Retention times of compound 8-La(III) and Macropa-(III) were confirmed (3.5 min and 4.95 min, respectively).

[0713] EXAMPLE 17

[0714] COMPLEXATION RATE OF COMPOUNDS 8, 12, 13, AND 15 WITH VARIOUS METALS IN COMPARISON WITH ACROPA

[0715] A complexation study of compounds 8, 12, 13, and 15 and Macropa with various metals was performed following a procedure similar to that described in Example 16, and the summary of results is presented in Table 2 below.

[0716] Table!:

[0717] A summary of complexation study of the compounds of the present disclosure with La, Tb, Lu,

[0718] Cu, and Sc compared to Macropa metal complexes

[0719] Compound La Tb Lu Cu Sc

[0720] 8 ** ** ** ** **

[0721] 12 ** ** ** *** **

[0722] 13 ** ** ** *** **

[0723] 15 * ** ** *** **

[0724]

[0725] ***: Much faster complexation rate compared to Macropa

[0726] **: Faster complexation rate compared to Macropa

[0727] *: Similar complexation rate compared to Macropa

[0728] EXAMPLE 18

[0729] STABILITY STUDY FOR COMPOUNDS 8, 12, 13, AND 15 WITH VARIOUS METALS Stability testing via DTP A (diethylenetriaminepentaacetic acid) challenge (500 times excess of DTP A) was performed to evaluate the kinetic stability of radiometal-chelator complexes. The following describes an experiment conducted with La(III). Experiments involving other radiometals were carried out using similar procedures.Complex formation of Compound 8 with La (III)

[0730] a) 4uL each of lOmM stock solution of Compound 8 and lOmM of LaC13 were added to 192uL of lOmM NaOAc buffer (pH= 4.7) to obtain 200uL of the mixture at 200uM.

[0731] b) The solution was incubated at 25°C for at least 1 hour.

[0732] c) UV / HPLC was used to confirm full conversion to the complex.

[0733] DTPA challenge and analysis

[0734] a) lOOuL each of Compound 8 (La) complex mixture and DTPA (lOOmM stock) were added to 800uL of H2O to obtain ImL of 20uM Compound 8(La) complex and lOmM DTPA (500x excess).

[0735] b) Inject 50uL to HPLC analysis at various time points (up to fifteen days).

[0736] c) Fractions of interest were collected and analyzed by ICP-MS to confirm La(III) presence.

[0737] HPLC conditions:

[0738] Mobile phase A: lOmM NaOAc, pH4.7

[0739] Mobile phase B: MeOH

[0740] Gradient: 5% to 95%B over 5min, 0.5min at 95%B, equilibrate to 5%B over 2.5min.

[0741] Flowrate: 0.65mL / min

[0742] Column: Restek Raptor Inert ARC-182.7um, 50 x 2.1mm

[0743] A stability study for compounds 8, 12, 13, and 15 with various metals was performed as described above and the summary of results is presented in Table 3 below.

[0744] Table 3:

[0745] A summary of stability study for the compounds of the present disclosure with La, Tb, Lu, Cu, and Sc

[0746] Compound Cu In La Lu Sc Tb

[0747] 8 15 days 15 days 15 days 24 h 12h 15 days 12 3 h Not 4 h 2 days Not stable Not performed performed 13 1 day 1 day 3 h 1 day Not 1 day performed

[0748] 15 3 h 1 day l h 1 day Not stable 1 day

[0749]

[0750] EXAMPLE 19

[0751] MINIMAL CONCENTRATION FOR COMPLEX FORMATION

[0752] A concentration-dependent metal-chelator complex formation was used to identify the minimum chelator concentration necessary for quantitative complexation with metals such as La, Lu, Tb, and others. High-performance liquid chromatography coupled with inductively coupled plasma mass spectrometry (HPLC / ICP / MS) was applied, where the complex was separated from free metal via HPLC, and ICP / MS enabled detection of the metals.

[0753] Serial dilution of compound 8:

[0754] A solution of 15 uL of 10 mM stock solution of the compound 8 was added to 1485 pL of 20 mM NaOAc with a pH=6 to obtain 1.5 mL at 100 pM. A serial dilute from 100 pM was done to obtain solutions with concentrations at 30 pM, 9pM, 2.7 uM, 810 nM, 273nM, 73 nM, and 22 nM.

[0755] Procedure for the reaction:

[0756] A 2.5 uL solution of La(III) (100 nM) was added to 250 uL of ligand at a given concentration.

[0757] The reaction was then incubated for 30 minutes at 25°C. A 30 pL sample was then injected using an Agilent 1260 Infinity Series HPLC coupled with an Agilent 7850 (G8422A) ICP-MS. The following HPLC conditions were used:

[0758] Mobile phase A: 0.01% formic acid, H2O

[0759] bMobile phase B: 0.01% formic acid, MeOH

[0760] Gradient: 4%B for 0.2min, 4% to 85%B over 3.5min, 0.5min at 85%B, equilibrate to 1%B over 1%B for 3.8min.

[0761] Flowrate: 0.5mL / min

[0762] Column: Restek Raptor Inert ARC-182.7um, 50 x 2.1mm

[0763] This procedure was repeated for each concentration tested and HPLC / ICP-MS data are presented in FIG. 3.

[0764] The procedure was repeated for DOTA incubated at 25°C as well as at 95°C.

[0765] As illustrated in FIG. 2A, Compound 8-La(III) exhibited a 50% complexation concentration that was substantially lower than that of DOTA, even when DOTA was assessed at elevatedtemperature (95 °C). These results indicate that compound 8 demonstrates greater efficiency in metal complexation and would enable higher molar activity when used with a radioactive metal. Minimum chelator concentrations for quantitative complexation with Lu and Tb were assessed using similar methods, with results shown in FIG. 2B and 2C.

[0766] EXAMPLE 20

[0767] COMPLEXATION RATE OF COMPOUNDS III-l - III-10 WITH VARIOUS METALS IN COMPARISON WITH ACROPA

[0768] A complexation study for compounds III-l - III-10 and Macropa with other metals was performed and the summary of results is presented in Table 4 below.

[0769] Table 4:

[0770] A summary of complexation study for the compounds of the present disclosure with La, Tb,

[0771] Lu, Cu, and Sc compared to Macropa metal complexes Compound La Tb Lu Cu Sc

[0772] III-l + N / C N / C ++ ++

[0773] III-2 + N / C N / C + ++

[0774] III-3 N / C N / C N / C +++ N / C

[0775] III-4 N / C N / C N / C ++ N / C

[0776] III-5 + N / C N / C + N / C

[0777] III-6 + N / C N / C + N / C

[0778] III-7 + N / C N / C + N / C

[0779] III-8 + N / C N / C + ++

[0780] III-9 ++ N / C N / C ++ ++

[0781] ID-10 + N / C N / C + ++

[0782]

[0783] +++: Faster complexation rate to Macropa

[0784] ++: Similar complexation rate to Macropa

[0785] +: Slower complexation rate to Macropa

[0786] N / C: No complex formed

Claims

CLAIMSWe claim:

1. A pyrrolidine containing chelating agent compound comprising a polydentate chelating ligand having the formula comprising one or more bidentate donor moieties having the chemical structure -X-P-Y- where X and Y are each independently oxygen or nitrogen and P is a pyrrolidine covalently bonded to X and Y, and wherein the pyrrolidine containing chelating agent chelates one or more metal or nonmetal ions.

2. The compound of claim 1, wherein a chelating agent has a chemical structurecharacterized by incorporating a pyrrolidine substructure selected fromin place of an ethylenediamine( ) substructure in a chelating agent. One or more pyrrolidine type bidentate moieties as described above can be introduced in a chelator.

3. The compound of claim 1 wherein a chelating agent has a chemical structurecharacterized by incorporating a pyrrolidine substructure selected fromin place of an aminoethanol () substructure in a chelating agent. One or more pyrrolidine type bidentate moieties as described above can be introduced in a chelator.

4. The compound in claim 1 wherein a chelating agent has a chemical structurecharacterized by incorporating a pyrrolidine substructure selected fromorin place of an ethylene glycol () substructure in a chelating agent. One or more pyrrolidine type bidentate moieties as described above can be introduced in a chelator.

5. The compound of claim 1, wherein the one or more bidentate donor moieties are independently selected from the group consisting of:

6. The compound of claim 5, wherein the pyrrolidine containing chelating agent comprises a single pyrrolidine bidentate donor moiety.

7. The compound of claim 1, comprising one or more of the following:a. at least one modified ethylene glycol moiety wherein the ethyl moiety of the modified ethylene glycol moiety is replaced by a pyrrolidine; orb. at least one ethylenediamine replaced by at least one modified ethylenediamine moiety wherein the ethyl moiety of the modified ethylenediamine moiety is replaced by a pyrrolidine; andc. at least one ethanolamine replaced by at least one modified ethanolamine moiety wherein the ethyl moiety of the modified ethanolamine moiety is replaced by a pyrrolidine.

8. The compound of claim 1, wherein the compound is a compound of Formula (I)(I),whereinXi isN;X2isN;Z1 is C(R24)(R25C);Ri, R2, and Re is a linker to a vector, a chelating pendant moiety, or hydrogen;R24 andR25c are each independently, NHRi, CO2R1, a linker to a vector, a chelating pendant moiety, or hydrogen;25a, R25b, R25d, and R25e are each independently hydrogen; or C1-C4 alkyl; and the one or more chelating pendant moieties are each independently carboxy, C1-C20 alkyl optionally substituted with one or more carboxy, amino or sulfur, or aryl or heteroaryl each optionally substituted with one or more carboxy, sulfur or amino or optionally substituted with C1-C4 alkyl optionally substituted with one or more carboxy, sulfur or amino,wherein each chelating pendant moiety is independently selected from the group consisting of but not limited to: -COOH, -(C1-C4 alkyl)COOH, -(C1-C4 alkyl)COOH,-(C1-C4 alkyl)-S-CH3, -(C1-C4 alkyl)-Py-COOH (wherein Py is pyridinyl), and -(C1-C4 alkyl)-Ph-COOH (wherein Ph in phenyl).

9. The compound of any one of claims 1-8, wherein the compound is a compound of Formula (I A) or Formula (IB):(IA), (IB), wherein Ri, R2, R3, and R7 are each independently a linker to a vector, a chelating pendant moiety, or hydrogen.

10. The compound of claim 9, wherein R2 and R7 are each chelating pendant moiety selected from the group consisting of but not limited to: -COOH, -(CH2)COOH:-(CH2)2COOH,11. The compound of claim 10, wherein the compound is a compound of formula (IA).

12. The compound of claim 10, wherein the compound is a compound of formula (IB).

13. The compound of claim 1, wherein the compound is a compound of F ormula (II)Z I N-fyV> *2(II),wherein:Xi is NR2 or 0;XiisNR? orO;Y 1 is NR3 or 0Y2 is NRe or 0;Ri, R2, R3 Re and R7 are each independently a linker to a vector, a chelating pendant moiety, or hydrogen;Z is - (CH2)2-X3-[(CH2)2-Y3]n-(CH2)2-, wherein:X3 is NR4, 0, methylene or a direct bond;Y3 is NR5, or 0; andn is 0 or 1; andR4 and R5 are each independently a linker to a vector, a chelating pendant moiety, or hydrogen.

14. The compound of claim 13 where ethylene moiety is replaced by a pyrrolidine moiety.

15. The compound of claim 12, whereinRi is hydrogen, a protecting group, a linker to a vector, C1-C4 alkyl optionally substituted with carboxy, amino or C1-C4 alkylamino; andR2, R3 and R4 are each independently C1-C4 alkyl optionally substituted with carboxy, amino or C1-C4 alkylamino.

16. The compound of any one of claims 13-14, wherein the compound is a compound selected from the group consisting of Formula IIA and Formula IIB:Z T N-fy Zx[ N-fyY2Y2X2(IIA), (IIB).

17. The compound of any one of claims 13-16, wherein Xi and X2 are each oxygen.

18. The chelating agent any one of claims 13-16, wherein Y i is NR3, and Y2 is NRe and R3 and Rfi are each a chelating pendant moiety independently selected from the group consisting of: -COOH, -(C1-C4 alkyl)COOH, -(C1-C4 alkyl)COOH, -(C1-C4 alkyl)-S-CH3, -(C1-C4 alkyl)-Py-COOH (wherein Py is pyridinyl), and -(C1-C4 alkyl)-Ph-COOH (wherein Ph in phenyl).

19. The compound of any one of claims 13-16, wherein Z is -(CH2)2-, -(CH2)3-, — (CH2)2-O— (CH2)2-, — (CH2)2-NH— (CH2)2-, or-(CH2)2-O-(CH2)2-O-(CH2)2-.

20. The compound of claim 1, wherein the compound is selected from the group containing:The compound of claim 1, wherein the compound is a compound of Formula (III):(HI)wherein:each ring A and ring B is independently a 6-10 membered aryl or a 5-10 membered heteroaryl, wherein each of ring A and ring B is optionally substituted with one or more substituents independently selected from the group consisting of halo, alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, -OR13, -SR13, -(CH2)PCOORi3, -OC(O)R13, -N(RB)2, -CON(R13)2, -NO2, -CN, -OC(O)N(R13)2, and X;Zi and Z2 are independently -(C(Ri2)2)m-, or -(CH2)n-C(Ri2)(X)-(CH2)n-;each X is independently -Li-Rn;each n is independently 0, 1, 2, 3, 4, or 5;each m is independently 1, 2, 3, 4 or 5;each p is independently 0 or 1;Li is absent or a linker;R11 is a nucleophilic moiety or an electrophilic moiety, or Rn comprises a targeting ligand;each R12 is independently hydrogen, alkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl;each R13 is independently hydrogen or alkyl;provided the chelator comprises at least one X and when X is present on ring A or ring B, then Li is a linker or R12 is not hydrogen; andRi is a linker to a vector, a chelating pendant moiety, or a group modulating physical chemical properties.

22. The compound of claim 21, wherein:Zi is -CH2- or -CHR12-;Z2 is -L1-R11 or -CR12-;ring A and ring B are each independently phenyl, or a 5-6 member heteroaryl optionally substituted with one or more carboxy;Li is absent or a linker;R11 is a nucleophilic moiety or an electrophilic moiety, or R11 comprises a targeting ligand, andeach R12 is independently hydrogen, -methyl, or ethyl, provided at least one R12 is methyl or ethyl.

23. The compound of claim 22, wherein ring A and ring B are each independently phenyl, diazole, pyridine, or pyrimidine, optionally substituted with one or more carboxy.

24. The compound of claim 23, wherein ring A and ring B are each25. The compound of any one of claims 21-24, wherein Zi is -CH2- or -CHR12- and Z2 is -CHLi-Rn.

26. The compound of any one of claims 21-25, wherein Li in Formula (III) is selected from the group consisting of:wherein n is an integer of 0 to 10, preferably an integer of 1 to 4, and m is an integer of 0 to 12, preferably an integer of 0 to 6.

27. The compound of claim 1, wherein the compound is28. The compound of claim 1, wherein the compound is29. The compound of claim 1, wherein the compound is30. The compound of claim 1, wherein the compound is31. The compound of claim 1, wherein the compound isHOHO32. The compound of claim 1, wherein the compound comprises a deprotected version of33. The compound of claim 1, wherein the compound comprises a deprotected version of34. The compound of claim 1, wherein the compound comprises a deprotected version of35. The compound of claim 1, wherein the compound comprises a deprotected version of36. The compound of claim 1, wherein the compound comprises a deprotected version of37. The compound of claim 1, wherein the compound comprises a deprotected version of38. The compound of claim 1, wherein the compound comprises a deprotected version of39. The compound of claim 1, wherein the compound comprises a deprotected version of40. The compound of claim 1, wherein the compound comprises a deprotected version of41. The compound of claim 1, wherein the compound comprises a deprotected version of42. The compound of claim 1, wherein the compound comprises:wherein * indicates either stereochemical orientation at each chiral center.

43. The compound of claim 1, wherein the compound comprisesOH44. The compound of claim 1, wherein the compound comprisesOOH45. The compound of any one of claims 1-44, further comprising a chelated metal.

46. The compound of claim 45, wherein the chelated metal is a metal element having atomic number ranging between 20 and 31, 39, 42, 43, 44, 49, and between 57 and 83.

47. The compound of claim 46, wherein the chelated metal is a radioisotope.

48. The compound of claim 47, wherein the radioisotope is selected from the group consisting of:203Pb,67Ga,68Ga,72As,111In,113In,90Y,97Ru,62Cu,64Cu,52Fe,52mMn,140La,175Yb,153Sm,166Ho,149Pm,177Lu,142Pr,159Gd,212Bi,47Sc,149Pm,67Cu,17tAg,199Au,161Tb and51Cr; or89Zr,44Sc,111In,90Y,66Ga,67Ga,68Ga,177Lu,99mTc,61Cu,62Cu,64Cu,67Cu,149Tb,152Tb,155Tb,161Tb,153Gd,155Gd,157Gd,213Bi,225Ac,230U,223Ra,165Er and52Fe; or111In,90Y,68Ga,64Cu,153Gd,155Gd,213Bi,225Ac,52Fe, and177Lu.

49. The compound of claim 47, wherein the radioisotope comprises one or more of177Lu,225Ac,211At,67Cu,161Tb,67Ga,203Pb,223Ra, and212Pb.

50. The compound of claim 47, wherein the radioisotope comprises177Lu or225Ac.

51. A pharmaceutical composition comprising a compound of any one of claims 1 -50, or a salt thereof, and a pharmaceutically acceptable carrier.

52. The composition of claim 51, wherein the composition further comprises an excipient.

53. A pyrrolidine containing chelating compound comprising one or more bidentate s-X— p— Y- - donor moieties having a chemical structure of5 5, wherein X and Y are each independently oxygen or nitrogen and P is a pyrrolidine covalently bonded to X and Y, and wherein the pyrrolidine containing chelating compound chelates to one or more metal or nonmetal ions.

54. A chelating ligand, comprising:one or more bidentate donor moieties having, provided that at least one of theelectron donor unit iswherein:the one or more bidentate donor moieties are covalently bonded by C1-C4 alkyl; R10is, each independently, H, a nitrogen protecting group, a linker to a vector, a chelating pendant moiety, or a vector;R12and R13are, each independently, H, Ci-Ce alkyl optionally substituted with aryl or heteroaryl, which are optionally substituted with -COOR16, -P(O)(OR16)2, or Ci-Ce heteroalkyl;Ri4a, R14b, R15a, R15b, and R16are, each independently, H, Ci-Cg alkyl, or Ci-Ce heteroalkyl; andthe chelating ligand has from 4 to 12 coordination sites for one or more metals or nonmetal ions to bind to the one or more bidentate donor moieties.

55. The chelating ligand of claim 54, wherein one or more bidentate donor moieties56. The chelating ligand of any one of claims 54-55, wherein R14a, R14b, R15a, R15b, and R16are H.

57. The chelating ligand of any one of claims 54-56, wherein the one or more bidentate donor moieties are independently selected from the group consisting of:

58. The chelating ligand of any one of claims 54-57, wherein the nitrogen protectinggroup of R10is BOC, Fmoc, Cbz, Ac, trifluoroacetamide, Bn, '" T", ■''T'or^ v Ou59. The chelating ligand of any one of claims 54-58, wherein R12and R13are H or Bn.

60. The chelating ligand of any one of claims 54-59, wherein Ci-Ce heteroalkyl of R12and R13is -CH2C(=O)OH, -(CH2)2C(=O)OH, -CH2C(=O)OtBu, -C(=O)OH, -C(=O)OtBu,61. The chelating ligand of any one of claims 54-60, wherein the one or more metals comprises203Pb,67Ga,68Ga,72As,111In,113In,90Y,97Ru,62Cu,64Cu,52Fe,52mMn,140La,175Yb,153Sm,166Ho,149Pm,177LU,142Pr,159Gd,212Bi,47Sc,149Pm,67Cu,lJtAg,199Au,161Tb or51Cr.

62. The chelating ligand of any one of claims 54-60, wherein the one or more metals comprises89Zr,44Sc,111In,90Y,66Ga,67Ga,68Ga,177Lu,99mTc,61Cu,62Cu,64Cu,67Cu,149Tb,152Tb,155Tb,161Tb,153Gd,155Gd,157Gd,213Bi,225Ac,230U,223Ra,165Er,or52Fe.

63. The chelating ligand of any one of claims 54-60, wherein the one or more metals comprises111In,90Y,68Ga,64Cu,153Gd,155Gd,213Bi,225Ac,52Fe, or177Lu.

64. The chelating ligand of any one of claims 54-60, wherein the one or more metals comprises177Lu,225Ac,211At,67Cu,161Tb,67Ga,203Pb,223Ra, or212Pb.

65. The chelating ligand of any one of claims 54-60, wherein the one or more metals comprises177Lu or225Ac.

66. The chelating ligand of claim 54, wherein the chelating ligand has one of the following structures:

67. The chelating ligand of claim 53, wherein the chelating ligand has a structure of Formula (III):— 0 0 —(AY-Z^N N~Z2~(B)S?(III)wherein:each ring A and ring B is independently a 6-10 membered aryl or a 5-10 membered heteroaryl, wherein each of ring A and ring B is optionally substituted with one or more substituents independently selected from the group consisting of halo, alkyl, alkenyl, cycloalkyl, cycloalkenyl, aryl, heterocyclyl, heteroaryl, -OR13, -SR13, -(CH2)pCOOR13, -OC(O)R13, -N(R13)2, -CON(R13)2, -NO2, -CN, and -OC(O)N(RI3)2;Zi and Z2 are independently -(C(Ri2)2)m-, or-(CH2)n-C(Ri2)(X)-(CH2)n-;each n is independently 0, 1, 2, 3, 4, or 5;each m is independently 1, 2, 3, 4 or 5;each p is independently 0 or 1;each R12 is independently hydrogen, alkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl;each R13 is independently hydrogen or alkyl; andRi is H, a linker to a vector, a chelating pendant moiety, a group modulating physical chemical properties, or a nitrogen protecting group.

68. The chelating ligand of claim 67, wherein:Zi is -CH2-;Z2 is -CH2-;ring A and ring B are each a 6 member heteroaryl substituted with -COOH or -COOH and -N(CH3)2; andRi is a nitrogen protecting group.

69. The chelating ligand of claim 67 or 68, wherein the one or more metals comprises203Pb,67Ga,68Ga,72As,111In,113In,90Y,97Ru,62Cu,64Cu,52Fe,52mMn,140La,175Yb,153Sm,166Ho,149Pm,177LU,142Pr,159Gd,212Bi,47Sc,149Pm,67Cu,177Ag,199Au,161Tb or51Cr.

70. The chelating ligand of any one of claims 67-69, wherein the one or more metals comprises89Zr,44Sc,111In,90Y,66Ga,67Ga,68Ga,177Lu,99mTc,61Cu,62Cu,64Cu,67Cu,149Tb,152Tb,155Tb,161Tb,153Gd,155Gd,157Gd,213Bi,225Ac,230U,223Ra,165Er,or52Fe.

71. The chelating ligand of any one of claims 67-70, wherein the one or more metals comprises111In,90Y,68Ga,64Cu,153Gd,155Gd,213Bi,225Ac,52Fe, or177Lu.

72. The chelating ligand of any one of claims 67-71, wherein the one or more metals comprises177Lu,225Ac,211At,67Cu,161Tb,67Ga,203Pb,223Ra, or212Pb.

73. The chelating ligand of any one of claims 67-72, wherein the one or more metals comprises177Lu or225Ac.

74. The compound of claim 67 or 68, wherein the compound has one of the following structures:

75. A pharmaceutical composition comprising the chelating ligand of any one of claims 54-74, or a salt thereof, and a pharmaceutically acceptable carrier.

76. A composition comprising the chelating ligand of any one of claims 54-74 and a therapeutic agent.

77. A method for administering a therapeutic agent to a patient in need thereof, the method comprising preparing or providing the composition of claim 75 or 76, and administering the composition to the patient.

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