2,4-dihyroxy-5-isopropylphenyl derivatives and pharmaceutical compositions thereof effective as extracellular HSP90, (EHSP90)-targeted radiopharmaceuticals useful in the treatment of cancer

Abstract

Radiopharmaceutical compounds of Formula (I) comprising a targeting moiety that specifically binds to Hsp90, in particular, extracellular Hsp90, a unique linker and a chelator. Also disclosed are pharmaceutical compositions and methods of treating cancer with the same.

Description

[0001] ,4-DIHYROXY-5-ISOPROPYLPHENYL DERIVATIVES AND PHARMACEUTICAL COMPOSITIONS THEREOF EFFECTIVE AS EXTRACELLULAR HSP90, (EHSP90)-TARGETED RADIOPHARMACEUTICALS USEFUL IN THE TREATMENT OF CANCER

[0002] BACKGROUND

[0003] [1] Heat shock protein 90 (HSP90) is a molecular chaperone that regulates protein folding to ensure correct conformation and translocation and to avoid protein aggregation. Many oncogenic proteins are HSP90 client proteins, such as epidermal growth factor receptor (EGFR) mutant, cyclin dependent kinase 4 (CDK4), hypoxia-inducible factor (HIF)-la, and matrix metallopeptidase 2 (MMP2).

[0004] [2] There is a fraction of HSP90 identified at the surface of a number of cell types, i.e., extracellular HSP90 (eHSP90), which can be a useful tumor antigen for eliciting a host immune response. Studies show that eHSP90 plays a crucial role in maintaining oncogenic protein homoeostasis and participates in the invasion and metastatic processes of various cancers including breast cancer, which make eHSP90 an attractive target for developing cancer therapy.

[0005] [3] HSP90 inhibition has been shown to have a significant direct impact on cell cycle and DNA repair mechanisms, thus offering great promise in the treatment of a wide variety of solid and hematological malignancies. However, early clinical trials have demonstrated that certain HSP90 inhibitors exhibited limited efficacy and various side effects.

[0006] [4] Therefore, there is a need for improved treatment of cancers with good efficacy and acceptable safety profiles by targeting certain proteins, e.g., eHSP90.

[0007] SUMMARY

[0008] [5] The present disclosure encompasses the insight that certain radiopharmaceuticals comprising a targeting moiety that specifically binds to HSP90, in particular, eHSP90, can be effective as HSP90 radioligand therapy (RLT) for treating cancers. Radioactive decay can cause direct physical damage (such as single or double-stranded DNA breaks) or indirect damage (such as by-stander or crossfire effects) to the biomolecules that constitute a cell. Drugs that deliver radionuclides to cancer cells, i.e., radiopharmaceuticals, provide a mechanism to generate DNA damage with anti-cancer therapeutic effect. The present disclosure provides certain radiopharmaceuticals, specifically, small molecule-based radiopharmaceuticals targeting HSP90- overexpressing tumors and using actinium-225 (225Ac), lutetium-177 (177Lu) or other suitable therapeutic radionuclides to target cancer cells to treat or ameliorate cancers such as lung cancer, sarcoma, pancreatic cancer, breast cancer, or colon cancer.

[0009] [6] In one aspect, the present disclosure provides compounds of Formula I, or a pharmaceutically acceptable salt thereof:

[0010] (I), wherein G is Ci-s alkyl, Ci-s heteroalkyl, C3-8 cycloalkyl, C2-8 heterocycloalkyl, aryl, or heteroaryl;

[0011] Q1and Q2each are, independently, N or C;

[0012] Q3and Q4each are, independently, NRaor O, in which Rais absent, H, C1-3 alkyl, or (C=O)Ci-3alkyl;

[0013] R is H or C1-6 alkyl;

[0014] L is a linker selected from the group consisting of C1-20 alkyl, C1-20 heteroalkyl, C3-10 cycloalkyl, C1-10 heterocycloalkyl, aryl, heteroaryl, C=O, (C=O)NR1, (C=S)NR1, NR1(C=O)NR1, NR1(C=S)NR1, and a combination thereof, in which each R1independently is H or C1-3 alkyl; or L is a hydrophilic space linker comprising one or more cyclic or acyclic polyhydroxy groups; and

[0015] W is a chelator, wherein the compound binds to HSP90, wherein the compound optionally further comprises a radionuclide chelated by the chelator thereof, and wherein each of alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is optionally substituted with one or more moieties selected from the group consisting of OH, halo, alkoxy, amino, oxo, C1-3 haloalkyl, C1-3 alkyl-OH, C1-3 alkyl-NH2, (C=O)Ci-3 alkyl, COOH, and CONH2. [7] In some embodiments, the compounds of this disclosure specifically bind to extracellular

[0016] HSP90 (eHSP90).

[0017] [8] In some embodiments, the compounds of this disclosure have the structure of Formula II:

[0018] (II), wherein X is CH or N;

[0019] Y is O or NR2, in which R2is H, C1-3 alkyl, (C=O)Ci-3 alkyl,

[0020] R is H or C1-6 alkyl;

[0021] L1is absent or C1-5 alkyl;

[0022] L2is Ci -20 alkyl, Ci -20 heteroalkyl, C3-10 cycloalkyl, C1-10 heterocycloalkyl, aryl, or heteroaryl; n is an integer of 1-5 (inclusive);

[0023] W is a chelator selected from the group consisting of DOTA, DOT AGA, NOTA, NOD AGA, NODASA, DTP A, TETA, EDTA, TRITA, CDTA, and DFO; and

[0024] Z1and Z2each are, independently, absent or a moiety selected from the group consisting of C=O, (C=O)NR3, NR3(C=O)NR3, NR3(C=S)NR3, (CH2CH2)NR3, an amino acid unit, and a combination thereof, in which each of R3, independently, is H or C1-3 alkyl.

[0025] [9] In some embodiments, referring to Formula II, the compounds of this disclosure have the following features:

[0026] X is CH, Y is NCOCH3;

[0027] L1is absent or C1-3 alkyl;

[0028] L2is C2-10 heteroalkyl or C2.2o polyethylene glycol; n is 1 or 2; and

[0029] Z1and Z2each are, independently, absent, or a moiety selected from the group consisting of (C=O)NH, NH(C=O)NH, (CH2CH2)NH, an amino acid unit, and a combination thereof wherein at least one of Z1and Z2is present.

[0010] In certain embodiments, L1is absent or -CH2CH2-; L2is C2-10 heteroalkyl or Ce-s polyethylene glycol; n is 1; Z1is absent, -(C=O)NH-, -NH(C=O)-, NH(C=O)NH, or (CH2CH2)NH; Z2is (C=O)NH, NH(C=O)NH, (CH2CH2)NH, or a combination thereof; and W is DOTA or DOTAGA.

[0030]

[0011] In some embodiments, at least one of Z1and Z2is an amino acid unit. In certain embodiments, the amino acid unit is formed from aspartic acid (Asp), glutamic acid (Glu), 2,4- diaminobutyric acid (Dab), 2,3-diaminopropionic acid (Dap), lysine (Lys), or arginine (Arg).

[0031]

[0012] In some embodiments, the compounds of this disclosure have the structure of Formula III: wherein X is CH or N;

[0032] Y is O or NR2, in which R2is H, C1-3 alkyl, (C=O)Ci-3alkyl, ,

[0033] R is H or C1-6 alkyl;

[0034] L1is absent or C1-3 alkyl; Z1and Z2each are, independently, absent or a moiety selected from the group consisting of C=O, (C=O)NR3, NR3(C=O)NR3, (CH2CH2)NR3, an amino acid unit, and a combination thereof, in which each of R3, independently, is H or C1-3 alkyl; n is an integer of 1-5 (inclusive);

[0035] W is a chelator selected from the group consisting of DOTA, DOT AGA, NOTA, NOD AGA, NODASA, DTP A, TETA, EDTA, TRITA, CDTA, and DFO;

[0036] X is absent or C1-5 alkyl;

[0037] A is absent, C=O, CONR3, heterocycle, or heterocycle-Ci-3 alkyl, R3being H or C1-5 alkyl;

[0038] Y is absent, C1-5 alkyl, or C1-10 heteroalkyl; and

[0039] B is a cyclic or acyclic polyhydroxy group, wherein the compound comprises a hydrophilic space linker comprising one or more cyclic or acyclic polyhydroxy groups.

[0040]

[0013] In some embodiments, referring to Formula III, L1, Z1, and Z2are absent, and n is 1-3.

[0041]

[0014] In some embodiments, referring to Formula III, Z1is (CEECE^NH or (C=O)NH(CH2CH2)NH, Z2is absent, and n is 1-3.

[0042]

[0015] In some embodiments, referring to Formula III, at least one of Z1and Z2is an amino acid unit. The amino acid unit can be formed from aspartic acid (Asp), glutamic acid (Glu), 2,4- diaminobutyric acid (Dab), 2,3-diaminopropionic acid (Dap), lysine (Lys), or arginine (Arg). polyethylene glycol, wherein indicates the attachment point to X, and “#” indicates the attachment point to Y.

[0043]

[0017] In some embodiments, referring to Formula III, B is a cyclic polyhydroxy group having the structure wherein “#” indicates the attachment point to Y, r is 1-2, s is 3-4, and each R, independently, is OH, -CH2OH, -CO2H, -CONH2, NH2, or NHCOCH3, wherein B comprises at least three OH groups.

[0044]

[0018] In some embodiments, referring to Formula III, B is an acyclic polyhydroxy group having the structure wherein “#” indicates the attachment point to Y; t is 2-5; each

[0045] R, independently, is OH, -CH2OH, -CO2H, -CONH2, NH2, or NHCOCH3; and Raand Rbeach are H or Raand Rbtogether form C=O, wherein B comprises at least three OH groups.

[0046]

[0019] In some embodiments, the compounds of this disclosure have the structure of Formula IV:

[0047] (IV), wherein X is CH or N;

[0048] Y is O or NR2, in which R2is H, C1-3 alkyl, (C=O)Ci-3alkyl, ,

[0049] R is H or C1-6 alkyl;

[0050] L1is absent or C1-3 alkyl;

[0051] Z1is absent or a moiety selected from the group consisting of C=O, (C=O)NR3, NR3(C=O)NR3, (CH2CH2)NR3, an amino acid unit, and a combination thereof, in which each of R3, independently, is H or C1-3 alkyl; m is an integer of 1-5 (inclusive), p is an integer of 1-4 (inclusive);

[0052] W is a chelator selected from the group consisting of DOTA, DOT AGA, NOTA, NOD AGA, NODASA, DTP A, TETA, EDTA, TRITA, CDTA, and DFO; and R in each occurrence, independently, is OH, -CH2OH, -CO2H, -CONH2, NH2, or NHCOCH3, wherein the compound comprises a hydrophilic space linker comprising at least three OH groups.

[0020] In some embodiments, referring to Formula IV, X is CH, Y is (C=O)CH3, m is 3-4, p is 1-2, and R is OH or -CH2OH.

[0053]

[0021] In some embodiments, the compounds of this disclosure have a structure selected from the group consisting of: wherein each of the variables G, L, and W is as defined for Formula I set forth above.

[0022] In some embodiments, the compounds of Formulas I-IV feature that each compound comprises a radionuclide selected from the group consisting of47Sc,55Co,60Cu,61Cu,62Cu,64Cu,67Cu,66Ga,67Ga,68Ga,82Rb,86Y,87Y,89Zr,90Y,97Ru, "Tc, "mTc,105Rh,109Pd,n iIn,117mSn,134Ce,134La,149Pm,149Tb,153Sm,166Ho,177Lu,186Re,188Re,198Au,199AU,201T1,203Pb,211At,212Pb,212Bi213Bi,223Ra,225Ac,227Th, and229Th.

[0054]

[0023] In certain embodiments, the radionuclide is selected from the group consisting of68Ga,89Zr,90Y,i nIn,177LU, and225AC.

[0055]

[0024] In certain embodiments, the radionuclide is177Lu or225Ac.

[0056]

[0025] In another aspect, the present disclosure also covers a pharmaceutical composition comprising one of the compounds set forth above and a pharmaceutically acceptable excipient.

[0057]

[0026] Still within the scope of this disclosure is a method of treating cancer or radiation treatment planning, the method comprising administering to a subject in need thereof one of the compounds set forth above or the above-described pharmaceutical composition.

[0058]

[0027] In some embodiments, the method of treating cancer comprises administering to the subject in need thereof a first dose of one of the compounds or the composition described above in an amount effective for radiation treatment planning, followed by administering subsequent doses of one of the compounds or the composition described above in a therapeutically effective amount.

[0059]

[0028] In some embodiments, the cancer is small-cell lung cancer, non-small-cell lung cancer, sarcoma, pancreatic cancer, breast cancer, or colon cancer.

[0060]

[0029] This disclosure also relates to any one of the compounds provided herein for use in a method of treating cancer.

[0061]

[0030] Further covered within this disclosure is use of any of the compounds provided herein in the manufacture of a medicament for the treatment of cancer.

[0062] BRIEF DESCRIPTION OF THE DRAWINGS

[0063]

[0031] Figure 1. In vitro Binding of177Lu-Compound A to HSP90. Plots represent average specific binding (± SD, n = 2) to either HSP90 Alpha (dotted line) or HSP90 Beta (solid line) recombinant protein.

[0032] Figure 2. In vitro Binding of177Lu-Compound B to HSP90. Plots represent average specific binding (± SD, n = 2) to either HSP90 Alpha (dotted line) or HSP90 Beta (solid line) recombinant protein.

[0064] DETAILED DESCRIPTION

[0065]

[0033] The present disclosure relates to radiopharmaceutical compounds comprising a targeting moiety that specifically binds to HSP90, in particular, extracellular HSP90 (eHSP90).

[0066]

[0034] Radio-labelled targeting molecules (also known as radiopharmaceuticals) are designed to target a protein or receptor (e.g., eHSP90) that is upregulated in a disease state and / or specific to diseased cells (e.g., tumor cells) to deliver a radioactive payload to damage and kill cells of interest. Radiopharmaceuticals targeting eHSP90 provided in this disclosure can be used for treating various cancers including, but not limited to, small-cell lung cancer, non-small-cell lung cancer, sarcoma, pancreatic cancer, breast cancer, and colon cancer.

[0067] Definitions

[0068] Chemical Terms

[0069]

[0035] The term “alkyl,” as used herein, is inclusive of both straight chain and branched chain saturated groups from 1 to 20 carbons represented as C1-20 alkyl (e.g., C1-6 alkyl having 1 to 6 carbons, C1-8 alkyl having 1 to 8 carbons, C1-10 alkyl having 1 to 10 carbons, or C1-12 alkyl having 1 to 12 carbons), unless otherwise specified. Alkyl groups are exemplified by methyl, ethyl, n- and iso-propyl, n-, sec-, iso- and tert-butyl, neopentyl, and the like, and may be optionally substituted with one, two, three, or, in the case of alkyl groups of two carbons or more, four substituents independently selected from the group consisting of: (1) C1-6 alkoxy; (2) C1-6 alkyl sulfinyl; (3) amino, as defined herein (e.g., unsubstituted amino (i.e., -NH2) or a substituted amino (i.e., -N(RN1)2, where RN1is as defined for amino); (4) Ce-io aryl-Ci-6 alkoxy; (5) azido;

[0070] (6) halo; (7) (C2-9 heterocyclyl)oxy; (8) hydroxy, optionally substituted with an ( -protecting group; (9) nitro; (10) oxo (e.g., carboxyaldehyde or acyl); (11) C1-7 spirocyclyl; (12) thioalkoxy;

[0071] (13) thiol; (14) -CC>2RA, optionally substituted with an (9-protecting group and where RAis selected from the group consisting of (a) C1-20 alkyl (e.g., C1-6 alkyl), (b) C2-20 alkenyl (e.g., C2-6 alkenyl), (c) Ce-io aryl, (d) hydrogen, (e) C1-6 alk-Ce-io aryl, (f) amino-Ci-20 alkyl, (g) polyethylene glycol of -(CH2)s2(OCH2CH2)si(CH2)s3OR’, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and R’ is H or C1-20 alkyl, and (h) amino-polyethylene glycol of -NRN1(CH2)S2(CH2CH2O)SI(CH2)S3NRN1, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and each RN1is, independently, hydrogen or optionally substituted C1-6 alkyl;

[0072] (15) -C(O)NRBRc, where each of RBand Rcis, independently, selected from the group consisting of (a) hydrogen, (b) C1-6 alkyl, (c) Ce-io aryl, and (d) C1-6 alk-Ce-io aryl; (16) -SO2RD’, where RDis selected from the group consisting of (a) C1-6 alkyl, (b) Ce-io aryl, (c) C1-6 alk-Ce-io aryl, and (d) hydroxy; (17) -SC>2NRERF, where each of REand REis, independently, selected from the group consisting of (a) hydrogen, (b) C1-6 alkyl, (c) Ce-io aryl and (d) C1-6 alk-Ce-io aryl;

[0073] (18) -C(O)RG, where RGis selected from the group consisting of (a) C1-20 alkyl (e.g., C1-6 alkyl), (b) C2-20 alkenyl (e.g., C2-6 alkenyl), (c) Ce-io aryl, (d) hydrogen, (e) C1-6 alk-Ce-io aryl, (f) amino- C1-20 alkyl, (g) polyethylene glycol of -(CH2)s2(OCH2CH2)si(CH2)s3OR’, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and R’ is H or C1-20 alkyl, and (h) amino-polyethylene glycol of -NRN1(CH2)S2(CH2CH2O)SI(CH2)S3NRN1, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and each RN1is, independently, hydrogen or optionally substituted C1-6 alkyl;

[0074] (19) -NRHC(O)R* , wherein RHis selected from the group consisting of (al) hydrogen and (bl) C1-6 alkyl, and R1is selected from the group consisting of (a2) C1-20 alkyl (e.g., C1-6 alkyl), (b2) C2-20 alkenyl (e.g., C2-6 alkenyl), (c2) Ce-io aryl, (d2) hydrogen, (e2) C1-6 alk-Ce-io aryl, (f2) amino-Ci-20 alkyl, (g2) polyethylene glycol of -(CH2)s2(OCH2CH2)si(CH2)s3OR’, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and R’ is H or Ci -20 alkyl, and (h2) amino-polyethylene glycol of - NRN1(CH2)S2(CH2CH2O)SI(CH2)S3NRN1, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and each RN1is, independently, hydrogen or optionally substituted C1-6 alkyl; (20) -NRJC(O)ORK, wherein RJis selected from the group consisting of (al) hydrogen and (bl) Ci-6 alkyl, and RKis selected from the group consisting of (a2) Ci-20 alkyl (e.g., Ci-6 alkyl), (b2) C2-20 alkenyl (e.g., C2-6 alkenyl), (c2) Ce-io aryl, (d2) hydrogen, (e2) C1-6 alk-Ce-io aryl, (f2) amino-Ci-20 alkyl, (g2) polyethylene glycol of - (CH2)s2(OCH2CH2)si(CH2)s3OR’, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and R’ is H or C1-20 alkyl, and (h2) amino-polyethylene glycol of -NRN1(CH2)s2(CH2CH2O)si(CH2)s3NRN1, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and each RN1is, independently, hydrogen or optionally substituted C1-6 alkyl; and (21) amidine. In some embodiments, each of these groups can be further substituted as described herein. For example, the alkylene group of a Ci-alkaryl can be further substituted with an oxo group to afford the respective aryloyl substituent.

[0075]

[0036] The terms “alkylene”, “alkylidene”, and the prefix “alk-,” as used herein, represent a saturated divalent hydrocarbon group derived from a straight or branched chain saturated hydrocarbon by the removal of two hydrogen atoms, and is exemplified by methylene, ethylene, isopropylene, and the like. The terms “Cx-y alkyl,” “Cx-yalkylene,” “Cx.yalkylidene,” and the prefix “Cx-yalk-” represent alkyl or alkylene groups having between x and y carbons. Exemplary values for x are 1, 2, 3, 4, 5, and 6, and exemplary values for y are 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, or 20 (e.g., C1-6, C1-10, C2-5, C2-8, C2-10, or C2-20 alkyl or alkylene). In some embodiments, the alkylene can be further substituted with 1, 2, 3, or 4 substituent groups as defined herein for an alkyl group.

[0076]

[0037] The term “alkenyl,” as used herein, represents monovalent straight or branched chain groups of, unless otherwise specified, from 2 to 20 carbons (e.g., from 2 to 6 or from 2 to 10 carbons) containing one or more carbon-carbon double bonds and is exemplified by ethenyl, 1 -propenyl,

[0077] 2 -propenyl, 2-methyl-l -propenyl, 1-butenyl, 2-butenyl, and the like. Alkenyls include both cis and trans isomers. Alkenyl groups may be optionally substituted with 1, 2, 3, or 4 substituent groups that are selected, independently, from amino, aryl, cycloalkyl, or heterocyclyl (e.g., heteroaryl), as defined herein, or any of the exemplary alkyl substituent groups described herein.

[0078]

[0038] The term “alkynyl,” as used herein, represents monovalent straight or branched chain groups from 2 to 20 carbon atoms (e.g., from 2 to 4, from 2 to 6, or from 2 to 10 carbons) containing a carbon-carbon triple bond and is exemplified by ethynyl, 1-propynyl, and the like. Alkynyl groups may be optionally substituted with 1, 2, 3, or 4 substituent groups that are selected, independently, from aryl, cycloalkyl, or heterocyclyl (e.g., heteroaryl), as defined herein, or any of the exemplary alkyl substituent groups described herein.

[0079]

[0039] The term “amino,” as used herein, represents -N(RN1)2, wherein each RN1is, independently, H, OH, NO2, N(RN2)2, SO2ORN2, SO2RN2, SORN2, an ^'-protecting group, alkyl, alkenyl, alkynyl, alkoxy, aryl, alkaryl, cycloalkyl, alkcycloalkyl, carboxyalkyl (e.g., optionally substituted with an (9-protecting group, such as optionally substituted arylalkoxycarbonyl groups or any described herein), sulfoalkyl, acyl (e.g., acetyl, trifluoroacetyl, or others described herein), alkoxycarbonylalkyl (e.g., optionally substituted with an (9-protecting group, such as optionally substituted arylalkoxycarbonyl groups or any described herein), heterocyclyl (e.g., heteroaryl), or alkheterocyclyl (e.g., alkheteroaryl), wherein each of these recited RN1groups can be optionally substituted, as defined herein for each group; or two RN1combine to form a heterocyclyl or an N- protecting group, and wherein each RN2is, independently, H, alkyl, or aryl. Amino groups can be unsubstituted amino (i.e., -NH2) or substituted amino (i.e., -N(RN1)2) groups. In a preferred embodiment, amino is -NH2 or -NHRN1, wherein RN1is, independently, OH, NO2, NH2, NRN22, SO2ORN2, SO2RN2, SORN2, alkyl, carboxyalkyl, sulfoalkyl, acyl (e.g., acetyl, trifluoroacetyl, or others described herein), alkoxycarbonylalkyl (e.g., t-butoxycarbonylalkyl) or aryl, and each RN2can be H, C1-20 alkyl (e.g., C1-6 alkyl), or Ce-io aryl.

[0080]

[0040] The term “amino acid,” as described herein, refers to a molecule having a side chain, an amino group, and an acid group (e.g., a carboxy group of -CO2H or a sulfo group of -SChH), wherein the amino acid is attached to the parent molecular group by the side chain, amino group, or acid group (e.g., the side chain). The term “amino acid” used herein includes both natural amino acids and unnatural amino acids. In some embodiments, the amino acid is attached to the parent molecular group by a carbonyl group, where the side chain or amino group is attached to the carbonyl group. Exemplary side chains include an optionally substituted alkyl, aryl, heterocyclyl, alkaryl, alkheterocyclyl, aminoalkyl, carbamoylalkyl, and carboxyalkyl. Exemplary amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, hydroxynorvaline, isoleucine, leucine, lysine, methionine, norvaline, ornithine, phenylalanine, proline, pyrrolysine, selenocysteine, serine, taurine, threonine, tryptophan, tyrosine, and valine. Amino acid groups may be optionally substituted with one, two, three, or, in the case of amino acid groups of two carbons or more, four substituents independently selected from the group consisting of: (1) Ci-6 alkoxy; (2) Ci-6 alkylsulfinyl; (3) amino, as defined herein (e.g., unsubstituted amino (i.e., -NEh) or a substituted amino (i.e., -N(RN1)2, where RN1is as defined for amino); (4) Ce-io aryl-Ci-6 alkoxy; (5) azido;

[0081] (6) halo; (7) (C2-9 heterocyclyl)oxy; (8) hydroxy; (9) nitro; (10) oxo (e.g., carboxyaldehyde or acyl); (11) C1-7 spirocyclyl; (12) thioalkoxy; (13) thiol; (14) -CC>2RA, where RAis selected from the group consisting of (a) C1-20 alkyl (e.g., C1-6 alkyl), (b) C2-20 alkenyl (e.g., C2-6 alkenyl), (c) Ce-io aryl, (d) hydrogen, (e) C1-6 alk-Ce-io aryl, (f) amino-Ci-20 alkyl, (g) polyethylene glycol of - (CH2)s2(OCH2CH2)si(CH2)s3OR’, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and R’ is H or C1-20 alkyl, and (h) amino-polyethylene glycol of -NRN1(CH2)S2(CH2CH2O)SI(CH2)S3NRN1, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and each RN1is, independently, hydrogen or optionally substituted C1-6 alkyl; (15) -C(O)NRBRc, where each of RBand Rcis, independently, selected from the group consisting of (a) hydrogen, (b) C1-6 alkyl, (c) Ce-io aryl, and (d) C1-6 alk-Ce-io aryl; (16) -SChR0, where RDis selected from the group consisting of (a) C1-6 alkyl, (b) Ce-io aryl, (c) C1-6 alk-Ce-io aryl, and (d) hydroxy; (17) - SC>2NRERF, where each of REand REis, independently, selected from the group consisting of (a) hydrogen, (b) C1-6 alkyl, (c) Ce-io aryl and (d) C1-6 alk-Ce-io aryl; (18) -C(O)RG, where RGis selected from the group consisting of (a) C1-20 alkyl (e.g., C1-6 alkyl), (b) C2-20 alkenyl (e.g., C2-6 alkenyl), (c) Ce-io aryl, (d) hydrogen, (e) C1-6 alk-Ce-io aryl, (f) amino-Ci-20 alkyl, (g) polyethylene glycol of -(CH2)s2(OCH2CH2)si(CH2)s3OR’, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and R’ is H or C1-20 alkyl, and (h) amino-polyethylene glycol of -NRN1(CH2)S2(CH2CH2O)SI(CH2)S3NRN1, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and each RN1is, independently, hydrogen or optionally substituted C1-6 alkyl; (19) -NRHC(O)RJ, wherein RHis selected from the group consisting of (al) hydrogen and (bl) C1-6 alkyl, and R1is selected from the group consisting of (a2) C1-20 alkyl (e.g., C1-6 alkyl), (b2) C2-20 alkenyl (e.g., C2-6 alkenyl), (c2) Ce-io aryl, (d2) hydrogen, (e2) Ci-6 alk-Ce-io aryl, (f2) amino-Ci-20 alkyl, (g2) polyethylene glycol of -(CH2)s2(OCH2CH2)si(CH2)s3OR’, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and R’ is H or C1-20 alkyl, and (h2) amino-polyethylene glycol of -NRN1(CH2)S2(CH2CH2O)SI(CH2)S3NRN1, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and each RN1is, independently, hydrogen or optionally substituted C1-6 alkyl; (20) - NRJC(O)ORK, wherein RJis selected from the group consisting of (al) hydrogen and (bl) C1-6 alkyl, and RKis selected from the group consisting of (a2) C1-20 alkyl (e.g., C1-6 alkyl), (b2) C2-20 alkenyl (e.g., C2-6 alkenyl), (c2) Ce-io aryl, (d2) hydrogen, (e2) C1-6 alk-Ce-io aryl, (f2) amino-Ci- 20 alkyl, (g2) polyethylene glycol of -(CH2)s2(OCH2CH2)si(CH2)s3OR’, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and R’ is H or C1-20 alkyl, and (h2) amino-polyethylene glycol of -NRN1(CH2)S2(CH2CH2O)SI(CH2)S3NRN1, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and each RN1is, independently, hydrogen or optionally substituted C1-6 alkyl; and (21) amidine. In some embodiments, each of these groups can be further substituted as described herein.

[0082]

[0041] The term “aryl,” as used herein, represents a mono-, bicyclic, or multicyclic carbocyclic ring system having one or two aromatic rings and is exemplified by phenyl, naphthyl, 1,2- dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, anthranyl, phenanthrenyl, fluorenyl, indanyl, indenyl, and the like, and may be optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from the group consisting of: (1) C1-7 acyl (e.g., carboxyaldehyde); (2) C1-20 alkyl (e.g., C1-6 alkyl, C1-6 alkoxy-Ci-6 alkyl, C1-6 alkyl sulfinyl -C 1-6 alkyl, amino-Ci-6 alkyl, azido-Ci-6 alkyl, (carboxyaldehyde)-Ci-6 alkyl, halo-Ci-6 alkyl (e.g., perfluoroalkyl), hydroxy-Ci- 6 alkyl, nitro-Ci-6 alkyl, or C1-6 thioalkoxy-Ci-6 alkyl); (3) C1-20 alkoxy (e.g., C1-6 alkoxy, such as perfluoroalkoxy); (4) C1-6 alkyl sulfinyl; (5) Ce-io aryl; (6) amino; (7) C1-6 alk-Ce-io aryl; (8) azido; (9) C3-8 cycloalkyl; (10) C1-6 alk-Cs-s cycloalkyl; (11) halo; (12) C1-12 heterocyclyl (e.g., C1-12 heteroaryl); (13) (C1-12 heterocyclyl)oxy; (14) hydroxy; (15) nitro; (16) C1-20 thioalkoxy (e.g., Ci-6 thioalkoxy); (17) -(CH2)qCC>2RA, where q is an integer from zero to four, and RAis selected from the group consisting of (a) Ci-6 alkyl, (b) Ce-io aryl, (c) hydrogen, and (d) Ci-6 alk- Ce-io aryl; (18) -(CH2)qC0NRBRc, where q is an integer from zero to four and where RBand Rcare independently selected from the group consisting of (a) hydrogen, (b) Ci-6 alkyl, (c) Ce-io aryl, and (d) Ci-6 alk-Ce-io aryl; (19) -(CH2)qSO2RD, where q is an integer from zero to four and where RDis selected from the group consisting of (a) alkyl, (b) Ce-io aryl, and (c) alk-Ce-io aryl; (20) -(CH2)qSC>2NRERF, where q is an integer from zero to four and where each of REand REis, independently, selected from the group consisting of (a) hydrogen, (b) Ci-6 alkyl, (c) Ce-io aryl, and (d) Ci-6 alk-Ce-io aryl; (21) thiol; (22) Ce-io aryloxy; (23) C3-8 cycloalkoxy; (24) Ce-io aryl-Ci-6 alkoxy; (25) C1-6 alk-Ci-12 heterocyclyl (e.g., C1-6 alk-Ci-12 heteroaryl); (26) C2-20 alkenyl; and (27) C2-20 alkynyl. In some embodiments, each of these groups can be further substituted as described herein. For example, the alkylene group of a Ci-alkaryl or a Ci- alkheterocyclylcan be further substituted with an oxo group to afford the respective aryloyl and (heterocyclyl)oyl substituent group.

[0083]

[0042] The term “acyl,” as used herein, represents an a radical of general formula -C(O)R, where R is an alkyl or aryl group. Examples of an “acyl” group include, but are not limited to, - C(O)CH3, -C(O)C2H5, and -C(O)Ph.

[0084]

[0043] The term “carbonyl,” as used herein, represents a C(O) group, which can also be represented as C=O.

[0085]

[0044] The term “carboxy,” as used herein, means -CO2H.

[0086]

[0045] The term “cycloalkyl,” as used herein represents a monovalent saturated or unsaturated nonaromatic cyclic hydrocarbon group from three to eight carbons represented as C3-8 cycloalkyl (e.g., C3-5 cycloalkyl having 3 to 5 carbons, C3-6 cycloalkyl having 3 to 6 carbons), unless otherwise specified. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicycle heptyl, and the like. When the cycloalkyl group includes one carbon-carbon double bond or one carbon-carbon triple bond, the cycloalkyl group can be referred to as a “cycloalkenyl” or “cycloalkynyl” group respectively. Exemplary cycloalkenyl and cycloalkynyl groups include cyclopentenyl, cyclohexenyl, cyclohexynyl, and the like. Cycloalkyl groups can be optionally substituted with: (1) C1-7 acyl (e.g., carboxyaldehyde); (2) C1-20 alkyl (e.g., C1-6 alkyl, C1-6 alkoxy-Ci-6 alkyl, C1-6 alkylsulfinyl- C1-6 alkyl, amino-Ci-6 alkyl, azido-Ci-6 alkyl, (carboxyaldehyde)-Ci-6 alkyl, halo-Ci-6 alkyl (e.g., perfluoroalkyl), hydroxy-Ci-6 alkyl, nitro-Ci-6 alkyl, or Ci-6thioalkoxy-Ci-6 alkyl); (3) C1-20 alkoxy (e.g., C1-6 alkoxy, such as perfluoroalkoxy); (4) C1-6 alkyl sulfinyl; (5) Ce-io aryl; (6) amino; (7) C1-6 alk-Ce-io aryl; (8) azido; (9) C3-8 cycloalkyl; (10) C1-6 alk-Cs-s cycloalkyl; (11) halo; (12) C1-12 heterocyclyl (e.g., C1-12 heteroaryl); (13) (C1-12 heterocyclyl)oxy; (14) hydroxy; (15) nitro; (16) Ci -20 thioalkoxy (e.g., C1-6 thioalkoxy); (17) -(CH2)qCC>2RA, where q is an integer from zero to four, and RAis selected from the group consisting of (a) C1-6 alkyl, (b) Ce-io aryl, (c) hydrogen, and (d) C1-6 alk-Ce-io aryl; (18) -(CH2)qCONRBRc, where q is an integer from zero to four and where RBand Rcare independently selected from the group consisting of (a) hydrogen, (b) Ce-io alkyl, (c) Ce-io aryl, and (d) C1-6 alk-Ce-io aryl; (19) -(CH2)qSC>2RD, where q is an integer from zero to four and where RDis selected from the group consisting of (a) Ce-io alkyl, (b) Ce-io aryl, and (c) C1-6 alk-Ce-io aryl; (20) -(CH2)qSC>2NRERF, where q is an integer from zero to four and where each of REand REis, independently, selected from the group consisting of (a) hydrogen, (b) Ce-io alkyl, (c) Ce-io aryl, and (d) C1-6 alk-Ce-io aryl; (21) thiol; (22) Ce-io aryloxy; (23) C3-8 cycloalkoxy; (24) Ce-io aryl-Ci-6 alkoxy; (25) C1-6 alk-Ci-12 heterocyclyl (e.g., C1-6 alk-Ci-12 heteroaryl); (26) oxo; (27) C2-20 alkenyl; and (28) C2-20 alkynyl. In some embodiments, each of these groups can be further substituted as described herein. For example, the alkylene group of a Ci-alkaryl or a Ci-alkheterocyclyl can be further substituted with an oxo group to afford the respective aryloyl and (heterocyclyl)oyl substituent group.

[0087]

[0046] The term “halo” or “halogen,” as used herein, represents a halogen selected from bromine, chlorine, iodine, or fluorine.

[0088]

[0047] The terms “heteroalkyl,” as used herein, refers to an alkyl group, as defined herein, in which one or more of the constituent carbon atoms have each been replaced by nitrogen, oxygen, or sulfur. For example, C1-20 heteroalkyl represents an alkyl group comprising one or more of nitrogen, oxygen, or sulfur in addition to 1 to 20 carbons (e.g., C1-6 heteroalkyl having 1 to 6 carbons and one or more of nitrogen, oxygen, or sulfur, C1-8 heteroalkyl having 1 to 8 carbons and one or more of nitrogen, oxygen, or sulfur, C1-10 heteroalkyl having 1 to 10 carbons and one or more of nitrogen, oxygen, or sulfur, or C1-12 heteroalkyl having 1 to 12 carbons and one or more of nitrogen, oxygen, or sulfur). A typical example of heteroalkyl is polyethylene glycol (PEG), wherein two more of the carbons have each been replaced by oxygen, e.g., C2-20 polyethylene glycol (e.g., C2-10 polyethylene glycol or Ce-8 polyethylene glycol). In some embodiments, the heteroalkyl group can be further substituted with 1, 2, 3, or 4 substituent groups as described herein for alkyl groups. The terms “heteroalkenyl” and heteroalkynyl,” as used herein refer to alkenyl and alkynyl groups, as defined herein, respectively, in which one or two of the constituent carbon atoms have each been replaced by nitrogen, oxygen, or sulfur. In some embodiments, the heteroalkenyl and heteroalkynyl groups can be further substituted with 1, 2, 3, or 4 substituent groups as described herein for alkyl groups.

[0089]

[0048] The terms “heterocycloalkyl,” as used herein, refers to a cycloalkyl group, as defined herein, in which one or more of the constituent carbon atoms have each been replaced by nitrogen, oxygen, or sulfur. For example, C1-20 heterocycloalkyl represents a cycloalkyl group comprising one or more of nitrogen, oxygen, or sulfur in addition to 1 to 20 carbons (e.g., C1-6 heterocycloalkyl, Ci-s heterocycloalkyl, C2-8 heterocycloalkyl, C1-10 heterocycloalkyl, or C1-12 heterocycloalkyl).

[0090]

[0049] The term “heteroaryl,” as used herein, represents that subset of heterocycles, as defined herein, which are aromatic: i.e., they contain 4 / ?+2 pi electrons within the mono- or multicyclic ring system. Exemplary unsubstituted heteroaryl groups are of 1 to 12 (e.g., 1 to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10, or 2 to 9) carbons. Examples of heteroaryl include, but are not limited to, furan, oxazole, thiophene, 1,2,3-triazole, 1,2,4-triazine, 1,2, 4 -tri azole, 1,2,5- thiadiazole 1,1-dioxide, 1,2,5-thiadiazole 1-oxide, 1,2,5-thiadiazole, 1,3, 4 -oxadi azole, 1,3,4- thiadiazole, 1,3,5-triazine, imidazole, isothiazole, isoxazole, pyrazole, pyridazine, pyridine, pyridine-N-oxide, pyrazine, pyrimidine, pyrrole, tetrazole, and thiazole. In some embodiment, the heteroaryl is substituted with 1, 2, 3, or 4 substituents groups as defined for a heterocyclyl group.

[0091]

[0050] The term “oxo” as used herein, represents =0.

[0092]

[0051] The term “polyethylene glycol,” as used herein, represents an alkoxy chain comprised of one or more monomer units, each monomer unit consisting of -OCH2CH2-. Polyethyelene glycol (PEG) is also sometimes referred to as polyethylene oxide (PEO) or polyoxyethylene (POE), and these terms may be considered interchangeable for the purpose of this disclosure. For example, a polyethylene glycol may have the structure, -(CH2)s2(OCH2CH2)si(CH2)s3O-, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), and each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10). Polyethylene glycol may also be considered to include an amino-polyethylene glycol of - NRN1(CH2)S2(CH2CH2O)SI(CH2)S3NRN1-, wherein si is an integer from 1 to 10 (e.g., from 1 to 6 or from 1 to 4), each of s2 and s3, independently, is an integer from 0 to 10 (e.g., from 0 to 4, from 0 to 6, from 1 to 4, from 1 to 6, or from 1 to 10), and each RN1is, independently, hydrogen or optionally substituted Ci-6 alkyl.

[0093]

[0052] The term “isomer,” as used herein, means any tautomer, stereoisomer, enantiomer, or diastereomer of any compound. It is recognized that the compounds can have one or more chiral centers and / or double bonds and, therefore, exist as stereoisomers, such as double-bond isomers (i.e., geometric E / Z isomers) or diastereomers, enantiomers (i.e., (+) or (-)) or cis / trans isomers). Unless otherwise noted, chemical structures depicted herein encompass all of the corresponding stereoisomers, that is, both the stereomerically pure form (e.g., geometrically pure, enantiomerically pure, or diastereomerically pure) and enantiomeric and stereoisomeric mixtures, e.g., racemates. Enantiomeric and stereoisomeric mixtures of compounds can typically be resolved into their component enantiomers or stereoisomers by well-known methods, such as chiral-phase gas chromatography, chiral-phase high performance liquid chromatography, crystallizing the compound as a chiral salt complex, or crystallizing the compound in a chiral solvent. Enantiomers and stereoisomers can also be obtained from stereomerically or enantiomerically pure intermediates, reagents, and catalysts by well-known asymmetric synthetic methods.

[0094]

[0053] The term “stereoisomer,” as used herein, refers to all possible different isomeric as well as conformational forms which a compound may possess (e.g., a compound of any formula described herein), in particular all possible stereochemically and conformationally isomeric forms, all diastereomers, enantiomers and / or conformers of the basic molecular structure. Some compounds may exist in different tautomeric forms, all of the latter being included within the scope of the present disclosure.

[0095]

[0054] The term “diastereomer,” as used herein means stereoisomers that are not mirror images of one another and are non-superimposable on one another.

[0096]

[0055] The term “enantiomer,” as used herein, means each individual optically active form of a compound, having an optical purity or enantiomeric excess (as determined by methods standard in the art) of at least 80% (i.e., at least 90% of one enantiomer and at most 10% of the other enantiomer), preferably at least 90% and more preferably at least 98%. Other terms

[0097]

[0056] As used herein, the term “about” or “approximately” refers to a ±10% variation from the recited quantitative value (and includes the recited quantitative value itself) unless otherwise indicated or inferred from the context. For example, unless otherwise stated or inferred from the context, a dose of about 100 kBq / kg indicates a dose range of 100±10% kBq / kg, i.e., from 90 kBq / kg to 110 kBq / kg, inclusive.

[0098]

[0057] As used herein, the term “administered in combination,” “combined administration,” or “coadministered” means that two or more agents are administered to a subject at the same time or within an interval such that there may be an overlap of an effect of each agent on the patient. Thus, two or more agents that are administered in combination need not be administered together. In some embodiments, they are administered within 90 days (e.g., within 80, 70, 60, 50, 40, 30, 20, 10, 5, 4, 3, 2, or 1 day(s)), within 28 days (e.g., with 14, 7, 6, 5, 4, 3, 2, or 1 day(s), within 24 hours (e.g., 12, 6, 5, 4, 3, 2, or 1 hour(s), or within about 60, 30, 15, 10, 5, or 1 minute of one another. In some embodiments, the administrations of the agents are spaced sufficiently closely together such that a combinatorial effect is achieved.

[0099]

[0058] As used herein, “administering” an agent to a subject includes contacting cells of said subject with the agent.

[0100]

[0059] The term “cancer” refers to any cancer caused by the proliferation of malignant neoplastic cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias, and lymphomas. A “solid tumor cancer” is a cancer comprising an abnormal mass of tissue, e.g., sarcomas, carcinomas, and lymphomas. A “hematological cancer” or “liquid cancer,” as used interchangeably herein, is a cancer present in a body fluid, e.g., lymphomas and leukemias.

[0101]

[0060] The term “chelate” as used herein, refers to an organic compound or portion thereof that can be bonded to a central metal or radiometal atom at two or more points.

[0102]

[0061] The term “conjugate,” as used herein, refers to a molecule that contains a chelating group or metal complex thereof, a linker group, and which optionally contains a therapeutic moiety or a targeting moiety.

[0103]

[0062] The term “therapeutic moiety” as used herein refers to any molecule or any part of a molecule that confers a therapeutic benefit. In some embodiments, the therapeutic moiety is a protein or polypeptide, e.g., an antibody, an antigen-binding fragment thereof. In some embodiments, the therapeutic moiety is a small molecule.

[0063] The term “targeting moiety” as used herein refers to any molecule or any part of a molecule that binds to a given target. In some embodiments, the targeting moiety is a protein or polypeptide such as an antibody or antigen binding fragment thereof, a nanobody, an affibody, or a consensus sequence from a Fibronectin type III domain. In some embodiments, the targeting moiety is a peptide or a small molecule.

[0104]

[0064] As used herein, the term “compound,” is meant to include all stereoisomers, geometric isomers, and tautomers of the structures depicted.

[0105]

[0065] The compounds described herein can be asymmetric (e.g., having one or more stereocenters). All stereoisomers, such as enantiomers and diastereomers, are intended unless otherwise indicated. Compounds of the present disclosure that contain asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods on how to prepare optically active forms from optically active starting materials are known in the art, such as by resolution of racemic mixtures or by stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, and the like can also be present in the compounds described herein, and all such stable isomers are contemplated in the present disclosure. Cis and trans geometric isomers of the compounds of the present disclosure are described and may be isolated as a mixture of isomers or as separated isomeric forms.

[0106]

[0066] Compounds of the present disclosure also include tautomeric forms. Tautomeric forms result from the swapping of a single bond with an adjacent double bond and the concomitant migration of a proton. Tautomeric forms include prototropic tautomers which are isomeric protonation states having the same empirical formula and total charge. Examples prototropic tautomers include ketone - enol pairs, amide - imidic acid pairs, lactam - lactim pairs, amide - imidic acid pairs, enamine - imine pairs, and annular forms where a proton can occupy two or more positions of a heterocyclic system, such as, 1H- and 3H-imidazole, 1H-, 2H- and 4H- 1,2,4- triazole, 1H- and 2H- isoindole, and 1H- and 2H-pyrazole. Tautomeric forms can be in equilibrium or sterically locked into one form by appropriate substitution.

[0107]

[0067] At various places in the present specification, substituents of compounds of the present disclosure are disclosed in groups or in ranges. It is specifically intended that the present disclosure include each and every individual sub-combination of the members of such groups and ranges. For example, the term “Ci-6 alkyl” is specifically intended to individually disclose methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and Ce alkyl. Herein a phrase of the form “optionally substituted X” (e.g., optionally substituted alkyl) is intended to be equivalent to “X, wherein X is optionally substituted” (e.g., “alkyl, wherein said alkyl is optionally substituted”). It is not intended to mean that the feature “X” (e.g., alkyl) per se is optional.

[0108]

[0068] As used herein, the terms “decrease,” “decreased,” “increase,” “increased,” or “reduction,” “reduced,” (e.g., in reference to therapeutic outcomes or effects) have meanings relative to a reference level. In some embodiments, the reference level is a level as determined by the use of said method with a control in an experimental animal model or clinical trial. In some embodiments, the reference level is a level in the same subject before or at the beginning of treatment. In some embodiments, the reference level is the average level in a population not being treated by said method of treatment.

[0109]

[0069] The term an “effective amount” of an agent (e.g., any of the foregoing compounds or conjugates), as used herein, is that amount sufficient to effect beneficial or desired results, such as clinical results, and, as such, an “effective amount” depends upon the context in which it is being applied.

[0110]

[0070] The term “pharmaceutical composition,” as used herein, represents a composition containing a compound described herein formulated with a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition is manufactured or sold with the approval of a governmental regulatory agency as part of a therapeutic regimen for the treatment of disease in a mammal. Pharmaceutical compositions can be formulated, for example, for oral administration in unit dosage form (e.g., a tablet, capsule, caplet, gelcap, or syrup); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate emboli and in a solvent system suitable for intravenous use); or in any other formulation described herein.

[0111]

[0071] A “pharmaceutically acceptable excipient,” as used herein, refers any ingredient other than the compounds described herein (for example, a vehicle capable of suspending or dissolving the active compound) and having the properties of being nontoxic and non-inflammatory in a patient. Excipients may include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, radioprotectants, sorbents, suspending or dispersing agents, sweeteners, or waters of hydration. Exemplary excipients include, but are not limited to: ascorbic acid, histidine, phosphate buffer, butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

[0112]

[0072] The term “pharmaceutically acceptable salt,” as use herein, represents those salts of the compounds described here that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, or allergic response. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Berge et al., J. Pharmaceutical Sciences 66: 1-19, 1977 and in Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. The salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting the free base group with a suitable organic acid.

[0113]

[0073] Compounds may have ionizable groups so as to be capable of preparation as pharmaceutically acceptable salts. These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of compounds, be prepared from inorganic or organic bases. Frequently, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases are well-known in the art, such as hydrochloric, sulphuric, hydrobromic, acetic, lactic, citric, or tartaric acids for forming acid addition salts, and potassium hydroxide, sodium hydroxide, ammonium hydroxide, caffeine, various amines for forming basic salts. Methods for preparation of the appropriate salts are well- established in the art.

[0114]

[0074] Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3 -phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate salts, among others. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium, as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine.

[0115]

[0075] The term “radiopharmaceutical” or “radioconjugate,” as used herein, refers to any compound or conjugate that includes a radioisotope or radionuclide, such as any of the radioisotopes or radionuclides described herein.

[0116]

[0076] As used herein, the term “radionuclide,” refers to an atom capable of undergoing radioactive decay (e.g.,3H,14C,15N,18F,35S,47Sc,55Co,60Cu,61Cu,62Cu,64Cu,67Cu,75Br,76Br ,77Br ,89Zr,86Y,87Y,90Y,97RU, "TC, "mTc,105Rh,109Pd,mIn,123I,124I,125I,131I,134Ce,134La,149Pm,149Tb,153Sm,166HO,177LU,186Re,188Re,198Au,199Au,203Pb,211At,212Pb ,212Bi,213Bi,223Ra,225Ac,227Th,229Th,66Ga,67Ga,68Ga,82Rb,117mSn, or2O1T1). The terms radioactive nuclide, radioisotope, or radioactive isotope may also be used to describe a radionuclide. Radionuclides may be used as detection agents. Exemplary radionuclides used in this disclosure include, but are not limited to,47Sc,55Co,60Cu,61Cu,62Cu,64Cu,67Cu,66Ga,67Ga,68Ga,82Rb,86Y,87Y,89Zr,90Y,97Ru,99Tc, "mTc,105Rh,109Pd,mIn,117mSn,149Pm,149Tb,153Sm,166Ho,177Lu,186Re,188Re,198Au,199Au,2O1T1,203Pb,211At,212Pb,212Bi,213Bi,223Ra,225Ac,227Th, and229Th.

[0117]

[0077] As used herein, and as well understood in the art, “to treat” a condition or “treatment” of the condition (e.g., the conditions described herein such as cancer) is an approach for obtaining beneficial or desired results, such as clinical results. Beneficial or desired results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions; diminishment of extent of disease, disorder, or condition; stabilized (i.e., not worsening) state of disease, disorder, or condition; preventing spread of disease, disorder, or condition; delay or slowing the progress of the disease, disorder, or condition; amelioration or palliation of the disease, disorder, or condition; and remission (whether partial or total), whether detectable or undetectable. In the context of cancer treatment, “ameliorating” may include, for example, reducing incidence of metastases, reducing tumor volume, reducing tumor vascularization and / or reducing the rate of tumor growth. “Palliating” a disease, disorder, or condition means that the extent and / or undesirable clinical manifestations of the disease, disorder, or condition are lessened and / or time course of the progression is slowed or lengthened, as compared to the extent or time course in the absence of treatment.

[0118] Chelators

[0119]

[0078] The compounds of Formula I comprise chelating moieties or chelators.

[0120]

[0079] As disclosed herein, the chelator can be selected from the group consisting of DOTA (l,4,7,10-tetraazacyclododecane-l,4,7,10-tetraacetic acid), DOTMA (lR,4R,7R,10R)-a, a’, a”, a’”-tetramethyl-l,4,7,10-tetraazacyclododecane-l,4,7,10-tetraacetic acid, DOTAM (1,4,7,10- tetrakis(carbamoylmethyl)-l,4,7, 10-tetraazacyclododecane), DOTPA (1,4,7, 10- tetraazacyclododecane-l,4,7,10-tetra propionic acid), DO3 AM-acetic acid (2-(4,7,10-tris(2- amino-2-oxoethyl)-l,4,7,10-tetraazacyclododecan-l-yl)acetic acid), DOTA-GA anhydride (2,2’,2”-(10-(2,6-dioxotetrahydro-2H-pyran-3-yl)-l,4,7,10-tetraazacyclododecane-l,4,7- triyl)triacetic acid, DOTP (l,4,7,10-tetraazacyclododecane-l,4,7,10-tetra(methylene phosphonic acid)), DOTMP (l,4,6,10-tetraazacyclodecane-l,4,7,10-tetramethylene phosphonic acid, DOTA- 4AMP (1,4,7, 10-tetraazacyclododecane- 1, 4,7, 10-tetrakis(acetamido-methylenephosphonic acid), CB-TE2A (l,4,8,l l-tetraazabicyclo[6.6.2]hexadecane-4,l l-diacetic acid), NOTA (1,4,7- triazacyclononane-l,4,7-triacetic acid), NODA-GA (l,4,7-triazacyclononane-4,7-diacetic acid-1- [2-glutaric acid]), NOTP (l,4,7-triazacyclononane-l,4,7-tri(methylene phosphonic acid), TETPA ( 1,4, 8, 11-tetraazacy cl otetradecane- 1,4, 8, 11 -tetrapropionic acid), TETA (1,4,8,11- tetraazacyclotetradecane-l,4,8,l l-tetra acetic acid), HEHA (1,4,7,10,13,16- hexaazacyclohexadecane-l,4,7,10,13,16-hexaacetic acid), PEPA (1,4,7,10,13- pentaazacyclopentadecane-N,N’,N”,N”’, N”” -pentaacetic acid), EUoctapa (N,N’-bis(6-carboxy- 2-pyridylmethyl)-ethylenediamine-N,N’-diacetic acid), EEdedpa (l,2-[[6-(carboxy)-pyridin-2- yl]-methylamino]ethane), Efcphospa (N,N’-(methylenephosphonate)-N,N’-[6- (methoxycarbonyl)pyridin-2-yl]-methyl-l,2-diaminoethane), TTHA (triethylenetetramine- N,N,N’,N”,N”’, N’” -hexaacetic acid), DO2P (tetraazacyclododecane dimethanephosphonic acid), HP-D03A (hydroxypropyltetraazacyclododecanetriacetic acid), EDTA (ethylenediaminetetraacetic acid), Deferoxamine, DTPA (diethylenetriaminepentaacetic acid), DTPA-BMA (diethylenetriaminepentaacetic acid-bismethylamide), and porphyrin.

[0121]

[0080] In certain embodiments, the chelator is selected from DOTA, DOTA-GA, NOTA, NODA- GA, NODA-SA, DTPA, TETA, EDTA, TRITA, CDTA, and DFO, which are defined as below:

[0122] DOTA stands for l,4,7,10-tetrazacyclododecane-l,4,7,10-tetraacetic acid,

[0123] DOTA-GA, or DOTAGA as used herein, stands for 1,4,7, 10-tetraazacyclododececane, 1- (glutaric acid)-4,7,10-triacetic acid,

[0124] NOTA stands for 1,4,7-triazacyclononanetriacetic acid,

[0125] NODA-GA, or NODAGA as used herein, stands for 1,4,7-triazacyclononane-N-glutaric acid-N',N" -diacetic acid,

[0126] NODA-SA, or NODASA as used herein, stands for 1,4,7- triazacyclononane -1-succinic acid-4, 7-diacetic acid,

[0127] DTPA stands for diethylenetriaminepentaacetic acid,

[0128] TETA stands for 1, 4, 8, 11 -tetraazacyclododecane- 1,4, 8,11 -tetraacetic acid,

[0129] EDTA stands for ethylenediamine-N,N'-tetraacetic acid,

[0130] TRITA stands for 1,4,7,10 tetraazacyclotridecane-l,4,7,10-tetraacetic acid,

[0131] CDTA stands for traw -l,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid,

[0132] DFO stands for the Desferal or Desferri oxamine type group of chelators, the chemical name of the non-limiting example is N-[5-({3-[5-(Acetyl-hydroxy-amino)-pentylcarbamoyl]- propionyl}-hydroxy-amino)-pentyl]-N'-(5-amino-pentyl)-N' -hydroxy-succinamide, and with the chemical structures thereof being as follows:

[0133]

[0081] In certain embodiments, the chelator is selected from DOTA, DOTAGA, NOTA, and

[0134] NODAGA.

[0135] Radionuclides

[0082] The present disclosure includes radiopharmaceuticals each comprising a radionuclide.

[0136] Examples of suitable radionuclides include, but are not limited to,47Sc,55Co,60Cu,61Cu,62Cu,64Cu,66Ga,67Ga,67Cu,68Ga,69Er,77As,82Rb,89Zr,86Y,87Y,90Y,97Ru, "Tc, "mTc,105Rh,109Pd,

[0137]

[0083] In some embodiments, the radionuclide is selected from the group consisting of64Cu,67Cu,68Ga,90Y,n iIn,149Tb,153Sm,177Lu,211At,212Bi,212Pb,213Bi,223Ra,225Ac, and227Th.

[0138]

[0084] In some embodiments, the radionuclide is68Ga,89Zr,90Y,U 1ln,177Lu, or225Ac. In certain embodiments, the radionuclide is177Lu or225Ac.

[0085] In some embodiments, the radionuclide used herein is a beta-emitting radionuclide such as177LU. In some embodiments, the radionuclide used herein is an alpha-emitting radionuclide such as225Ac.

[0139] Linkers

[0140]

[0086] The compounds of the present disclosure comprise a linker as shown within the structure of Formula I, wherein the linker is selected from the group consisting of C1-20 alkyl, C1-20 heteroalkyl, C3-10 cycloalkyl, C1-10 heterocycloalkyl, aryl, heteroaryl, C=O, (C=O)NR1, (C=S)NR1, NR1(C=O)NR1, NR1(C=S)NR1, and a combination thereof, in which each R1independently is H or C1-3 alkyl; alternatively, the linker is a hydrophilic space linker comprising one or more cyclic or acyclic polyhydroxy groups.

[0141]

[0087] In some embodiments, the linker can be featured wherein L1is absent or C1-5 alkyl; L2is C1-20 alkyl, C1-20 heteroalkyl, C3-10 cycloalkyl, C1-10 heterocycloalkyl, aryl, or heteroaryl; n is an integer of 1-5 (inclusive); and Z1and Z2each are, independently, absent or a moiety selected from the group consisting of C=O, (C=O)NR3, NR3(C=O)NR3, NR3(C=S)NR3, (CH2CH2)NR3, an amino acid unit, and a combination thereof, in which each of R3, independently, is H or C1-3 alkyl.

[0142]

[0088] In some embodiments, the linker can be featured wherein L1is absent or C1-3 alkyl; L2is C2-10 heteroalkyl or C2-20 polyethylene glycol; n is 1 or 2; and Z1and Z2each are, independently, absent, or a moiety selected from the group consisting of (C=O)NH,

[0143] NH(C=O)NH, (CH2CH2)NH, an amino acid unit, and a combination thereof wherein at least one of Z1and Z2is present.

[0144]

[0089] In some embodiments, the linker can be featured wherein L1is absent or -CH2CH2-; L2is C2-10 heteroalkyl or Ce-s polyethylene glycol; n is 1; Z1is absent, - (C=O)NH-, -NH(C=O)-, NH(C=O)NH, or (CH2CH2)NH; and Z2is (C=O)NH, NH(C=O)NH, (CH2CH2)NH, or a combination thereof.

[0145]

[0090] In some embodiments, the linker can be featured wherein at least one of Z1and Z2is an amino acid unit. Examples of the amino acid unit include, but are not limited to, aspartic acid (Asp), glutamic acid (Glu), 2,4-diaminobutyric acid (Dab), 2,3- diaminopropionic acid (Dap), lysine (Lys), and arginine (Arg).

[0146]

[0091] The term “amino acid unit” described herein refers to an organic moiety formed from an amino acid (e.g., a natural amino acid or an unnatural amino acid). Typically, an amino acid unit is formed as shown below: amino acid amino acid unit where R represents an alkyl, cycloalkyl, aryl, or heteroaryl, each of which can be optionally substituted with a suitable substituent described herein; alternatively, R together with the -NH2 group within the amino acid form a heterocycle. The amino acid or amino acid unit can be in either D- or L-form (i.e., D-enantiomer or L-enantiomer).

[0092] In some embodiments, an amino acid unit is formed from aspartic acid (Asp), glutamic acid

[0147] (Glu), 2,4-diaminobutyric acid (Dab), 2,3-diaminopropionic acid (Dap), lysine (Lys), or arginine (Arg), with their structures shown below:

[0148]

[0149] Aspartic Acid (Asp, D) Glutamic Acid pu, EJ 2,4-diaminobutyric acid (Dab)

[0150] 2,3-diaminopropionic acid (Dap) Lysine flys, IK)

[0151]

[0093] In some embodiments, at least one of Z1and Z2is an amino acid unit formed from Asp.

[0094] In some embodiments, both Z1and Z2are absent.

[0152]

[0095] In some embodiments, the linker can be featured as , wherein the linker comprises one of the following:

[0153] 29

[0154] RECTIFIED SHEET (RULE 91.1 ) which can be optionally substituted with a suitable substituent described herein.

[0155]

[0096] In some embodiments, the linker can be featured wherein a representative exemplary compound is one of the following:

[0156]

[0157]

[0158]

[0097] In some embodiments, the linker is a hydrophilic space linker comprising one or more cyclic or acyclic polyhydroxy groups, including without limitation sugars, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, inositols, and the like.

[0159]

[0098] In some embodiments, the linker can be featured wherein:

[0160] L1is absent or C1-3 alkyl;

[0161] Z1and Z2each are, independently, absent or a moiety selected from the group consisting of C=O, (C=O)NR3, NR3(C=O)NR3, (CH2CH2)NR3, an amino acid unit, and a combination thereof, in which each of R3, independently, is H or C1-3 alkyl; n is an integer of 1-5 (inclusive);

[0162] X is absent or C1-5 alkyl;

[0163] A is absent, C=O, CONR3, heterocycle, or heterocycle-Ci-3 alkyl, R3being H or C1-5 alkyl;

[0164] Y is absent, C1-5 alkyl, or C1-10 heteroalkyl; and

[0165] B is a cyclic or acyclic polyhydroxy group, wherein the linker is a hydrophilic space linker comprising one or more cyclic or acyclic polyhydroxy groups.

[0099] In some embodiments, the linker can be featured wherein

[0166] L1, Z1, and Z2are absent, and n is 1-3; alternatively, wherein Z1is (CH2CH2)NH or (C=O)NH(CH2CH2)NH, Z2is absent, and n is 1-3; alternatively, wherein at least one of Z1and Z2is an amino acid unit, in which the amino acid unit can be formed from aspartic acid (Asp), glutamic acid (Glu), 2,4-diaminobutyric acid (Dab), 2,3 -diaminopropionic acid (Dap), lysine

[0167] (Lys), or arginine (Arg).

[0168]

[0100] In some embodiments, the linker can be featured wherein X is C1-3 alkyl, and Y is absent, C1-5 alkoxyl, or C2-8 polyethylene glycol, wherein indicates the attachment point to X, and “#” indicates the attachment point to Y.

[0169]

[0101] In some embodiments, the linker can be featured wherein B is a cyclic polyhydroxy group having the structure wherein “#” indicates the attachment point to Y, r is 1-2, s is 3-4, and each R, independently, is OH, -CH2OH, -CO2H, -CONH2, NH2, or NHCOCH3, wherein B comprises at least three OH groups. In certain embodiments, B is one of the following, or a stereoisomer thereof:

[0170]

[0171]

[0102] In some embodiments, the linker can be featured wherein B is an acyclic polyhydroxy group having the structure wherein “#” indicates the attachment point to Y; t is 2-5; each R, independently, is OH, -CH2OH, -CO2H,

[0172] -CONH2, NH2, or NHCOCH3; and Raand Rbeach are H or Raand Rbtogether form C=O, wherein B comprises at least three OH groups. In certain embodiments, B is one of the following, or a stereoisomer thereof:

[0173]

[0103] In some embodiments, the linker can be featured wherein a representative exemplary compound is one of the following:

[0174]

[0104] In some embodiments, the linker can be featured wherein: L1is absent or C1-3 alkyl; Z1is absent or a moiety selected from the group consisting of C=O, (C=O)NR3,

[0175] NR3(C=O)NR3, (CH2CH2)NR3, an amino acid unit, and a combination thereof, in which each of R3, independently, is H or C1-3 alkyl; m is an integer of 1-5 (inclusive), p is an integer of 1-4 (inclusive); and

[0176] R in each occurrence, independently, is OH, -CH2OH, -CO2H, -CONH2, NH2, or NHCOCH3, wherein the linker is a hydrophilic space linker comprising at least three OH groups.

[0177]

[0105] In some embodiments, the linker can be featured wherein both L1and Z1are absent, m is 3-4, p is 1-2, and R is OH or -CH2OH.

[0106] In some embodiments, the linker can be featured wherein a representative exemplary compound is one of the following:

[0178]

[0107] In some embodiments, the compounds of this disclosure have a structure selected from one of the following, each of which can attach the chelator “W” to the HSP90 target binder via the linker “L” disclosed herein:

[0179] with all variables as described herein: Subjects

[0180]

[0108] In some disclosed methods, a therapy (e.g., comprising a therapeutic agent) is administered to a subject. In some embodiments, the subject is a mammal, e.g., a human.

[0181]

[0109] In some embodiments, the subject has cancer or is at risk of developing cancer. For example, the subject may have been diagnosed with cancer. The cancer may be a primary cancer or a metastatic cancer. Subjects may have any stage of cancer, e.g., stage I, stage II, stage III, or stage IV with or without lymph node involvement and with or without metastases. Provided compositions may prevent or reduce further growth of the cancer and / or otherwise ameliorate the cancer (e.g., prevent or reduce metastases). In some embodiments, the subject does not have cancer but has been determined to be at risk of developing cancer, e.g., because of the presence of one or more risk factors such as environmental exposure, presence of one or more genetic mutations or variants, family history, etc. In some embodiments, the subject has not been diagnosed with cancer.

[0182] [HO] In some embodiments, the cancer is small-cell lung cancer, non-small-cell lung cancer, sarcoma, pancreatic cancer, breast cancer, or colon cancer.

[0183] Administration and dosage

[0184] Effective doses

[0185] [Hl] The present disclosure provides methods of using a compound of Formula I for treating clinical indications expressing HSP90, with the compound administered to a subject (e.g., a human) in an amount therapeutically effective for such treatment.

[0186]

[0112] This disclosure also covers combination therapies in which the amounts of each therapeutic may or may not be, on their own, therapeutically effective. In some embodiments, therapeutic combinations as disclosed herein are administered to a subject in a manner (e.g., dosing amount and timing) sufficient to cure or at least partially arrest the symptoms of the disorder and its complications. In the context of a single therapy (a “monotherapy”), an amount adequate to accomplish this purpose is defined as a “therapeutically effective amount,” an amount of a compound sufficient to substantially improve at least one symptom associated with the disease or a medical condition. The “therapeutically effective amount” typically varies depending on the therapeutic. For known therapeutic agents, the relevant therapeutically effective amounts may be known to or readily determined by those of skill in the art.

[0113] For example, in the treatment of cancer, an agent or compound that decreases, prevents, delays, suppresses, or arrests any symptom of the disease or condition would be therapeutically effective. A therapeutically effective amount of an agent or compound is not required to cure a disease or condition but will provide a treatment for a disease or condition such that the onset of the disease or condition is delayed, hindered, or prevented, or the disease or condition symptoms are ameliorated, or the term of the disease or condition is changed or, for example, is less severe or recovery is accelerated in an individual. For example, a treatment may be therapeutically effective if it causes a cancer to regress or to slow the cancer’s growth.

[0187]

[0114] The dosage regimen (e.g., amounts of each therapeutic, relative timing of therapies, etc.) that is effective for these uses may depend on the severity of the disease or condition and the weight and general state of the subject. For example, the therapeutically effective amount of a particular composition comprising a therapeutic agent applied to mammals (e.g., humans) can be determined by the person of ordinary skill in the art with consideration of individual differences in age, weight, and the condition of the mammal. Because certain conjugates of the present disclosure exhibit an enhanced ability to target cancer cells and residualize, the dosage of these compounds can be lower than (e.g., less than or equal to about 90%, 75%, 50%, 40%, 30%, 20%, 15%, 12%, 10%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% of) the equivalent dose of required for a therapeutic effect of the unconjugated agent. Therapeutically effective and / or optimal amounts can also be determined empirically by those of skill in the art. Thus, lower effective doses can also be determined by those of skill in the art.

[0188]

[0115] Single or multiple administrations of a radiopharmaceutical or a composition (e.g., a pharmaceutical composition comprising a therapeutic agent or a radiopharmaceutical) can be carried out with dose levels and pattern being selected by the treating physician. The dose and administration schedule can be determined and adjusted based on the severity of the disease or condition in the subject, which may be monitored throughout the course of treatment according to the methods commonly practiced by clinicians or those described herein.

[0189]

[0116] As provided above, a radiopharmaceutical compound of this disclosure may be administered in combination with another therapeutic agent. In the disclosed combination therapy methods, the first and second therapies may be administered sequentially or concurrently to a subject. For example, a first composition comprising a first therapeutic agent and a second composition comprising a second therapeutic agent may be administered sequentially or concurrently to a subject. Alternatively, a composition comprising a combination of a first therapeutic agent and a second therapeutic agent may be administered to the subject.

[0190]

[0117] In some embodiments, the radiopharmaceutical is administered in a single dose. In some embodiments, the radiopharmaceutical is administered more than once, i.e., multiple doses.

[0191] When the radiopharmaceutical is administered more than once, the dose of each administration may be the same or different.

[0192]

[0118] In some embodiments, compositions (such as compositions comprising radiopharmaceuticals) are administered for radiation treatment planning or diagnostic purposes. When administered for radiation treatment planning or diagnostic purposes, compositions may be administered to a subject in a diagnostically effective dose and / or an amount effective to determine the therapeutically effective dose. In some embodiments, a first dose of disclosed conjugate or a composition (e.g., pharmaceutical composition) thereof is administered in an amount effective for radiation treatment planning, followed administration of a combination therapy including a conjugate as disclosed herein and another therapeutic.

[0193]

[0119] Pharmaceutical compositions comprising one or more agents (e.g., radiopharmaceuticals) can be formulated for use in accordance with disclosed methods and systems in a variety of drug delivery systems. One or more physiologically acceptable excipients or carriers can also be included in the composition for proper formulation. Examples of suitable formulations are found in Remington ’s Pharmaceutical Sciences, Mack Publishing Company, Philadelphia, PA, 17th ed., 1985. For a brief review of methods for drug delivery, see, e.g., Langer Science 249: 1527- 1533, 1990).

[0194] Formulations

[0195]

[0120] Pharmaceutical compositions may be formulated for parenteral, intranasal, topical, oral, or local administration, such as by a transdermal means, for prophylactic and / or therapeutic treatment. Pharmaceutical compositions can be administered parenterally (e.g., by intravenous, intramuscular, or subcutaneous injection), or by oral ingestion, or by topical application or intraarticular injection at areas affected by the vascular or cancer condition. Examples of additional routes of administration include intravascular, intra-arterial, intratumor, intraperitoneal, intraventricular, intraepidural, as well as nasal, ophthalmic, intrascleral, intraorbital, rectal, topical, or aerosol inhalation administration. Also specifically contemplated are sustained release administration, by such means as depot injections or erodible implants or components. Suitable compositions include compositions comprising include agents (e.g., compounds as disclosed herein) dissolved or suspended in an acceptable carrier, preferably an aqueous carrier, e.g., water, buffered water, saline, or PBS, among others, e.g., for parenteral administration. Compositions may contain pharmaceutically acceptable auxiliary substances to approximate physiological conditions, such as pH adjusting and buffering agents, tonicity adjusting agents, wetting agents, or detergents, among others. In some embodiments, compositions are formulated for oral delivery; for example, compositions may contain inert ingredients such as binders or fillers for the formulation of a unit dosage form, such as a tablet or a capsule. In some embodiments, compositions are formulated for local administration; for example, compositions may contain inert ingredients such as solvents or emulsifiers for the formulation of a cream, an ointment, a gel, a paste, or an eye drop.

[0196]

[0121] Compositions may be sterilized, e.g., by conventional sterilization techniques, or sterile filtered. Aqueous solutions may be packaged for use as is, or lyophilized, the lyophilized preparation being combined with a sterile aqueous carrier prior to administration. The pH of the preparations typically will be between 3 and 11, more preferably between 5 and 9 or between 6 and 8, and most preferably between 6 and 7, such as 6 to 6.5. In some embodiments, compositions in solid form are packaged in multiple single dose units, each containing a fixed amount of the above-mentioned agent or agents, such as in a sealed package of tablets or capsules. In some embodiments, compositions in solid form are packaged in a container for a flexible quantity, such as in a squeezable tube designed for a topically applicable cream or ointment.

[0197] Analytical Assays

[0198]

[0122] Provided below are analytical assays for characterizing the compounds of Formula I.

[0199]

[0123] Actinium-225 (225Ac) was supplied by the U.S. Department of Energy Isotope Program in the Office of Science for Nuclear Physics or the Canadian Nuclear Laboratories. Lutetium-177 (177LU) was received from ITM Pharma Solutions GmbH or Monrol. Indium- 111 (n iIn) was received from BWXT Medical.

[0124] RadioTLC was performed with Bioscan AR-2000 Imaging Scanner, carried out on iTLC- SG glass microfiber chromatography paper (Agilent Technologies, SGI0001) or iTLC-SA glass microfiber chromatography paper (Agilent Technologies, A120B12).

[0200]

[0125] Radioactive HPLC was performed using a Waters system comprised of a Waters 1525 Binary HPLC pump, a Waters 2489 UV / Visible Detector (monitoring at 254 and 214 nm), a Bioscan Flow Count radiodetector (FC-3300) and a reverse phase (Cl 8) column, or an Agilent 1260 Infinity II LC system comprised of a G711 IB Quat Pump, a G7129A Vial sampler, a G1364F analytical scale fraction collector, a G7114A variable wavelength detector (monitoring at 254 nm), a G7116A 1260 MCT column unit, a Bioscan Flow Count radiodetector (FC-3300), and a reverse phase (Cl 8) column.

[0201]

[0126] Analytical HPLC-MS is performed using a Waters Acquity HPLC-MS system comprised of a Waters Acquity Binary Solvent Manager, a Waters Acquity Sample Manager, a Water Acquity Column Manager (column temperature 30 °C), a Waters Acquity Photodiode Array Detector (monitoring at 254 nm and 214 nm), a Waters Acquity TQD with electrospray ionization and a Waters Acquity BEH C18, 2.1 x 50 mm (1.7 pm) column or using a Waters UPLC-MS-Xevo TQS -Cronos system. Preparative HPLC is performed using a Waters HPLC system comprised of a Waters 1525 Binary HPLC pump, a Waters 2489 UV / Visible Detector (monitoring at 254 nm and 214 nm) and a Waters XBridge Prep C18 19 x 100 mm (5 pm) column or Waters XBridge Prep Phenyl 19 x 100 mm (5 pm).

[0202]

[0127] HPLC elution method 1 : Waters Acquity BEH C18 2.1 x 50 mm (1.7 pm) column; mobile phase A: H2O (0.1% v / v TFA); mobile phase B: acetonitrile (0.1% v / v TFA); flow rate = 0.3 mL / min; wavelength = 214, 254 nm; initial = 90% A, 8 min = 0% A, 10 min = 0% A, 11 min = 90% A, 12 min = 90% A.

[0203]

[0128] HPLC elution method 2: Waters Acquity BEH C18 2.1 x 50 mm (1.7 pm) column; mobile phase A: H2O (0.1% v / v TFA); mobile phase B: acetonitrile (0.1% v / v TFA); flow rate = 0.3 mL / min; wavelength = 214, 254 nm; initial = 90% A, 3 min = 0% A, 3.5 min = 0% A, 4 min = 90% A, 5 min = 90% A.

[0204]

[0129] HPLC elution method 3: Waters Acquity BEH C18 2.1 x 50 mm (1.7 pm) column; mobile phase A: H2O (0.1% v / v Formic Acid); mobile phase B: acetonitrile (0.1% v / v Formic Acid); flow rate = 0.3 mL / min; wavelength = 214, 254 nm; initial = 90% A, 8 min = 0% A, 10 min = 0% A, 11 min = 90% A, 12 min = 90% A.

[0130] HPLC elution method 4: Waters Acquity BEH C18 2.1 x 50 mm (1.7 pm) column; mobile phase A: H2O (0.1% v / v Formic Acid); mobile phase B: acetonitrile (0.1% v / v Formic Acid); flow rate = 0.3 mL / min; wavelength = 214, 254 nm; initial = 90% A, 3 min = 0% A, 3.5 min = 0% A, 4 min = 90% A, 5 min = 90% A.

[0205] EXAMPLES

[0206] Example 1. Synthesis of Radiopharmaceuticals Comprising Compounds of Formula I

[0207]

[0131] Compounds of Formula I comprise conjugates targeting HSP90, which can be radiolabeled with a radionuclide such as Indium-111 (i nIn), Lutetium-177 (177Lu), or Actinium- 225 (225AC) to form radionuclide-chelated radiopharmaceuticals. The synthesis of compounds of Formula I, or their radionuclide-chelated radiopharmaceuticals, can be referred to WO 2020 / 205948. Below are the synthetic schemes or protocols that can be followed to synthesize the corresponding compounds. A person of ordinary skill in the art would have been able, based on the disclosures provided herein and the knowledge in the field, to prepare the depicted compounds and their structurally similar analogs (e.g., compounds with different chelators such as DOTA, NOTA, and NODA-GA).

[0208]

[0132] Scheme 1: Synthesis of 2,2',2"-(10-(2-((2-(4-(4-((l-Acetylpiperidin-4-yl)methyl)phenyl)- 5-(2,4-dihydroxy-5-isopropylphenyl)-4J / -l,2,4-triazole-3-carboxamido)ethyl)amino)-2- oxoethyl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (Compound A).

[0209]

[0210] Step 1: Synthesis of 4-(4-((l-Acetylpiperidin-4-yl)methyl)phenyl)-N-(2-aminoethhours5-(2,4- bis(benzyloxy)-5-isopropylphenyl)-4H-l,2,4-triazole-3-carboxamide (Intermediate 1-A): To ethyl 4-(4-((l-acetylpiperidin-4-yl)methyl)phenyl)-5-(2,4-bis(benzyloxy)-5-isopropylphenyl)-

[0211] 4 / / - 1 , 2, 4-triazole-3 -carboxylate (106.0 mg, 0.15 mmol, 1 equiv.), in a reaction vial was added ethane- 1,2-diamine (1.0 mL, 926.2 mg, 15.4 mmol, 100 equiv.) at room temperature and the reaction was stirred for 1.5 hour. The reaction was stopped by removal of ethane- 1,2-diamine under vacuum and the crude product was purified by reverse phase column chromatography to afford Intermediate 1-A (118 mg, 93%, purity: 99%) as white solid, as TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 4.68 min; MS (positive ESI): found m / z 701.1 [M+H]+; C42H48N6O4 (calc. 701.4).1H NMR (700 MHz, DMSO- d6) d 9.12 (t, J= 5.9 Hz, 1H), 7.83 (s, 3H), 7.42 - 7.36 (m, 6H), 7.36 - 7.31 (m, 2H), 7.30 - 7.27 (m, 2H), 7.12 - 7.08 (m, 2H), 7.06 - 7.02 (m, 2H), 7.00 (s, 1H), 6.79 (s, 1H), 5.09 (s, 2H), 4.97 (s, 2H), 4.36 - 4.30 (m, 1H), 3.78 - 3.72 (m, 1H), 3.44 (q, J= 6.1 Hz, 2H), 3.11 (p, J= 6.9 Hz, 1H), 3.02 - 2.83 (m, 3H), 2.55 - 2.46 (m, 1H), 2.43 (td, J= 12.8, 2.9 Hz, 1H), 1.95 (s, 3H), 1.76 - 1.68 (m, 1H), 1.57 - 1.51 (m, 1H), 1.50 - 1.45 (m, 1H), 1.09 (qd, J= 12.4, 4.4 Hz, 1H), 1.02 (app d, J= 6.9 Hz, 6H), 0.95 (qd, J= 12.3, 4.3 Hz, 1H).

[0212] Step 2: Synthesis of tri-tert-Butyl 2,2',2"-(10-(2-((2-(4-(4-((l-acetylpiperidin-4-yl)methyl)phenyl)~ 5-(2, 4-bis(benzyloxy)-5-isopropylphenyl)-4H-l, 2, 4-triazole-3-carboxamido)ethyl)amino)-2- oxoethyl)-l,4, 7,10-tetraazacyclododecane-l,4, 7-triyl)triacetate (Intermediate 1-B): To a 20 mL scintillation vial with a stir bar was added DOTA-tris( / Bu)ester; 2-{4,7,10-tris[2-(tert-butoxy)-2- oxoethyl]-l,4,7,10-tetraazacyclododecan-l-yl} acetic acid, (20.7 mg, 0.04 mmol, 1 equiv.) and 3 mL DMF. The reaction mixture was cooled to 0 °C and DIPEA (25.2 pL, 18.7 mg, 0.14 mmol, 4 equiv.), followed by HBTU (15.4 mg, 0.04 mmol, 1.1 equiv.) were added and then stirred for 10 min at 0 °C. The solution was brought to room temperature and stirred for 15 min and then 4-(4- ((l-acetylpiperidin-4-yl)methyl)phenyl)-7V-(2-aminoethyl)-5-(2,4-bis(benzyloxy)-5- isopropylphenyl)-4J / -l,2,4-triazole-3-carboxamide (Intermediate 1-A, 30 mg, 0.04 mmol, 1 equiv.) was added as a solid. The reaction was stirred at room temperature and monitored by HPLC-MS. The reaction was stopped after 6h and the solvent was evaporated under vacuum to give the crude product, which was purified by RP chromatography to afford Intermediate 1-B (44 mg, 77%, purity: 94%) as a white solid as the TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 5.51 min; MS (positive ESI): found m / z 1255.1 [M+H]+; C70H99N10O11 (calc. 1255.7). 'H NMR* (700 MHz, DMSO- d6) d 9.09 (t, J = 6.3 Hz, 1H), 8.70 (s, 1H), 7.44 - 7.32 (m, 10H), 7.29 (d, J= 7.5 Hz, 2H), 7.11 (d, J= 8.0 Hz, 2H), 7.02 (d, J= 7.8 Hz, 2H), 6.96 (s, 1H), 6.79 (s, 1H), 5.09 (s, 2H), 4.98 (s, 2H), 4.36 - 4.31 (m, 1H), 4.19 (s, 2H), 3.84 (s, 2H), 3.75 (d, J = 13.5 Hz, 1H), 3.66 - 3.60 (m, 2H), 3.46 (s, 2H), 3.29 - 3.19 (m, 3H), 3.13 - 3.06 (m, 2H), 3.03 (s, 4H), 2.93 (td, J= 13.2, 2.8 Hz, 1H), 2.82 - 2.80 (m, 2H), 2.50 - 2.39 (m, 2H), 1.95 (s, 3H), 1.75 - 1.67 (m, 1H), 1.57 - 1.52 (m, 1H), 1.49 (s, 8H), 1.48 - 1.44 (m, 2H), 1.39 (s, 17H), 1.12 - 1.05 (m, 1H), 1.00 (app. d, J = 6.9 Hz, 6H), 0.98 - 0.90 (m, 1H). (*Protons obscured by H2O not reported).

[0213] Step 3: Synthesis of tri-tert-Butyl 2,2',2"-(10-(2-((2-(4-(4-((l-acetylpiperidin-4- yl)methyl)phenyl)-5-(2, 4-dihydroxy-5-isopropylphenyl)-4H-l, 2, 4-triazole-3- carboxamido)ethyl)amino)-2-oxoethyl)-l, 4, 7, 10-tetraazacyclododecane-l, 4, 7 -triyl) triacetate (Intermediate 1-C): To a flask containing a solution of Intermediate 1-B (42 mg , 31.8 pmol) in methanol (5 mL) was added 10% Pd / C (11.8 mg, 11.1 pmol, 0.4 equiv.) at room temperature. The reaction mixture was degassed under vacuum and subjected to hydrogen gas via a balloon. The reaction was stirred overnight and then monitored by HPLC-MS. The reaction was stopped after 15 hours, filtered through an Acrodisc One (0.2 pm PTFE) syringe filter and the solvent was evaporated under vacuum. The crude product was purified by RP chromatography to afford Intermediate 1-C (29 mg, 69%, purity: 98%) as a white solid as the TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 2; retention time: 2.27 min; MS (positive ESI): found m / z 1075.3 [M+H]+; CseHsyNioOn (calc. 1075.7).

[0214] Step 4: Synthesis of2,2\2"-(10-(2-((2-(4-(4-((l-Acetylpiperidin-4-yl)methyl)phenyl)-5-(2,4- dihydroxy-5-isopropylphenyl)-4H-l,2,4-triazole-3-carboxamido)ethyl)amino)-2-oxoethyl)- 1,4, 7, 10-tetraazacyclododecane-l, 4, 7 -triyl) triacetic acid (Compound A): To a 20 mL scintillation vial with a stir bar was added Intermediate 1-C (18 mg , 10.73 pmol) and 2.5 mL of deprotection cocktail TFA:TIPS:H2O (95:2.5:2.5) at room temperature. The reaction was brought to 37 °C by placing on a pre-heated oil bath at 37 °C and monitored by HPLC-MS. The reaction was stopped after 4h and the TFA was removed under a stream of air. The crude product was purified by preparative HPLC to afford Compound A (12 mg, 97 %, purity: 98%) as a white solid as the TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 2.73 min; MS (positive ESI): found m / z 907.7 [M+H]+; C44H62N10O11 (calc. 907.5). 'HNMR* (700 MHz, DMSO- d6) d 9.81 (s, 1H), 9.05 (t, J= 6.2 Hz, 1H), 8.62 (s, 1H), 7.29 - 7.24 (m, 4H), 6.58 (s, 1H), 6.35 (s, 1H), 4.38 - 4.32 (m, 1H), 4.15 - 3.01 (m, 14H), 2.98 - 2.87 (m, 2H), 2.60 - 2.51 (m, 1H), 2.44 (td, J= 12.8, 2.9 Hz, 1H), 1.98 (s, 3H), 1.79 - 1.71 (m, 1H), 1.66 - 1.61 (m, 1H), 1.60 - 1.55 (m, 1H), 1.14 (qd, J= 12.5, 4.2 Hz, 1H), 1.00 (qd, J =

[0215] 12.5, 4.4 Hz, 1H), 0.83 (app. dd, J= 6.9, 1.2 Hz, 6H). (*Protons obscured by H2O not reported).

[0216]

[0133] Scheme 2: Synthesis of 2,2',2"-(10-(l-(4-(4-((l-Acetylpiperidin-4-yl)methyl)phenyl)-5- (2,4-dihydroxy-5-isopropylphenyl)-47 / - l ,2,4-triazol-3-yl)- l , 18-dioxo-5,8, 11, 14-tetraoxa-2, 17- diazanonadecan-19-yl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (Compound B).

[0217] Step 1: Synthesis of 4-(4-((l-Acetylpiperidin-4-yl)methyl)phenyl)-N-(14-amino-3,6,9,12- tetraoxatetradecyl)-5-(2,4-bis(benzyloxy)-5-isopropylphenyl)-4H-l,2,4-triazole-3-carboxamide (Intermediate 2-A): To 3,6,9, 12-tetraoxatetradecane-l,14-diamine (262 mg, 1.09 mmol), in a reaction vial was added acetonitrile (1.6 mL), followed by ethyl 4-(4-((l-acetylpiperidin-4- yl)methyl)phenyl)-5-(2,4-bis(benzyloxy)-5-isopropylphenyl)-4J / -l,2,4-triazole-3-carboxylate (50 mg, 0.07 mmol, 1 equiv.). The reaction was brought to 60 °C and monitored by HPLC-MS. After 2 hours the reaction was brought to 45 °C and stirred overnight. The reaction was stopped, and the crude reaction mixture was directly purified by RP column chromatography to afford Intermediate 2-A as clear film (52.4 mg, 68%). An aliquot was analyzed by HPLC-MS using elution method 1; retention time: 4.80 min; MS (positive ESI): found m / z 877.1 [M+H]+; C5OH65N608(calc. 877.5).

[0218] Step 2: Synthesis of tri-tert-butyl 2,2',2"-(10-(l-(4-(4-((l-acetylpiperidin-4-yl)methyl)phenyl)-5- (2, 4-bis(benzyloxy)-5-isopropylphenyl)-4H-l,2, 4-triazol-3-yl)-l, 18-dioxo-5, 8, 11, 14-tetraoxa- 2, 17-diazanonadecan-19-yl)-l, 4, 7, 10-tetraazacyclododecane-l, 4, 7 -triyl) triacetate (Intermediate 2-B): To a 20 mL scintillation vial with a stir bar was added DOTA-tris^^ester; 2-{4,7,10-tris[2-(tert-butoxy)-2-oxoethyl]-l,4,7,10-tetraazacyclododecan-l-yl } acetic acid, (18.9 mg, 33.0 pmol, 1.5 equiv.), HBTU, (17.2 mg, 44.0 pmol, 2 equiv.) and anhydrous THF (1 mL). The reaction mixture was stirred at room temperature for 5 min, and then a solution of Intermediate 2-A (26 mg, 22.0 pmol, 1 equiv.) in THF (1 mL) was added, followed by DIPEA (19.2 pL, 14.2 mg, 110.1 pmol, 5 equiv.). The reaction was stirred at room temperature for 1 h and then anhydrous DMF (1 mL) was added to improve the solubility of reaction mixture. The reaction was stopped after 2 h and the solvent evaporated under vacuum. The crude product was purified by RP chromatography to afford Intermediate 2-B (33.4 mg, 87%, purity: 97%) as a clear film as the TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 5.59 min; MS (positive ESI): found m / z 1431.4 [M+H]+; C78H115N10O15 (calc.

[0219] 1431.8).

[0220] Step 3: Synthesis of tri-tert-Butyl 2,2',2"-(10-(l-(4-(4-((l-acetylpiperidin-4-yl)methyl)phenyl)-5- (2, 4-dihydroxy-5-isopropylphenyl)-4H-l, 2, 4-triazol-3-yl)-l, 18-dioxo-5, 8, 11, 14-tetraoxa-2, 17- diazanonadecan-19-yl)-l ,4, 7,10-tetraazacyclododecane-l,4, 7-triyl)triacetate (Intermediate 2- C): To a flask containing a solution of Intermediate 2-B (28 mg, 16.0 pmol) in methanol (5 mL) was added 10% Pd / C (5 mg, 4.8 pmol, 0.3 equiv.) at room temperature. The reaction mixture was degassed under vacuum and subjected to hydrogen gas via a balloon. The reaction was stirred overnight and was monitored by LC-MS. Reaction was incomplete after overnight, thus additional 10% Pd / C (2 mg, 1.6 pmol, 0.1 equiv.) was added and the stirring was continued under hydrogen atmosphere. The reaction was stopped after 21 hours, filtered through an Acrodisc One (0.2 pm PTFE) syringe filter and the solvent was evaporated under vacuum, followed by lyophilization to afford Intermediate 2-C (26 mg, quant., purity: 95%) as a white solid as the TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 4.32 min; MS (positive ESI): found m / z 1251.4 [M+H]+; C64H103N10O15 (calc.

[0221] 1251.8).

[0222] Step 4: Synthesis of 2,2',2"-(10-(l-(4-(4-((l-Acetylpiperidin-4-yl)methyl)phenyl)-5-(2,4- dihydroxy-5-isopropylphenyl)-4H-l, 2, 4-triazol-3-yl)-l, 18-dioxo-5, 8,11, 14-tetraoxa-2, 17- diazanonadecan-19-yl)-l ,4, 7,10-tetraazacyclododecane-l,4, 7-triyl)triacetic acid (Compound B): To a 20 mL scintillation vial with a stir bar was added Intermediate 2-C (20.5 mg, 11.8 pmol) and 2.5 mL of deprotection cocktail TFA:TIPS:H2O (95:2.5:2.5) at room temperature. The reaction was brought to 37 °C by placing in a pre-heated oil bath at 37 °C and monitored by HPLC-MS. The reaction was stopped after 3h and the TFA was removed under a stream of air. The crude product was purified by preparative HPLC to afford Compound B (12.2 mg, 88%, purity: 98%) as a white solid as the TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 2.93 min; MS (positive ESI): found m / z 1083.3 [M+H]+; C52H79N10O15 (calc. 1083.6). *H NMR* (700 MHz, DMSO- d6) A 9.80 (s, 1H), 8.84 (t, J= 5.9 Hz, 1H), 8.56 (s, 1H), 7.29 - 7.23 (m, 4H), 6.58 (s, 1H), 6.35 (s, 1H), 4.37 - 4.32 (m, 1H), 4.05 (s, 2H), 3.88 (s, 2H), 3.79 (d, J= 13.7 Hz, 1H), 3.60 (s, 3H), 3.54 - 3.49 (m, 12H), 3.45 (q, J =

[0223] 6.3 Hz, 4H), 3.35 (s, 6H), 3.31 (q, J= 6.0 Hz, 2H), 3.27 (q, J= 5.7 Hz, 2H), 3.18 - 3.02 (m, 8H), 2.99 - 2.88 (m, 2H), 2.88 (s, 1H), 2.56 (qd, J= 13.3, 7.2 Hz, 2H), 2.44 (td, J= 12.8, 2.9 Hz, 1H), 1.98 (s, 3H), 1.80 - 1.72 (m, 1H), 1.63 (d, J= 12.8 Hz, 1H), 1.58 (d, J= 12.2 Hz, 1H), 1.14 (qd, J = UA, 4.2 Hz, 1H), 1.00 (qd, J= 12.4, 4.4 Hz, 1H), 0.82 (app. d, J= 6.9 Hz, 6H). (*Protons obscured by H2O not reported).

[0224]

[0134] Scheme 3: Synthesis of (7?)-2,2',2"-(10-(l-(4-(4-((l-Acetylpiperidin-4-yl)methyl)phenyl)- 5-(2,4-dihydroxy-5-isopropylphenyl)-4Z7-l,2,4-triazol-3-yl)-26-carboxy-l,6,23-trioxo- 10, 13, 16, 19-tetraoxa-2,5,7,22-tetraazahexacosan-26-yl)-l,4,7, 10-tetraazacyclododecane-l,4,7- triyl)triacetic acid (Compound C).

[0225] Intermediate 3-A

[0226]

[0227] Compound C

[0228] Step 1: Synthesis of tri-tert-Butyl 2,2',2"-(10-(l-(4-(4-((l-acetylpiperidin-4-yl)methyl)phenyl)-5- (2, 4-bis(benzyloxy)-5-isopropylphenyl)-4H-l,2, 4-triazol-3-yl)-29, 29-dimethyl-l, 6, 23, 27- tetraoxo-10, 13, 16, 19, 28-pentaoxa-2, 5, 7, 22-tetraazatriacontan-26-yl)-l, 4, 7, 10- tetraazacyclododecane- 1 ,4, 7 -triyl) (R) -triacetate (Intermediate 3- A):

[0229] To a solution of tri-tert-Butyl 2,2',2"-(10-(l-amino-22,22-dimethyl-16,20-dioxo-3,6,9,12,21- pentaoxa-15-azatricosan-19-yl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)(7?)-triacetate (50 mg, 0.04 mmol, 1 equiv.) in anhydrous THF (1 mL) with a stir bar was added DIPEA (28.7 pL, 0.16 mmol, 4 equiv.) and the mixture was cooled to 0 °C. Lastly, 4-nitrophenylchloroformate (8.5 mg, 0.04 mmol, 1 equiv.) was added at 0 °C in one portion. Following the addition, the reaction was brought to room temperature, purged with argon and monitored by HPLC-MS. After 30 min complete conversion to the nitrophenyl intermediate was observed by HPLC-MS. A solution of Intermediate 1-A (38.5 mg, 0.04 mmol, 1.1 equiv.) in anhydrous DMF (1 mL) was added to the reaction followed by DIPEA (7.2 pL, 0.04 mmol, 1 equiv.) under argon. The stirring was continued at room temperature and additional DIPEA (14.4 pL, 2 equiv.) was added after 2.5h as the reaction had stalled. After an additional 45 min, the reaction was stopped and concentrated under vacuum. The crude product was purified by RP chromatography to afford Intermediate 3- A (32 mg, 40%) as a white solid as the TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 2; retention time: 2.70 min; MS (positive ESI): found m / z 1645.4 [M+H]+; C88H133N12O18 (calc. 1646.0).

[0230] Step 2: Synthesis of tri-tert-Butyl 2,2',2"-(10-(l-(4-(4-((l-acetylpiperidin-4-yl)methyl)phenyl)-5- (2, 4-dihydroxy-5-isopropylphenyl)-4H-l, 2, 4-triazol-3-yl)-29, 29-dimethyl-l, 6, 23, 27-tetraoxo- 10, 13, 16, 19, 28-pentaoxa-2, 5, 7, 22-tetraazatriacontan-26-yl)-l, 4, 7, 10-tetraazacyclododecane- 1, 4, 7 -triy I) (R) -triacetate (Intermediate 3-B): Intermediate 3-A (26.6 mg, 13.2 pmol, 1 equiv.) was subjected to hydrogenolysis conditions in methanol (5.3 mL) with 10% Pd / C (5.8 mg, 5.4 pmol, 0. 4 equiv.) along with a stir bar. The reaction mixture was degassed under vacuum and subjected to hydrogen gas via a balloon (cycled 3X). The reaction was stirred overnight at room temperature (21 °C) and was monitored by HPLC-MS. After 21 h the reaction was re-dosed with 10% Pd / C (5.8 mg, 5.4 pmol, 0.4 equiv.), MeOH (1 mL) and degassed under vacuum and subjected to hydrogen gas via a balloon (cycled 3X). The reaction was stopped after an additional 2 hours, filtered through a Acrodisc One (0.2 pm PTFE) syringe filter, and concentrated under vacuum. The crude product was then purified by RP chromatography to afford Intermediate 3-B (20.2 mg, 84%) as a white solid as the TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 4.33 min; MS (positive ESI): found m / z 1465.3 [M+H]+; C74H121N12O18 (calc. 1465.9).

[0231] Step 3: Synthesis of (R)-2,2',2"-(10-(l-(4-(4-((l-Acetylpiperidin-4-yl)methyl)phenyl)-5-(2,4- dihydroxy-5-isopropylphenyl)-4H-l, 2, 4-triazol-3-yl)-26-carboxy-l , 6,23-trioxo-10, 13, 16, 19- tetraoxa-2, 5, 7, 22-tetraazahexacosan-26-yl)-l, 4, 7,10-tetraazacyclododecane-l, 4, 7 -triyl) triacetic acid (Compound C): To a 20 mL scintillation vial with a stir bar was added Intermediate C-2 (14.3 mg, 8.36 pmol) and 2.5 mL of deprotection cocktail TFA:TIPS:H2O (95:2.5:2.5) at room temperature. The reaction was brought to 37 °C by placing in a pre-heated oil bath at 37 °C and monitored by HPLC-MS. The reaction was stopped after 3h and the TFA was removed under a stream of air. The crude product was purified by preparative HPLC to afford Compound C (8 mg, 72%, purity: 99%) as a white solid as the TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 2.82 min; MS (positive ESI): found m / z 1241.2 [M+H]+; C58H89N12O18 (calc. 1241.6). *H NMR* (700 MHz, DMSO- d6) 3 9.79 (s, 1H), 8.92 (t, J= 5.6 Hz, 1H), 7.94 (s, 1H), 7.27 - 7.24 (m, 4H), 6.58 (s, 1H), 6.35 (s, 1H), 6.09 (t, J= 5.9 Hz, 1H), 5.98 (app s, 1H), 4.37 - 4.32 (m, 1H), 4.13 - 3.71 (m, 2H), 3.53 - 3.47 (m, 14H), 3.41 - 3.34 (m, 4H), 3.26 - 3.07 (m, 7H), 2.98 - 2.86 (m, 2H), 2.59 - 2.52 (m, 2H), 2.50 - 2.47 (m, 1H), 2.44 (td, J= 12.8, 2.9 Hz, 1H), 2.41 - 2.38 (m, 3H), 1.98 (s, 3H), 1.90 - 1.85 (m, 2H), 1.80 - 1.72 (m, 1H), 1.66 - 1.56 (m, 2H), 1.14 (qd, J= 12.5, 4.2 Hz, 1H), 1.00 (qd, J= 12.6, 4.3 Hz,

[0232] 1H), 0.83 - 0.80 (m, 6H). (*Protons obscured by H2O not reported).

[0233]

[0135] Scheme 4: Synthesis of 2,2',2"-(10-(l-(4-(4-((l-Acetylpiperidin-4-yl)methyl)phenyl)-5- (2,4-dihydroxy-5-isopropylphenyl)-47 / - l ,2,4-triazol-3-yl)- l ,6,22-trioxo-9, l 2, l 5, l 8-tetraoxa- 2,5,21-triazatricosan-23-yl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid

[0234] (Compound D).

[0235]

[0236]

[0136] Scheme 5: Synthesis of 2,2',2"-(10-(l-(4-(4-((l-Acetylpiperidin-4-yl)methyl)phenyl)-5-

[0237] (2,4-dihydroxy-5-isopropylphenyl)-4 / 7-l,2,4-triazol-3-yl)-l,6,22,27-tetraoxo-9,l2,l5,l8- tetraoxa-2,5,21,23,26-pentaazaoctacosan-28-yl)-l,4,7,10-tetraazacyclododecane-l,4,7- triyl)triacetic acid (Compound E).

[0238]

[0137] Scheme 6: Synthesis of (7?)-2,2',2"-(10-(l-(4-(4-((l-Acetylpiperidin-4-yl)methyl)phenyl)- 5-(2,4-dihydroxy-5-isopropylphenyl)-4Z7-l,2,4-triazol-3-yl)-7-benzyl-l,6,9,19-tetraoxo-12,15- dioxa-2,5,8,18-tetraazaicosan-20-yl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (Compound F).

[0239] Compound F

[0138] Scheme 7: Synthesis of 2,2',2",2"'-(2-(4-(3-(2-(4-(4-((l-Acetylpiperidin-4- yl)methyl)phenyl)-5-(2,4-dihydroxy-5-isopropylphenyl)-4JH-l,2,4-triazole-3- carboxamido)ethyl)thioureido)benzyl)- 1 ,4,7, 10-tetraazacyclododecane- 1 ,4,7,10- tetrayl)tetraacetic acid (Compound G).

[0240]

[0139] Scheme 8: Synthesis of 2,2',2"-(10-(2-(((5)-l-((2-(4-(4-((l-Acetylpiperidin-4- yl)methyl)phenyl)-5-(2,4-dihydroxy-5-isopropylphenyl)-4JH-l,2,4-triazole-3- carboxamido)ethyl)amino)-3-(l-((25',3A,45',5A)-l-amino-3,4,5,6-tetrahydroxy-l-oxohexan-2-yl)- IT / -l,2,3-triazol-4-yl)-l-oxopropan-2-yl)amino)-2-oxoethyl)-l,4,7, IO-tetraazacyclododecane- l,4,7-triyl)triacetic acid (Compound H).

[0241] Step 1

[0242] Intermediate 8-A

[0243] Intermediate 7-A

[0244] Compound H

[0140] Scheme 9: Synthesis of 2,2',2"-(10-((75,105)-l-(4-(4-((l-Acetylpiperidin-4- yl)methyl)phenyl)-5-(2,4-dihydroxy-5-isopropylphenyl)-4JH-l,2,4-triazol-3-yl)-7,10-bis((l- ((25',3^,45',57?)-l-amino-3,4,5,6-tetrahydroxy-l-oxohexan-2-yl)-lJH-l,2,3-triazol-4-yl)methyl)- l,6,9,12-tetraoxo-2,5,8,l l-tetraazatridecan-13-yl)-l,4,7,10-tetraazacyclododecane-l,4,7- triyl)triacetic acid (Compound I).

[0245] Intermediate 9-A

[0246] Intermediate 7-A

[0247] Intermediate 9-B

[0248] Compound-I

[0249]

[0141] Scheme 10: Synthesis of 2,2',2"-(10-(2-(((5)-l-((2-(4-(4-((l-Acetylpiperidin-4- yl)methyl)phenyl)-5-(2,4-dihydroxy-5-isopropylphenyl)-4JH-l,2,4-triazole-3- carboxamido)ethyl)amino)-l-oxo-3-(l-(2-(2-(2-(2-(((2A,3 ?,45',55',6A)-3,4,5-trihydroxy-6-

[0250] (hydroxymethyl)tetrahydro-2JH-pyran-2-yl)oxy)ethoxy)ethoxy)ethoxy)ethyl)-lJH-l,2,3-triazol-4- yl)propan-2-yl)amino)-2-oxoethyl)-l,4,7,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid

[0251] (Compound J).

[0252] Intermediate 7- A Intermediate 10 - A

[0253] Scheme 11: Synthesis of 2,2',2"-(10-(2-((2-(5-(2,4-Dihydroxy-5-isopropylphenyl)-4-(4-((l- (methylsulfonyl)piperidin-4-yl)methyl)phenyl)-4 / / -l,2,4-triazole-3-carboxamido)ethyl)aniino)- 2 -oxoethyl)- 1,4, 7 ,10-tetraazacyclododecane-l,4,7-triyl)triacetic acid (Compound K)

[0254] 66

[0255] RECTIFIED SHEET (RULE 91.1 )

[0256]

[0257] Intermediate 11-C

[0258] Intermediate 11-D

[0259] Compound K

[0260] Step 1: Synthesis of Ethyl 5-(2,4-bis(benzyloxy)-5-isopropylphenyl)-4-(4-((l- (methylsulfonyl)piperidin-4-yl)methyl)phenyl)-4H-l, 2, 4-triazole-3-carboxylate (Intermediate 11-A)

[0261] To a solution of ethyl 5-[2,4-bis(benzyloxy)-5-(propan-2-yl)phenyl]-4-{4-[(piperidin-4- yl)methyl]phenyl}-4J / -l, 2, 4-triazole-3 -carboxylate (200.0 mg , 0.29 mmol, 1 equiv.), DMAP( 3.55 mg , 0.03 mmol, 0.1 equiv.) and triethylamine (202.52 pL, 147.03 mg , 1.44 mmol, 5 equiv.) in DCM (5 mL) at 0° C, under argon was added methanesulfonyl chloride (44.98 pL , 66.58 mg , 0.58 mmol, 2 equiv.). The mixture was brought to room temperature after 10 min and stirred at room temperature. After 4h20 min the reaction was stopped by adding methanol and the solvent evaporated under vacuum. The crude product was purified by filtration through a normal phase column with DCM to obtain Intermediate 11-A as a white solid (172 mg, 81%, purity: 98%). An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 6.21 min; MS (positive ESI): found m / z 723.5 [M+H]+; C42H48N6O4 (calc. 723.3). 'H NMR (700 MHz, DMSO-tC) b 7.42 - 7.39 (m, 4H), 7.39 - 7.36 (m, 2H), 7.36 - 7.31 (m, 2H), 7.30 - 7.28 (m, 2H), 7.17 - 7.12 (m, 2H), 7.12 - 7.07 (m, 2H), 7.06 (s, 1H), 6.79 (s, 1H), 5.09 (s, 2H), 5.00 (s, 2H), 4.19 (q, J= 7.1 Hz, 2H), 3.54 - 3.49 (m, 2H), 3.33 (s, 1H), 3.10 (hept, J= 6.9 Hz, 1H), 2.81 (s, 3H), 2.61 (td, J= 12.0, 2.3 Hz, 2H), 2.55 (d, J= 6.8 Hz, 2H), 1.63 - 1.56 (m, 3H), 1.24 - 1.15 (m, 2H), 1.11 (t, J= 7.1 Hz, 3H), 1.02 (s, 3H), 1.01 (s, 3H). Step 2: Synthesis ofN-(2-Aminoethyl)-5-(2,4-bis(benzyloxy)-5-isopropylphenyl)-4-(4-((l- (methylsulfonyl)piperidin-4-yl)methyl)phenyl)-4H-l, 2, 4-triazole-3-carboxamide (Intermediate 11-B)

[0262] To ethyl 5-(2,4-bis(benzyloxy)-5-isopropylphenyl)-4-(4-((l-(methylsulfonyl)piperidin-4- yl)methyl)phenyl)-4J / -l, 2, 4-triazole-3 -carboxylate (43.0 mg, 0.06 mmol, 1 equiv.), in a reaction vial with a stir bar was added ethane- 1,2-diamine (79.44 pL, 71.50 mg , 1.19 mmol, 20 equiv.) at room temperature and the reaction was stirred for 50 min. The reaction was stopped by removal of ethane- 1,2-diamine under vacuum. Acetonitrile was added to reaction mixture and subjected to evaporation again to afford Intermediate 11-B as white solid (42.9 mg, 93%, purity: 95%). An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 4.97 min; MS (positive ESI): found m / z 737.3 [M+H]+; C41H49N6O5S (calc. 737.4).

[0263] Step 3: Synthesis of tri-tert-Butyl 2,2',2"-(10-(2-((2-(5-(2,4-bis(benzyloxy)-5-isopropylphenyl)-4- ( 4-( ( 1 -(methylsulfonyl)piperidin-4-yl)methyl)phenyl)-4H-l, 2, 4-triazole-3- carboxamido)ethyl)amino)-2-oxoethyl)-l, 4, 7, 10-tetraazacyclododecane-l, 4, 7 -triyl) triacetate (Intermediate 11-C): To a 20 mL scintillation vial with a stir bar was added DOTA- tris^ujester; 2-{4,7,10-tris[2-(te / 7-butoxy)-2-oxoethyl]-l,4,7,10-tetraazacyclododecan-l-yl } acetic acid (15.18 mg, 0.03 mmol, 1.1 equiv.) and 3 mL DMF. The reaction mixture was cooled to 0 °C and DIPEA (17.61 pL, 13.07 mg, 0.10 mmol, 4 equiv.), followed by HBTU (10.76 mg, 0.03 mmol, 1.1 equiv.) were added and the reaction was stirred for 10 min at 0 °C. The solution was brought to room temperature and stirred for 15 min, followed by the addition of N-(2- aminoethyl)-5-(2,4-bis(benzyloxy)-5-isopropylphenyl)-4-(4-((l-(methylsulfonyl)piperidin-4- yl)methyl)phenyl)-4J / -l, 2, 4-triazole-3 -carboxamide (Intermediate 11-B) (21.0 mg , 0.03 mmol, 1 equiv.) as a solid. The reaction was stirred at room temperature and monitored by HPLC-MS. After 3.5h an additional 0.5 eq of HBTU and DIPEA were added and then the reaction was stopped after 21h by evaporation of the solvent under vacuum. The crude product was purified by RP chromatography to afford Intermediate 11-C (31.5 mg, 80.4 %, purity: 98%) as a white solid as the TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 5.67 min; MS (positive ESI): found m / z 1291.8 [M+H]+; C69H99N10O12S (calc. 1291.7). 'HNMR* (700 MHz, DMSO-tfc) b 9.09 (d, J= 6.4 Hz, 1H), 8.70 (s, 1H), 7.42 (s, 2H), 7.41 - 7.38 (m, 4H), 7.38 - 7.36 (m, 2H), 7.36 - 7.32 (m, 2H), 7.31 - 7.28 (m, 2H), 7.12 (d, J = 8.1 Hz, 2H), 7.02 (d, J= 8.0 Hz, 2H), 6.97 (s, 1H), 6.79 (s, 1H), 5.09 (s, 2H), 4.98 (s, 2H), 4.19 (s, 2H), 3.84 (s, 2H), 3.65 - 3.60 (m, 2H), 3.55 - 3.50 (m, 2H), 3.46 (s, 2H), 3.22 (d, J= 5.9 Hz, 2H), 3.10 (h, J= 6.4, 5.9 Hz, 1H), 3.03 (s, 4H), 2.82 (s, 3H), 2.61 (td, J= 11.9, 2.4 Hz, 2H), 2.53 (d, = 7.0 Hz, 1H), 1.63 - 1.56 (m, 3H), 1.49 (s, 9H), 1.39 (s, 18H), 1.23 - 1.14 (m, 2H), 1.01 (app. D, J= 6.9 Hz, 6H). (*Protons obscured by H2O not reported)

[0264] Step 4:Synthesis of tri-tert-Butyl 2,2',2"-(10-(2-((2-(5-(2,4-dihydroxy-5-isopropylphenyl)-4-(4- ((1 -(methylsulfonyl)piperidin-4-yl)methyl)phenyl)-4H-l, 2, 4-triazole-3- carboxamido)ethyl)amino)-2-oxoethyl)-l, 4, 7, 10-tetraazacyclododecane-l, 4, 7 -triyl) triacetate (Intermediate 11-D): To a flask containing a solution of Intermediate 11-C (28 mg, 20 pmol, 1 equiv.) in methanol (4 mL) was added 10% Pd / C (8.64 mg, 8.2 pmol, 0.4 equiv.) at room temperature. The reaction mixture was degassed under vacuum and subjected to hydrogen gas via a balloon. The reaction was stirred at room temperature and stopped after 5 hours. The reaction mixture was filtered through an Acrodisc One (0.2 pm PTFE) syringe filter and the solvent was evaporated under vacuum to give the crude product, Intermediate 11-D (27 mg, 95%, purity: 85%) as a white film as the TFA salt. The crude product was used for the next step without purification An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 4.56 min; MS (positive ESI): found m / z 1112.7 [M+H]+; C55H87N10O12S (calc. 1111.6).

[0265] Step 5: Synthesis of2,2 2''-(10-(2-((2-(4-(4-((l-Acetylpiperidin-4-yl)methyl)phenyl)-5-(2,4- dihydroxy-5-isopropylphenyl)-4H-l,2,4-triazole-3-carboxamido)ethyl)amino)-2-oxoethyl)- 1,4, 7, 10-tetraazacyclododecane-l, 4, 7 -triyl) triacetic acid (Compound K): To a 20 mL scintillation vial with a stir bar was added Intermediate 11-D (27 mg , 18 pmol) and 2.5 mL of deprotection cocktail TFA:TIPS:H2O (95:2.5:2.5) at room temperature. The reaction was brought to 37 °C by placing in a pre-heated oil bath at 37 °C and monitored by HPLC-MS. The reaction was stopped after 3h and the TFA was removed under a stream of air. The crude product was purified by preparative HPLC to afford Compound K (20 mg, 94%, purity: 97%) as white solid as the TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 3.00 min; MS (positive ESI): found m / z 943.5 [M+H]+; C43H63N10O12S (calc. 943.4). 'H NMR* (700 MHz, DMSO-tL) b 12.93 (s, 2H), 9.84 (s, 1H), 9.04 (t, J= 6.3 Hz, 1H), 8.62 (s, 1H), 7.29 - 7.26 (m, 4H), 6.58 (s, 1H), 6.35 (s, 1H), 4.03 (s, 3H), 3.75 (s, 1H), 3.64 - 3.45 (m, 3H), 3.31 (q, J= 6.7, 6.1 Hz, 4H), 3.18 (q, J= 6.0 Hz, 3H), 3.09 (s, 5H), 2.91 (p, J= 6.9 Hz, 1H), 2.84 (s, 3H), 2.64 (td, J= 11.9, 2.5 Hz, 2H), 2.58 (d, J= 6.9 Hz, 2H), 1.71 - 1.67 (m, 2H), 1.66 - 1.62 (m, 1H), 1.25 (qd, J= 12.3, 4.1 Hz, 3H), 0.83 (app. D, J= 6.9 Hz, 6H). (*Protons obscured by H2O not reported).

[0266] Scheme 12: Synthesis of 2,2',2"-(10-(2-((4-((2-(4-(4-((l-Acetylpiperidin-4-yl)methyl)phenyl)-5- (2,4-dihydroxy-5-isopropylphenyl)-4Z / -l,2,4-triazole-3- carboxamido)ethyl)carbamoyl)benzyl)amino)-2-oxoethyl)- 1 ,4,7, 10-tetraazacyclododecane- 1,4,7- triyl)triacetic acid (Compound L)

[0267] Intermediate 12-C

[0268] Compound L

[0269] Step l:Synthesis of (9H-Fluoren-9-yl)methyl (4-((2-(4-(4-((l-acetylpiperidin-4- yl)methyl)phenyl)-5-(2, 4-bis(benzyloxy)-5-isopropylphenyl)-4H-l, 2, 4-triazole-3- carboxamido) ethyl) carbamoyl benzyl) carbamate (Intermediate 12-A): To a 20 mL scintillation vial with a stir bar was added 4-(((((97 / -fluoren-9-yl)methoxy)carbonyl)amino)methyl)benzoic acid (15.1 mg , 0.04 mmol, 1.1 equiv.) and 3 mL DMF. The reaction mixture was cooled to 0 °C and DIPEA (26.8 pL, 19.9 mg, 0.15 mmol, 4 equiv.), followed by HBTU, (16.4 mg, 0.04 mmol, 1.1 equiv.) was added and the reaction was stirred for 10 min at 0 °C. The solution was brought to room temperature and stirred for 15 min and then 4-(4-((l-acetylpiperidin-4- yl)methyl)phenyl)-7V-(2-aminoethyl)-5-(2,4-bis(benzyloxy)-5-isopropylphenyl)-4J / -l,2,4- triazole-3 -carboxamide (Intermediate 1-A, 32 mg, 0.04 mmol, 1 equiv.) was added as solid. The reaction was stirred at room temperature and was monitored by HPLC-MS. The reaction was stopped after 2h20m and the solvent was evaporated under vacuum to give the crude product, which was purified by normal phase chromatography using 0-10% MeOH in DCM to afford Intermediate 12-A (37 mg, 89%, purity: 98%) as a white solid. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 6.46 min; MS (positive ESI): found m / z 1056.9 [M+H]+; C65H66N7O7 (calc. 1056.5). 'HNMR* (700 MHz, DMSO-tL) d 9.08 (t, J= 5.5 Hz, 1H), 8.48 (t, J= 5.5 Hz, 1H), 7.93 - 7.88 (m, 3H), 7.79 - 7.76 (m, 2H), 7.72 - 7.69 (m, 2H), 7.44 - 7.38 (m, 6H), 7.37 - 7.28 (m, 7H), 7.27 - 7.23 (m, 2H), 7.03 (dd, J= 6.6, 1.9 Hz, 3H), 7.00 - 6.96 (m, 2H), 6.77 (s, 1H), 5.09 (s, 2H), 4.94 (s, 2H), 4.37 (d, J= 6.9 Hz, 2H), 4.31 (dt, J= 12.7, 2.1 Hz, 1H), 4.26 - 4.22 (m, 3H), 3.75 - 3.70 (m, 1H), 3.41 - 3.36 (m, 4H), 3.10 (hept, J= 6.9 Hz, 1H), 2.93 - 2.86 (m, 1H), 2.47 (d, J= 7.2 Hz, 2H), 2.40 (td, J= 12.8, 2.9 Hz, 1H), 1.94 (s, 3H), 1.72 - 1.64 (m, 1H), 1.53 - 1.48 (m, 1H), 1.48 - 1.44 (m, 1H), 1.10 - 1.03 (m, 1H), 1.02 (app d, J= 6.9 Hz, 6H), 0.93 (qd, J= 12.5, 4.3 Hz, 1H). (*1 labile NH proton not observed).

[0270] Step 2: Synthesis of (9H-Fluoren-9-yl)methyl (4-((2-(4-(4-((l-acetylpiperidin-4- yl)methyl)phenyl)-5-(2, 4-dihydroxy-5-isopropylphenyl)-4H-l, 2, 4-triazole-3- carboxamido) ethyl) carbamoylfbenzyl) carbamate (Intermediate 12-B): To a flask containing Intermediate 12-A (30 mg , 26.7 pmol), methanol (2 mL) and ethyl acetate (2 mL) were added followed by 10% Pd / C (9.0 mg , 8.01 pmol, 0.3 equiv.) at room temperature. The reaction mixture was degassed under vacuum and subjected to hydrogen gas via a balloon. The reaction was stirred overnight and was monitored by HPLC-MS. The reaction was stopped after 45 hours, filtered through an Acrodisc One (0.2 pm PTFE) syringe filter and the solvent was evaporated under vacuum to give the crude product. The crude product was resubjected to hydrogenation as above but using methanol and ethanol (1: 1) as the solvent. The reaction was stopped after 18 hours and filtered through an Acrodisc One (0.2 pm PTFE) syringe filter. The solvent was evaporated under vacuum to give the crude Intermediate 12-B (26 mg, 79%, purity: 71%) as a colorless film, which was used without purification for the next step. The crude product was analyzed by HPLC-MS using elution method 2; retention time: 5.01 min; MS (positive ESI): found m / z 876.5 [M+H]+; C5iH54N7O7 (calc. 876.4).

[0271] Step 3: Synthesis of 4-(4-((l-Acetylpiperidin-4-yl)methyl)phenyl)-N-(2-(4- (aminomethyl)benzamido)ethyl)-5-(2, 4-dihydroxy-5-isopropylphenyl)-4H-l, 2, 4-triazole-3- carboxamide (Intermediate 12-C): To a vial containing Intermediate 12-B (25 mg , 28.5 pmol, 1 equiv.) in DMF was added piperidine (0.3 ml , 2.85 mmol, 100 equiv.) at room temperature to make a 15% piperidine solution. The reaction mixture was stirred at room temperature and was monitored by HPLC-MS. The reaction was stopped after 70 mins and the solvent was evaporated under vacuum to give the crude product. The crude was purified by reverse phase chromatography to afford Intermediate 12-C (16 mg, 62%, purity: 85%) as a white solid, as TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 2.98 min; MS (positive ESI): found m / z 654.3 [M+H]+; C36H44N7O5 (calc. 654.3).

[0272] Step 4: Synthesis of 2,2',2"-(10-(2-((4-((2-(4-(4-((l -Acetylpiperidin-4-yl)methyl)phenyl)-5-(2, 4- dihydroxy-5-isopropylphenyl)-4H-l,2,4-triazole-3-carboxamido)ethyl)carbamoyl)benzyl)amino)- 2 -oxoethyl) -1,4, 7,10-tetraazacyclododecane-l,4, 7-triyl)triacetic acid (Compound L): To a vial with stir bar was added 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-l-yl)oxy)-2-oxoethyl)-l,4,7,10- tetraazacyclododecane-l,4,7-triyl)triacetic acid (NHS-DOTA, 10.7 mg, 13.2 pmol, 1.5 equiv.), Intermediate 12-C (8.5 mg , 8.9 p mol, 1 equiv.) followed by DMF (1.5 mL). The reaction mixture was cooled to 0 °C and DIPEA (9.27 pL, 6.87 mg, 53.2 pmol, 6 equiv.) was added. The reaction was stirred at 0 °C for 10 min and then brought to room temperature and was monitored by HPLC-MS. The reaction was stopped after 2h 45mins and the solvent was evaporated under vacuum to give the crude product. The crude product was resubjected to the above condition using NHS-DOTA (6 mg, 0.75 equiv.), DIPEA (5 pL, 3 equiv.) which were added at 0 °C and then the reaction was brought to room temperature after 30 min. The reaction was stopped after 2h30mins and the solvent was evaporated under vacuum to give the crude product, which was purified by reverse phase chromatography to afford Compound L (6.7 mg, 57.8%, purity: 97%) as a white solid, as TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 2.89 min; MS (positive ESI): found m / z 1040.8 [M+H]+; C52H70N11O12 (calc. 1040.5). ‘HNMR* (700 MHz, DMSO-tL) 69.72 (s, 1H), 8.98 (t, J= 5.5 Hz, 1H), 8.44 (s, 1H), 7.76 - 7.70 (m, 2H), 7.31 (d, J= 8.0 Hz, 2H), 7.21 - 7.14 (m, 4H), 6.52 (s, 1H), 6.48 (s, 2H), 6.28 (s, 1H), 4.33 (d, J= 5.8 Hz, 2H), 4.29 - 4.24 (m, 1H), 3.85 (s, 5H), 3.71 (d, J= 13.6 Hz, 1H), 3.56 (s, 5H), 3.31 - 3.26 (m, 2H), 3.02 (s, 9H), 2.94 - 2.85 (m, 1H), 2.87 - 2.79 (m, 1H), 2.54 - 2.46 (m, 2H), 2.37 (td, J= 12.8, 2.9 Hz, 1H), 1.92 (d, J= 3.6 Hz, 1H), 1.90 (s, 3H), 1.68 (dtt, J= 11.1, 7.4, 3.6 Hz, 1H), 1.61 - 1.54 (m, 1H), 1.53 - 1.48 (m, 1H), 1.07 (qd, J= 12.4, 4.2 Hz, 1H), 0.93 (qd, J= 12.6, 4.3 Hz, 1H), 0.77 - 0.73 (app m, 6H). (*Protons obscured by H2O not reported).

[0273] Scheme 13: Synthesis of 2,2',2"-(10-(2-((3-(5-(2,4-Dihydroxy-5-isopropylphenyl)-A-methyl-4- (4-((l-(methylsulfonyl)piperidin-4-yl)methyl)phenyl)-4J / -l,2,4-triazole-3- carboxamido)propyl)(methyl)amino)-2-oxoethyl)-l,4,7,10-tetraazacyclododecane-l,4,7- triyl)triacetic acid (Compound M)

[0274]

[0275] Intermediate 11-A

[0276] Intermediate 13-A

[0277] Compound M Step 1: Synthesis of 5-(2,4-bis(Benzyloxy)-5-isopropylphenyl)-N-methyl-N-(3- (methylamino)propyl)-4-( 4-( ( 1 -(methylsulfonyl)piperidin-4-yl)methyl)phenyl)-4H-l, 2, 4-triazole- 3-carboxamide (Intermediate 13-A).

[0278] To ethyl 5-(2,4-bis(benzyloxy)-5-isopropylphenyl)-4-(4-((l-(methylsulfonyl)piperidin-4- yl)methyl)phenyl)-4J / -l, 2, 4-triazole-3 -carboxylate (Intermediate 11-A) (35.0 mg, 0.05 mmol, 1 equiv.), in a reaction vial with a stir bar was added 7V1,7V3-dimethylpropane-l,3-diamine (123.6 pL, 100.9 mg, 0.96 mmol, 20 equiv.) at room temperature, and the reaction was stirred for 28 hours. The reaction was stopped by removal of N1,A3-dimethylpropane-l,3-diamine under vacuum. The crude product was purified by reverse phase chromatography to afford Intermediate 13-A as white solid (20.7 mg, 52.7%, purity: 95%). HPLC-MS elution using elution method 1; retention time: 5.03 min; MS (positive ESI): found m / z 779.5 [M+H]+; C44H55N6O5S (calc. 779.4).

[0279] Step 2: Synthesis of tri-tert-Butyl 2,2',2"-(10-(2-((2-(5-(2,4-bis(benzyloxy)-5-isopropylphenyl)-4- ( 4-( ( 1 -(methylsulfonyl)piperidin-4-yl)methyl)phenyl)-4H-l, 2, 4-triazole-3- carboxamido)ethyl)amino)-2-oxoethyl)-l, 4, 7, 10-tetraazacyclododecane-l, 4, 7 -triyl) triacetate (Intermediate 13-B): To a 20 mL scintillation vial with a stir bar was added 2-{4,7,10-tris[2- ( / c / 7-butoxy)-2-oxoethyl]- l ,4,7,10-tetraazacyclododecan-l -yl } acetic acid, (DOTA-tris('Bu)ester, 11.4 mg , 19.66 pmol, 1.1 equiv.) and 3 mL DMF. The reaction mixture was cooled to 0 °C and DIPEA (12.5 pL, 9.24 mg, 71.49 pmol, 4 equiv.), followed by HBTU, (7.61 mg, 19.66 pmol, 1.1 equiv.) was added and the reaction was stirred for 10 min at 0 °C. The solution was brought to room temperature and stirred for 15 min, followed by addition of Intermediate 13-A (16.8 mg, 17.87 pmol, 1 equiv.) as solid. The reaction was stirred at room temperature and monitored by HPLC-MS. After 4h an additional 0.5 eq of DOTA-tris( / Bu)ester, HBTU and DIPEA were all added. After an additional 1.5h the reaction was stopped and the solvent was evaporated under vacuum to give the crude product, which was purified by RP chromatography to afford Intermediate 13-B (20 mg, 70.9 %, purity: 99%) as a white solid as the TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 3; retention time: 4.74 min; MS (positive ESI): found m / z 1334.9 [M+H]+; C72H105N10O12S (calc. 1333.8). *HNMR* (700 MHz, DMSO-tL) b 7.45 - 7.39 (m, 4H), 7.39 - 7.30 (m, 4H), 7.24 - 7.19 (m, 2H), 7.15 - 7.09 (m, 3H), 7.01 - 6.88 (m, 2H), 6.82 - 6.78 (m, 1H), 5.11 (s, 2H), 4.96 - 4.86 (m, 2H), 4.18 (s, 4H), 3.60 - 3.29 (m, 9H), 3.26 - 3.09 (m, 4H), 3.04 (s, 1H), 3.00 (s, 1H), 2.93 - 2.88 (m, 1H), 2.90 - 2.85 (m, 1H), 2.82 (s, 3H), 2.83 - 2.79 (m, 1H), 2.73 (s, 1H), 2.64 - 2.58 (m, 2H), 2.56 - 2.52 (m, 1H), 1.91 - 1.84 (m, 1H), 1.81 - 1.72 (m, 1H), 1.69 - 1.64 (m, 1H), 1.63 - 1.54 (m, 3H), 1.51 - 1.46 (m, 9H), 1.46 - 1.36 (m, 19H), 1.22 - 1.13 (m, 2H), 1.08 - 1.00 (m, 6H). (*Protons obscured by H2O not reported).

[0280] Step 3: Synthesis of Tri-tert-butyl 2,2',2"-(10-(2-((3-(5-(2,4-dihydroxy-5-isopropylphenyl)-N- methyl-4-( 4-( ( 1 -(methylsulfonyl)piperidin-4-yl)methyl)phenyl)-4H-l, 2, 4-triazole-3- carboxamido)propyl)(methyl)amino)-2-oxoethyl)-l, 4, 7, 10-tetraazacyclododecane-l, 4, 7- triy I) triacetate (Intermediate 13-C): To a flask containing Intermediate 13-B (20 mg , 12.68 pmol, 1 equiv.), methanol (4 mL) was added, followed by 10% Pd / C (5.4 mg , 5.07 pmol, 0.4 equiv.) at room temperature. The reaction mixture was degassed under vacuum and subjected to hydrogen gas via a balloon. The reaction was stirred at room temperature and monitored by HPLC-MS. The reaction was stopped after 4hl5min, filtered through an Acrodisc One (0.2 pm PTFE) syringe filter and the solvent was evaporated under vacuum to give the crude product Intermediate 13-C (17 mg, quantitative). The crude produce was analyzed via HPLC-MS using elution method 1; retention time: 3.67 min; MS (positive ESI): found m / z 1153.7 [M+H]+; C58H93N10O12S (calc. 1153.7).

[0281] Step 4: Synthesis of 2,2',2"-(10-(2-((3-(5-(2,4-Dihydroxy-5-isopropylphenyl)-N-methyl-4-(4-((l- (methylsulfonyl)piperidin-4-yl)methyl)phenyl)-4H-l,2, 4-triazole-3- carboxamido)propyl)(methyl)amino)-2-oxoethyl)-l, 4, 7, 10-tetraazacyclododecane-l, 4, 7- triy I) triacetic acid (Compound M): To a 20 mL scintillation vial with a stir bar was added Intermediate 13-C (17 mg , 11.69 pmol) and 2.0 mL of deprotection cocktail TFA:TIPS:H2O (95:2.5:2.5) at room temperature. The reaction was brought to 37 °C by placing on a pre-heated heating block at 37 °C and monitored by HPLC-MS. The reaction was stopped after 2h20min and the reaction mixture was evaporated under vacuum to remove TFA. The crude product was purified by preparative HPLC to afford Compound M (11.4 mg, 79%, purity: 98%) as white solid as the TFA salt. HPLC-MS analysis was done using elution method 1; retention time: 2.79 min; MS (positive ESI): found m / z 985.5 [M+H]+; C46H69N10O12S (calc. 985.5). 'H NMR* (700 MHz, DMSO-tL) b 12.95 (s, 2H), 9.79 (s, 1H), 7.56 (s, 2H), 7.26 - 7.17 (m, 4H), 6.61 - 6.56 (m, 1H), 6.33 - 6.30 (m, 1H), 4.26 - 3.93 (m, 6H), 3.60 (s, 2H), 3.49 - 3.44 (m, 2H), 3.41 - 3.36 (m, 1H), 3.35 - 3.28 (m, 9H), 3.27 - 3.23 (m, 1H), 3.15 - 3.08 (m, 1H), 3.02 (s, 1H), 3.01 - 2.97 (m, 1H), 2.96 (s, 1H), 2.90 - 2.83 (m, 2H), 2.82 - 2.79 (m, 1H), 2.78 - 2.75 (m, 4H), 2.73 (s, 1H), 2.56 (td, J= 12.0, 2.3 Hz, 2H), 2.54 - 2.50 (m, 2H), 1.83 - 1.76 (m, 1H), 1.70 - 1.65 (m, 1H), 1.62 - 1.57 (m, 3H), 1.59 - 1.51 (m, 1H), 1.21 - 1.13 (m, 2H), 0.82 - 0.72 (m, 6H). (*Protons obscured by H2O not reported).

[0282] Scheme 14: Synthesis of [4,7-Bis(carboxymethyl)-10-{[(14-{[5-(2,4-dihydroxy-5- isopropylphenyl)-4-[4-(morpholin-4-ylmethyl)phenyl]-l,2-oxazol-3-yl]formamido}-3,6,9,12- tetraoxatetradecan-l-yl)carbamoyl]methyl}-l,4,7,10-tetraazacyclododecan-l-yl]acetic acid

[0283] (Compound N)

[0284]

[0285] Step 1: Synthesis of tert-Butyl 2-[4-({[14-({5-[2,4-bis(benzyloxy)-5-isopropylphenyl]-4-[4- (morpholin-4-ylmethyl)phenyl] -1, 2-oxazol-3-yl}formamido)-3, 6,9,12 -tetraoxatetradecan- 1- yl]carbamoyl}methyl)-7, 10-bis[2-(tert-butoxy)-2-oxoethyl] -1 , 4, 7, 10-tetraazacyclododecan-l- yl]acetate (Intermediate 14 - A): A 20 mL scintillation vial with a stir bar was charged with 5- [2,4-bis(Benzyloxy)-5-(propan-2-yl)phenyl]-4-{4-[(morpholin-4-yl)methyl]phenyl}-l,2-oxazole- 3-carboxylic acid (J. Med.Chem. 2020, 63, 5421) (17.0 mg, 0.03 mmol, 1 equiv.), anhydrous acetonitrile (0.5 mL), PyBOP (17.5 mg, 0.03 mmol, 1.2 equiv.), and DIPEA (48.3 pL, 0.27 mmol, 10 equiv.), and the reaction was stirred for 5 min at room temperature (21.5 °C). Lastly, a solution of tert-Butyl 2-(7-{[(14-amino-3,6,9,12-tetraoxatetradecan-l-yl)carbamoyl]methyl}-4,10-bis[2- (tert-butoxy)-2-oxoethyl]-l,4,7,10-tetraazacyclododecan-l-yl)acetate (28.0 mg, 0.03 mmol, 1 equiv.) was added and the resulting solution was stirred overnight at room temperature. The reaction was monitored by HPLC-MS and after 18.5 h the reaction was re-dosed with PyBOP (10 mg, 0.02 mmol, 0.7 equiv.). Following an additional 3h stirring at room temperature the reaction was found to have gone to completion and was concentrated under vacuum. The crude product was purified by reverse phase-C18 column chromatography to afford two fractions of Intermediate 14 - A (12.9 mg, 29%, purity: 99%, and 20.5 mg, 39%, purity: 85%) as clear films, TFA salts. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 5.20 min; MS (positive ESI): found m / z 1391.4 [M+H]+; CveHmNsOie (calc. 1391.8). Step 2: Synthesis of tert-Butyl 2-{4, 7-bis[2-(tert-butoxy)-2-oxoethyl]-10-{[(14-{[5-(2,4- dihydroxy-5-isopropylphenyl)-4-[ 4-(morpholin-4-ylmethyl)phenyl -l, 2-oxazol-3-ylformamido}- 3, 6, 9, 12-tetraoxatetradecan-l-yl)carbamoyl]methyl}-l, 4, 7, 10-tetraazacyclododecan-l- yl}acetate (Intermediate 14 - B): To a solution of Intermediate 14 - A (20.5 mg, 0.011 mmol, 1 equiv.) in MeOH (2 mL) in a microwave vial was added a stir bar and Pd / C, 10% Pd basis (4.6 mg, 0.0043 mmol, 0.4 equiv.). The reaction mixture was degassed and subjected to H2 atmosphere via a balloon (3X). The reaction was then stirred at room temperature (22 °C) and monitored by HPLC-MS. After 21 h the reaction was filtered through an Acrodisc One (0.2 pm PTFE) syringe filter into a 20 mL scintillation vial and the reaction vial was rinsed with MeOH (2 x 2 mL). The crude reaction was then concentrated crude under vacuum to afford 20 mg of a clear film. The crude product was purified by reverse phase-C18 column chromatography to afford two fractions of Intermediate 14 - B (1.7 mg, 11%, purity: 98% and 11.4 mg, 48%, purity: 65%) as clear films, TFA salts. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 3.91 min; MS (positive ESI): found m / z 1212.0 [M+H]+; C62H99N8Oi6 (calc. 1211.7). *H NMR (700 MHz, DMSO-tL + -10% D2O) 87.41 (d, J= 7.0 Hz, 2H), 7.29 (d, J= 7.0 Hz, 2H), 6.71 (s, 1H), 6.38 (s, 1H), 4.27 (s, 2H), 3.61-2.71 (multiplets, 53H), 1.42 (br s, 9H), 1.34 (br s, 18H), 0.87 (d, J =7.0 Hz, 6H).

[0286] Step 3: Synthesis of [4, 7-bis(Carboxymethyl)-10-{[(14-{[5-(2,4-dihydroxy-5-isopropylphenyl)-4- [ 4-(morpholin-4-ylmethyl)phenyl -l, 2-oxazol-3-yl]formamido}-3, 6, 9, 12-tetraoxatetradecan-l- yl) carbamoyl] methyl}- 1,4, 7, 10-tetraazacyclododecan-l-yl] acetic acid (Compound N): To a 20 mL vial was added Intermediate 14 - B (11.4 mg, 6.4 pmol, 1 equiv., 65% purity) and a stir bar, followed by 2 mL of TFA / TIPS / H2O (95:2.5:2.5 v / v / v). The resulting clear solution was placed in a 37 °C oil bath and stirred, and the reaction was monitored by HPLC-MS. After 2.5 h the reaction had gone to completion and was concentrated under a stream of air. The crude product was purified by reverse phase-C18 column chromatography to afford Compound N (4.0 mg, 60%, purity: >96%) as a white solid, TFA salt. An aliquot was analyzed by HPLC-MS elution using elution method 1; retention time: 2.60 min; HRMS (positive ESI): found m / z 1043.5302 [M+H]+; C5oH75N8Oi6 (calc. 1043.5296). *HNMR (700 MHz, DMSO-tL + -10% D2O) 6 7.79 (s, 0.7 H), 7.48 (d, J= 7.0 Hz, 1.4 H), 7.41 (d, J= 14.0 Hz, 2H), 7.31 (d, J= 14.0 Hz, 1.4 H), 7.29 (d, J= 7.0 Hz, 2H), 6.96 (s, 0.7 H), 6.72 (s, 1H), 6.38 (s, 1H), 4.30 (s, 1.4 H), 4.26 (s, 2H), 4.1 - 2.9 (multiplets overlapping with H2O, 90H), 1.18 (app d, J= 7.0 Hz, 4.2 H), 0.87 (d, J= 7.0 Hz, 6H). -0.7: 1 ratio of atropisomers observed.

[0287] Example 2. Method for Radiolabeling Disclosed Compounds

[0142] Prior to radiolabeling, a stock solution of each Compound was made by dissolving the appropriate quantity of Compound in sodium acetate buffer.

[0288]

[0143] A 1.5 mL Eppendorf tube was charged with the desired Compound followed by sodium acetate buffer to raise the total volume as desired (0.05-0.5 mL). Next, a solution of [177Lu]LuC13 or [n iIn]InC13 or [225Ac] in hydrochloric acid solution was added and the mixture heated. After heating the reaction for a desired amount of time, iTLC analysis of the reaction mixture (solid phase: Silica gel (SG) or silicic acid (SA); mobile phase: 0.02 M citrate buffer with 5 % MeOH) indicated a radiochemical conversion (RCC) of >95 %. To the reaction mixture, sodium acetate buffer solutions of sodium / .-ascorbate and diethylenetriamine-pentaacetic acid calcium trisodium salt hydrate (DTP A) were added. iTLC and reverse phase HPLC at end-of-synthesis (EOS) indicated the formation of the desired radiolabelled products (>95 % radiochemical purity).

[0289]

[0144] Table 1: Radiolabeling of Compounds with [177Lu]LuCh

[0290]

[0145] Table 2: Radiolabeling of Compounds with [11 Un InCh

[0146] Scheme 15: Sample radiolabeling of a Compound featuring a DOTA chelator with [177LU]LUC13, [H lIn]InCl3, or [225Ac],

[0291]

[0147] Scheme 16: Sample radiolabeling of Compound featuring a DOTAGA chelator with

[0292] [177LU]LUC13, [H lIn]InCl3, or [225Ac], Example 3. In Vitro HSP90 Binding Studies of Radiopharmaceuticals Comprising Compounds of Formula I

[0293]

[0148] HSP90 binding is determined using a cell-free binding affinity assay, where human recombinant HSP90 protein is coated onto a Reacti-Bind® microtiter plate. Plates are incubated at 4 °C overnight and then washed 3 times with cold PBS containing 0.05% Tween-20 (PBS-T). Plates are equilibrated to room temperature, blocked for 1 hour on ice with PBS-T containing 0.2% gelatin, followed by 3 washes with PBS-T. Increasing concentrations of Radiolabeled (Lu- 177) conjugates are incubated in duplicate in the absence (total binding) or presence (nonspecific binding) of unlabeled conjugate and incubated at 37 °C for 1 hour with gentle shaking. Wells are then rinsed three times with PBS-T and stripped with a solution of 20 mM sodium acetate, pH 3.0 for up to 20 minutes at room temperature, followed by neutralization with an equal amount of 0. IN NaOH solution. Samples are transferred into gamma counting tubes and analyzed for Lu-177 content by gamma counter. Results are plotted relative to molar concentration of labeled conjugate, where specific binding affinity (Kd) is calculated using Graph Pad Prism, as shown in Figures 1-2. Binding affinity (Kd) results are summarized in Table 3, where 0 nM < A < 3 nM, and B > 3 nM.

[0294]

[0149] Table 3. Binding affinities (Kd) of selected compounds to recombinant Hsp90a or P isoforms

[0295] Example 4. Biodistribution Studies of Radiopharmaceuticals Comprising Compounds of Formula I

[0296]

[0150] In an animal model, Lutetium-177 (Lu-177) or Indium-111 (In-111) accumulation is measured in tumor, blood and healthy tissues of NCI-H460 tumor-bearing mice. Tumor bearing mice are dosed with 20 pCi of Lu-177 (or In-111) conjugates. At the time points indicated (ranging from Ih to 72h), mice are sacrificed and tumor, liver, kidneys and blood are isolated. All tissues are analyzed for Lu- 177 (or In-111) content by gamma counter, and Lu- 177 (or In- 111) uptake can be determined as percentage injected dose per gram of tissue (%ID / g). Average radioligand uptake (%ID / g, n = 3) is summarized in Table 4.

[0297]

[0151] Table 4 Biodistribution of selected compounds in tumor-bearing mice.

[0298] Example 5. In Vivo Studies of Radiopharmaceuticals Comprising Compounds of Formula I

[0299]

[0152] Using an in vivo model, overall survival and tumor growth regression in HSP90 expressing NCI-H460 tumor-bearing mice will be evaluated to determine the in vivo efficacy of the compounds of Formula I. Mice implanted with HSP90 expressing NCI-H460 xenograft tumors will be treated with various concentrations and doses of Ac-225 labelled HSP90 targeting conjugates (ranging from 0.1 to 3 pCi of activity) of Formula I administered intravenously via tail vein injection to evaluate in vivo efficacy. Tumor measurements will be taken 2-3 times per week for at least 28 days with vernier calipers in two dimensions to evaluate tumor growth, and body weight, along with overall body condition and general behaviour will be assessed daily. Tumor volumes (V; in mm3) will be calculated as V = 0.5 x L (length) x W2(width) and expressed as relative tumor volumes (tumour volume on day X divided by tumor volume on day of dosing). If necessary, the effectiveness of single and multiple doses of the compounds of Formula I will be evaluated. Pre-administration or co-admini strati on of non-radioconjugated HSP90 targeting conjugates with the compounds of Formula I and the effect on tumor regression and overall survival of HSP90 expressing xenograft tumour bearing mice can also be assessed.

[0300] OTHER EMBODIMENTS

[0301]

[0153] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments described herein. Such equivalents are intended to be encompassed by the following claims.

Claims

WHAT IS CLAIMED IS:

1. A compound of Formula I, or a pharmaceutically acceptable salt thereof:(I), whereinG is Ci-8 alkyl, Ci-8 heteroalkyl, C3-8 cycloalkyl, C2-8 heterocycloalkyl, aryl, or heteroaryl;Q1and Q2each are, independently, N or C;Q3and Q4each are, independently, NRaor O, in which Rais absent, H, C1-3 alkyl, or (C=O)Ci-3alkyl;R is H or C1-6 alkyl;L is a linker selected from the group consisting of C1-20 alkyl, C1-20 heteroalkyl, C3-10 cycloalkyl, C1-10 heterocycloalkyl, aryl, heteroaryl, C=O, (C=O)NR1, (C=S)NR1, NR1(C=O)NR1, NR1(C=S)NR1, and a combination thereof, in which each R1independently is H or C1-3 alkyl; or L is a hydrophilic space linker comprising one or more cyclic or acyclic polyhydroxy groups; andW is a chelator, wherein the compound binds to HSP90, wherein the compound optionally further comprises a radionuclide chelated by the chelator thereof, and wherein each of alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is optionally substituted with one or more moieties selected from the group consisting of OH, halo, alkoxy, amino, oxo, C1-3 haloalkyl, C1-3 alkyl-OH, C1-3 alkyl-NH2, (C=O)Ci-3 alkyl, COOH, and CONH2.

2. The compound of claim 1, having the structure of Formula II:(II), whereinX is CH or N;Y is O or NR2, in which R2is H, C1-3 alkyl, (C=O)Ci-3 alkyl,,R is H or C1-6 alkyl;L1is absent or C1-5 alkyl;L2is Ci -20 alkyl, Ci -20 heteroalkyl, C3-10 cycloalkyl, C1-10 heterocycloalkyl, aryl, or heteroaryl; n is an integer of 1-5 (inclusive);W is a chelator selected from the group consisting of DOTA, DOT AGA, NOTA, NOD AGA, NODASA, DTP A, TETA, EDTA, TRITA, CDTA, and DFO; andZ1and Z2each are, independently, absent or a moiety selected from the group consisting of C=O, (C=O)NR3, NR3(C=O)NR3, NR3(C=S)NR3, (CH2CH2)NR3, an amino acid unit, and a combination thereof, in which each of R3, independently, is H or C1-3 alkyl.

3. The compound of claim 2, whereinX is CH, Y is NCOCH3;L1is absent or C1-3 alkyl;L2is C2-10 heteroalkyl or C2.2o polyethylene glycol; n is 1 or 2; andZ1and Z2each are, independently, absent, or a moiety selected from the group consisting of (C=O)NH, NH(C=O)NH, (CH2CH2)NH, an amino acid unit, and a combination thereof wherein at least one of Z1and Z2is present.

4. The compound of claim 3, whereinL1is absent or -CH2CH2-;L2is C2-10 heteroalkyl or Ce-s polyethylene glycol; n is 1;Z1is absent, -(C=O)NH-, -NH(C=O)-, NH(C=O)NH, or (CH2CH2)NH;Z2is (C=O)NH, NH(C=O)NH, (CH2CH2)NH, or a combination thereof; and W is DOTA or DOTAGA.

5. The compound of any one of claims 2-4, wherein at least one of Z1and Z2is an amino acid unit.

6. The compound of claim 5, wherein the amino acid unit is formed from aspartic acid (Asp), glutamic acid (Glu), 2,4-diaminobutyric acid (Dab), 2,3 -diaminopropionic acid (Dap), lysine (Lys), or arginine (Arg).

7. The compound of any one of claims 1-6, wherein the compound is one of the following, or a stereoisomer thereof:

8. The compound of claim 1, having the structure of Formula III:whereinX is CH or N;Y is O or NR2, in which R2is H, C1-3 alkyl, (C=O)Ci-3 alkyl,,R is H or C1-6 alkyl;L1is absent or C1-3 alkyl;Z1and Z2each are, independently, absent or a moiety selected from the group consisting of C=O, (C=O)NR3, NR3(C=O)NR3, (CH2CH2)NR3, an amino acid unit, and a combination thereof, in which each of R3, independently, is H or C1-3 alkyl; n is an integer of 1-5 (inclusive);W is a chelator selected from the group consisting of DOTA, DOT AGA, NOTA, NOD AGA, NODASA, DTP A, TETA, EDTA, TRITA, CDTA, and DFO;X is absent or C1-5 alkyl;A is absent, C=O, CONR3, heterocycle, or heterocycle-Ci-3 alkyl, R3being H or C1-5 alkyl;Y is absent, C1-5 alkyl, or C1-10 heteroalkyl; and B is a cyclic or acyclic polyhydroxy group,wherein the compound comprises a hydrophilic space linker comprising one or more cyclic or acyclic polyhydroxy groups.

9. The compound of claim 8, wherein L1, Z1, and Z2are absent, and n is 1-3.

10. The compound of claim 8, wherein Z1is (CH2CH2)NH or(C=O)NH(CH2CH2)NH, Z2is absent, and n is 1-3.

11. The compound of claim 8, wherein at least one of Z1and Z2is an amino acid unit.

12. The compound of claim 11, wherein the amino acid unit is formed from aspartic acid (Asp), glutamic acid (Glu), 2,4-diaminobutyric acid (Dab), 2,3 -diaminopropionic acid (Dap), lysine (Lys), or arginine (Arg).

13. The compound of any one of claims 8-12, wherein X is C1-3 alkyl, A isN-N,N-N,N“N, orN“N, and Y is absent, C1-5 alkoxyl, or C2-8 polyethylene glycol, wherein indicates the attachment point to X, and “#” indicates the attachment point to Y.

14. The compound of any one of claims 8-13, wherein B is a cyclic polyhydroxy group having the structurewherein “#” indicates the attachment point to Y, r is 1-2, s is 3-4, and each R, independently, is OH, -CH2OH, -CO2H, -CONH2, NH2, orNHCOCH3, wherein B comprises at least three OH groups.

15. The compound of any one of claims 8-14, wherein B is one of the following, or a stereoisomer thereof:

16. The compound of any one of claims 8-13, wherein B is an acyclic polyhydroxyR ' rjD group having the structure of R , whereinindicates the attachment point to Y; t is 2-5; each R, independently, is OH, -CH2OH, -CO2H, -CONH2, NH2, or NHCOCH3; and Raand Rbeach are H or Raand Rbtogether form C=O, wherein B comprises at least three OH groups.

17. The compound of any one of claims 8-13 and 16, wherein B is one of the following, or a stereoisomer thereof:

18. The compound of any one of claims 8-17, wherein W is DOTA or DOTAGA.

19. The compound of any one of claims 1 and 8-18, wherein the compound is one of20. The compound of claim 1, having the structure of Formula IV:(IV), whereinX is CH or N;Y is O or NR2, in which R2is H, C1-3 alkyl, (C=O)Ci-3alkyl,,R is H or Ci-6 alkyl;L1is absent or C1-3 alkyl;Z1is absent or a moiety selected from the group consisting of C=O, (C=O)NR3, NR3(C=O)NR3, (CH2CH2)NR3, an amino acid unit, and a combination thereof, in which each of R3, independently, is H or C1-3 alkyl; m is an integer of 1-5 (inclusive), p is an integer of 1-4 (inclusive);W is a chelator selected from the group consisting of DOTA, DOTAGA, NOTA, NOD AGA, NODASA, DTP A, TETA, EDTA, TRITA, CDTA, and DFO; andR in each occurrence, independently, is OH, -CH2OH, -CO2H, -CONH2, NH2, or NHCOCH3, wherein the compound comprises a hydrophilic space linker comprising at least three OH groups.

21. The compound of claim 20, wherein X is CH, Y is (C=O)CH3, m is 3-4, p is 1-2, and R is OH or -CH2OH.

22. The compound of claim 20 or 21, wherein W is DOTA or DOTAGA.

23. The compound of any one of claims 1 and 20-22, wherein the compound is one of the following, or a stereoisomer thereof:

24. The compound of claim 1, having a structure selected from the group consisting of:

25. The compound of any one of claims 1-24, wherein the compound comprises a radionuclide selected from the group consisting of47Sc,55Co,60Cu,61Cu,62Cu,64Cu,67Cu,66Ga,67Ga,68Ga,82Rb,86Y,87Y,89Zr,90Y,97Ru, "Tc, "mTc,105Rh,109Pd,mIn,117mSn,149Pm,149Tb,153Sm,166HO,177LU,186Re,188Re,198Au,199AU,201T1,203Pb,211At,212Pb,212Bi,213Bi,223Ra,225Ac,227Th, and229Th.

26. The compound of claim 25, wherein the radionuclide is selected from the group consisting of68Ga,89Zr,90Y,U 1ln,177Lu, and225Ac.

27. The compound of claim 26, wherein the radionuclide is225Ac.

28. A pharmaceutical composition comprising a compound of any one of claims 1-27 and a pharmaceutically acceptable excipient.

29. A method of treating cancer, wherein the method comprises administering to a subject in need thereof a compound of any one of claims 1-27 or the composition of claim 28 in a therapeutically effective amount.

30. The method of claim 29, wherein the cancer is small-cell lung cancer, non-smallcell lung cancer, sarcoma, pancreatic cancer, breast cancer, or colon cancer.

31. A compound of any one of claims 1-27, or the pharmaceutical composition of claim 28, for use in a method of treating cancer.

32. Use of a compound of any one of claims 1-27, or the pharmaceutical composition of claim 28, in the manufacture of a medicament for the treatment of cancer.

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