HSP90-binding conjugates and preparations thereof

Abstract

Conjugates have been designed in which an active agent is attached to at least one targeting moiety, such as an HSP90-binding moiety, via a linker. Such conjugates can provide improved spatiotemporal delivery of the active agent, improved biodistribution and tumor penetration, and / or reduced toxicity. Methods for making the conjugates and formulations thereof are provided. Methods for administering the formulations to a subject in need thereof, for example, to treat or prevent cancer, are provided.

Description

[Technical Field]

[0001] The present invention relates to the use of molecules that target heat shock proteins, including heat shock protein 90 (HSP90), for example in the treatment of cancer. [Background technology]

[0002] Heat shock protein 90 (HSP90) is a molecular chaperone important for maintaining the stability and function of numerous client proteins. It is considered a major therapeutic target for anti-cancer drug development. Summary of the Invention

[0003] The present application provides conjugates comprising an active agent coupled to an HSP90 targeting moiety by a linker, and pharmaceutical compositions comprising such conjugates. Methods of making and using such conjugates are also provided. [Brief explanation of the drawings]

[0004] [Figure 1]

[0023] Figure 1 shows biodistribution data for radioactive analogs of CMP51 (including Lu177) in the NCI-H460 tumor model. CMP51 shows selective tumor exposure, excluding the kidney. [Figure 2] 1 shows biodistribution data for non-radioactive analogs of CMP6 (including Lu175) in the NCI-H460 tumor model. CMP6 shows selective tumor retention over kidney and liver. DETAILED DESCRIPTION OF THE INVENTION

[0005] The present applicants have designed HSP90-targeted conjugates containing active agents. Such targeting can, for example, improve the amount of active agent at the site and reduce active agent toxicity to the subject. The HSP90-targeted conjugates of the present invention have deep and rapid tumor penetration. The high accumulation and long residence time of the HSP90-targeted conjugates enable the use of cytotoxic and non-cytotoxic payloads, such as radionuclides, chemotherapeutic agents, kinase inhibitors, or immuno-oncological modifiers.

[0006] As used herein, "toxicity" refers to the ability of a substance or composition to be harmful or toxic to a cell, tissue, organism, or cellular environment. Reduced toxicity refers to a reduced ability of a substance or composition to be harmful or toxic to a cell, tissue, organism, or cellular environment. Such reduced or low toxicity can be relative to a standard measurement, relative to a treatment, or relative to the absence of treatment.

[0007] Toxicity may also be measured based on a subject's weight loss, where a weight loss of more than 15%, 20%, or 30% of body weight is indicative of toxicity. Other measures of toxicity may also be measured, such as measures of patient symptomatology, including lethargy and general malaise. Neutropenia or thrombocytopenia may also be measures of toxicity.

[0008] Pharmacological indicators of toxicity include elevated AST / ALT levels, neurotoxicity, renal damage, and GI disorders. In addition, the toxicity of a conjugate comprising an HSP90 targeting moiety linked to an active agent to cells that do not overexpress HSP90 is expected to be reduced compared to the toxicity of the active agent alone. Without being bound to any particular theory, Applicants believe this characteristic is due to the reduced ability of the conjugated active agent to remain in normal cells compared to tumor cells.

[0009] In some embodiments, the active agent and the targeting moiety, when joined by a linker to form a conjugate, have a synergistic effect: the efficacy of the conjugate is greater than that of the active agent and / or the targeting moiety alone.

[0010] In some embodiments, the potency of the active agent is reduced when it is linked to the targeting moiety by a cleavable linker: upon cleavage of the linker at the target site, such as a tumor site, the active agent is released and full potency is restored.

[0011] It is an object of the present invention to provide improved compounds, compositions and formulations for spatiotemporal drug delivery. It is a further object of the present invention to provide methods for making improved compounds, compositions and formulations for spatiotemporal drug delivery.

[0012] It is also an object of the present invention to provide methods of administering the improved compounds, compositions, and formulations to individuals in need thereof. I. Conjugates The conjugate comprises an active agent or a prodrug thereof attached by a linker to a targeting moiety, e.g., a molecule capable of binding to HSP90. The conjugate can be a conjugate between a single active agent and a single targeting moiety, e.g., a conjugate having the structure XYZ, where X is the targeting moiety, Y is a linker, and Z is the active agent.

[0013] In some embodiments, the conjugate contains two or more targeting moieties, two or more linkers, two or more active agents, or any combination thereof. The conjugate can have any number of targeting moieties, linkers, and active agents. The conjugate has the structure XYZYX, (XY) n -Z, X-(YZ) n , X n -YZ, XYZ n , (XYZ) n , (XYZY) n-Z, where X is a targeting moiety, Y is a linker, Z is an active agent, and n is an integer from 1 to 50, from 2 to 20, e.g., from 1 to 5. X, Y, and Z can be the same or different at each occurrence, e.g., the conjugate can contain more than one targeting moiety, more than one linker, and / or more than one active agent.

[0014] The conjugate may contain two or more targeting moieties attached to a single active agent. For example, the conjugate may contain an active agent to which multiple targeting moieties are attached, each via a different linker. The conjugate may have the structure XYZYX, where each X is a targeting moiety that may be the same or different, each Y is a linker that may be the same or different, and Z is an active agent.

[0015] The conjugate may contain two or more active agents attached to a single targeting moiety. For example, the conjugate may include a targeting moiety to which multiple active agents are each attached via a different linker. The conjugate may have the structure ZYXYZ, where X is a targeting moiety, each Y is a linker that may be the same or different, and each Z is an active agent that may be the same or different.

[0016] A. Active Agent The conjugates described herein contain at least one active agent (first active agent). The conjugates may contain two or more active agents, which may be the same as or different from the first active agent. The active agent may be a therapeutic, prophylactic, diagnostic, or nutritional agent. A variety of active agents are known in the art, and they or their analogs and derivatives may be used in the conjugates described herein. The active agent may be a protein or peptide, a small molecule, a nucleic acid or nucleic acid molecule, a lipid, a sugar, a glycolipid, a glycoprotein, a lipoprotein, or a combination thereof. In some embodiments, the active agent is an antigen, an adjuvant, a radioactive substance, an imaging agent (e.g., a fluorescent moiety), or a polynucleotide. In some embodiments, the active agent is an organometallic compound or a radioactive element. The active agent has chemical functionality for covalent attachment to a linker or is modified into an analog or derivative for covalent attachment to a linker.

[0017] In certain embodiments, the active agent of the conjugate comprises a predetermined molar weight percentage of about 1% to about 10%, or about 10% to about 20%, or about 20% to about 30%, or about 30% to about 40%, or about 40% to about 50%, or about 50% to about 60%, or about 60% to about 70%, or about 70% to about 80%, or about 80% to about 90%, or about 90% to about 99%, where the sum of the molar weight percentages of the components of the conjugate equals 100%. The amount of one or more active agents of the conjugate may also be expressed as a ratio relative to one or more targeting ligands. For example, the present teachings provide active agent to ligand ratios of about 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4; 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10.

[0018] radioactive material In some embodiments, the active agent Z is a radioactive substance or chemical moiety, e.g., a metal chelating group, that binds to a radionuclide (such as a radioisotope). Various radionuclides have emission characteristics, including α, β, γ, and Auger emissions, and can be used for therapeutic and / or diagnostic purposes. For example, the active agent Z may comprise a radioisotope such as Y-90, Y-86, 1-311, Re-186, Re-188, Y-90, Bi-212, At-211, Zr-89, Sr-89, Ho-166, Sm-153, Cu-67, Cu-64, Lu-177, Ac-225, Pb-203, Bi-213, Th-227, Pb-212, Ra-223, P-32, Sc-47, Br-77, Rh-105, Pd-103, Ag-111, Pr-142, Pm-149, Gd-159, Ir-194, and Pt-199.

[0019] In some embodiments, the active agent comprises an imaging probe such as a radioactive label (e.g., a radioisotope). Non-limiting examples of radioisotopes for imaging include I-124, I-131, In-111, Re-186, Re-188, Y-90, Bi-212, At-211, Sr-89, Ho-166, Sm-153, Cu-60, Cu-67, Cu-64, Lu-177, Ac-225, Bi-213, Th-227, Pb-212, Ra-223, P-32, Sc-47, These include Br-76, Br-77, Rh-105, Pd-103, Ag-111, Pr-142, Pm-149, Gd-159, In-111, Ir-194, Pt-199, Tc-99m, Co-57, Ga-66, Ga-67, Ga-68, Kr-81m, Rb-82, Sr-92, Tl-201, Y-86, Zr-89, C-11, N-13, O-15, and F-18.

[0020] In some embodiments, the active agent Z comprises a radioactive material, a chelator, or a radioactive material bound to a chelator. Conjugates comprising a radioactive material (e.g., a radioisotope) bound to a chelator are radioactive analogs of conjugates with the chelator alone or with the chelator bound to a non-radioactive isotope.

[0021] The chelating agent can be a metal chelating agent that binds to metals, including metal nuclides. The chelating agent can also be a moiety that binds to non-metal active agents. The chelating agent can be acyclic or macrocyclic. Non-limiting examples of chelating agents include 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA); DOTA derivatives: DO3A; diethylenetriamine-N,N,N',N",N"-pentaacetic acid (DTPA); DTPA derivatives: 2-(p-SCN-Bz)-6-methyl-DTPA, CHX-A"-DTPA, and cyclic anhydride of DTPA (CA-DTPA); 1,4,7-triazacyclononane-1,4-7-triacetic acid (NOTA); NOTA derivatives (e.g., BCNOTA, p-NCS-Bz-NOTA, BCNOT); 6-hydrazinonicotinamine EDTA (HYNIC); Ethylenediaminetetraacetic acid (EDTA); N,N'-Ethylene-di-L-cysteine; N,N'-Bis(2,2-dimethyl-2-mercaptoethyl)ethylenediamine-N,N'-diacetic acid (6SS); 1-(4-Carboxymethoxybenzyl)-N,N'-bis[(2-mercapto-2,2-dimethyl)ethyl]-1,2-ethylenediene-N,N'-diacetic acid (B6SS); Deferoxamine (DFO); 1,1,1-Tris(aminomethyl)ethane (TAME); Tris(aminomethyl)ethane-N,N,N',N',N",N"-hexaacetic acid (TAME Hex); O-hydroxybenzyliminodiacetic acid; 1,4,7-triazacyclononane (TACN); 1,4,7,10-thretraazacyclododecane (cyclene); 1,4,7-triazacyclononane-1-succinic acid-4,7-diacetic acid (NODASA); 1-(1-carboxy-3-carboxypropyl)-4,7-bis-(carboxymethyl)-1,4,7-triazacyclononane (NODAGA); 1,4,7-tris(2-mercaptoethyl)-1,4,7-triazacyclononane (triazacyclononane-TM); 1,4,7-triazacyclononane-N,N',N"-tris(methylenephosphonic) acid (NOTP); 1,4,8,11-tetraazacyclotetradecane-N,N',N",N"'-tetraacetic acid (TETA);These include 1,4,7,10,13-pentaazacyclopentadecane-N,N',N",N"',N""-pentaacetic acid (PEPA), 1,4,7,10,13,16-hexaazacyclohexadecane-N,N',N",N"',N"",N""-hexaacetic acid (HEHA); 1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetraazacyclododecane (TCMC); and derivatives or analogs thereof.

[0022] In some embodiments, the chelating agent is a polyaminocarboxylate agent such as ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 1,4,7,10-tetra-azacyclododecane-N,N',N",N'"-tetraacetic acid (DOTA), or a derivative thereof. They can coordinate with metals such as Fe, In, Ga, Zr, Y, Bi, Pb, or Ac.

[0023] [ka]

[0024] In some embodiments, the quenching agent is a macrocycle: 1,4,7-triazacyclononane-N,N',N"-triacetic acid (NOTA), 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid (TETA), 1,4,7,10,13-pentaazacyclopentadecane-N,N',N",N"',N""-pentaacetic acid (PEPA), 1,4,7,10,13,16-hexaazacyclohexadecane-N,N',N",N"',N"",N""'-hexaacetic acid (HEHA), or a derivative thereof.

[0025] Non-limiting examples of DTPA and its derivatives are:

[0026] [ka]

[0027] Non-limiting examples of DOTA and its derivatives are:

[0028] [ka]

[0029] In some embodiments, the conjugate of the present disclosure comprises DOTA, DOTAGA, or any derivative / analog thereof as a chelating agent. Any of the chelating agents disclosed in Eisenwiener et al., Bioorg Med Chem Lett. vol. 1.10(18):2133 (2000), the entire contents of which are incorporated herein by reference, such as 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, α-(2-carboxyethyl) (DOTAGA) or 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid, 10-(1,2-dicarboxyethyl) (DOTASA) can be used as the chelating agent.

[0030] [ka]

[0031] Other non-limiting examples of chelating agents are:

[0032] [ka]

[0033] [ka]

[0034] B. Linker The conjugate contains one or more linkers connecting the active agent and the targeting moiety. The conjugate is formed by linking the linker Y to one or more active agents and one or more targeting ligands. The linker Y is attached to the targeting moiety X and the active agent Z by a functional group independently selected from an ester bond, disulfide, amide, acylhydrazone, ether, carbamate, carbonate, sulfonamide, alkyl, aryl, heteroaryl, thioether, and urea. Alternatively, the linker can be attached to either the targeting ligand or the active agent by a group such as that provided by a bond between a thiol and a maleimide, or an azide and an alkyne. In some embodiments, the linker is a small molecule. In some embodiments, the linker is independently selected from the group consisting of alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl, wherein each of the alkyl, alkenyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl groups is optionally substituted with one or more groups each independently selected from halogen, cyano, nitro, hydroxyl, carboxyl, carbamoyl, ether, alkoxy, aryloxy, amino, amido, carbamate, alkyl, alkenyl, alkynyl, aryl, arylalkyl, cycloalkyl, heteroaryl, and heterocyclyl. wherein each of the carboxyl, carbamoyl, ether, alkoxy, aryloxy, amino, amido, carbamate, alkyl, alkenyl, alkynyl, aryl, arylalkyl, cycloalkyl, heteroaryl, or heterocyclyl is optionally substituted with one or more groups each independently selected from halogen, cyano, nitro, hydroxyl, carboxyl, carbamoyl, ether, alkoxy, aryloxy, amino, amido, carbamate, alkyl, alkenyl, alkynyl, aryl, arylalkyl, cycloalkyl, heteroaryl, and heterocyclyl.

[0035] In some embodiments, the linker comprises a cleavable functional group that is cleavable. The cleavable functional group can be hydrolyzed in vivo or can be designed to be enzymatically hydrolyzed, for example, by cathepsin B. As used herein, a "cleavable" linker refers to any linker that can be physically or chemically cleaved. Examples of physical cleavage can be cleavage by light, radiation, or heat, while examples of chemical cleavage include cleavage by redox reactions, hydrolysis, pH-dependent cleavage, or enzymatic cleavage. For example, the cleavable functional group can be a disulfide bond or a carbamate bond.

[0036] In some embodiments, the alkyl chain of the linker may be optionally interrupted by one or more atoms or groups selected from -O-, -C(=O)-, -NR, -OC(=O)-NR-, -S-, and -SS-. The linker may be selected from dicarboxylic acid derivatives of succinic acid, glutaric acid, or diglycolic acid. In some embodiments, the linker Y is selected from X'-R 1 -Y'-R 2 -Z', and the conjugate has formula Ia:

[0037] [ka]

[0038] wherein X is a targeting moiety as defined above; Z is an active agent; X′, R 1 , Y', R 2 and Z' is as defined herein. X' is either absent or independently selected from carbonyl, amido, urea, amino, ester, aryl, arylcarbonyl, aryloxy, arylamino, one or more natural or unnatural amino acids, thio, or succinimide; R 1 and R 2is either absent or composed of alkyl, substituted alkyl, aryl, substituted aryl, polyethylene glycol (2-30 units); Y' is absent or substituted or unsubstituted 1,2-diaminoethane, polyethylene glycol (2-30 units), or amide; Z' is either absent or independently selected from carbonyl, amido, urea, amino, ester, aryl, arylcarbonyl, aryloxy, arylamino, thio, or succinimide. In some embodiments, the linker may allow for linking one active agent molecule to two or more ligands, or one ligand to two or more active agent molecules.

[0039] In some embodiments, the linker Y is A m and the conjugate has formula Ib:

[0040] [ka]

[0041] wherein A is as defined herein and m=0-20. A in Formula Ia is a spacer unit and is either absent or independently selected from the following substituents: For each substituent, the dashed line represents the point of substitution with another independently selected unit of X, Z, or A, where X, Z, or A may be attached to either side of the substituent:

[0042] [ka]

[0043] where z=0-40, R is H or an optionally substituted alkyl group, and R′ is any side chain found in any natural or unnatural amino acid. In some embodiments, the conjugate has formula Ic:

[0044] [ka]

[0045] wherein A is defined above, m=0-40, n=0-40, x=1-5, y=1-5, and C is a branching element as defined herein. C in Formula Ic is a branched unit containing 3 to 6 functional groups selected from amines, carboxylic acids, thiols, or succinimides, including amino acids such as lysine, 2,3-diaminopropanoic acid, 2,4-diaminobutyric acid, glutamic acid, aspartic acid, and cysteine, for covalent attachment of a spacer unit, ligand, or active drug.

[0046] C.HSP90 targeting moiety As used herein, a targeting ligand (also referred to as a targeting moiety) includes any molecule capable of binding to one or more HSP90 proteins. Such targeting ligands may be peptides, antibody mimetics, nucleic acids (e.g., aptamers), polypeptides (e.g., antibodies), glycoproteins, small molecules, carbohydrates, or lipids.

[0047] The targeting moiety X can be, but is not limited to, natural compounds (e.g., geldanamycin and radicicol), and synthetic compounds, such as the geldanamycin analog 17-AAG (i.e., 17-allylaminogeldanamycin), the prince scaffold HSP90 inhibitor series, e.g., PU24FC1 (He H. et al., J. Med. Chem. 49:381, 2006, the contents of which are incorporated herein by reference in their entirety), BIIB021 (Lundgren K. et al., Mol. Cancer Ther. 8:4:921, 2009, the contents of which are incorporated herein by reference in their entirety), 4,5-diarylpyrazoles (Cheung K.M. et al., Mol. Cancer Ther. 8:4:921, 2009, the contents of which are incorporated herein by reference in their entirety), and the 4,5-diarylpyrazoles (Cheung K.M. et al., Mol. Cancer Ther. 8:4:921, 2009, the contents of which are incorporated herein by reference in their entirety). KM et al., Bioorg. Med. Chem. Lett., 15:3338, 2005, the contents of which are incorporated herein by reference in their entirety), 3-aryl,4-carboxamidopyrazoles (Brough PA et al., Bioorg. Med. Chem. Lett., 15:5197, 2005, the contents of which are incorporated herein by reference in their entirety), 4,5-diarylisoxazoles (Brough PA et al., Bioorg. Med. Chem. Lett., 15:5197, 2005, the contents of which are incorporated herein by reference in their entirety), PA) et al., J. Med. Chem., Vol. 51, p. 196, 2008, the contents of which are incorporated herein by reference in their entirety), 3,4-diarylpyrazole resorcinol derivatives (Dymock BW et al., J. Med. Chem., Vol. 48, p.4212, 2005, the contents of which are incorporated herein by reference in their entirety), thieno[2,3-d]pyrimidine (WO 2005034950 to VERNALIS et al., the contents of which are incorporated herein by reference in their entirety), aryltriazole derivatives of Formula I of EP 2655345 to Giannini et al., the contents of which are incorporated herein by reference in their entirety, or any other example of an HSP90 binding ligand or derivative / analog thereof.

[0048] In some embodiments, the HSP90 binding moiety may be a heterocyclic derivative containing three heteroatoms. WO 2009134110 to MATULIS et al., the contents of which are incorporated herein by reference in their entirety, discloses 4,5-diarylthiadiazoles that demonstrate good HSP90 binding affinity. Although this results in rather modest cell growth inhibition, it may be used as an HSP90 binding moiety in the conjugates of the invention. Another class of azaheterocyclic adducts, namely, triazole derivatives or analogs thereof, may also be used as an HSP90 binding moiety in the conjugates of the invention. For example, 1,2,4-triazole scaffolds have been well documented to have HSP90 inhibitory properties. WO 2009139916 to BURLISON et al. (Synta Pharmaceuticals Corp.), the contents of which are incorporated herein by reference in their entirety, discloses tricyclic 1,2,4-triazole derivatives that inhibit HSP90 at high micromolar concentrations. Any of the tricyclic 1,2,4-triazole derivatives disclosed in WO 2009139916 or derivatives / analogs thereof may be used as the HSP90-binding moiety in the conjugates of the invention. Any of the trisubstituted 1,2,4-triazole derivatives or derivatives / analogues thereof disclosed in WO 2010017479 and WO 2010017545 (Synta Pharmaceuticals Corp.), the contents of which are incorporated herein by reference in their entireties, may be used as the HSP90 binding moiety in the conjugates of the invention.In another example, the triazolone-containing HSP90 inhibitor designated ganetespib (formerly known as STA-9090, or its highly soluble phosphate prodrug STA-1474), disclosed in WO 2006055760 (Synta Pharmaceuticals Corp.), the contents of which are incorporated herein by reference in their entirety, or derivatives / analogs thereof, may be used as the HSP90 binding moiety in the conjugates of the invention.

[0049] [ka]

[0050] In some embodiments, ganetespib or a derivative / analog thereof may be used as the targeting moiety. Non-limiting examples of ganetespib derivatives / analogs are shown below.

[0051] [ka]

[0052] [ka]

[0053] [ka]

[0054] In some embodiments, onarespib (AT13387) or a derivative / analog thereof may be used as a targeting moiety in the conjugates of the invention. Non-limiting examples of onarespib and onarespib derivatives / analogs are provided below.

[0055] [ka]

[0056] In some embodiments, the targeting moiety comprises AUY-922, or an analog / derivative / fragment thereof. In one embodiment, the targeting moiety has the following structure:

[0057] [ka]

[0058] Any HSP90 ligand or HSP90 inhibitor disclosed in WO2013158644, WO2015038649, WO2015066053, WO2015116774, WO2015134464, WO2015143004, WO2015184246 (the contents of which are incorporated by reference in their entirety) or a derivative / analogue thereof may be used as the HSP90 binding moiety in the conjugates of the invention, for example: Formula I

[0059] [ka]

[0060] wherein R1 may be alkyl, aryl, halide, carboxamide, or sulfonamide; R2 may be alkyl, cycloalkyl, aryl, or heteroaryl, where when R2 is a 6-membered aryl or heteroaryl, R2 is substituted at the 3- and 4-positions relative to the point of attachment on the triazole ring through which the linker L is attached; and R3 may be SH, OH, -CONHR4, aryl, or heteroaryl, where when R3 is a 6-membered aryl or heteroaryl, R3 is substituted at the 3- or 4-position; Formula II

[0061] [ka]

[0062] wherein R1 can be alkyl, aryl, halo, carboxamido, sulfonamido; and R2 can be optionally substituted alkyl, cycloalkyl, aryl, or heteroaryl. Examples of such compounds include 5-(2,4-dihydroxy-5-isopropylphenyl)-N-(2-morpholinoethyl)-4-(4-(morpholinomethyl)phenyl)-4H-1,2,4-triazole-3-carboxamide and 5-(2,4-dihydroxy-5-isopropylphenyl)-4-(4-(4-methylpiperazin-1-yl)phenyl)-N-(2,2,2-trifluoroethyl)-4H-1,2,4-triazole-3-carboxamide; Formula III

[0063] [ka]

[0064] wherein X, Y, and Z may independently be CH, N, O, or S (with appropriate substitution and satisfying the valence of the corresponding atoms and aromaticity of the rings); R1 may be alkyl, aryl, halide, carboxamide, or sulfonamide; R2 may be substituted alkyl, cycloalkyl, aryl, or heteroaryl, where the linker L is directly attached or attached to an extended substitution on these rings; R3 may be SH, OH, NR4R5, and -CONHR6, to which an effector moiety may be attached; R4 and R5 may independently be H, alkyl, aryl, or heteroaryl; and R6 may be alkyl, aryl, or heteroaryl having at least one functional group to which an effector moiety may be attached; or Formula IV

[0065] [ka]

[0066] wherein R1 can be alkyl, aryl, halo, carboxamido, or sulfonamido; R2 and R3 independently are C1-C5 hydrocarbyl groups optionally substituted with one or more of hydroxy, halogen, C1-C2 alkoxy, amino, mono- and di-C1-C2 alkylamino; 5- to 12-membered aryl or heteroaryl groups; or R2 and R3 together with the nitrogen atom to which they are attached form a 4- to 8-membered monocyclic heterocyclic group, of which up to five ring members are selected from O, N, and S. An example of such a compound is AT-13387.

[0067] The HSP90 targeting moiety may be ganetespib, luminespib (AUY-922, NVP-AUY922), Devio-0932, MPC-3100, onarespib (AT-13387), SNX-2112, 17-amino-geldanamycin hydroquinone, PU-H71, or a derivative / analogue thereof.

[0068] [ka]

[0069] [ka]

[0070] The HSP90 targeting moiety may be SNX5422 (PF-04929113), or any of the compounds described in U.S. Pat. No. 8,080,556 (Pfizer), WO 2008096218 (Pfizer), WO 2006117669 (Pfizer), WO 2008059368 (Pfizer), WO 2008053319 (Pfizer), WO 2006117669 (Pfizer), European Patent No. The HSP90 inhibitor may be any other HSP90 inhibitor disclosed in EP 85701 (Novartis), EP 1776110 (Novartis), EP 2572709 (Novartis), WO 2012131413 (Debiopharm), or WO 2012131468 (Debiopharm), the contents of each of which are incorporated herein by reference in their entirety.

[0071] [ka]

[0072] HSP90 targeting moieties have also been used for PET imaging. 124 The compound may be a radiolabeled HSP90 inhibitor, PU-H71, or a derivative / analogue thereof. A conjugate comprising SNX-2112, 17-amino-geldanamycin hydroquinone, PU-H71, or AT13387 may have the following structure:

[0073] [ka]

[0074] In some embodiments, the targeting moiety comprises an imaging probe such as a radioactive label (e.g., a radioisotope). Non-limiting examples of radioisotopes include I-131, Re-186, Re-188, Y-90, Bi-212, At-211, Sr-89, Ho-166, Sm-153, Cu-67, Cu-64, Lu-177, Ac-225, Bi-213, Th-227, Pb-212, Ra-223, P-32, S C-47, Br-77, Rh-105, Pd-103, Ag-111, Pr-142, Pm-149, Gd-159, Ir-194, Pt-199, Tc-99m, Co-57, Ga-67, Kr-81m, Rb-82, Sr-92, Tl-201, C-11, N-13, O-15 and F-18.

[0075] In some embodiments, a conjugate of the present disclosure comprises two or more targeting moieties, for example, a conjugate can comprise two, three, four, or five HSP90 targeting moieties.

[0076] Extracellular HSP90 (eHSP90) In normal cells, HSP90 secretion occurs when cells are exposed to environmental stresses such as heat, drugs, cytokines, UV, and / or gamma radiation. The primary function of extracellular HSP90 (eHSP90) is to aid in tissue repair by promoting migration of cells at the edge of damaged tissue into the damaged area. However, in tumors, constitutively activated oncogenes trigger HSP90 secretion even in the absence of environmental stress. Tumor-secreted Hsp90, eHSP90α, promotes migration of both tumor cells and tumor stromal cells during invasion and metastasis. The extracellular stimulatory function of HSP90α depends on a 115-amino acid fragment (F-5) located on the surface of HSP90 (Li et al., Int Rev Cell Mol Biol., vol. 303:203-235 (2013), the entire contents of which are incorporated herein by reference). It has been shown that eHSP90 is present on the surface of tumor cells and can also be internalized (Crowe et al., ACS Chem. Biol., vol. 12:1047-1055 (2017)). Therefore, surface expression of eHSP90 on tumor cells represents a target for selectively delivering therapeutics to tumors over healthy cells. Therefore, eHSP90 (especially eHSP90α) may be an excellent target for treating tumors.

[0077] In some embodiments, the targeting moiety selectively binds to eHSP90. In some embodiments, the targeting moiety binds to the F-5 region of eHSP90. In some embodiments, the targeting moiety has low cell permeability and preferentially binds to cell surface eHSP90. In some embodiments, the targeting moiety is cell impermeable and binds exclusively to eHSP90. In some embodiments, the conjugate comprising the targeting moiety has low cell permeability or is cell impermeable.

[0078] In some embodiments, the targeting moiety comprises HS-23, HS-131 (disclosed in Crowe et al., ACS Chem. Biol., vol. 12:1047-1055 (2017), the contents of which are incorporated herein by reference in their entirety), or DMAG-N-oxide (a cell-impermeable version of 17-AAG disclosed in Tsutsumi et al., Oncogene, vol. 27(17):2478-2487 (2008), the contents of which are incorporated herein by reference in their entirety), or an analog / derivative thereof, the structures of which are shown below.

[0079] [ka]

[0080] In certain embodiments, the one or more targeting moieties of the conjugate are present in a molar weight percentage of about 0.1% to about 10%, or about 1% to about 10%, or about 10% to about 20%, or about 20% to about 30%, or about 30% to about 40%, or about 40% to about 50%, or about 50% to about 60%, or about 60% to about 70%, or about 70% to about 80%, or about 80% to about 90%, or about 90% to about 99%, such that the sum of the molar weight percentages of the components of the conjugate equals 100%. The amount of targeting moiety of the conjugate can also be expressed as a ratio to the active agent, e.g., a ligand to active agent ratio of about 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4; 1:5, 1:6, 1:7, 1:8, 1:9, or 1:10.

[0081] D. Pharmacokinetic Modulation Unit The conjugate of the present invention may further comprise at least one exolinker bonded to a reactive group that reacts with a functional group of a protein or artificial protein or its derivative / analog / mimetics, or may comprise at least one exolinker bonded to a pharmacokinetics-modulating unit (PMU). The exolinker connecting the conjugate to the reactive group or the pharmacokinetics-modulating unit may be a cleavable linker that allows the conjugate to be released. Thus, the conjugate can be separated from the protein or the pharmacokinetics-modulating unit as needed.

[0082] Any reactive group or PMU (such as a polymer-containing PMU) disclosed in WO 2017 / 197241, the entire contents of which are incorporated herein by reference, can be attached to a conjugate of the present disclosure.

[0083] In some embodiments, the conjugate comprises a protein-binding reactive group attached to its active agent. In some embodiments, the conjugate comprises a protein-binding reactive group attached to its targeting moiety. In some embodiments, the conjugate comprises a protein-binding reactive group attached to its linker. The reactive group binds reversibly or irreversibly to the protein. The protein can be a naturally occurring protein, such as a serum or plasma protein, or a fragment thereof. Specific examples include fetal Fc receptor (FcRn), thyroxine-binding protein, transthyretin, alpha 1-acid glycoprotein (AAG), transferrin, fibrinogen, albumin, immunoglobulins, alpha 2-macroglobulin, lipoproteins, or fragments thereof. The reactive group can bind to such proteins through covalent or non-covalent interactions, such as hydrogen bonds, ionic bonds, van der Waals interactions, and hydrophobic bonds.

[0084] In some embodiments, the protein-binding reactive group can bind to serum proteins through non-covalent interactions. For example, the reactive group can bind to serum proteins with weak affinity (10 -4 ~10 -5The reactive group may be a saturated fatty acid that binds to albumin at the cleavage site (M). Non-limiting examples of such fatty acids include myristic acid (a fatty acid having 14 carbon atoms) and palmitic acid (a fatty acid having 16 carbon atoms). Other non-limiting examples of reactive groups include a naphthalene acylsulfonamide group, a diphenylcyclohexanol phosphate ester group, a 6-(4-(4-iodophenyl)butanamido)hexanoate group ("Albu" tag), and a series of peptides having the core sequence DICLPRWGCLW, including SA21 (a cyclic peptide having the 18 amino acids Ac-RLIEDICLPRWGCLWEDD-NH2) disclosed in Dennis et al., in. J. Biol. Chem., vol. 277:35035 (2002), the contents of which are incorporated herein by reference in their entirety.

[0085] The protein binding reactive group may include the following structure:

[0086] [ka]

[0087] In some embodiments, the protein-binding reactive group may include any of the peptide-fatty acid albumin-binding ligands disclosed in Zorzi et al., Nature Communications, vol. 8:16092, (2017) (the entire contents of which are incorporated herein by reference). These peptide-fatty acid albumin-binding ligands include a fatty acid attached to a short peptide, e.g., a heptapeptide, via an amino acid side chain. The fatty acid may be attached to the short peptide via its carboxylic acid group to a lysine side chain. The fatty acid binds to albumin with micromolar affinity, and the short peptide increases the affinity by forming additional contact points with albumin. The peptide-fatty acid ligand may have the following general structure:

[0088] [ka]

[0089] where X = any amino acid (such as Gly or Ser), K = Lys, and n = 12 (myristic acid), 14 (palmitic acid), or 16 (stearic acid). In some embodiments, any albumin-binding functional group disclosed in U.S. Pat. No. 9,670,482 (Bicyclic Therapeutic Agents), the entire contents of which are incorporated herein, can be used as the protein-binding reactive group herein. In some embodiments, the protein-binding reactive group comprises a fluorene ring and binds non-covalently and / or reversibly to albumin. As a non-limiting example, the protein-binding reactive group comprises a fluorenylmethyloxycarbonyl (FMOC) group. Optionally, the protein-binding reactive group comprises at least one amino acid linked to FMOC, such as Lys, Trp, Gly, or Phe. For example, the small molecule may comprise Fmoc-Lys-, Fmoc-Gly-, Fmoc-Phe-, Fmoc-GGSGD-, Fmoc-FGGGD-, Fmoc-FGSGD-, Fmoc-WGSGD-, Fmoc-WGGGA, or Fmoc-Trp-GGG.

[0090] [ka]

[0091] Non-limiting examples of conjugates In some embodiments, the conjugate comprises at least one HSP90 targeting moiety attached to a radioactive chelator with a linker. The HSP90 targeting moiety can be a ganetespib analog or derivative (such as TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, or TM14), an onarespib analog or derivative (such as TM6 or TM7), or TM15. The radioactive material can be lutetium-177 (Lu177 or 177 The conjugate may contain any radioactive isotope, such as lutetium isotope 177 ( 177Lu) confers radioactivity to the conjugate. The chelating agent can be any suitable chelating agent, such as DOTA or DOTAGA. The molecular weight of the conjugate can be less than 5000 Da, for example, from about 1000 Da to about 3000 Da, or from about 1500 Da to 2500 Da.

[0092] In some embodiments, the conjugate has formula X:

[0093] [ka]

[0094] where TM(s) refers to at least one targeting moiety that binds to HSP90. TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15. In some embodiments, the conjugate has one targeting moiety. In some embodiments, the conjugate has two targeting moieties. In some embodiments, the conjugate has three targeting moieties. In some embodiments, the conjugate has four targeting moieties.

[0095] In some embodiments, the conjugate has the formula X10:

[0096] [ka]

[0097] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15.

[0098] In some embodiments, the conjugate has the formula X1:

[0099] [ka]

[0100] wherein TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15. Non-limiting examples of conjugates within Formula X1 include CMP24, CMP26, and CMP27.

[0101] In some embodiments, the conjugate has the formula X20:

[0102] [ka]

[0103] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15.

[0104] In some embodiments, the conjugate has the formula X2:

[0105] [ka]

[0106] wherein TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15. Non-limiting examples of conjugates within formula X2 include CMP22, CMP8, CMP9, CMP10, CMP11, CMP12, CMP17, CMP18, CMP19, CMP20, CMP21, CMP25, CMP28, T25, T26, T30, T31, T40, and T43.

[0107] Non-limiting examples of conjugates of the present disclosure within Formula X include CMP7, CMP8, CMP9, CMP10, CMP11, CMP12, CMP17, CMP18, CMP19, CMP20, CMP21, CMP22, CMP24, CMP25, CMP26, CMP27, CMP28, T18, T19, T20, T21, T25, T26, T30, T31, T43, or T40:

[0108] [Table 1A-1]

[0109] [Table 1A-2]

[0110] [Table 1A-3]

[0111] [Table 1A-4]

[0112] [Table 1A-5]

[0113] [Table 1A-6]

[0114] [Table 1A-7]

[0115] [Table 1A-8]

[0116] [Table 1A-9]

[0117] [Table 1A-10]

[0118] [Table 1A-11]

[0119] [Table 1A-12]

[0120] Radioactive-free conjugate analog structures are included in Table 1. The conjugates can include any radioactive isotope, such as Y-90, Y-86, I-131, Re-186, Re-188, Y-90, Bi-212, At-211, Zr-89, Sr-89, Ho-166, Sm-153, Cu-67, Cu-64, Lu-177, Ac-225, Pb-203, Bi-213, Th-227, Pb-212, Ra-223, P-32, Sc-47, Br-77, Rh-105, Pd-103, Ag-111, Pr-142, Pm-149, Gd-159, Ir-194, and Pt-199.

[0121] [Table 1B-1]

[0122] [Table 1B-2]

[0123] [Table 1B-3]

[0124]

Table 1B-4

[0125]

Table 1B-5

[0126]

Table 1B-6

[0127]

Table 1B-7

[0128]

Table 1B-8

[0129]

Table 1B-9

[0130]

Table 1B-10

[0131]

Table 1B-11

[0132]

Table 1B-12

[0133] In some embodiments, the conjugate comprises at least one TM1 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises one TM1 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises two TM1 as ligands. Non-limiting examples of conjugates include CMP11 and T18.

[0134] In some embodiments, the conjugate comprises at least one TM2 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises one TM2 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises two TM2 as ligands. A non-limiting example of a conjugate includes CMP12.

[0135] In some embodiments, the conjugate comprises at least one TM3 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises one TM3 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises two TM3 as ligands. A non-limiting example of a conjugate includes CMP10.

[0136] In some embodiments, the conjugate comprises at least one TM5 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises one TM5 as a ligand, such as CMP22. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises two TM5 as ligands. Non-limiting examples of conjugates include CMP8, CMP17, CMP18, CIMP19, CMP20, CMP21, CMP26, T20, T25, T26, T30, T31, and T43. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises three TM5 as ligands. Non-limiting examples of conjugates include T40. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises four TM5 as ligands. Non-limiting examples of conjugates include CMP9.

[0137] In some embodiments, the conjugate comprises at least one TM9 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises one TM9 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises two TM9 as ligands. Non-limiting examples of conjugates include CMP24, CMP25, CMP27, and CMP28.

[0138] In some embodiments, the conjugate comprises at least one TM10 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises one TM10 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises two TM10 as ligands. Non-limiting examples of conjugates include CMP7 and T21.

[0139] In some embodiments, the conjugate comprises at least one TM14 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises one TM14 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and comprises two TM14 as ligands. Non-limiting examples of conjugates include T19.

[0140] Amino Acid Spacer In some embodiments, the conjugate comprises at least one HSP90 targeting moiety bound to a radioactive chelator with a linker. The HSP90 targeting moiety can be a ganetespib analog or derivative (such as TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, or TM14), an onarespib analog or derivative (such as TM6 or TM7), or TM15. The linker can include a spacer consisting of at least one amino acid or analog thereof, for example, two amino acids or analogs thereof, three amino acids or analogs thereof, four amino acids or analogs thereof, or five amino acids or analogs thereof. The amino acid or analog thereof can be a D-amino acid. The amino acid or analog thereof can be anionic (e.g., DGlu), cationic (e.g., DLys), or uncharged (e.g., Sar, where Sar = N-methylglycine). The spacer can be DGlu-DGlu-DLys, DLys-DLys-DGlu, DGlu-DGlu-DGlu, DLys-DLys-DLys, Sar-DLys-Sar, Sar-Sar-Sar, Sar-DGlu-Sar, Ala-Asp-D-Ser, Ala-Asp-L-Ser, or Glu. Without wishing to be bound by any theory, the spacer may affect the biodistribution of the conjugate and reduce hepatic uptake of the conjugate. The binding affinity for HSP90 is maintained regardless of the charge present on the spacer.

[0141] In some embodiments, the conjugate has Formula A:

[0142] [ka]

[0143] where TM represents at least one targeting moiety that binds to HSP90. In some embodiments, the conjugate has two targeting moieties. In some embodiments, the conjugate has three targeting moieties. In some embodiments, the conjugate has four targeting moieties. Non-limiting examples of conjugates encompassed by Formula A1 include T1, T10, T27, T28, CMP13, CMP16, T2, T4, T5, T6, T8, T9, T29, T39, CMP14, CMP15, CMP23, CMP37, CMP38, CMP43, CMP44, CMP45, CMP46, CMP47, CMP48, CMP49, and CMP50.

[0144] In some embodiments, the conjugate has formula A10:

[0145] [ka]

[0146] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15.

[0147] In some embodiments, the conjugate has formula A1:

[0148] [ka]

[0149] wherein TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15. Non-limiting examples of conjugates encompassed by Formula A1 include CMP23, CMP37, and CMP38.

[0150] In some embodiments, the conjugate has formula A20:

[0151] [ka]

[0152] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15.

[0153] In some embodiments, the conjugate has formula A2:

[0154] [ka]

[0155] wherein TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15. Non-limiting examples of conjugates encompassed by Formula A2 include T10, T27, T28, CMP13, CMP16, T2, T4, T5, T6, T8, T9, T29, T39, CMP14, CMP15, CMP43, CMP44, CMP45, CMP46, CMP47, CMP48, CMP49, and CMP50.

[0156] Non-limiting examples of conjugates of the present disclosure encompassed by Formula A include T1, T2, T4, T5, T6, T8, T9, T10, T27, T28, T29, T39, CMP13, CMP14, CMP15, CMP16, CMP23, CMP37, CMP38, CMP43, CMP44, CMP45, CMP46, CMP47, CMP48, CMP49, or CMP50:

[0157] [Table 2A-1]

[0158] [Table 2A-2]

[0159] [Table 2A-3]

[0160] [Table 2A-4]

[0161] [Table 2A-5]

[0162] [Table 2A-6]

[0163] [Table 2A-7]

[0164] [Table 2A-8]

[0165] [Table 2A-9]

[0166] [Table 2A-10]

[0167] [Table 2A-11]

[0168] [Table 2A-12]

[0169] [Table 2A-13]

[0170] Radioactive-free conjugate analog structures are included in Table 2'. The conjugates can contain any radioactive isotope, such as Y-90, Y-86, I-131, Re-186, Re-188, Y-90, Bi-212, At-211, Zr-89, Sr-89, Ho-166, Sm-153, Cu-67, Cu-64, Lu-177, Ac-225, Pb-203, Bi-213, Th-227, Pb-212, Ra-223, P-32, Sc-47, Br-77, Rh-105, Pd-103, Ag-111, Pr-142, Pm-149, Gd-159, Ir-194, and Pt-199.

[0171] [Table 2B-1]

[0172] [Table 2B-2]

[0173]

Table 2B-3

[0174]

Table 2B-4

[0175]

Table 2B-5

[0176]

Table 2B-6

[0177]

Table 2B-7

[0178]

Table 2B-8

[0179]

Table 2B-9

[0180]

Table 2B-10

[0181]

Table 2B-11

[0182]

Table 2B-12

[0183]

Table 2B-13

[0184] In some embodiments, the conjugate comprises at least one amino acid spacer and at least one TM1 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and one TM1 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and two TM1 as ligands. Non-limiting examples include T10, T27, and T28.

[0185] In some embodiments, the conjugate comprises at least one amino acid spacer and at least one TM2 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and one TM2 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and two TM2 as ligands. Non-limiting examples include CMP13 and CMP16.

[0186] In some embodiments, the conjugate comprises at least one amino acid spacer and at least one TM5 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and one TM5 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and two TM5 as ligands. Non-limiting examples include T1, T2, T4, T5, T8, T9, T39, CMP14, CMP15, CMP43, CMP44, CMP45, CMP46, CMP47, CMP48, CMP49, and CMP50.

[0187] In some embodiments, the conjugate comprises at least one amino acid spacer and at least one TM10 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and one TM10 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and two TM10 as ligands. Non-limiting examples include T6, T29, CMP37, and CMP38.

[0188] In some embodiments, the conjugate comprises at least one amino acid spacer and at least one TM15 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and one TM15 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer and two TM10 as ligands. Non-limiting examples include CMP23.

[0189] PEG spacer In some embodiments, the conjugate comprises an HSP90 targeting moiety bound to a radioactive chelator with a linker. The HSP90 targeting moiety can be a ganetespib analog or derivative (such as TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, or TM14), an onarespib analog or derivative (such as TM6 or TM7), or TM15. The linker can include a spacer comprising polyethylene glycol (PEG). The PEG spacer can be constructed from (PEG)n, where n is an integer from 1 to 20. In some embodiments, the PEG spacer is (PEG)4. In some embodiments, the PEG spacer is (PEG)12. The binding affinity for HSP90 is maintained regardless of the charge present on the spacer.

[0190] In some embodiments, the conjugate has formula B:

[0191] [ka]

[0192] wherein TM represents at least one targeting moiety that binds to HSP90. In some embodiments, the conjugate has two targeting moieties. In some embodiments, the conjugate has three targeting moieties. In some embodiments, the conjugate has four targeting moieties.

[0193] In some embodiments, the conjugate has formula B10:

[0194] [ka]

[0195] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM6, TM7, or TM14.

[0196] In some embodiments, the conjugate has formula B1:

[0197] [ka]

[0198] wherein TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM6, TM7, or TM14. Non-limiting examples of conjugates encompassed by Formula B1 include CMP1, CMP29, CMP30, CMP31, CMP32, CMP33, CMP34, CMP35, CMP36, CMP39, CMP40, CMP41, and CMP42.

[0199] In some embodiments, the conjugate has formula B20:

[0200] [ka]

[0201] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM6, TM7, or TM14.

[0202] In some embodiments, the conjugate has formula B2:

[0203] [ka]

[0204] wherein TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM6, TM7, or TM14. Non-limiting examples of conjugates encompassed by Formula B2 include CMP2.

[0205] Non-limiting examples of conjugates of the present disclosure encompassed by Formula B include CMP1, CMP2, CMP29, CMP30, CMP31, CMP32, CMP33, CMP34, CMP35, CMP36, CMP39, CMP40, CMP41, CMP42, CMP52.

[0206] [Table 3A-1]

[0207] [Table 3A-2]

[0208] [Table 3A-3]

[0209] [Table 3A-4]

[0210] Radioactive-free conjugate analog structures are included in Table 3. The conjugates can contain any radioactive isotope, such as Y-90, Y-86, I-131, Re-186, Re-188, Y-90, Bi-212, At-211, Zr-89, Sr-89, Ho-166, Sm-153, Cu-67, Cu-64, Lu-177, Ac-225, Pb-203, Bi-213, Th-227, Pb-212, Ra-223, P-32, Sc-47, Br-77, Rh-105, Pd-103, Ag-111, Pr-142, Pm-149, Gd-159, Ir-194, and Pt-199.

[0211] [Table 3B-1]

[0212] [Table 3B-2]

[0213] [Table 3B-3]

[0214] [Table 3B-4]

[0215] [Table 4A]

[0216] Radioactive-free conjugate analog structures are included in Table 4. The conjugates can contain any radioactive isotope, such as Y-90, Y-86, I-131, Re-186, Re-188, Y-90, Bi-212, At-211, Zr-89, Sr-89, Ho-166, Sm-153, Cu-67, Cu-64, Lu-177, Ac-225, Pb-203, Bi-213, Th-227, Pb-212, Ra-223, P-32, Sc-47, Br-77, Rh-105, Pd-103, Ag-111, Pr-142, Pm-149, Gd-159, Ir-194, and Pt-199.

[0217] [Table 4B]

[0218] In some embodiments, the conjugate comprises at least one (PEG) spacer. Non-limiting examples include CMP1, CMP2, CMP29, CMP30, CMP31, CMP39, CMP40, and CMP35.

[0219] In some embodiments, the conjugate comprises at least one (PEG)12 spacer. Non-limiting examples include CMP32, CMP33, CMP34, CMP36, CPM41, and CMP42.

[0220] Albumin-binding pharmacokinetics control unit In some embodiments, the conjugate comprises an HSP90 targeting moiety bound to a radioactive chelator with a linker and at least one pharmacokinetic modulating unit (PMU). The PMU can be any group that binds to albumin. The HSP90 targeting moiety can be a ganetespib analog or derivative (TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, or TM14), an onarespib analog or derivative (such as TM6 or TM7), or TM15. In some embodiments, the linker can include a spacer comprising polyethylene glycol (PEG), e.g., (PEG)4 or (PEG)12. In some embodiments, the linker can include at least one amino acid. HSP90 binding affinity is maintained regardless of the charge present on the spacer or PMU.

[0221] In some embodiments, the PMU comprises:

[0222] [ka]

[0223] It contains a functional group that binds to albumin, such as (4-(4-iodophenyl)butanamide group). In some embodiments, the conjugate has formula C:

[0224] [ka]

[0225] wherein TM represents at least one targeting moiety that binds to HSP90. In some embodiments, the conjugate has two targeting moieties. In some embodiments, the conjugate has three targeting moieties. In some embodiments, the conjugate has four targeting moieties. In some embodiments, the linker comprises a PEG spacer. In some embodiments, the linker comprises an amino acid spacer.

[0226] In some embodiments, the conjugate has formula C10:

[0227] [ka]

[0228] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15.

[0229] In some embodiments, the conjugate has formula C1:

[0230] [ka]

[0231] wherein TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15. Non-limiting examples of conjugates encompassed by Formula C1 include CMP3, CMP4, CMP5, CMP6.

[0232] In some embodiments, the conjugate has formula C20:

[0233] [ka]

[0234] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15.

[0235] In some embodiments, the conjugate has formula C2:

[0236] [ka]

[0237] wherein TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15. Non-limiting examples of conjugates encompassed by Formula C1 include T3, T7, T11, T12, T13, T14, T15, T16, T17, T22, T23, T32, T24, T33, T34, T35, T36, T37, T38, T41, and T42.

[0238] Non-limiting examples of conjugates of the present disclosure encompassed by Formula C include CMP3, CMP4, CMP5, CMP6, T3, T7, T11, T12, T13, T14, T15, T16, T17, T22, T23, T32, T24, T33, T34, T35, T36, T37, T38, T41, or T42:

[0239] [Table 5A-1]

[0240] [Table 5A-2]

[0241] [Table 5A-3]

[0242] [Table 5A-4]

[0243] [Table 5A-5]

[0244]

Table 5A-6

[0245]

Table 5A-7

[0246]

Table 5A-8

[0247]

Table 5A-9

[0248]

Table 5A-10

[0249]

Table 5A-11

[0250]

Table 5A-12

[0251]

Table 5A-13

[0252]

Table 5A-14

[0253]

Table 5A-15

[0254] [Table 5A-16]

[0255] [Table 5A-17]

[0256] [Table 5A-18]

[0257] [Table 5A-19]

[0258] [Table 5A-20]

[0259] [Table 5A-21]

[0260] Radioactive-free conjugate analog structures are included in Table 5. The conjugates can include any radioactive isotope, such as Y-90, Y-86, I-131, Re-186, Re-188, Y-90, Bi-212, At-211, Zr-89, Sr-89, Ho-166, Sm-153, Cu-67, Cu-64, Lu-177, Ac-225, Pb-203, Bi-213, Th-227, Pb-212, Ra-223, P-32, Sc-47, Br-77, Rh-105, Pd-103, Ag-111, Pr-142, Pm-149, Gd-159, Ir-194, and Pt-199.

[0261] [Table 5B-1]

[0262]

Table 5B-2

[0263]

Table 5B-3

[0264]

Table 5B-4

[0265]

Table 5B-5

[0266]

Table 5B-6

[0267]

Table 5B-7

[0268]

Table 5B-8

[0269]

Table 5B-9

[0270]

Table 5B-10

[0271]

Table 5B-11

[0272]

Table 5B-12

[0273]

Table 5B-13

[0274]

Table 5B-14

[0275]

Table 5B-15

[0276]

Table 5B-16

[0277]

Table 5B-17

[0278]

Table 5B-18

[0279]

Table 5B-19

[0280]

Table 5B-20

[0281] In some embodiments, the conjugate comprises at least one albumin-binding PMU and at least one TM1 as a ligand. In some embodiments, the conjugate comprises at least one albumin-binding PMU and one TM1 as a ligand. In some embodiments, the conjugate comprises at least one albumin-binding PMU and two TM1 as ligands, such as T17.

[0282] In some embodiments, the conjugate comprises at least one albumin-binding PMU and at least one TM2 as a ligand. In some embodiments, the conjugate comprises at least one albumin-binding PMU and one TM2 as a ligand. In some embodiments, the conjugate comprises at least one albumin-binding PMU and two TM2 as ligands, such as T13.

[0283] In some embodiments, the conjugate comprises at least one albumin-binding PMU and at least one TM5 as a ligand. In some embodiments, the conjugate comprises at least one albumin-binding PMU and one TM5 as a ligand, such as T7. In some embodiments, the conjugate comprises at least one albumin-binding PMU and two TM5 as ligands. Non-limiting examples include T3, T14, T15, T16, T22, T23, T32, T24, T33, T34, T35, T36, T37, T38, T41, and T42.

[0284] In some embodiments, the conjugate comprises at least one albumin-binding PMU and at least one TM9 as a ligand, hi some embodiments, the conjugate comprises at least one albumin-binding PMU and at least one TM9 as a ligand, such as CMP3 and CMP4.

[0285] In some embodiments, the conjugate comprises at least one albumin-binding PMU and at least one TM10 as a ligand. In some embodiments, the conjugate comprises at least one albumin-binding PMU and one TM10 as a ligand, such as CMP5, CMP6, and T11. In some embodiments, the conjugate comprises at least one amino acid spacer and two TM10 as ligands, such as T12.

[0286] Conjugate T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12, T13, T14, T15, T16, T17, T18, T19, T20, T21, T22, T23, T24, T25, T26, T27, T28, T29, T30, T 31, T32, T33, T34, T35, T36, T37, T38, T39, T40, T41, T42, T43, CMP1, CMP2, CMP3, CMP4, CMP5, CMP6, CMP7, CMP8, CMP9, CMP10, CMP11, CMP12, CMP13, Lutetium (Lu) in CMP14, CMP15, CMP16, CMP17, CMP18, CMP19, CMP20, CMP21, CMP22, CMP23, CMP24, CMP25, CMP26, CMP27, CMP28, CMP30, CMP31, CMP32, CMP33, CMP34, CMP35, CMP36, CMP37, CMP38, CMP39, CMP40, CMP41, CMP42, CMP43, CMP44, CMP45, CMP46, CMP47, CMP48, CMP49, CMP50, or CMP52 is replaced with Lu177( 177Lu) or other radioactive isotopes (such as Y-90, Y-86, I-131, Re-186, Re-188, Y-90, Bi-212, At-211, Zr-89, Sr-89, Ho-166, Sm-153, Cu-67, Cu-64, Ac-225, Pb-203, Bi-213, Th-227, Pb-212, Ra-223, P-32, Sc-47, Br-77, Rh-105, Pd-103, Ag-111, Pr-142, Pm-149, Gd-159, Ir-194, and Pt-199) can be substituted to provide radioactive analogs of the conjugate.

[0287] II. Formulation In some embodiments, the composition is administered to a human, human patient, or subject. For purposes of this disclosure, the phrase "active ingredient" generally refers to a conjugate as described herein.

[0288] Although the description of pharmaceutical compositions provided herein primarily relates to pharmaceutical compositions suitable for administration to humans, one skilled in the art will understand that such compositions are generally suitable for administration to any other animal, e.g., non-human animals, e.g., non-human mammals. Modifications of pharmaceutical compositions suitable for administration to humans to make them suitable for administration to a variety of animals are well understood, and a veterinary pharmacologist of ordinary skill can design and / or implement such modifications with no more than routine experimentation, if any. Subjects to which the pharmaceutical compositions are intended for administration include, but are not limited to, humans and / or other primates; mammals, e.g., commercially relevant mammals such as cows, pigs, horses, sheep, cats, dogs, mice, and / or rats; and / or birds, e.g., commercially relevant birds such as poultry, chickens, ducks, geese, and / or turkeys.

[0289] The formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of bringing into association the active ingredient(s) with an excipient and / or one or more other accessory ingredients, and then, as necessary and / or desirable, dividing, shaping, and / or packaging the formulation into the desired single or multiple dosage units.

[0290] Pharmaceutical compositions according to the invention may be prepared, packaged, and / or sold in bulk, as a single unit dose, and / or as a plurality of single unit doses. As used herein, a "unit dose" is a discrete amount of a pharmaceutical composition comprising a predetermined amount of an active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient that would be administered to a subject and / or a convenient fraction of such a dosage, such as one-half or one-third of such a dosage.

[0291] The relative amounts of active ingredient, pharmaceutically acceptable excipient, and / or any additional ingredients in a pharmaceutical composition of the present invention may vary depending on the identity, size, and / or condition of the subject being treated, and further depending on the route by which the composition is administered. By way of example, the composition may comprise from 0.1% to 100%, e.g., 0.5 to 50%, 1 to 30%, 5 to 80%, at least 80% (w / w) active ingredient.

[0292] The conjugates of the present invention can be formulated with one or more excipients to (1) increase stability; (2) enable sustained or delayed release (e.g., from a monomaleimide depot formulation); (3) alter biodistribution (e.g., target the monomaleimide compound to specific tissues or cell types); or (4) alter the release profile of the monomaleimide compound in vivo. Non-limiting examples of excipients include any solvent, dispersion medium, diluent, or other liquid vehicle, dispersing or suspending aid, surfactant, isotonicity agent, thickening or emulsifying agent, and preservative. Excipients of the present invention can also include, without limitation, lipidoids, liposomes, lipid nanoparticles, polymers, lipoplexes, core-shell nanoparticles, peptides, proteins, hyaluronidase, nanoparticle mimics, and combinations thereof. Thus, the formulations of the present invention can include one or more excipients, each in an amount that, taken together, increases the stability of the monomaleimide compound.

[0293] excipients Pharmaceutical formulations can further comprise pharmaceutically acceptable excipients, which, as used herein, include any solvents, dispersion media, diluents, or other liquid vehicles, dispersing or suspending aids, surfactants, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, and the like, as appropriate for the particular dosage form desired. Remington's The Science and Practice of Pharmacy, 21st Edition, A.R. Gennaro (Lippincott, Williams & Wilkins, Baltimore, MD, 2006; incorporated herein by reference in its entirety), discloses various excipients used in formulating pharmaceutical compositions and known techniques for their preparation. Except insofar as any conventional excipient medium is incompatible with a substance or its derivatives, such as by the production of any undesired biological effect or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutical composition, its use is contemplated within the scope of the present invention.

[0294] In some embodiments, the pharmaceutically acceptable excipient is at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% pure. In some embodiments, the excipient is approved for human and veterinary use. In some embodiments, the excipient is approved by the U.S. Food and Drug Administration. In some embodiments, the excipient is pharmaceutical grade. In some embodiments, the excipient meets the specifications of the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia, and / or the International Pharmacopoeias.

[0295] Pharmaceutically acceptable excipients used in the manufacture of pharmaceutical compositions include, but are not limited to, inert diluents, dispersing and / or granulating agents, surfactants and / or emulsifying agents, disintegrating agents, binders, preservatives, buffers, lubricants, and / or oils. Such excipients may optionally be included in the pharmaceutical composition.

[0296] Exemplary diluents include, but are not limited to, calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and the like, and / or combinations thereof.

[0297] Exemplary granulating and / or dispersing agents include, but are not limited to, potato starch, corn starch, tapioca starch, sodium starch glycolate, clay, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose and wood products, natural sponge, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethylcellulose, cross-linked sodium carboxymethylcellulose (croscarmellose), methylcellulose, pregelatinized starch (starch 1500), microcrystalline starch, water-insoluble starch, calcium carboxymethylcellulose, magnesium aluminum silicate (VEEGUM®), sodium lauryl sulfate, quaternary ammonium compounds, and the like, and / or combinations thereof.

[0298] Exemplary surfactants and / or emulsifiers include, but are not limited to, natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, wax, and lecithin), colloidal clays (e.g., bentonite [aluminum silicate] and VEEGUM® [magnesium aluminum silicate]), long-chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxypolymethylene, polyacrylic acid, acrylic acid polymers, and carboxyvinyl polymers), carrageenan, cellulose derivatives (e.g., sodium carboxymethylcellulose, powdered cellulose, hydroxymethylcellulose, hydroxypropyl ... propyl methylcellulose, methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate [TWEEN® 20], polyoxyethylene sorbitan [TWEEN® 60], polyoxyethylene sorbitan monooleate [TWEEN® 80], sorbitan monopalmitate [SPAN® 40], sorbitan monostearate, [SPAN® 60], sorbitan tristearate [SPAN® 65], glyceryl monooleate, sorbitan monooleate [SPAN® 80]), polyoxyethylene esters (e.g., polyoxyethylene monostearate [MYRJ® 45], polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate,and Kolliphor (SOLUTOL®), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g., CREMOPHOR®), polyoxyethylene ethers (e.g., polyoxyethylene lauryl ether [BRIJ® 30]), poly(vinylpyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, PLUORINC® F 68, POLOXAMER® 188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium, and / or combinations thereof.

[0299] Exemplary binders include, but are not limited to, starches (e.g., cornstarch and starch paste); gelatin; sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol); natural and synthetic gums (e.g., acacia, sodium alginate, Irish moss extract, breadwort gum, ghatti gum, isapol shell mucilage, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinylpyrrolidone), magnesium aluminum silicate (Veegum®), and larch arabinogalactan); alginic acid; polyethylene oxide; polyethylene glycol; inorganic calcium salts; silicic acid; polymethacrylates; waxes; water; alcohols, etc.; and combinations thereof.

[0300] Exemplary preservatives include, but are not limited to, antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives, and / or other preservatives. Exemplary antioxidants include, but are not limited to, alpha-tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and / or sodium sulfite. Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA), citric acid monohydrate, disodium edetate, dipotassium edetate, edetic acid, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid, and / or trisodium edetate. Exemplary antimicrobial preservatives include, but are not limited to, benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and / or thimerosal. Exemplary antifungal preservatives include, but are not limited to, butylparaben, methylparaben, ethylparaben, propylparaben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and / or sorbic acid. Exemplary alcohol preservatives include, but are not limited to, ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoates, and / or phenylethyl alcohol. Exemplary acidic preservatives include, but are not limited to, vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and / or phytic acid.Other preservatives include, but are not limited to, tocopherol, tocopheryl acetate, deteroxime mesylate, cetrimide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, GLYDANT PLUS®, PHENONIP®, methylparaben, GERMALL® 115, GERMABEN® II, NEOLONE™, KATHON™, and / or EUXYL®.

[0301] Exemplary buffering agents include, but are not limited to, citrate buffer, acetate buffer, phosphate buffer, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, calcium hydrogen phosphate, phosphoric acid, tribasic calcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic sodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, and the like, and / or combinations thereof.

[0302] Exemplary lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behanate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and the like, and combinations thereof.

[0303] Exemplary oils include, but are not limited to, almond oil, apricot kernel oil, avocado oil, babassu oil, bergamot oil, black current seed oil, borage oil, cade oil, chamomile oil, canola oil, caraway oil, carnauba, castor oil, cinnamon bark oil, cocoa butter, coconut oil, cod liver oil, coffee oil, corn oil, cottonseed oil, emu oil, eucalyptus oil, evening primrose oil, fish oil, linseed oil, geraniol oil, gourd oil, grapeseed oil, hazelnut oil, hyssop oil, isopropyl myristate, jojoba oil, kukui nut oil, lavandin oil, lavender oil, lemon oil, litsea cubeba oil, macadamia nut oil, and the like. nut oil, mallow oil, mango seed oil, meadowfoam seed oil, mink oil, nutmeg oil, olive oil, orange oil, orange roughy oil, palm oil, palm kernel oil, peach kernel oil, peanut oil, poppy seed oil, pumpkin seed oil, rapeseed oil, rice bran oil, rosemary oil, safflower oil, sandalwood oil, sasquana oil, savory oil, sea buckthorn oil, sesame oil, shea butter, silicone oil, soybean oil, sunflower oil, tea plant oil, thistle oil, camellia oil, vetiver oil, walnut oil, and wheat germ oil. Exemplary oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and / or combinations thereof.

[0304] Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening, flavoring, and / or perfuming agents can also be present in the composition, according to the judgment of the formulator. Administration The conjugates of the present invention may be administered by any route that produces a therapeutically effective result. Such routes include, but are not limited to, enteral, gastrointestinal, epidural, oral, transdermal, epidural (around the dura), intracerebral (into the cerebrum), intraventricular (into the ventricles of the brain), epicutaneous (application on top of the skin), intradermal (into the skin itself), subcutaneous (under the skin), nasal administration (through the nose), intravenous (into a vein), intraarterial (into an artery), intramuscular (into a muscle), intracardiac (into the heart), intraosseous injection (into the bone marrow), intrathecal (into the spinal canal), intraperitoneal (infusion or injection into the peritoneum), intravesical instillation, intravitreal (through the eye), intracavernosal injection (into the base of the penis), intravaginal administration, intrauterine, extra-amniotic administration, transdermal (diffusion through intact skin for systemic distribution), transmucosal (diffusion through mucous membranes), insufflation (inhalation through the nose), sublingual, sublabial, enema, eye drops (on the conjunctiva), or ear drops. In specific embodiments, the composition may be administered in a manner that allows it to cross the blood-brain barrier, blood-vessel barrier, or other epithelial barrier.

[0305] The formulations described herein contain an effective amount of the conjugate in a pharmaceutical carrier suitable for administration to an individual in need thereof. The formulations may be administered parenterally (e.g., by injection or infusion). The formulations or variations thereof may be administered in any manner, including enterally, topically (e.g., to the eye), or by pulmonary administration. In some embodiments, the formulations are administered topically.

[0306] Dose determination The present invention provides methods comprising administering a conjugate as described herein to a subject in need thereof. The conjugate as described herein can be administered to a subject in any amount and using any route of administration effective for preventing or treating or imaging a disease, disorder, and / or condition (e.g., a disease, disorder, and / or condition associated with working memory deficit). The exact amount required may vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the disease, the particular composition, its mode of administration, its mode of activity, etc.

[0307] The compositions of the present invention are typically formulated into dosage unit forms for ease of administration and uniformity of dosage.However, it will be understood that the total daily use amount of the compositions of the present invention can be determined by the attending physician within the scope of sound medical judgment.The specific therapeutically effective dose, prophylactically effective dose, or appropriate imaging dose level for any particular patient can depend on various factors, including the disorder under treatment and the severity of the disorder; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health, sex, and diet; the administration timing, administration route, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination or simultaneously with the specific compound used; and similar factors well known in the medical field.

[0308] In some embodiments, the composition of the present invention is administered daily from about 0.0001 mg / kg to about 100 mg / kg, from about 0.001 mg / kg to about 0.05 mg / kg, from about 0.005 mg / kg to about 0.05 mg / kg, from about 0.001 mg / kg to about 0.005 mg / kg, from about 0.05 mg / kg to about 0.5 mg / kg, from about 0.01 mg / kg to about 50 mg / kg, from about 0.1 mg / kg to about 40 mg / kg, from about 0.5 mg / kg to about 30 mg / kg, from about 0.01 mg / kg to about 10 mg / kg, or from about 0.1 mg / kg to about 10 The desired therapeutic, diagnostic, prophylactic, or imaging effect can be achieved by administration once or more times daily at a dosage level sufficient to deliver about 1 mg / kg to about 25 mg / kg, about 25 mg / kg to about 50 mg / kg, about 50 mg / kg to about 100 mg / kg, about 100 mg / kg to about 125 mg / kg, about 125 mg / kg to about 150 mg / kg, about 150 mg / kg to about 175 mg / kg, about 175 mg / kg to about 200 mg / kg, or about 200 mg / kg to about 250 mg / kg of the subject's body weight. The desired dosage may be delivered three times a day, twice a day, once a day, every other day, every three days, every week, every two weeks, every three weeks, or every four weeks. In some embodiments, the desired dosage may be delivered using multiple administrations (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or more administrations). When multiple administrations are utilized, split-dosing regimens such as those described herein may be used.

[0309] The concentration of the conjugate in the pharmaceutical composition can be about 0.01 mg / mL to about 50 mg / mL, about 0.1 mg / mL to about 25 mg / mL, about 0.5 mg / mL to about 10 mg / mL, or about 1 mg / mL to about 5 mg / mL.

[0310] As used herein, a "split dose" refers to the division of a single unit dose or total daily dose into two or more doses, for example, two or more administrations of a single unit dose. As used herein, a "single unit dose" refers to a dose of any therapeutic agent administered in one dose / once / single route / single point of contact, i.e., a single administration event. As used herein, a "total daily dose" refers to the amount given or prescribed in a 24-hour period. This may be administered as a single unit dose.

[0311] Dosage form The pharmaceutical compositions described herein can be formulated into dosage forms described herein, such as topical, intranasal, intratracheal, or injectable (e.g., intravenous, intraocular, intravitreal, intramuscular, intracardiac, intraperitoneal, subcutaneous) dosage forms.

[0312] VI. How to Use the Conjugate Conjugates as described herein can be administered to treat any hyperproliferative disease, metabolic disease, infectious disease, or cancer, as appropriate. Formulations may be administered by injection, orally, or topically, typically to mucosal surfaces (pulmonary, nasal, oral, buccal, sublingual, intravaginal, rectal) or to the eye (intrachitically or ocularly).

[0313] In various embodiments, methods of treating a subject having cancer are provided, the methods comprising administering to a subject having, suspected of having, or predisposed to cancer a therapeutically effective amount of a conjugate as described herein, or a salt form thereof. According to the present invention, cancer includes any disease or condition characterized by uncontrolled cell proliferation, e.g., hyperproliferation. Cancer may be characterized by a tumor, e.g., a solid tumor, or any neoplasm.

[0314] In some embodiments, the cancer is a solid tumor. Large drug molecules have limited penetration into solid tumors. Penetration of large drug molecules is slow. On the other hand, small molecules, such as the conjugates of the present invention, can penetrate solid tumors quickly and more deeply. With regard to drug penetration depth, larger molecules have more durable pharmacokinetics but penetrate less. Small molecules, such as the conjugates of the present invention, penetrate deeper. Dreher et al. (Dreher et al., Journal of the National Cancer Institute (JNCI), Vol. 98, No. 5, p. 335, 2006, the contents of which are incorporated herein by reference in their entirety) studied the penetration of dextrans of various sizes into tumor xenografts.

[0315] In one embodiment, the conjugate of the invention penetrates from the tumor's vascular surface to at least about 25 μm, about 30 μm, about 35 μm, about 40 μm, about 45 μm, about 50 μm, about 75 μm, about 100 μm, about 150 μm, about 200 μm, about 250 μm, about 300 μm, about 400 μm, about 500 μm, about 600 μm, about 700 μm, about 800 μm, about 900 μm, about 1000 μm, about 1100 μm, about 1200 μm, about 1300 μm, about 1400 μm, or about 1500 μm into a solid tumor. A distance of zero is defined as the tumor's vascular surface, and any distance greater than zero is defined as the distance measured in three dimensions to the nearest vascular surface.

[0316] In another embodiment, the conjugate of the present invention penetrates into the center of a tumor. As used herein, the "center" of a tumor refers to the central area of ​​the tumor. The distance from any part of the central area of ​​a tumor to the surface of the tumor's blood vessels is about 30% to about 50% of the length or width of the tumor. The distance from any part of the central area of ​​a tumor to the center point of the tumor is less than about 20% of the length or width of the tumor. The central area of ​​a tumor is approximately the central one-third of the tumor.

[0317] In another embodiment, the conjugate of the present invention penetrates to the middle of a solid tumor. As used herein, the "middle" of a tumor refers to the middle area of ​​the tumor. The distance from any part of the middle area of ​​the tumor to the surface of the tumor's blood vessels is about 15% to about 30% of the length or width of the tumor. The distance from any part of the middle area of ​​the tumor to the center point of the tumor is about 20% to about 35% of the length or width of the tumor. The middle area of ​​a tumor is approximately between the central 1 / 3 of the tumor and the outer 1 / 3 of the tumor.

[0318] In some embodiments, the subject may not otherwise be a candidate for treatment with the conjugate. In some embodiments, the method involves the use of cancer cells, including but not limited to mammalian cancer cells. In some examples, the mammalian cancer cells are human cancer cells.

[0319] In some embodiments, the conjugates of the present teachings have been found to inhibit cancer and / or tumor growth. They also reduce cell proliferation, invasion, and / or metastasis, thus making them useful in the treatment of cancer.

[0320] In some embodiments, conjugates of the present teachings can be used to prevent the growth of tumors or cancers and / or prevent the metastasis of tumors or cancers, hi some embodiments, compositions of the present teachings can be used to shrink or destroy cancers.

[0321] In some embodiments, the conjugates provided herein are useful for inhibiting cancer cell proliferation. In some embodiments, the conjugates provided herein are useful for inhibiting cell proliferation, e.g., inhibiting the rate of cell proliferation, preventing cell proliferation, and / or inducing cell death. Generally, the conjugates as described herein can inhibit cell proliferation of cancer cells, or can both inhibit cancer cell proliferation and / or induce cell death. In some embodiments, cell proliferation is reduced by at least about 25%, about 50%, about 75%, or about 90% after treatment with a conjugate of the invention compared to untreated cells. In some embodiments, the cell cycle arrest marker phosphohistone H3 (PH3 or PHH3) is increased by at least about 50%, about 75%, about 100%, about 200%, about 400%, or about 600% after treatment with a conjugate of the invention compared to untreated cells. In some embodiments, the cellular apoptosis marker cleaved caspase 3 (CC3) is increased by at least 50%, about 75%, about 100%, about 200%, about 400%, or about 600% after treatment with a conjugate of the invention compared to untreated cells.

[0322] Furthermore, in some embodiments, the conjugates of the invention are effective in inhibiting tumor growth in multiple tumor types, whether measured as net size (weight, surface area, or volume) or as rate over time.

[0323] In some embodiments, the size of a tumor is reduced by about 60% or more after treatment with a conjugate of the invention, hi some embodiments, the size of a tumor is reduced by at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100%, as measured by weight, and / or area and / or volume.

[0324] Cancers treatable by the methods of the present teachings generally occur in mammals, including, for example, humans, non-human primates, dogs, cats, rats, mice, rabbits, ferrets, guinea pigs, horses, pigs, sheep, goats, and cows. In various embodiments, cancers include, but are not limited to, acoustic neuroma, acute leukemia, acute lymphocytic leukemia, acute myeloid leukemia (monocytic, myeloblastic, adenocarcinoma, hemangiosarcoma, astrocytoma, myelomonocytic, and promyelocytic), acute T-cell leukemia, basal cell carcinoma, bile duct carcinoma, bladder cancer, brain cancer, breast cancer, bronchogenic carcinoma, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid (granulocytic) leukemia, chronic myeloid leukemia, and ovarian cancer. Intestinal cancer, colorectal cancer, craniopharyngioma, cystadenocarcinoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, hyperproliferative changes (dysplasia and metaplasia), embryonal carcinoma, endometrial cancer, endothelial sarcoma, ependymoma, epithelial carcinoma, erythroleukemia, esophageal cancer, estrogen receptor-positive breast cancer, essential thrombocythemia, Ewing's tumor, fibrosarcoma, follicular lymphoma, testicular germ cell carcinoma, glioma, heavy chain disease, hemangioblastoma, hepatoma, hepatocellular carcinoma, hormone-insensitive prostate cancer, leiomyosarcoma, adipose tissue Sarcoma, lung cancer, lymphangioendotheliosarcoma, lymphangiosarcoma, lymphoblastic leukemia, lymphoma (Hodgkin's and non-Hodgkin's), malignant tumors and hyperproliferative disorders of the bladder, breast, colon, lung, ovary, pancreas, prostate, skin, and uterus, lymphoid malignancies of T-cell or B-cell origin, leukemia, lymphoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myeloid leukemia, myeloma, myxosarcoma, myeloma These include eosinophilic sarcoma, non-small cell lung cancer, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary adenocarcinoma, papillary carcinoma, pinealoma, polycythemia vera, prostate cancer, rectal cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, sebaceous gland carcinoma, seminoma, skin cancer, small cell lung cancer, solid tumors (carcinoma and sarcoma), small cell lung cancer, gastric cancer, squamous cell carcinoma, synovium, sweat gland carcinoma, thyroid cancer, Waldenstrom's macroglobulinemia, testicular cancer, uterine cancer, and Wilms' tumor.Other cancers include primary cancer, metastatic cancer, oropharyngeal cancer, hypopharyngeal cancer, liver cancer, gallbladder cancer, bile duct cancer, small intestine cancer, urinary tract cancer, kidney cancer, urothelial cancer, female reproductive organ cancer, uterine cancer, gestational trophoblastic disease, male reproductive organ cancer, seminal vesicle cancer, testicular cancer, germ cell tumor, endocrine gland tumor, thyroid cancer, adrenal cancer, pituitary cancer, hemangioma, sarcoma derived from bone and soft tissue, Kaposi's sarcoma, nerve cancer, eye cancer, meningeal cancer solid tumors derived from hematopoietic malignancies such as glioblastoma, neuroma, neuroblastoma, schwannoma, and leukemia, metastatic melanoma, recurrent or persistent ovarian epithelial cancer, fallopian tube cancer, primary peritoneal cancer, gastrointestinal stromal tumor, colorectal cancer, gastric cancer, melanoma, glioblastoma multiforme, non-squamous non-small cell lung cancer, malignant glioma, epithelial ovarian cancer, primary serous peritoneal cancer, metastatic liver cancer, neuroendocrine cancer, refractory malignancies, triple-negative breast cancer, HER2-amplified breast cancer, and nasopharyngeal carcinoma These include oral cancer, biliary tract cancer, hepatocellular carcinoma, squamous cell carcinoma of the head and neck (SCCHN), non-medullary thyroid cancer, recurrent glioblastoma multiforme, neurofibromatosis type 1, CNS cancer, liposarcoma, leiomyosarcoma, salivary gland cancer, mucosal melanoma, acral / lentiginous melanoma, paraganglioma, pheochromocytoma, advanced metastatic cancer, solid tumors, triple-negative breast cancer, colorectal cancer, sarcoma, melanoma, renal cancer, endometrial cancer, thyroid cancer, rhabdomysarcoma, multiple myeloma, ovarian cancer, glioblastoma, gastrointestinal stromal tumor, mantle cell lymphoma, and refractory malignancies.

[0325] In one embodiment, a conjugate as described herein or a formulation comprising a conjugate as described herein is used to treat small cell lung cancer. Approximately 12% to 15% of patients with lung cancer have small cell lung cancer. The survival rate for metastatic small cell lung cancer is low; the survival rate is less than 5% five years after diagnosis. The incidence of small cell lung cancer in the United States is approximately 26,000 to 30,000 cases.

[0326] In some embodiments, a conjugate as described herein or a formulation comprising a conjugate as described herein is used to treat a patient having a tumor that expresses or overexpresses HSP90.

[0327] A characteristic of the conjugates of the present invention is that they have relatively low toxicity to an organism while maintaining efficacy in inhibiting, e.g., slowing or halting, tumor growth. As used herein, "toxicity" refers to the ability of a substance or composition to be harmful or toxic to a cell, tissue, organism, or cellular environment. Low toxicity refers to a reduced ability of a substance or composition to be harmful or toxic to a cell, tissue, organism, or cellular environment. Such reduced or low toxicity can be relative to a standard measurement, relative to treatment, or relative to the absence of treatment. For example, the conjugates of the present invention can have lower toxicity than the active agent moiety Z administered alone. For conjugates including DM1, the toxicity is lower than DM1 administered alone.

[0328] Toxicity may also be measured based on a subject's weight loss, where a weight loss of more than 15%, more than 20%, or more than 30% of body weight is indicative of toxicity. Other measures of toxicity may also be measured, such as measures of patient symptomatology, including lethargy and general malaise. Neutropenia, thrombocytopenia, white blood cell (WBC) count, and complete blood count (CBC) count may also be measures of toxicity. Pharmacological indicators of toxicity include elevated aminotransferase (AST / ALT) levels, neurotoxicity, renal damage, GI disorders, and the like. In one embodiment, the conjugates of the present invention do not cause a significant change in a subject's weight. After treatment with the conjugates of the present invention, a subject loses less than about 30%, about 20%, about 15%, about 10%, or about 5% of their body weight. In another embodiment, the conjugates of the present invention do not cause a significant increase in a subject's AST / ALT levels. After treatment with a conjugate of the invention, the subject's AST or ALT levels increase by less than about 30%, about 20%, about 15%, about 10%, or about 5%. In yet another embodiment, the conjugate of the invention does not cause a significant change in the subject's CBC or WBC count after treatment with a conjugate of the invention. After treatment with a conjugate of the invention, the subject's CBC or WBC levels decrease by less than about 30%, about 20%, about 15%, about 10%, or about 5%.

[0329] Combination therapy In some embodiments, the conjugate or particle of the present invention is combined with at least one additional active agent. The active agent may be any suitable drug. The conjugate and the at least one additional active agent may be administered simultaneously, sequentially, or in any order. The conjugate and the at least one additional active agent may be administered at different doses, at different dosing frequencies, or via different routes, as appropriate.

[0330] In some embodiments, the additional active agent affects the biodistribution (i.e., tissue distribution) of the conjugates of the present invention. For example, radioactive materials can accumulate in the kidney, potentially causing radiotoxicity problems in the kidney and surrounding organs. The additional active agent can reduce kidney accumulation or retention time. Preferably, kidney update of the conjugate is reduced, but tumor uptake of the conjugate is not affected. The kidney and surrounding organs are protected without reducing the efficacy of the conjugate. In one non-limiting example, the conjugates of the present invention can be administered in combination with at least one amino acid or analog thereof. The amino acid or analog can be a positively charged basic amino acid, such as lysine (L-lysine or D-lysine) or arginine, or a combination thereof. In another non-limiting example, the conjugates of the present invention can be administered in combination with an active agent that binds to HSP90, such as an HSP90 inhibitor. Any of the ligands described in the "HSP90 Targeting Moiety" section, such as ganetespib or its derivatives / analogs, can be used. In another non-limiting example, the conjugates of the present invention may be administered in combination with monosodium glutamate (MSG) or glutamic acid. In yet another non-limiting example, the conjugates of the present invention may be administered in combination with amifostine (Ethyol, WR-2721), a bovine gelatin-containing solution, gelofusin, or an albumin fragment. The albumin fragment may have a molecular weight of 3 to 50 kDa.

[0331] The additional active agent may also be a cancer symptom-relieving agent. Non-limiting examples of symptom-relieving agents include octreotide or lanreotide; interferon, cypoheptadine, or any other antihistamine. In some embodiments, the conjugate of the present invention does not have drug interactions with the additional active agent. In one embodiment, the conjugate of the present invention does not inhibit cytochrome P450 (CYP) isoenzymes. CYP isoenzymes may include CYP3A4 midazolam, CYP3A4 testosterone, CYP2C9, CYP2D6, CYP1A2, CYP2C8, CYP2B6, and CYP2C19. The additional active agent may be administered simultaneously with the conjugate of the present invention.

[0332] In another example, the conjugates of the invention may be combined with moderate doses of chemotherapeutic agents such as mitomycin C, vinblastine, and cisplatin (see Ellis et al., Br J Cancer, 71(2), 366-370 (1995), the contents of which are incorporated herein by reference in their entirety).

[0333] In yet another example, a patient may first receive a pharmaceutically effective amount of an unconjugated active agent, followed by a pharmaceutically effective amount of a conjugate comprising the same active agent. In some embodiments, a non-radioactive conjugate of the present invention can be combined with a radioactive analog of the conjugate. For example, the non-radioactive conjugate can be administered before the radioactive analog. In another example, a subject can receive a mixture of the non-radioactive conjugate and its radioactive analog. In yet another example, a subject can first receive non-radioactive conjugate treatment, followed by a mixture of the non-radioactive conjugate and its radioactive analog.

[0334] In some embodiments, a conjugate of the present invention comprising one radiolabel may be combined with at least one other conjugate of the present invention comprising one or more different radiolabels. For example, a conjugate comprising an imaging radiolabel may be combined with a conjugate comprising a non-imaging radiolabel. In one embodiment, a conjugate comprising lutetium (Lu) may be combined with a conjugate comprising gallium (Ga).

[0335] The conjugates as described herein or formulations containing the conjugates as described herein can be used to deliver therapeutic, prophylactic, or diagnostic agents to tissue-selectively to individuals or patients in need thereof. For example, the conjugates of the present invention are used to deliver radioactive materials to selective tissues. These tissues can be tumor tissues. Dosage regimens can be adjusted to provide the optimum desired response (e.g., therapeutic or prophylactic response). For example, a single bolus can be administered, or several divided doses can be administered over time, or the dose can be proportionally reduced or increased as dictated by the exigencies of the therapeutic situation. Dosage unit form, as used herein, refers to physically discrete units suited as unitary dosages for the mammalian subject to be treated; each unit containing a predetermined quantity of active compound calculated to yield the desired therapeutic agent.

[0336] V. Kits and Devices The present invention provides various kits and devices for conveniently and / or effectively carrying out the methods of the present invention. Typically, kits may contain components in amounts and / or numbers sufficient to allow a user to perform multiple treatments of one or more subjects and / or to perform multiple experiments.

[0337] In one embodiment, the present invention provides a kit for inhibiting tumor cell growth in vitro or in vivo comprising a conjugate of the present invention or a combination of conjugates of the present invention, optionally in combination with any other active agent.

[0338] The kit may further include packaging and instructions and / or a delivery agent for forming a formulation composition. The delivery agent may include saline, a buffer solution, or any of the delivery agents disclosed herein. The amount of each component may be varied to achieve a consistent and reproducible high-concentration saline or simple buffer formulation. The components may also be varied to increase the stability of the conjugate in the buffer solution over a period of time and / or under various conditions.

[0339] The present invention provides devices that can incorporate the conjugates of the present invention. These devices include stable formulations available for immediate delivery to a subject, e.g., a human patient, in need thereof. In some embodiments, the subject has cancer.

[0340] Non-limiting examples of devices include pumps, catheters, needles, transdermal patches, pressurized olfactory delivery devices, iontophoresis devices, and multilayer microfluidic devices. These devices can be used to deliver the conjugates of the present invention in single, multiple, or divided dose regimens. The devices can be used to deliver the conjugates of the present invention through biological tissue, intradermally, subcutaneously, or intramuscularly.

[0341] VI.Definitions The term "compound," as used herein, is intended to include all stereoisomers, geometric isomers, tautomers, and isotopes of the depicted structure. In this application, compound is used interechangably with conjugate. Thus, conjugate, as used herein, is also intended to include all stereoisomers, geometric isomers, tautomers, and isotopes of the depicted structure.

[0342] The compounds described herein can be asymmetric (e.g., have one or more stereocenters). Unless otherwise indicated, all stereoisomers, such as enantiomers and diastereomers, are intended. Compounds of the present disclosure containing 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, may also exist in the compounds described herein, and all such stable isomers are contemplated in this disclosure. Cis and trans geometric isomers of the compounds of the present disclosure are described and can be isolated as a mixture of isomers or as separate isomeric forms.

[0343] The compounds of the present disclosure also include tautomeric forms. Tautomeric forms result from the exchange of a single bond with an adjacent double bond, accompanied by the transfer of a proton. Tautomeric forms include prototropic tautomers, which are isomeric protonation states with the same empirical formula and total charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, amide-imidic acid pairs, enamine-imine pairs, and cyclic forms in which protons can occupy more than one position in a heterocyclic ring 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 may be in equilibrium or sterically locked into one form by appropriate substitution.

[0344] The compounds of the present disclosure also include any isotopes of atoms present in the intermediates or final compounds. "Isotopes" refer to atoms having the same atomic number but different mass numbers as a result of different numbers of neutrons in the nucleus. For example, isotopes of hydrogen include tritium and deuterium.

[0345] The compounds and salts of the present disclosure may be prepared by combining with a solvent or water molecules to form solvates and hydrates in a conventional manner. The terms "subject" or "patient," as used herein, refer to any living organism to which a conjugate can be administered, e.g., for experimental, therapeutic, diagnostic, and / or prophylactic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, guinea pigs, cows, pigs, sheep, horses, dogs, cats, hamsters, llamas, non-human primates, and humans).

[0346] The terms "treating" or "preventing," as used herein, can include preventing a disease, disorder, or condition from occurring in an animal that may be predisposed to the disease, disorder, and / or condition but has not yet been diagnosed with the disease, disorder, or condition; inhibiting a disease, disorder, or condition, e.g., preventing its progression; and alleviating a disease, disorder, or condition, e.g., causing regression of the disease, disorder, and / or condition. Treating a disease, disorder, or condition can include ameliorating at least one symptom of a particular disease, disorder, or condition even if the underlying pathophysiology is unaffected, such as treating pain in a subject by administering an analgesic agent even though such an agent does not treat the cause of the pain.

[0347] "Target," as used herein, is intended to mean the site to which a targeted construct binds. Targets may be in vivo or in vitro. In certain embodiments, targets may be cancer cells found in leukemias or tumors (e.g., brain, lung (small cell and non-small cell), ovarian, prostate, breast, and colon tumors, as well as other carcinomas and sarcomas). In yet other embodiments, targets may refer to molecular structures to which a targeting moiety or ligand binds, such as haptens, epitopes, receptors, dsDNA fragments, carbohydrates, or enzymes. Targets may also be certain tissues, such as nervous tissue, intestinal tissue, pancreatic tissue, liver, kidney, prostate, ovary, lung, bone marrow, or breast tissue.

[0348] "Target cells" that can be targeted by the present methods or conjugates are generally animal cells, e.g., mammalian cells. The present methods can be used to alter the cellular function of living cells in vitro, i.e., in cell culture, or in vivo, where the cells form part of animal tissue or are otherwise present. Thus, target cells can include, for example, blood, lymphatic tissue, cells lining the digestive tract, such as the oral and pharyngeal mucosa, cells forming the villi of the small intestine, cells lining the large intestine, cells lining the respiratory system (nasal passages / lungs) of an animal (which can be contacted by inhalation in the present invention), dermal / epidermal cells, cells of the vagina and rectum, cells of visceral organs, including placental cells, and the so-called blood-brain barrier. Generally, target cells express at least one HSP90. In some embodiments, target cells can be cells that express HSP90 and are in the vicinity of cells targeted by the conjugates described herein and affected by the release of the active agent of the conjugates. For example, HSP90-expressing blood vessels in the vicinity of a tumor may be targeted, while active agents released at that site will affect the tumor.

[0349] The term "therapeutic effect" is art-recognized and refers to a local or systemic effect in animals, particularly mammals, and more particularly humans, caused by a pharmacologically active substance. Thus, the term refers to any substance intended for use in the diagnosis, cure, mitigation, treatment, or prevention of a disease, disorder, or condition in an animal, such as a human, in the enhancement of desirable physical or mental development and status.

[0350] The term "modulation" is art-recognized and refers to the upregulation (i.e., activation or stimulation), downregulation (i.e., inhibition or suppression) of a response, or the two in combination or separately. Modulation is generally compared to a baseline or standard, which may be internal or external to the entity being treated.

[0351] "Parenteral administration," as used herein, means administration by any method other than through the digestive tract (enteral) or a non-invasive topical route. For example, parenteral administration can include administration to a patient intravenously, intradermally, intraperitoneally, intrapleurally, intratracheally, intraossiously, intracerebrally, intrathecally, intramuscularly, subcutaneously, subjunctivally, by injection, and by infusion.

[0352] "Topical administration," as used herein, refers to non-invasive administration to the skin, orifices, or mucous membranes. Topical administration can be delivered locally, i.e., the therapeutic agent can have a local effect in the area of ​​delivery with no or minimal systemic exposure. Some topical formulations can have a systemic effect, for example, by adsorption into an individual's bloodstream. Topical administration can include, but is not limited to, dermal and transdermal administration, buccal administration, intranasal administration, intravaginal administration, intravesical administration, ocular administration, and rectal administration.

[0353] "Enteral administration," as used herein, means administration by absorption through the gastrointestinal tract. Enteral administration can include oral and sublingual administration, gastric administration, or rectal administration. "Pulmonary administration," as used herein, means administration into the lungs by inhalation or intratracheal administration. As used herein, the term "inhalation" refers to taking in air into the alveoli of the lungs. Air intake can occur through the mouth or nose.

[0354] The terms "sufficient" and "effective," as used interchangeably herein, refer to the amount (e.g., mass, volume, dosage, concentration, and / or time) necessary to achieve one or more desired results. A "therapeutically effective amount" is at least the minimum concentration required to produce a measurable improvement or prevention of at least one symptom or a particular condition or disorder, produce a measurable improvement in life expectancy, or generally improve a patient's quality of life. Thus, the therapeutically effective amount will depend on the specific biologically active molecule and the specific condition or disorder being treated. Therapeutically effective amounts of many active agents, such as antibodies, are known in the art. Therapeutically effective amounts of the compounds and compositions described herein, for example, for treating a particular disorder, can be determined by techniques well within the skill of a physician or other skilled artisan.

[0355] The terms "bioactive agent" and "active agent," as used interchangeably herein, include, without limitation, physiologically or pharmacologically active substances that act locally or systemically in the body. A bioactive agent is a substance used in the treatment (e.g., therapeutic agents), prevention (e.g., prophylactic agents), diagnosis (e.g., diagnostic agents), cure, or mitigation of a disease or condition; a substance that affects the structure or function of the body; or a prodrug that becomes biologically active or more active after being placed in a defined physiological environment.

[0356] The term "prodrug" refers to an agent, including a small organic molecule, peptide, nucleic acid, or protein, that is converted in vitro and / or in vivo into a biologically active form. Prodrugs can be useful in some situations because they may be easier to administer than the parent compound (the active compound). For example, a prodrug may be oral bioavailable, but the parent compound is not. A prodrug may also have improved solubility in pharmaceutical compositions compared to the parent drug. A prodrug may also have less toxicity than the parent. Prodrugs may be converted into the parent drug by various mechanisms, including enzymatic processes and metabolic hydrolysis. Harper, NJ, 1962, Drug Latentiation, Jucker (ed.), Progress in Drug Research, Vol. 4, pp. 221-294; Morozowich et al., 1977, Application of Physical Organic Principles to Prodrug Design, E.B. Roche (ed.), Design of Biopharmaceutical Properties through Prodrugs and Analogs, APhA; Acad. Pharm. Sci.; E.B. Roche (ed.), 1977, Bioreversible Carriers in Drug in Drug Design, Theory and Application. Application), American Pharmaceutical Association (APhA); H. BandgeardBundgaard (ed.), 1985, Design of Prodrugs, Elsevier; Wang et al., 1999, Prodrug approaches to the improved delivery of peptide drugs, Curr. Pharm. Design., Vol. 5, No. 4, pp. 265-287; Pauletti et al., 1997, Improvement in peptide bioavailability: Peptidomimetics and Prodrug Strategies, Adv. Drug. Delivery Rev., Vol. 27, pp. 235-256; Mizen et al., 1998, The use of esters as prodrugs for oral delivery of β-lactam antibiotics. Use of Esters as Prodrugs for Oral Delivery of β-Lactam Antibiotics, Pharm. Biotech., Vol. 11, pp. 345-365; Gaignault et al., 1996, Designing Prodrugs and Bioprecursors I. Carrier Prodrugs, Pract. Med. Chem., pp. 671-696; M. Asgharnejad, 2000, Improving Oral Drug Transport Via Prodrugs, G.L. Amidon, P.I. Lee, and E.M. TopTopp (ed.), Transport Processes in Pharmaceutical Systems, Marcell Dekker, pp. 185-218; Balant et al., 1990, Prodrugs for the improvement of drug absorption via different routes of administration, Eur. J. Drug Metab. Pharmacokinet., Vol. 15, No. 2, pp. 143-53; Balimane and Sinko, 1999, Involvement of multiple transporters in the oral absorption of nucleoside analogues, Adv. Drug Delivery Reviews Rev., Vol. 39, No. 1-3, pp. 183-209; Browne, 1997, Fosphenytoin (Cerebyx), Clin. Neuropharmacol., Vol. 20, No. 1, pp. 1-12; Bundgaard, 1979, Bioreversible derivatization of drugs—principle and applicability to improve the therapeutic effects of drugs, Arch. Pharm. Chemi., Vol. 86, No. 1, pp. 1-39; H. Bundgaard, 1979, Bioreversible derivatization of drugs—principle and applicability to improve the therapeutic effects of drugs, Arch. Pharm. Chemi., Vol. 86, No. 1, pp. 1-39; H. Bundgaard, 1980, Bioreversible derivatization of drugs—principle and applicability to improve the therapeutic effects of drugs, Arch. Pharm. Chemi., Vol. 86, No. 1, pp. 1-39.Bundgaard, ed., 1985, Design of Prodrugs, New York: Elsevier; Fleisher et al., 1996, Improved oral drug delivery: solubility limitations overcome by the use of prodrugs, Adv. Drug Delivery Rev., Vol. 19, No. 2, pp. 115-130; Fleisher et al., 1985, Design of prodrugs for improved gastrointestinal absorption by intestinal enzyme targeting, Methods Enzymol., Vol. 112, pp. 360-361; Farquhar, D. D. et al., 1983, Biologically Reversible Phosphate-Protective Groups, J. Pharm. Sci., Vol. 72, No. 3, pp. 324-325; Han, HK. et al., 2000, Targeted prodrug design to optimize drug delivery, AAPS Pharm. Sci., Vol. 2, No. 1, p. E6; Sadzuka Y., 2000, Effective prodrug liposome and conversion to active metabolite, Curr. Drug Metabolism Metab.), Vol. 1, No. 1, pp. 31-48; D.M. Lambert (DMLambert, 2000, Rationale and applications of lipids as prodrug carriers, Eur. J. Pharm. Sci., Vol. 11, Suppl. 2, pp. S15-27; Wang, W. et al., 1999, Prodrug approaches to the improved delivery of peptide drugs, Curr. Pharm. Des., Vol. 5, No. 4, pp. 265-87.

[0357] The term "biocompatible," as used herein, refers to a material that is generally non-toxic to and does not cause any significant adverse effects to the recipient, along with any metabolic or breakdown products thereof. Generally speaking, a biocompatible material is one that does not elicit a significant inflammatory or immune response when administered to a patient.

[0358] The term "biodegradable," as used herein, generally refers to a material that breaks down or erodes under physiological conditions into smaller units or chemical species that can be metabolized, excreted, or excreted by a subject. Degradation time is a function of composition and morphology. Degradation time can range from a few hours to a few weeks.

[0359] The term "pharmaceutically acceptable," as used herein, refers to compounds, materials, compositions, and / or dosage forms that are suitable for use in contact with the tissues of humans and animals without undue toxicity, irritation, allergic response, or other problem or complication, within the scope of sound medical judgment and commensurate with a reasonable risk-to-benefit ratio in accordance with guidelines from agencies such as the U.S. Food and Drug Administration. A "pharmaceutically acceptable carrier," as used herein, refers to all components of a pharmaceutical formulation that facilitate delivery of the composition in vivo. Pharmaceutically acceptable carriers include, but are not limited to, diluents, preservatives, binders, lubricants, disintegrants, swelling agents, fillers, stabilizers, and combinations thereof.

[0360] The term "molecular weight," as used herein, generally refers to the mass or average mass of a material. In the case of a polymer or oligomer, molecular weight may refer to the relative average chain length or relative chain mass of the bulk polymer. In practice, the molecular weight of polymers and oligomers can be estimated or characterized in a variety of ways, including gel permeation chromatography (GPC) or capillary viscometry. GPC molecular weight is the number average molecular weight (M n ), as opposed to the weight average molecular weight (M w Capillary viscometry provides an estimate of molecular weight as the intrinsic viscosity determined from a dilute polymer solution using a specific set of concentration, temperature, and solvent conditions.

[0361] The term "small molecule," as used herein, generally refers to an organic molecule having a molecular weight of less than 2000 g / mol, less than 1500 g / mol, less than 1000 g / mol, less than 800 g / mol, or less than 500 g / mol. Small molecules are non-polymeric and / or non-oligomeric.

[0362] The term "hydrophilic," as used herein, refers to a substance that has strongly polar groups that readily interact with water. The term "hydrophobic" as used herein refers to a substance that lacks an affinity for water; that repels water and tends not to absorb water and not to dissolve or mix in water.

[0363] The term "lipophilic," as used herein, refers to a compound that has an affinity for lipids. The term "amphiphilic," as used herein, refers to a molecule that combines hydrophilic and lipophilic (hydrophobic) properties. "Amphiphilic material," as used herein, refers to a material that contains a hydrophobic or highly hydrophobic oligomer or polymer (e.g., a biodegradable oligomer or polymer) and a hydrophilic or highly hydrophilic oligomer or polymer.

[0364] The term "targeting moiety," as used herein, refers to a moiety that binds to or localizes to a particular location. The moiety can be, for example, a protein, a nucleic acid, a nucleic acid analog, a carbohydrate, or a small molecule. The location can be a tissue, a particular cell type, or a subcellular compartment. In some embodiments, the targeting moiety can specifically bind to a molecule of choice.

[0365] The term "reactive coupling group," as used herein, refers to any chemical functional group capable of reacting with a second functional group to form a covalent bond. The selection of a reactive coupling group is within the capabilities of one skilled in the art. Examples of reactive coupling groups include primary amines (-NH) and amine-reactive linking groups, such as isothiocyanates, isocyanates, acyl azides, NHS esters, sulfonyl chlorides, aldehydes, glyoxals, epoxides, oxiranes, carbonates, aryl halides, imidoesters, carbodiimides, anhydrides, and fluorophenyl esters. Many of these are conjugated to amines by either acylation or alkylation. Examples of reactive coupling groups include aldehydes (-COH) and aldehyde-reactive linking groups, such as hydrazides, alkoxyamines, and primary amines. Examples of reactive coupling groups include thiol groups (-SH) and sulfhydryl-reactive groups, such as maleimides, haloacetyls, and pyridyl disulfides. Examples of reactive coupling groups include photoreactive coupling groups, such as aryl azides or diazirines. The coupling reaction may involve the use of a catalyst, heat, a pH buffer, light, or a combination thereof.

[0366] The term "protecting group," as used herein, refers to a functional group that can be added to and / or substituted with another desired functional group to protect the desired functional group from certain reaction conditions, and can be selectively removed and / or substituted to deprotect or expose the desired functional group. Protecting groups are known to those skilled in the art. Suitable protecting groups include those described by Greene and Wuts, Protective Groups in Organic Synthesis, 1991. Acid-sensitive protecting groups include dimethoxytrityl (DMT), tert-butylcarbamate (tBoc), and trifluoroacetyl (tFA). Base-sensitive protecting groups include 9-fluorenylmethoxycarbonyl (Fmoc), isobutyryl (iBu), benzoyl (Bz), and phenoxyacetyl (pac). Other protecting groups include acetamidomethyl, acetyl, tert-amyloxycarbonyl, benzyl, benzyloxycarbonyl, 2-(4-biphenylyl)-2-propyloxycarbonyl, 2-bromobenzyloxycarbonyl, tert-butyloxycarbonyl, 1-carbobenzoxamido-2,2,2-trifluoroethyl, 2,6-dichlorobenzyl, 2-(3,5-dimethoxyphenyl)-2-propyloxycarbonyl, 2,4- Examples include dinitrophenyl, dithiasuccinyl, formyl, 4-methoxybenzenesulfonyl, 4-methoxybenzyl, 4-methylbenzyl, o-nitrophenylsulfenyl, 2-phenyl-2-propyloxycarbonyl, α-2,4,5-tetramethylbenzyloxycarbonyl, p-toluenesulfonyl, xanthenyl, benzyl ester, N-hydroxysuccinimide ester, p-nitrobenzyl ester, p-nitrophenyl ester, phenyl ester, p-nitrocarbonate, p-nitrobenzyl carbonate, trimethylsilyl, and pentachlorophenyl ester.

[0367] The term "activated ester," as used herein, refers to an alkyl ester of a carboxylic acid in which the alkyl is a good leaving group and renders the carbonyl susceptible to nucleophilic attack by molecules bearing an amino group. Activated esters are therefore susceptible to aminolysis and react with amines to form amides. Activated esters contain a carboxylic acid ester group, -COR, where R is a leaving group.

[0368] The term "alkyl" refers to the radical of saturated aliphatic groups including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups.

[0369] In some embodiments, a straight chain or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., C1-C for a straight chain). 30 , C3 to C for branched chains 30 ), 20 or fewer, 12 or fewer, or 7 or fewer. Likewise, in some embodiments, cycloalkyls have from 3-10 carbon atoms in their ring structure, such as for example, 5, 6 or 7 carbons in the ring structure. The term "alkyl" (or "lower alkyl") as used throughout the specification, examples, and claims, is intended to include both "unsubstituted alkyls" and "substituted alkyls," the latter of which refers to an alkyl moiety having one or more substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone. Such substituents include, but are not limited to, halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as thioester, thioacetate, or thioformate), alkoxyl, phosphoryl, phosphate, phosphonate, a hosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties.

[0370] Unless the number of carbons is otherwise specified, "lower alkyl," as used herein, means an alkyl group, as defined above, but having from 1 to 10 carbons, or 1 to 6 carbon atoms, in its backbone structure. Similarly, "lower alkenyl" and "lower alkynyl" have similar chain lengths. In some embodiments, alkyl groups are lower alkyls. In some embodiments, a substituent designated herein as alkyl is a lower alkyl.

[0371] Those skilled in the art will understand that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate. For example, substituted alkyl substituents can include halogen, hydroxy, nitro, thiols, amino, azido, imino, amido, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamides, sulfamoyl, and sulfonates), and silyl groups, as well as ethers, alkylthios, carbonyls (including ketones, aldehydes, carboxylates, and esters), -CF3, -CN, and the like. Cycloalkyls can also be similarly substituted.

[0372] The term "heteroalkyl" as used herein refers to a linear or branched chain, or cyclic carbon-containing radical, or combinations thereof, containing at least one heteroatom. Suitable heteroatoms include, but are not limited to, O, N, Si, P, Se, B, and S, where the phosphorous and sulfur atoms are optionally oxidized, and the nitrogen heteroatom is optionally quaternized. Heteroalkyls may be substituted as defined above for alkyl groups.

[0373] The term "alkylthio" refers to an alkyl group, as defined above, having a sulfur radical attached thereto. In some embodiments, the "alkylthio" moiety is represented by one of -S-alkyl, -S-alkenyl, and -S-alkynyl. Representative alkylthio groups can include methylthio and ethylthio. The term "alkylthio" also encompasses cycloalkyl groups, alkene and cycloalkene groups, and alkyne groups. "Arylthio" refers to an aryl or heteroaryl group. An alkylthio group may be substituted as defined above for an alkyl group.

[0374] The terms "alkenyl" and "alkynyl" refer to unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond, respectively.

[0375] The term "alkoxyl" or "alkoxy," as used herein, refers to an alkyl group, as defined above, having an oxygen radical attached thereto. Representative alkoxyl groups include methoxy, ethoxy, propyloxy, and tert-butoxy. An "ether" is two hydrocarbons covalently linked by an oxygen. Thus, an alkyl substituent that renders the alkyl ether is or resembles an alkoxyl, such as may be represented by one of -O-alkyl, -O-alkenyl, and -O-alkynyl. Aroxy may be represented by -O-aryl or -O-heteroaryl, where aryl and heteroaryl are defined below. Alkoxy and aroxy groups may be substituted as described above for alkyl.

[0376] The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines, for example, those of the general formula:

[0377] [ka]

[0378] [In the formula, R9, R 10 , and R' 10 are each independently hydrogen, alkyl, alkenyl, -(CH2) m -R8, or R9 and R 10 together with the N atom to which they are attached complete a heterocycle having 4 to 8 atoms in the ring structure; R represents aryl, cycloalkyl, cycloalkenyl, heterocycle, or polycycle; and m is 0 or an integer ranging from 1 to 8. In some embodiments, R or R 10 Only one of R and R may be a carbonyl. 10 and the nitrogen together do not form an imide. In still other embodiments, the term "amine" does not include amides, e.g., R and R 10 In a further embodiment, one of R and R represents carbonyl. 10 (and optionally R' 10 ) each independently represent hydrogen, alkyl or cycloalkly, alkenyl or cycloalkenyl, or alkynyl. Thus, the term "alkylamine," as used herein, means an amine group to which is appended an alkyl, as defined above, which may be substituted or unsubstituted (as described above for alkyl), i.e., R and R 10 At least one of the groups is an alkyl group.

[0379] The term "amide" is art recognized as an amino-substituted carbonyl and has the general formula:

[0380] [ka]

[0381] [Wherein R9 and R 10 is as defined above. "Aryl," as used herein, refers to a C5-C 10"aryl" refers to a 5-membered aromatic, heterocyclic, fused aromatic, fused heterocyclic, biaromatic, or biheterocyclic ring system. In its broadest sense, "aryl" as used herein includes 5-, 6-, 7-, 8-, 9-, and 10-membered monocyclic aromatic groups that may contain 0 to 4 heteroatoms, such as benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine. These aryl groups with heteroatoms in the ring structure may also be referred to as "aryl heterocycles" or "heteroaromatics." The aromatic ring may be substituted at one or more ring positions with one or more substituents including, but not limited to, halogen, azido, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino (or quaternized amino), nitro, sulfhydryl, imino, amide, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moiety, -CF3, -CN; and combinations thereof.

[0382] The term "aryl" also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjacent rings (i.e., "fused rings"), where at least one of the rings is, for example, aromatic and the other cyclic ring(s) may be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, and / or heterocycles.Examples of heterocycles include, but are not limited to, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3 b] tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl , naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathinyl, phenoxazinyl, phthalazinyl, piperazinyl piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclinyl thiadinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, and xanthenyl.One or more of these rings may be substituted as defined above for "aryl."

[0383] The term "aralkyl," as used herein, refers to an alkyl group substituted with an aryl group (e.g., an aromatic or heteroaromatic group). The term "carbocycle," as used herein, refers to an aromatic or non-aromatic ring in which each atom of the ring is carbon.

[0384] "Heterocycle" or "heterocyclic" as used herein refers to a heterocyclic ring containing 3 to 10 ring atoms, e.g., 5 to 6 ring atoms, each of which is selected from the group consisting of carbon, non-peroxide oxygen, sulfur, and N(Y), where Y is absent or selected from the group consisting of H, O, (C1-C 10) alkyl, phenyl, or benzyl, and optionally containing 1 to 3 double bonds and optionally substituted by one or more substituents. Examples of heterocycles include, but are not limited to, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydroquinolinyl, 2H,6H ... Furo[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl , oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxepanyl, oxetanyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl , purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydropyranyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2,Examples of heterocyclic groups include 5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, and xanthenyl. Heterocyclic groups may optionally be substituted at one or more positions with one or more substituents as defined above for alkyl and aryl, such as halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amide, phosphate, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moiety, -CF3, and -CN.

[0385] The term "carbonyl" is art-recognized and has the general formula:

[0386] [ka]

[0387] wherein X is a bond or represents oxygen or sulfur, and R 11 represents hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, or alkynyl; R' 11 represents hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, or alkynyl, wherein X is oxygen and R 11 or R' 11 Where X is oxygen and R is not hydrogen, the formula represents an "ester". 11 or as defined above, this moiety is referred to herein as a carboxyl group, and in particular R 11 Where X is an oxygen and R' is a hydrogen, the formula represents a "carboxylic acid". 11is hydrogen, the formula represents a "formate." In general, where the oxygen atom of the above formula is replaced with sulfur, the formula represents a "thiocarbonyl" group. 11 or R' 11 Where X is a sulfur and R is not hydrogen, the formula represents a "thioester." 11 Where X is a sulfur and R' is hydrogen, the formula represents a "thiocarboxylic acid." 11 is hydrogen, the formula represents a "thioformate." On the other hand, where X is a bond and R 11 Where X is a bond, and R is not hydrogen, the above formula represents a "ketone" group. 11 Where is hydrogen, the above formula represents an "aldehyde" group.

[0388] The term "monoester" as used herein refers to an analog of a dicarboxylic acid, where one of the carboxylic acids is functionalized as an ester and the other carboxylic acid is the free carboxylic acid or a salt of the carboxylic acid. Examples of monoesters include, but are not limited to, the monoesters of succinic acid, glutaric acid, adipic acid, suberic acid, sebacic acid, azelaic acid, oxalic acid, and maleic acid.

[0389] The term "heteroatom," as used herein, means an atom of any element other than carbon or hydrogen. Examples of heteroatoms are boron, nitrogen, oxygen, phosphorus, sulfur, and selenium. Other useful heteroatoms include silicon and arsenic.

[0390] As used herein, the term "nitro" means -NO2; the term "halogen" refers to -F, -Cl, -Br, or -I; the term "sulfhydryl" means -SH; the term "hydroxyl" means -OH; and the term "sulfonyl" means -SO2-.

[0391] The term "substituted," as used herein, refers to all permissible substituents of the compounds described herein. In the broadest sense, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. Exemplary substituents include, but are not limited to, halogens, hydroxyl groups, or any other organic group containing any number of carbon atoms, e.g., 1 to 14 carbon atoms, and optionally one or more heteroatoms such as oxygen, sulfur, or nitrogen groups in a linear, branched, or cyclic structural format. Representative substituents include alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxyl, alkoxy, substituted alkoxy, phenoxy, substituted phenoxy, aroxy, substituted aroxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amido, substituted amido, sulfonyl, substituted sulfonyl, sulfonic acid, phosphoryl, substituted phosphoryl, phosphonyl, substituted phosphonyl, polyaryl, substituted polyaryl, C3-C 20 cyclic, substituted C3~C 20 Included are cyclic, heterocyclic, substituted heterocyclic, amino acid, peptide, and polypeptide groups.

[0392] Heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valences of the heteroatoms. It is understood that "substituted" or "substituted" includes the implicit proviso that such substitution is in accordance with the permissible valences of the replacing atom and substituents, and that the substitution results in a stable compound, i.e., a compound that does not spontaneously undergo transformation by, for example, rearrangement, cyclization, or elimination.

[0393] In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. Illustrative substituents include, for example, those described herein. The permissible substituents for appropriate organic compounds can be one or more and can be the same or different. Heteroatoms, such as nitrogen, can have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valences of the heteroatoms.

[0394] In various embodiments, the substituents are selected from alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amido, amino, aryl, arylalkyl, carbamate, carboxy, cyano, cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl, heterocyclyl, hydroxyl, ketone, nitro, phosphate, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thioketone, each of which is optionally substituted with one or more suitable substituents. In some embodiments, the substituents are selected from alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amido, amino, aryl, arylalkyl, carbamate, carboxy, cycloalkyl, ester, ether, formyl, haloalkyl, heteroaryl, heterocyclyl, ketone, phosphate, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thioketone, wherein each of the alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amido, amino, aryl, arylalkyl, carbamate, carboxy, cycloalkyl, ester, ether, formyl, haloalkyl, heteroaryl, heterocyclyl, ketone, phosphate, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thioketone may be further substituted with one or more suitable substituents.

[0395] Examples of substituents include, but are not limited to, halogen, azido, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino, amide, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, thioketone, ester, heterocyclyl, -CN, aryl, aryloxy, perhaloalkoxy, aralkoxy, heteroaryl, heteroaryloxy, heteroarylalkyl, heteroaralkyl, and the like. Examples include koxy, azido, alkylthio, oxo, acylalkyl, carboxy esters, carboxamido, acyloxy, aminoalkyl, alkylaminoaryl, alkylaryl, alkylaminoalkyl, alkoxyaryl, arylamino, aralkylamino, alkylsulfonyl, carboxamidoalkylaryl, carboxamidoaryl, hydroxyalkyl, haloalkyl, alkylaminoalkylcarboxy, aminocarboxamidoalkyl, cyano, alkoxyalkyl, perhaloalkyl, arylalkyloxyalkyl, etc. In some embodiments, the substituent is selected from cyano, halogen, hydroxyl, and nitro.

[0396] The term "copolymer," as used herein, generally refers to a single polymeric material composed of two or more different monomers. The copolymer may be in any form, such as a random, block, or graft copolymer. The copolymer may have any end group, including capped or acidic end groups.

[0397] The terms "polypeptide," "peptide," and "protein" generally refer to a polymer of amino acid residues. As used herein, the terms also apply to amino acid polymers in which one or more amino acids are chemical analogues or modified derivatives of corresponding naturally occurring amino acids or are unnatural amino acids. The term "protein," as generally used herein, refers to a polymer of amino acids linked to each other by peptide bonds to form a polypeptide of sufficient chain length to generate tertiary and / or quaternary structure. The term "protein," by definition, excludes small peptides, which lack the requisite higher-order structure necessary to be considered a protein.

[0398] The terms "nucleic acid," "polynucleotide," and "oligonucleotide" are used interchangeably to refer to deoxyribonucleotide or ribonucleotide polymers in linear or circular conformation and in either single- or double-stranded form. These terms should not be construed as limiting the length of the polymer. These terms can encompass known analogs of natural nucleotides as well as nucleotides modified in the base, sugar, and / or phosphate moieties (e.g., phosphorothioate backbones). Generally, unless otherwise specified, analogs of a particular nucleotide have the same base-pairing specificity; i.e., an analog of A can base pair with T. The term "nucleic acid" is a term of art that refers to a stretch of at least two base-sugar-phosphate monomer units. Nucleotides are the monomer units of a nucleic acid polymer. This term includes deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) in the form of messenger RNA, antisense, plasmid DNA, portions of plasmid DNA, or genetic material derived from viruses. Antisense nucleic acids are polynucleotides that interfere with the expression of DNA and / or RNA sequences. The term nucleic acid refers to a sequence of at least two base-sugar phosphate combinations. Natural nucleic acids have a phosphate backbone. Artificial nucleic acids may contain other types of backbones, but may contain the same bases as natural nucleic acids. The term also includes PNA (peptide nucleic acid), phosphorothioates, and other modifications of the phosphate backbone of natural nucleic acids.

[0399] A "functional fragment" of a protein, polypeptide, or nucleic acid is a protein, polypeptide, or nucleic acid whose sequence is not identical to that of the full-length protein, polypeptide, or nucleic acid, but which still retains at least one function of the full-length protein, polypeptide, or nucleic acid. A functional fragment can have more, fewer, or the same number of residues as the corresponding native molecule and / or can contain one or more amino acid or nucleotide substitutions. Methods for determining the function of a nucleic acid (e.g., coding function, ability to hybridize with another nucleic acid) are well known in the art. Similarly, methods for determining protein function are well known. For example, the DNA binding function of a polypeptide can be determined, for example, by filter binding, electrophoretic mobility shift, or immunoprecipitation assays. DNA cleavage can be assayed by gel electrophoresis. The ability of a protein to interact with another protein can be determined, for example, by co-immunoprecipitation, two-hybrid assays, or genetic or biochemical complementation, for example. See, for example, Fields et al., 1989, Nature 340:245-246; U.S. Pat. No. 5,585,245; and WO 98 / 44350.

[0400] As used herein, the term "linker" refers to a carbon chain that contains heteroatoms (e.g., nitrogen, oxygen, sulfur, etc.) and can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 atoms in length. Linkers may be substituted with various substituents, including, but not limited to, hydrogen atoms, alkyl, alkenyl, alkynyl, amino, alkylamino, dialkylamino, trialkylamino, hydroxyl, alkoxy, halogen, aryl, heterocyclic, aromatic heterocyclic, cyano, amido, carbamoyl, carboxylic acid, ester, thioether, alkylthioether, thiol, and ureido groups. Those skilled in the art will recognize that each of these groups may also be substituted. Examples of linkers include, but are not limited to, pH-sensitive linkers, protease-cleavable peptide linkers, nuclease-sensitive nucleic acid linkers, lipase-sensitive lipid linkers, glycosidase-sensitive carbohydrate linkers, hypoxia-sensitive linkers, photocleavable linkers, thermolabile linkers, enzyme-cleavable linkers (e.g., esterase-cleavable linkers), ultrasound-sensitive linkers, and X-ray-cleavable linkers.

[0401] The term "pharmaceutically acceptable counterion" refers to a pharmaceutically acceptable anion or cation. In various embodiments, the pharmaceutically acceptable counterion is a pharmaceutically acceptable ion. For example, pharmaceutically acceptable counterions are selected from citrate, malate, acetate, oxalate, chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1′-methylene-bis-(2-hydroxy-3-naphthoate)). In some embodiments, the pharmaceutically acceptable counterion is selected from chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, citrate, malate, acetate, oxalate, acetate, and lactate. In particular embodiments, the pharmaceutically acceptable counterion is selected from chloride, bromide, iodide, nitrate, sulfate, bisulfate, and phosphate.

[0402] The term "pharmaceutically acceptable salts" refers to salts of acidic or basic groups that may be present in compounds used in the present compositions. Compounds included in the present compositions that are basic in nature are capable of forming a wide variety of salts with various inorganic and organic acids. Acids that can be used to prepare pharmaceutically acceptable acid addition salts of such basic compounds are those that form non-toxic acid addition salts, i.e., salts containing pharmacologically acceptable anions, including, but not limited to, sulfate, citrate, malate, acetate, oxalate, chloride, bromide, iodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, oleate, tannate, and paclitaxel. The salts of the present compositions include anthraquinone, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)). Compounds contained in the present compositions that contain an amino moiety can form pharmaceutically acceptable salts with various amino acids in addition to the acids mentioned above. Compounds contained in the present compositions that are acidic in nature are capable of forming base salts with various pharmacologically acceptable cations. Examples of such salts include alkali metal or alkaline earth metal salts, particularly calcium, magnesium, sodium, lithium, zinc, potassium, and iron salts.

[0403] When a compound described herein is obtained as an acid addition salt, the free base can be obtained by basifying a solution of the acid salt. Conversely, when the product is a free base, an addition salt, particularly a pharmaceutically acceptable addition salt, can be made by dissolving the free base in a suitable organic solvent and treating the solution with an acid, according to conventional procedures for preparing acid addition salts from base compounds. Those skilled in the art will recognize various synthetic methods that can be used to prepare non-toxic, pharmaceutically acceptable addition salts.

[0404] Pharmaceutically acceptable salts include 1-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, and ethanesulfonic acid. , formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isethionic acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, pantothenic acid, phosphoric acid, proprionic acid, pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tartaric acid, thiocyanic acid, toluenesulfonic acid, trifluoroacetic acid, and undecylenic acid.

[0405] The term "bioavailable" is art-recognized and refers to an aspect of the invention that allows it, or a portion of a dose, to be absorbed, taken up, or otherwise physiologically available by the subject or patient to whom it is administered.

[0406] It will be understood that the following examples are intended to illustrate, not limit, the invention. Various other embodiments and modifications of the foregoing description and examples will be apparent to those skilled in the art after reading this disclosure without departing from the spirit and scope of the invention, and it is intended that all such embodiments or modifications be included within the scope of the appended claims. All publications and patents cited herein are hereby incorporated by reference in their entirety.

[0407] It will be understood that in the following examples, some conjugates have been prepared and characterized using non-radioactive metals such as Lu-175. It will be apparent to those skilled in the art that the corresponding radioactive Lu-177 analogs can be readily prepared using known methods, and the distribution data for the Lu-175 conjugates can be representative of the Lu-177 analogs.

[0408] Example Example 1: Synthesis of the conjugate The conjugates of the present invention can be prepared using any convenient method. In a rational approach, the conjugate is assembled from its individual components, the targeting moiety, optionally a linker, and an active drug moiety. These components can be covalently linked to each other via functional groups, as known in the art, where such functional groups may be present on the components or may be introduced onto the components using one or more steps, such as oxidation, reduction, cleavage, etc. Functional groups that can be used to covalently link components together to prepare pharmaceutical conjugates include hydroxy, sulfhydryl, amino, etc. The specific portions of the various components modified to provide covalent linkages will be selected so as not to adversely interfere with the desired binding activity of the components; for example, the active drug moiety will be modified in a region that does not affect target binding activity to ensure sufficient desired drug activity. As necessary and / or desired, particular moieties on the components may be protected using blocking groups as known in the art, see, for example, Green & Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991.

[0409] Alternatively, conjugates can be produced using known combinatorial methods to produce a large library of candidate conjugates, and then the library can be screened to identify bifunctional molecules with pharmacokinetic profiles.Alternatively, conjugates can be produced using medicinal chemistry and the known structure-activity relationship between targeting moiety and active drug moiety.In particular, this approach will provide insight into where to connect the two moieties to the linker.

[0410] Peptide synthesis Peptide 1 was synthesized on a Liberty Blue peptide synthesizer starting with Fmoc-beta-alanine (1.33 g, 0.75 mmol / g loading, 1.00 mmol) loaded onto 2-chlorotrityl resin and followed by the addition of Fmoc-D-Glu(OtBu) (x3), Fmoc-AEEA, Fmoc-Lys(Boc), and 4-(p-iodophenyl)butyric acid. The crude peptide was cleaved by treating the resin with 2% TFA in dichloromethane, and purified by preparative HPLC to give 624 mg (0.483 mmol, 48% yield) of peptide 1.

[0411] [ka]

[0412] Peptide 2 was synthesized on a Liberty Blue peptide synthesizer starting with Fmoc-beta-alanine (4.00 g, 0.75 mmol / g loading, 3.00 mmol) loaded onto 2-chlorotrityl resin and followed by the addition of Fmoc-D-Glu(OtBu) (x3) and Fmoc-AEEA. The crude peptide was cleaved by treating the resin with 2% TFA in dichloromethane, and purified by preparative HPLC to give 1.28 mg (1.26 mmol, 42% yield) of peptide 2. Peptides 3 and 4 were synthesized in a similar manner.

[0413] [ka]

[0414] Peptide 5 was synthesized on a Liberty Blue peptide synthesizer starting with Fmoc-beta-alanine (0.4 g, 0.5 mmol / g loading, 0.2 mmol) loaded onto 2-chlorotrityl resin, followed by the addition of Fmoc-D-Glu(OtBu) (x3), Fmoc-AEEA, Dde-Lys(Fmoc), (R)-tert-Bu4-DOTAGA, followed by treatment with 5% hydrazine in DMF, and then the addition of 4-(p-iodophenyl)butyric acid. The crude peptide was cleaved by treating the resin with 2% TFA in dichloromethane, and purified by preparative HPLC to give 50 mg (0.027 mmol, 13% yield) of peptide 5. Peptides 6 and 7 were synthesized in a similar manner.

[0415] [ka]

[0416] Peptide 8 was synthesized on a Liberty Blue peptide synthesizer starting with Fmoc-beta-alanine (0.5 g, 0.75 mmol / g loading, 0.375 mmol) loaded onto 2-chlorotrityl resin, followed by the addition of Fmoc-D-Glu(OtBu) (×3), Fmoc-AEEA, Fmoc-Dde-Lys(Dde), 4-(p-iodophenyl)butyric acid, and treatment with 5% hydrazine in DMF, followed by Fmoc-Gly, Fmoc-Val, Fmoc-Met, and Fmoc-Gly. The crude peptide was cleaved from the resin with 2% TFA in dichloromethane, and all solvent was removed. The crude peptide was dissolved in acetonitrile (5 mL) and triethylamine (2 mL), and the solution was heated at 70 °C for 1 h. All solvents were removed under vacuum, and the crude product was treated with (R)-tert-Bu4-DOTAGA NHS ester (172 mg, 0.215 mmol) in DMF (5 mL) and diisopropylethylamine (0.5 mL). The solution was stirred at 50 °C for 1 h and then purified by preparative HPLC to give 8 (124 mg, 55.9 mmol, 15% yield). Peptides 9–16 were prepared in a similar manner.

[0417] [ka]

[0418] [ka]

[0419] [ka]

[0420] To a vial containing the HCl salt of TM5 (170 mg, 0.340 mmol) and Boc-glutamic acid (42 mg, 0.170 mmol), a solution of HATU (128 mg, 0.340 mmol) in DMF (3 mL) was added, followed by diisopropylethylamine (0.60 mL). The solution was stirred at 50 °C for 1 h and then purified by preparative HPLC. The product-containing fractions were concentrated to dryness, and then trifluoroacetic acid (2 mL) was added. The solution was stirred at room temperature for 15 min, and then excess trifluoroacetic acid was removed under vacuum. The remaining material was redissolved in 1:1 acetonitrile:water (10 mL), frozen, and lyophilized to give 17 as the trifluoroacetate salt (192 mg, 0.138 mmol, 81% yield). Compounds 18 and 19 were prepared in a similar manner.

[0421] [ka]

[0422] A vial was charged with the HCl salt of T-1951 (460 mg, 0.893 mmol) and Fmoc-DGlu-bAla (200 mg, 0.454 mmol). To this was added a solution of HATU (343 mg, 0.909 mmol) in DMF (5 mL). Diisopropylethylamine (0.50 mL) was added, and the solution was stirred at 50 °C for 1 h, followed by the addition of DBU (0.50 mL). The reaction was stirred at 50 °C for an additional 1 h. The reaction was then acidified by the addition of 1 mL of TFA, and the reaction mixture was purified by preparative HPLC to give 20 as the trifluoroacetate salt (443 mg, 0.300 mmol, 65% yield). Compounds 21–32 were prepared in a similar manner.

[0423] [ka]

[0424] [ka]

[0425] [ka]

[0426] To a vial containing the trifluoroacetate salt of 20 (64.9 mg, 43.8 μmol), (R)-tert-Bu4-DOTAGA (74.9 mg, 109 μmol), and HATU (31.9 mg, 84.7 μmol), DMF (2 mL) and diisopropylethylamine (0.20 mL) were added. The solution was stirred at 50 °C for 1 h and then purified by preparative HPLC to give 33 (43 mg, 23.6 μmol), 53% yield. Compounds 34–46 were prepared in a similar manner.

[0427] [ka]

[0428] [ka]

[0429] [ka]

[0430] [ka]

[0431] [ka]

[0432] [ka]

[0433] A vial was charged with the trifluoroacetate salt of 17 (120 mg, 86.8 μmol) and 1 (131 mg, 101 μmol). To this was added a solution of HATU (36.0 mg, 95.5 μmol) in DMF (4 mL). Diisopropylethylamine (0.40 mL) was added, and the reaction was stirred at 50 °C for 1 h. The reaction mixture was purified by preparative HPLC to give 47 (38.0 mg, 16.4 μmol, 18% yield). Compounds 48–53 were prepared in a similar manner.

[0434] [ka]

[0435] [ka]

[0436] [ka]

[0437] [ka]

[0438] [ka]

[0439] A vial was charged with 47 (18.0 mg, 7.78 μmol) and TFA (1 mL) was added. The reaction was stirred at 50 °C for 1 h, and then the TFA was removed under vacuum. To the remaining material was added a solution of (R)-tert-Bu4-DOTAGA NHS ester (18.6 mg, 23.3 μmol) in DMF (3 mL). Diisopropylethylamine (0.5 mL) was added, and the solution was stirred at 50 °C for 30 min, then purified by preparative HPLC to give 54 (11.2 mg, 4.11 μmol, 52% yield). Compounds 55–60 were prepared in a similar manner.

[0440] [ka]

[0441] [ka]

[0442] [ka]

[0443] [ka]

[0444] [ka]

[0445] A vial was charged with 2 (41.0 mg, 40.5 μmol), the trifluoroacetate salt of 27 (43.8 mg, 31.7 μmol), and HATU (15.5 mg, 41.0 μmol). DMF (2 mL) and diisopropylethylamine (0.2 mL) were added, and the reaction was stirred at 50 °C for 1 h. DBU (0.2 mL) was then added, and the reaction was stirred at 50 °C for an additional 10 min. The reaction mixture was purified by preparative HPLC to give the trifluoroacetate salt of 61 (23.0 mg, 10.6 μmol, 33% yield). Compounds 62–67 were prepared in a similar manner.

[0446] [ka]

[0447] [ka]

[0448] [ka]

[0449] [ka]

[0450] A vial was charged with the trifluoroacetate salt of 61 (29.0 mg, 14.5 μmol) and a solution of (R)-tert-Bu4-DOTAGA NHS ester (16.3 mg, 20.4 μmol) in DMF (2 mL) was added. Diisopropylethylamine (0.2 mL) was added and the reaction was stirred at room temperature for 18 h, then purified by preparative HPLC to give 68 (18.0 mg, 7.01 μmol, 48% yield). Compounds 69–74 were prepared in a similar manner.

[0451] [ka]

[0452] [ka]

[0453] [ka]

[0454] [ka]

[0455] [ka]

[0456] [ka]

[0457] [ka]

[0458] A vial was charged with 5 (17.7 mg, 9.45 μmol) and the trifluoroacetate salt of 28 (17.2 mg, 11.6 μmol), and a solution of HATU (4.6 mg, 12.3 μmol) in DMF (2 mL) was added. Diisopropylethylamine (0.2 mL) was then added, and the reaction was stirred at 50 °C for 1 h and purified by preparative HPLC to give 75 (10.7 mg, 3.57 μmol, 37% yield). Compounds 76–87 were prepared in a similar manner.

[0459] [ka]

[0460] [ka]

[0461] [ka]

[0462] [ka]

[0463] [ka]

[0464] [ka]

[0465] [ka]

[0466] [ka]

[0467] [ka]

[0468] [ka]

[0469] [ka]

[0470] [ka]

[0471] [ka]

[0472] [ka]

[0473] [ka]

[0474] [ka]

[0475] Typical procedure for labeling with cold ruthenium: 33 (22.0 mg, 12.1 μmol) was dissolved in trifluoroacetic acid, and the solution was heated at 50°C for 1 h. Toluene (5 mL) was added, and all solvent was removed under vacuum. To the remaining residue was added a solution of lutetium(III) chloride (6.25 mg, 22.2 μmol) in pH 4.5 HCl / acetate buffer (1 mL). 0.2 N sodium acetate (1 mL) was added, and the solution was heated at 90°C for 20 min. The solution was then cooled to room temperature and purified by preparative HPLC to give T-2743 (17.3 mg, 9.78 μmol, 80% yield).

[0476] The table below lists the starting materials used in the general lutetium conjugation procedure above, the final products, and the major ions detected in the mass spectra.

[0477] [Table 6-1]

[0478] [Table 6-2]

[0479] Example 2: Biodistribution studies Lutetium accumulation was measured in tumors, plasma, and healthy tissues of NCI-H460 tumor-bearing mice (lung cancer). NCI-H460 tumor-bearing mice were administered 0.5 mg / kg of lutetium conjugate. At the designated time points, mice were sacrificed and tumors, livers, kidneys, and plasma were collected. All tissues were analyzed for lutetium content by ICP-MS, and lutetium uptake in %ID / g was determined using the following formula: %ID / g = ((tissue lutetium in ppb) / 175) × (molecular weight of conjugate)) / (0.5 × (mouse weight in grams) × 10) The biodistribution at 24 and 72 hours is shown in the table below.

[0480] [Table 7]

[0481] The biodistribution and tumor / kidney ratio (T / K), tumor / liver ratio (T / L), and tumor / plasma ratio (T / P) at 24 hours are shown in the table below.

[0482] [Table 8-1]

[0483] [Table 8-2]

[0484] Another study using the NCI-H460 tumor model investigated and compared the distribution of a radioactive (hot) analog of CMP51 (177Lu) and a non-radioactive (cold) analog of CMP51 (175Lu) at 24 and 72 hours. The radioactive analog exhibited similar levels of tumor uptake and retention as the cold analog. As shown in Figure 1, kidney levels of hot CMP51 were higher than tumor levels. High tumor / tissue ratios were observed in other tissues.

[0485] In a further study using the NCI-H460 tumor model, the distribution of a cold analog of CMP6 was measured at 24 and 72 hours. As shown in Figure 2, CMP6 showed significantly reduced kidney uptake and retention. Liver uptake was also significantly reduced. High tumor-to-liver and tumor-to-kidney ratios were observed. These radioactivity levels were enhanced at 72 hours.

[0486] Example 3: In vitro HSP90 binding studies using conjugates HSP90 binding was determined by a competitive fluorescence polarization assay using purified N-terminal HSP90α. Serial dilutions of test compounds were prepared in 10% DMSO in assay buffer, and 10 μl of each dilution was added to 100 μl of the reaction mixture, resulting in a final DMSO concentration of 1% in all reactions. Reactions were performed at room temperature for 3 hours in 100 μl mixtures containing assay buffer, 5 nM FITC-labeled geldanamycin, 350 ng of N-terminal HSP90α, and test compound. Fluorescence intensity was measured at 485 nm excitation and 530 nm emission using a Tecan Infinite M1000 microplate reader. Fluorescence intensity was converted to fluorescence polarization using Tecan Magellan6 software. Fluorescence polarization data were analyzed using computer software, Graphpad Prism. Fluorescence polarization (FPt) in the absence of compound in each data set was defined as 100% activity. Fluorescence polarization (FPb) values ​​in the absence of protein and compound in each data set were defined as 0% activity. The percent activity in the presence of compound was calculated according to the following formula: % activity = (FP - FPb) / (FPt - FPb) x 100%, where FP = fluorescence polarization in the presence of compound. HSP90 binding data are as follows:

[0487] [Table 9]

[0488] Example 4: Determination of the permeability of the conjugate To test the ability of the conjugates to enter cells, an artificial membrane permeability assay ("PAMPA") is used. PAMPA is a useful tool for predicting in vivo drug permeability for drugs that enter cells by passive transport mechanisms. LC / MS is used in conjunction with the PAMPA assay to determine the ability of the conjugates to penetrate cells.

[0489] Pre-coated PAMPA plates are allowed to warm to room temperature for at least 30 minutes before adding assay components. Prepare stock solutions using the conjugates to be tested. To create working solutions, add 50 μL of the 100 μM stock in DMSO + 950 μL of PBS or 50 μL of the 200 μM stock to a 96-deep-well plate for a final concentration of 5 μM or 10 μM, respectively. Add 300 μL of working solution containing each conjugate to be tested to the appropriate wells of the donor PAMPA plate. Add 200 μL of PBS to the corresponding wells of the acceptor PAMPA plate.

[0490] Lower the acceptor plate onto the donor plate and incubate for 5 hours. After 5 hours, remove a 50 µL aliquot from each well of each plate and add it to a new 96-deep-well plate.

[0491] 100 μL of methanol containing a given internal standard control compound is added to each aliquot and analyzed by LC / MS. The permeability of each conjugate is calculated. Example 5: Synthesis of conjugates CMP1 to CMP50 The conjugates of the present invention can be prepared using any convenient method. In a streamlined approach, the conjugate is assembled from its individual components, the targeting moiety, optionally a linker, and their active agent moiety or precursor. In some embodiments, the linker or precursor thereof is attached to the targeting moiety or precursor thereof, followed by coupling with the active agent moiety or precursor thereof to provide the conjugate. In some embodiments, the linker or precursor thereof is attached to the active agent moiety or precursor thereof, followed by coupling with the targeting moiety or precursor thereof to provide the conjugate. These components can be covalently linked to each other via functional groups, as known in the art, where such functional groups may be present on the components or may be introduced onto the components using one or more steps, such as oxidation, reduction, cleavage, etc. Functional groups that can be used to covalently link the components together to create pharmaceutical conjugates include hydroxy, sulfhydryl, amino, etc. The specific portions of the various components modified to provide covalent linkages will be selected so as not to adversely interfere substantially with the desired binding activity of the component; for example, for active drug moieties, regions that do not affect target binding activity will be modified to ensure sufficient desired drug activity. If necessary and / or desired, specific portions on the components may be protected using blocking groups as known in the art; see, for example, Green & Wuts, "Protective Groups in Organic Synthesis," John Wiley & Sons, 1991.

[0492] Alternatively, conjugates can be generated using known combinatorial methods to generate large libraries of candidate conjugates, which can then be screened to identify bifunctional molecules with desirable pharmacokinetic profiles. Alternatively, conjugates can be generated using medicinal chemistry and known structure-activity relationships for the targeting moiety and active agent moiety. Specifically, this approach will provide insight into where the two moieties should be joined to the linker. Conjugates CMP1-CMP50 can also be synthesized as shown in Example 1. The targeting moiety, linker, and active agent moieties of conjugates CMP1-CMP50 can also be synthesized as shown in Example 1.

[0493] The PEG spacers of the present disclosure can be obtained commercially or synthesized by one skilled in the art using conventional organic chemistry. In a rational approach, the PEG spacer can be constructed from polyethylene glycol (PEG). The PEG spacer can be (PEG) n where n is an integer from 1 to 20. (PEG) n can be covalently attached to a functional group as known in the art, such functional groups being (PEG) n (PEG) n If necessary and / or desired, (PEG) nCertain of the above moieties may be protected using blocking groups as known in the art; see, for example, Green & Wuts, "Protective Groups in Organic Synthesis," John Wiley & Sons, 1991. PEG linkers of the present disclosure can be homobifunctional or heterobifunctional. For example, in some embodiments, a PEG linker can have a carboxylic acid group at one end and an amino group at the other end. PEG linkers can be synthesized using individual monodisperse or polydisperse PEG reagents. Peptide / peptide linkers of the present disclosure can be synthesized according to the method set forth in Example 1.

[0494] CMP1-CMP50 can be similarly synthesized according to the conjugate synthesis method shown in Example 1. CMP2 is synthesized by combining the targeting moiety TM10 with 100, followed by coupling with (R)-tert-Bu4-DOTAGA and labeling with cold ruthenium, as shown below. CMP1, CMP22, CMP29-CMP34, and CMP39-CMP42 are made in a similar manner.

[0495] [ka]

[0496] CMP3 is synthesized by combining the linker with the appropriate DOTA reagent to generate 101, followed by coupling to the targeting moiety TMb and labeling with cold ruthenium as shown below. CMP4-CMP6 and CMP23 are made in a similar manner.

[0497] [ka]

[0498] CMP10 is synthesized by reacting the targeting moiety TM3 with 102, followed by coupling with (R)-tert-Bu4-DOTAGA and labeling with cold ruthenium, as shown below. CMP7-CMP9, CMP11, CMP12, CMP17-CMP21, and CMP24-CMP28 are prepared in a similar manner.

[0499] [ka]

[0500] CMP13 is synthesized by coupling the targeting moiety TM2 to 104 to generate 105. 105 is then coupled to peptide 106, synthesized according to the method set forth in Example 1, followed by coupling with (R)-tert-Bu4-DOTAGA and labeling with cold ruthenium, as shown below. CMPs 14-16, CMP35-38, and CMP43-CMP50 are prepared in a similar manner.

[0501] [ka]

Claims

1. A conjugate comprising an active agent attached via a linker to at least one targeting moiety (TM), wherein the TM binds to HSP90, and the active agent comprises a radioactive material or a chelator of a radioactive material, the conjugate having the formula X10: 【Chemical 1】 Or formula X20: 【Chemistry 2】 having the structure The TMs include ganetespib, luminespib (TM15, AUY-922, NVP-AUY922), Debio-0932, MPC-3100, onarespib (AT-13387), SNX-2112, 17-amino-geldanamycin hydroquinone, PU-H71, SNX5422 (PF-04929113), TM1, and TM2. , TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15, and TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15 is represented by the following formula: 【Chemistry 3-1】 【Chemistry 3-2】 【Chemistry 3-3】 【Chemistry 3-4】 The chelating agent in X10 is 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), and the chelating agent in X20 is 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, α-(2-carboxyethyl) (DOTAGA), the radioactive material comprises Y-90, Y-86, I-131, Re-186, Re-188, Y-90, Bi-212, At-211, Zr-89, Sr-89, Ho-166, Sm-153, Cu-67, Cu-64, Lu-177, Ac-225, Pb-203, Bi-213, Th-227, Pb-212, Ra-223, P-32, Sc-47, Br-77, Rh-105, Pd-103, Ag-111, Pr-142, Pm-149, Gd-159, Ir-194, or Pt-199; and (i) the linker comprises a spacer, and the spacer comprises polyethylene glycol (PEG); 【Chemistry 4】 does not contain, or (ii) the conjugate comprises an active agent linked to at least two targeting moieties (TMs) via a linker, each of the TMs binding to HSP90, the active agent comprising a radioactive material or a chelator of a radioactive material, the conjugate having the structure of Formula X10 or Formula X20, and (1) the linker comprises a spacer, the spacer comprising at least two amino acids, or (2) the conjugate comprises at least one pharmacokinetic modulating unit (PMU); The PMU includes: 【Chemistry 5】 A conjugate having the structure:

2. The conjugate of claim 1 , wherein the linker comprises a spacer, and the spacer comprises at least one amino acid.

3. 3. The conjugate of claim 2, wherein the spacer comprises two amino acids, three amino acids, four amino acids, or five amino acids.

4. Formula A10: 【Chemistry 6】 Or formula A20: 【Chemistry 7】 3. The conjugate of claim 2 having the structure:

5. The conjugate according to any one of claims 2 to 4, wherein the TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15 represented by the following formula: 【Chemistry 8-1】 【Chemistry 8-2】 【Chemistry 8-3】 【Chemistry 8-4】

6. The conjugates include T1, T2, T4, T5, T6, T8, T9, T10, T27, T28, T29, T39, CMP13, CMP14, CMP15, CMP16, CMP23, CMP37, CMP38, CMP43, CMP44, CMP45, CMP46, CMP47, CMP48, CMP49, CMP50, T1', T2', T4', T5', T6', T8 3. The conjugate of claim 2, which is CMP13', CMP14', CMP15', CMP16', CMP23', CMP37', CMP38', CMP43', CMP44', CMP45', CMP46', CMP47', CMP48', CMP49', CMP50', or a pharmaceutically acceptable salt thereof. 【Chemistry 9-1】 【Chemistry 9-2】 【Chemistry 9-3】 【Chemistry 9-4】 【Chemistry 9-5】 【Chemistry 9-6】 【Chemistry 9-7】 【Chemistry 9-8】 【Hua 9-9】 【Chemistry 9-10】 【9-11】 【Chemistry 9-12】 【Chemistry 9-13】 【Chemistry 9-14】 【Chemistry 9-15】 【Chemistry 9-16】 【Chemistry 9-17】 【Chemistry 9-18】 【Chemistry 9-19】 【Chemistry 9-20】 【Chemistry 9-21】 【Chemistry 9-22】 【Chemistry 9-23】 【Chemistry 9-24】 【Chemistry 9-25】 【Chemistry 9-26】

7. 2. The conjugate of claim 1, wherein the linker comprises a spacer, and the spacer comprises polyethylene glycol (PEG).

8. The conjugate of claim 7, wherein the spacer is (PEG)4 or (PEG)12.

9. Formula B10: 【Chemistry 10】 Or formula B20: 【Chemistry 11】 8. The conjugate of claim 7 having the structure:

10. The conjugate according to any one of claims 7 to 9, wherein the TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15 represented by the following formula: 【Chemistry 12-1】 【Chemistry 12-2】 【Chemistry 12-3】 【Chemistry 12-4】

11. The conjugate according to claim 7, which is CMP1, CMP2, CMP29, CMP30, CMP31, CMP32, CMP33, CMP34, CMP35, CMP36, CMP39, CMP40, CMP41, CMP42, CMP52, CMP1', CMP2', CMP29', CMP30', CMP31', CMP32', CMP33', CMP34', CMP35', CMP36', CMP39', CMP40', CMP41', CMP42', CMP52' represented by the following formula, or a pharmaceutically acceptable salt thereof: 【Chemistry 13-1】 【Chemistry 13-2】 【Chemistry 13-3】 【Chemistry 13-4】 【Chemistry 13-5】 【Chemistry 13-6】 【Chemistry 13-7】 【Chemistry 13-8】

12. The conjugate of claim 1 , comprising at least one pharmacokinetic modulating unit (PMU).

13. The conjugate of claim 12 , wherein the PMU binds to albumin.

14. The PMU 【Chemistry 14】 14. The conjugate of claim 13, comprising:

15. Formula C10: 【Chemistry 15】 Or formula C20: 【Chemistry 16】 13. The conjugate of claim 12 having the structure:

16. The conjugate according to any one of claims 12 to 15, wherein the TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15 represented by the following formula: 【Chemistry 17-1】 【Chemistry 17-2】 【Chemistry 17-3】 【Chemistry 17-4】

17. The conjugates include CMP3, CMP4, CMP5, CMP6, T3, T7, T11, T12, T13, T14, T15, T16, T17, T22, T23, T32, T24, T33, T34, T35, T36, T37, T38, T41, T42, CMP3', CMP4', CMP5', CMP 13. The conjugate of claim 12, wherein the conjugate is T6', T3', T7', T11', T12', T13', T14', T15', T16', T17', T22', T23', T32', T24', T33', T34', T35', T36', T37', T38', T41', T42', or a pharmaceutically acceptable salt thereof. 【Chemistry 18-1】 【Chemistry 18-2】 【Chemistry 18-3】 【Chemistry 18-4】 【Chemistry 18-5】 【Chemistry 18-6】 【Chemistry 18-7】 【Chemistry 18-8】 【Chemistry 18-9】 【Chemistry 18-10】 【Chemistry 18-11】 【Chemistry 18-12】 【Chemistry 18-13】 【Chemistry 18-14】 【Chemistry 18-15】 【Chemistry 18-16】 【Chemistry 18-17】 【Hua 18-18】 【Chemistry 18-19】 【Chemistry 18-20】 【Chemistry 18-21】 【Chemistry 18-22】 【Chemistry 18-23】 【Chemistry 18-24】 【Chemistry 18-25】 【Chemistry 18-26】 【Chemistry 18-27】 【Chemistry 18-28】 【Chemistry 18-29】 【18-30】 【Chemistry 18-31】 【Chemistry 18-32】 【Chemistry 18-33】 【Chemistry 18-34】 【18-35】 【Chemistry 18-36】 【Chemistry 18-37】 【18-38】

18. 2. The conjugate of claim 1, wherein TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7, or TM15 represented by the following formula: 【Chemistry 19-1】 【Chemistry 19-2】 【Chemistry 19-3】 【Chemistry 19-4】

19. The conjugates include CMP7, CMP8, CMP9, CMP10, CMP11, CMP12, CMP17, CMP18, CMP19, CMP20, CMP21, CMP22, CMP24, CMP25, CMP26, CMP27, CMP28, T18, T19, T20, T21, T25, T26, T30, T31, T43, T40, CMP7', CMP8', CMP9', CMP1 2. The conjugate of claim 1, wherein the conjugate is CMP11', CMP12', CMP17', CMP18', CMP19', CMP20', CMP21', CMP22', CMP24', CMP25', CMP26', CMP27', CMP28', T18', T19', T20', T21', T25', T26', T30', T31', T43', T40', or a pharmaceutically acceptable salt thereof. 【Chemistry 20-1】 【Chemistry 20-2】 【Chemistry 20-3】 【Chemistry 20-4】 【Chemistry 20-5】 【Chemistry 20-6】 【Chemistry 20-7】 【Chemistry 20-8】 【Chemistry 20-9】 【Chemistry 20-10】 【Chemistry 20-11】 【Chemistry 20-12】 【Chemistry 20-13】 【Chemistry 20-14】 【Chemistry 20-15】 【20-16】 【20-17】 【20-18】 【20-19】 【20-20】 【20-21】

20. A pharmaceutical composition comprising the conjugate of any one of claims 1 to 19 and at least one pharmaceutically acceptable excipient.

21. 20. The conjugate of any one of claims 1 to 19 for use in the manufacture of a medicament for reducing cell proliferation.

22. 22. The conjugate of claim 21, wherein the cell is a cancer cell.

23. 23. The conjugate of claim 22, wherein the cancer cell is a small cell lung cancer cell, a non-small cell lung cancer cell, a sarcoma cell, a pancreatic cancer cell, a breast cancer cell, or a colon cancer cell.

24. 21. The pharmaceutical composition of claim 20 for use in the manufacture of a medicament for treating cancer.

25. 25. The pharmaceutical composition of claim 24, wherein the cancer is small cell lung cancer, non-small cell lung cancer, sarcoma, pancreatic cancer, breast cancer, or colon cancer.

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