HSP90 conjugates and their preparations

Abstract

Conjugates of active substances have been designed, linking them via linkers to a target moiety (such as at least one HSP90 binding moiety). Such conjugates can provide improved spatiotemporal delivery of the active substance; improved biodistribution and penetration into tumors; and / or reduced toxicity. Methods for preparing such conjugates and formulations thereof are provided. Methods for administering such formulations to subjects in need, for example, for the treatment or prevention of cancer, are provided.

Description

[0001] Citation of relevant applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 62 / 986,245, filed March 6, 2020, entitled "HSP90-TARGETING CONJUGATES AND FORMULATIONS THEREOF", and U.S. Provisional Patent Application No. 62 / 828,645, filed April 3, 2019, entitled "HSP90-TARGETING CONJUGATES AND FORMULATIONS THEREOF", the contents of which are incorporated herein by reference in their entirety. Invention Field

[0003] This invention relates to the use of molecules targeting heat shock proteins (including heat shock protein 90 (HSP90)) for example, in the treatment of cancer. Background Technology

[0004] 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 the development of anticancer drugs. Summary of the Invention

[0005] This invention provides a conjugate comprising an active substance coupled to an HSP90 targeting moiety via a linker, and a pharmaceutical composition comprising such a conjugate. Methods for preparing and using such conjugates are also provided.

[0006] Brief description of the attached figures

[0007] Figure 1 Biodistribution data of radioanalytes of CMP51 (containing Lu177) in NCI-H460 tumors are shown. CMP51 demonstrates selective tumor exposure excluding the kidneys.

[0008] Figure 2 Biodistribution data for non-radioactive analogs of CMP6 (containing Lu175) in NCI-H460 tumors are shown. CMP6 demonstrates selective tumor retention relative to the kidneys and liver. Invention Details

[0010] The applicant has designed HSP90-targeting conjugates containing active substances. This targeting can, for example, improve the amount of active substance at the site and reduce the toxicity of the active substance to the subject. The HSP90-targeting conjugates of the present invention exhibit deep and rapid tumor penetration. The high accumulation and long retention time of the HSP90-targeting conjugates enable the use of cytotoxic and non-cytotoxic payloads, such as radionuclides, chemotherapeutic agents, kinase inhibitors, or immuno-oncology modulators.

[0011] As used herein, “toxicity” refers to the ability of a substance or composition to be harmful or toxic to cells, tissues, organisms, or the cellular environment. Low toxicity means a reduced ability of a substance or composition to be harmful or toxic to cells, tissues, organisms, or the cellular environment. Such reduced toxicity or low toxicity can be relative to a standard measurement, relative to a treatment, or relative to the absence of a treatment.

[0012] Toxicity can be further measured relative to the subject's weight loss, where a weight loss exceeding 15%, 20%, or 30% of body weight indicates toxicity. Other toxicity measures, such as patient performance measures, including drowsiness and malaise, can also be measured. Neutropenia or thrombocytopenia can also be measures of toxicity.

[0013] Pharmacological toxicity indicators include elevated AST / ALT levels, neurotoxicity, kidney damage, and GI damage.

[0014] Furthermore, it is predicted that conjugates containing an HSP90-targeting moiety linked to the active substance will exhibit reduced toxicity to cells that do not express HSP90 compared to the active substance alone. Without being bound by any particular theory, the applicant believes this characteristic is due to the reduced ability of the conjugated active substance to remain in normal cells relative to tumor cells.

[0015] In some implementations, when the active substance and the targeting moiety are linked together to form a conjugate, the active substance and the targeting moiety have a synergistic effect. The efficacy of the conjugate is superior to that of the active substance and / or the targeting moiety alone.

[0016] In some implementations, the potency of the active substance is reduced when it is attached to the target site via a cleavable linker. Upon cleavage of the linker at the target site, such as a tumor site, the active substance is released and its full potency is restored.

[0017] One object of the present invention is to provide compounds, compositions and formulations for improved spatiotemporal drug delivery.

[0018] Another object of the present invention is to provide methods for preparing improved compounds, compositions and formulations for spatiotemporal drug delivery.

[0019] Another objective of this invention is to provide a method for administering the improved compound, composition, and formulation to individuals in need.

[0020] I. Conjugates

[0021] Conjugates include active substances or prodrugs thereof that are linked to a target moiety (e.g., a molecule capable of binding to HSP90) via a linker. Conjugates can be conjugates between a single active substance and a single target moiety, such as conjugates having the structure XYZ, where X is the target moiety, Y is the linker, and Z is the active substance.

[0022] In some embodiments, the conjugate contains more than one targeting moiety, more than one linker, more than one active substance, or any combination thereof. The conjugate may have any number of targeting moieties, linkers, and active substances. The conjugate may have the structure XYZYX, (XY) n -Z, X-(YZ) n X n -YZ、XYZ n (XYZ) n (XYZY) n -Z, where X is the target moiety, Y is the linker, Z is the active substance, and n is an integer between 1 and 50, 2 and 20, for example, 1 and 5. X, Y, and Z may be the same or different each time they appear; for example, the conjugate may contain more than one type of target moiety, more than one type of linker, and / or more than one type of active substance.

[0023] Conjugates may contain more than one targeting moiety linked to a single active substance. For example, a conjugate may include an active substance and multiple targeting moieties, each linked via different linkers. A conjugate may have a structure XYZYX, where each X is a target moiety that may be the same or different, each Y is a linker that may be the same or different, and Z is the active substance.

[0024] Conjugates may contain more than one active substance linked to a single target moiety. For example, a conjugate may include a target moiety and multiple active substances, each linked via different linkers. A conjugate may have a structure ZYXYZ, where X is the target moiety, each Y is a linker that may be the same or different, and each Z is an active substance that may be the same or different.

[0025] A. Active substances

[0026] The conjugates described herein contain at least one active substance (the first active substance). A conjugate may contain more than one active substance, which may be the same as or different from the first active substance. The active substance may be a therapeutic agent, a preventative agent, a diagnostic agent, or a nutrient. Many active substances are known in the art, and they, or their analogues and derivatives, may be used in the conjugates described herein. The active substance 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 substance is an antigen, an adjuvant, a radioactive agent, an imaging agent (e.g., a fluorescent portion), or a polynucleotide. In some embodiments, the active substance is an organometallic compound or a radioactive element. The active substance has chemical functional groups for covalently linking to a linker, or is modified to be an analogue or derivative for covalent linking to a linker.

[0027] In some embodiments, the active substance of the conjugate constitutes 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%, such that the sum of the molar weight percentages of the components of the conjugate is 100%. The amount of the active substance of the conjugate may also be expressed as a ratio relative to the targeting ligand. For example, the ratio of active substance to ligand provided in this teaching is 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.

[0028] radioactive materials

[0029] In some embodiments, the active substance Z is a radioactive material or chemical moiety, such as a metal chelate group, bound to a radionuclide (such as a radioisotope). Many radionuclides possess emission properties, including α, β, γ, and Auger emission, and can be used for therapeutic and / or diagnostic purposes. For example, active substance Z may contain 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, 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.

[0030] In some embodiments, the active material comprises an imaging probe, such as a radiolabel (e.g., a radioactive isotope). Non-limiting examples of radioactive isotopes used 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, and Sc-47. 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.

[0031] In some embodiments, the active substance Z includes a radioactive substance, a chelating agent, or a radioactive substance linked to a chelating agent. A conjugate containing a radioactive substance (e.g., a radioactive isotope) linked to a chelating agent is a radioactive analogue having only a chelating agent or a conjugate having a chelating agent linked to a non-radioactive isotope.

[0032] Chelating agents can be metal chelating agents that bind to metals (including metal nuclides). Chelating agents can also be the portion linked to a nonmetallic active substance. Chelating agents 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 the cyclic anhydride of DTPA (CA-DTPA); 1,4,7-triazacyclononane-1,4-7-triacetic acid (NOTA); and NOTA derivatives (e.g., BCNOTA, p-NCS-Bz-NOTA, BCN...). OT); 6-Hydroxynicotinamide (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)-NN′-Bis[(2-mercapto-2,2-dimethyl)ethyl]-1,2-ethylenediamine-N,N′-diacetic acid (B6SS); Deferroamine (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-tetraazacyclododecane (cyclen); 1,4,7-triazacyclononane-1-succinic-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″ - Tri(methylenephosphonic acid) (NOTP); 1,4,8,11-tetraazacyclotetradecane-N,N′,N″,N″′-tetraacetic acid (TETA); 1,4,7,10,13-pentazacyclopentadecane-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-tetra(carbamoylmethyl)-1,4,7,10-tetraazacyclododecane (TCMC); and their derivatives or analogs.

[0033] In some embodiments, the chelating agent is a polyaminocarboxylic acid, such as ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), 1,4,7,10-tetraazacyclododecane-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.

[0034]

[0035] In some embodiments, the chelating agent is a macrocyclic substance: 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-pentaazacyclopentadecanane-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.

[0036] Non-limiting examples of DTPA and its derivatives are:

[0037]

[0038]

[0039] Non-limiting examples of DOTA and its derivatives are:

[0040]

[0041] In some embodiments, the conjugates disclosed herein contain DOTA, DOTAGA, or any derivative / analyte of the present invention as a chelating agent. Any chelating agent disclosed in Eisenwiener et al., Bioorg Med Chem Lett., vol. 10(18):2133(2000) (the contents of which are incorporated herein by reference in their entirety) may be used as a chelating agent, 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).

[0042]

[0043] DOTASA n=1

[0044] DOTAGA n=2

[0045] Other non-limiting examples of chelating agents are:

[0046]

[0047]

[0048] B. Connector

[0049] The conjugate comprises one or more linkers connecting the active substance and the targeting moiety. Linker Y is bound to one or more active substances and one or more targeting ligands to form the conjugate. Linker Y is connected to the targeting moiety X and the active substance Z via functional groups independently selected from ester bonds, disulfides, amides, hydrazones, ethers, carbamates, carbonates, sulfonamides, alkyl groups, aryl groups, heteroaryl groups, thioethers, and ureas. Alternatively, the linker may be connected to the targeting ligand or active drug substance via groups provided by conjugation between a thiol and 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: alkyl, cycloalkyl, heterocyclic, aryl, and heteroaryl, wherein each of the alkyl, alkenyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups is optionally substituted by one or more groups, each group being independently selected from: halogen, cyano, nitro, hydroxyl, carboxyl, carbamoyl, ether, alkoxy, aryloxy, amino, amide, carbamate, alkyl, alkenyl, alkynyl, aryl, arylalkyl, cycloalkyl, heteroaryl, and heterocyclic groups, wherein each of the carboxyl, carbamoyl, ether, alkoxy, aryloxy, amino, amide, carbamate, alkyl, alkenyl, alkynyl, arylalkyl, cycloalkyl, heteroaryl, or heterocyclic groups is optionally substituted by one or more groups, each group being independently selected from: halogen, cyano, nitro, hydroxyl, carboxyl, carbamoyl, ether, alkoxy, aryloxy, amino, amide, carbamate, alkyl, alkenyl, alkynyl, aryl, arylalkyl, cycloalkyl, heteroaryl, and heterocyclic groups.

[0050] In some embodiments, the linker comprises a cleavable functional group. The cleavable functional group can be hydrolyzed in vivo or can be programmed for enzymatic hydrolysis, for example, by cathepsin B. As used herein, a “cleavable” linker means any linker that can be physically or chemically cleaved. Examples of physical cleavage include cleavage by light, radioactivity, 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.

[0051] In some embodiments, the alkyl chain of the linker may optionally be interrupted by one or more atoms or groups selected from –O-, -C(=O)-, -NR, -OC(=O)-NR-, -S-, and -SS-. The linker may be a dicarboxylic acid ester derivative of succinic acid, glutaric acid, or diethanolic acid. In some embodiments, the linker Y may be X'-R. 1 -Y'-R 2 -Z', and the conjugate can be a compound according to formula Ia:

[0052]

[0053] Where X is the target portion as defined above; Z is the active substance; X', R 1 、Y'、R 2 And Z' as defined in this article.

[0054] X' is absent or independently selected from carbonyl, amide, urea, amino, ester, aryl, arylcarbonyl, aryloxy, arylamino, one or more natural or non-natural amino acids, thio or succinimide; R 1 and R 2 Y' is absent or contains alkyl, substituted alkyl, aryl, substituted aryl, polyethylene glycol (2-30 units); Y' is absent, or is substituted or unsubstituted 1,2-diaminoethane, polyethylene glycol (2-30 units), or amide; Z' is absent or independently selected from carbonyl, amide, urea, amino, ester, aryl, arylcarbonyl, aryloxy, arylamino, thio, or succinimide. In some embodiments, the linker may link one active substance molecule to two or more ligands, or one ligand to two or more active substance molecules.

[0055] In some implementations, the connector Y can be A m Furthermore, the conjugate can be a compound according to formula Ib:

[0056]

[0057] In this paper, A is defined as m = 0-20.

[0058] In Formula Ia, A is a spacer unit that is absent or independently selected from the following substituents. For each substituent, the dashed line indicates a site substituted by X, Z, or another independently selected A unit, where X, Z, or A can be attached to either side of the substituent:

[0059]

[0060] Where z = 0-40, R is H or an optionally substituted alkyl group, and R' is any side chain present in natural or non-natural amino acids.

[0061] In some embodiments, the conjugate can be a compound according to formula Ic:

[0062]

[0063] Where A is defined above, m = 0-40, n = 0-40, x = 1-5, y = 1-5, and C is the branch element defined in this paper.

[0064] In Formula Ic, C is a branch unit containing 3 to 6 functional groups for covalently linking spacer units, ligands, or active pharmaceutical ingredients. These functional groups are selected from amines, carboxylic acids, thiols, or succinimides, including amino acids such as lysine, 2,3-diaminopropionic acid, 2,4-diaminobutyric acid, glutamic acid, aspartic acid, and cysteine.

[0065] C.HSP90 Target Part

[0066] Targeting ligands (also referred to as targeting moieties) as described herein include any molecule capable of binding to one or more HSP90 proteins. Such targeting ligands can be peptides, antibody mimics, nucleic acids (e.g., aptamers), polypeptides (e.g., antibodies), glycoproteins, small molecules, carbohydrates, or lipids.

[0067] The target moiety X can be any HSP90 binding moiety, such as, but not limited to, natural compounds (e.g., geldanamycin and radicicol) and synthetic compounds such as the geldanamycin analog 17-AAG (i.e., 17-allylaminogeldanamycin), a series of purine skeleton HSP90 inhibitors including PU24FC1 (He H. et al., J. Med. Chem., vol. 49:381 (2006), the contents of which are incorporated herein by reference in their entirety), BIIB021 (Lundgren K. et al., Mol. Cancer Ther., vol. 8(4):921 (2009), the contents of which are incorporated herein by reference in their entirety), 4,5-diarylpyrazole (Cheung KM et al., Bioorg. Med. Chem. Lett., vol. 15:3338 (2005), the contents of which are incorporated herein by reference in their entirety; 3-aryl,4-carboxamide pyrazole (Brough P.A. et al., Bioorg. Med. Chem. Lett., vol. 15:5197 (2005), the contents of which are incorporated herein by reference in their entirety); 4,5-diarylisoazole (Brough PA et al., J. Med. Chem., vol. 51:196 (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.48:4212 (2005), the contents of which are incorporated herein by reference in their entirety, thieno[2,3-d]pyrimidine (WO2005034950 of VERNALIS et al., the contents of which are incorporated herein by reference in their entirety), Giannini et al., EP2655345 (the contents of which are incorporated herein by reference in their entirety), aryl triazole derivatives of formula I, or any other instance of an HSP90 binding ligand or a derivative / analogue thereof.

[0068] In some embodiments, the HSP90 binding moiety can be a heterocyclic derivative containing three heteroatoms. WO2009134110 by MATULIS et al. (the contents of which are incorporated herein by reference in their entirety) discloses 4,5-diarylthiadiazoles, which exhibit good HSP90 binding affinity. Even though they have fairly modest cell growth inhibition, they can be used as the HSP90 binding moiety in the conjugates of this invention. Another class of aza-heterocyclic adducts, namely triazole derivatives or analogues, can be used as the HSP90 binding moiety in the conjugates of this invention. For example, the 1,2,4-triazole skeleton has been extensively documented as having HSP90 inhibitory properties. WO2009139916 by BURLISON et al. (Synta Pharmaceuticals Corp., the contents of which are incorporated herein by reference in their entirety) discloses a tricyclic 1,2,4-triazole derivative that inhibits HSP90 at high micromolar concentrations. Any tricyclic 1,2,4-triazole derivative or its derivatives / analogues disclosed in WO2009139916 may be used as the HSP90 binding moiety in the conjugates of the present invention. Any trisubstituted 1,2,4-triazole derivative or its derivatives / analogues disclosed in WO 2010017479 and WO 2010017545 (Synta Pharmaceuticals Corp.) (the contents of which are incorporated herein by reference in their entirety) may be used as the HSP90 binding moiety in the conjugates of the present invention. In another instance, WO2006055760 (Synta Pharmaceuticals Corp., the contents of which are incorporated herein by reference in their entirety) discloses a triazolone-containing HSP90 inhibitor, ganedespib (formerly known as STA-9090, or its highly soluble phosphate prodrug STA-1474), or its derivatives / analogues may be used as the HSP90 binding moiety in the conjugates of the present invention.

[0069]

[0070] In some implementations, ganetespib or its derivatives / analogs can be used as the targeting moiety. Non-limiting examples of ganetespib derivatives / analogs are shown below.

[0071]

[0072]

[0073]

[0074] In some embodiments, onalespib (AT13387) or its derivatives / analogs can be used as the targeting portion in the conjugates of the present invention. Non-limiting examples of onalespib and onalespib derivatives / analogs are shown below.

[0075]

[0076] In some embodiments, the targeting portion comprises AUY-922 or its analogues / derivatives / fragments. In one embodiment, the targeting portion has the following structure:

[0077]

[0078] Any HSP90 ligand or HSP90 inhibitor or its derivative / analyte disclosed in WO2013158644, WO2015038649, WO2015066053, WO2015116774, WO2015134464, WO2015143004, and WO2015184246 (the contents of which are incorporated herein by reference in their entirety) may be used as the HSP90 binding moiety in the conjugates of this invention, such as:

[0079] Formula I R1 can be alkyl, aryl, halide, formamide, or sulfonamide; R2 can be alkyl, cycloalkyl, aryl, or heteroaryl, wherein when R2 is a 6-membered aryl or heteroaryl, R2 is substituted at the 3- and 4-positions relative to the linking point on the triazole ring, and the linker L is linked through this linking point; and R3 can be SH, OH, -CONHR4, aryl, or heteroaryl, wherein when R3 is a 6-membered aryl or heteroaryl, R3 is substituted at the 3- or 4-position.

[0080] Formula II R1 can be alkyl, aryl, halogenated, formamide, or sulfinamide; and R2 can be optionally substituted alkyl, cycloalkyl, aryl, or heteroaryl. Examples of such compounds include 5-(2,4-dihydroxy-5-isopropylphenyl)-N-(2-morpholinylethyl)-4-(4-(morpholinylmethyl)phenyl)-4H-1,2,4-triazol-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-triazol-3-carboxamide;

[0081] Formula III Where X, Y, and Z can independently be CH, N, O, or S (with appropriate substitutions and satisfying the valence of the corresponding atoms and the aromaticity of the ring); R1 can be alkyl, aryl, halide, formamide, or sulfinamide; R2 can be a substituted alkyl, cycloalkyl, aryl, or heteroaryl, wherein the linker L is directly attached to these rings or to extended substitutions attached to these rings; R3 can be SH, OH, NR4R5, and -CONHR6, to which the effector moiety can be attached; R4 and R5 can independently be H, alkyl, aryl, or heteroaryl; and R6 can be alkyl, aryl, or heteroaryl, having at least one functional group that can attach the effector moiety; or

[0082] Formula IV R1 can be alkyl, aryl, halogenated, formamide, or sulfinamide; R2 and R3 are independently C1-C5 hydrocarbon groups optionally substituted with one or more hydroxyl, halogen, C1-C2 alkoxy, amino, mono- or di-C1-C2 alkylamino groups; 5 to 12-membered aryl or heteroaryl; or R2 and R3 together with the nitrogen atom to which they are attached form a 4 to 8-membered monocyclic heterocyclic group, wherein up to 5 ring atoms are selected from O, N, and S. Examples of such compounds include AT-13387.

[0083] The target portion of HSP90 can be Ganetespib, Luminespib (AUY-922, NVP-AUY922), Debio-0932, MPC-3100, onaspib (AT-13387), SNX-2112, 17-amino-galdmycin hydroquinone, PU-H71, or their derivatives / analyses.

[0084]

[0085]

[0086] The HSP90 targeting portion may be SNX5422 (PF-04929113), or any other HSP90 inhibitor disclosed in the following literature: US 8080556 (Pfizer), WO2008096218 (Pfizer), WO2006117669 (Pfizer), WO2008059368 (Pfizer), WO2008053319 (Pfizer), WO2006117669 (Pfizer), EP1885701 (Novartis), EP1776110 (Novartis), EP2572709 (Novartis), WO2012131413 (Debiopharm), or WO2012131468 (Debiopharm), the contents of which are incorporated herein by reference in their entirety.

[0087]

[0088] The HSP90 targeting component can also be PU-H71 (an HSP90 inhibitor that is...). 124 I. Radiolabeled (for PET imaging) or its derivatives / analogs.

[0089] Conjugates containing SNX-2112, 17-amino-gerdemycin hydroquinone, PU-H71, or AT13387 can have the following structures:

[0090]

[0091] In some implementations, the targeting portion includes an imaging probe, such as a radiolabel (e.g., a radioactive isotope). Non-limiting examples of radioactive isotopes 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, Sc-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.

[0092] In some embodiments, the conjugates of the present invention comprise more than one targeting portion. For example, the conjugates may comprise 2, 3, 4, or 5 HSP90 targeting portions.

[0093] Extracellular HSP90 (eHSP90)

[0094] In normal cells, HSP90 secretion occurs when cells are exposed to environmental stresses such as heat, drugs, cytokines, UV, and / or gamma rays. The primary function of extracellular HSP90 (eHSP90) is to aid tissue repair by promoting cell migration from the periphery of damaged tissue to the damaged area. However, in tumors, constitutively activated oncogenes trigger HSP90 secretion even without any environmental stress. Tumor-secreted Hsp90 eHSP90α promotes the migration of tumor and tumor stromal cells during invasion and metastasis. The extracellular promoting function of HSP90α depends on the 115-amino acid fragment (F-5) on the surface of HSP90 (Li et al., Int Rev Cell Mol Biol., vol. 303:203-235 (2013), the contents of which are incorporated herein by reference in their entirety). It has been shown that eHSP90 exists on the surface of tumor cells and can also be internalized (Crowe et al., ACSChem.Biol., vol. 12: 1047-1055 (2017)). Therefore, the surface expression of eHSP90 in tumor cells indicates the ability to selectively target therapies to tumor cells rather than healthy cells. Thus, eHSP90 (especially eHSP90α) may be a good target for tumor treatment.

[0095] In some implementations, the targeting portion selectively binds to the eHSP90. In some implementations, the targeting portion binds to the F-5 region of the eHSP90.

[0096] In some embodiments, the targeting portion has low cell permeability and preferably binds to eHSP90 on the cell surface. In some embodiments, the targeting portion is cell-impermeable and binds only to eHSP90. In some embodiments, the conjugate containing the targeting portion has low cell permeability or is cell-impermeable.

[0097] In some embodiments, the targeting portion comprises HS-23, HS-131 (disclosed in Crowe et al., ACSChem.Biol., vol. 12: 1047-1055 (2017), the contents of which are incorporated herein by reference in their entirety) or DMAG-N-oxide (which is cell-impermeable to 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 analogues / derivatives thereof, the structures of which are shown below.

[0098]

[0099]

[0100] In some embodiments, one or more targeting portions of the conjugate are present in a predetermined 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 is 100%. The amount of the target portion of the conjugate can also be expressed as a ratio to (one or more) active substances, for example, as a ligand-to-active-substance 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.

[0101] D. Pharmacokinetic Regulation Unit

[0102] The conjugates of the present invention may further comprise at least one external linker attached to a reactive group that reacts with a functional group on a protein or engineered protein or its derivative / analysis / mimic, or comprise at least one external linker attached to a pharmacokinetic regulating unit (PMU). The external linker connecting the conjugate and the reactive group or PMU may be a cleavable linker that allows for the release of the conjugate. Therefore, the conjugate can be separated from the protein or PMU as needed.

[0103] Any reactive group or PMU (such as a polymer-containing PMU) disclosed in WO2017 / 197241 may be linked to the conjugates of the present invention, the contents of which are incorporated herein by reference in their entirety.

[0104] In some embodiments, the conjugate comprises a protein-binding reactive group linked to its active substance. In some embodiments, the conjugate comprises a protein-binding reactive group linked to its target moiety. In some embodiments, the conjugate comprises a protein-binding reactive group linked to its linker. The reactive group binds to the protein reversibly or irreversibly. The protein may be a naturally occurring protein, such as a serum or plasma protein, or a fragment thereof. Specific examples include Fc neonatal receptor (FcRn), thyroxine-binding protein, transthyroxine, α1-acid glycoprotein (AAG), transferrin, fibrinogen, albumin, immunoglobulin, α-2-macroglobulin, lipoprotein, or fragments thereof. The reactive group may bind to such a protein via covalent or non-covalent interactions, such as hydrogen bonds, ionic bonds, van der Waals interactions, and hydrophobic interactions.

[0105] In some implementations, the protein-binding reactive group can bind to serum proteins through non-covalent interactions. For example, the reactive group can be one with weak affinity (10⁻⁶). -4 Up to 10 -5 M) saturated fatty acids bound to albumin. Non-limiting examples of such fatty acids may 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 naphthylsulfonylamino groups, diphenylcyclohexanol phosphate groups, 6-(4-(4-iodophenyl)butyrylamino)hexanoate groups ('Albu'-tag), and a series of peptides including SA21 with the core sequence DICLPRWGCLW disclosed by Dennis et al. in J. Biol. Chem., vol. 277:35035 (2002) (the contents of which are incorporated herein by reference in their entirety).

[0106] Protein-binding reactive groups may include the following structures:

[0107] (Myristoyl),

[0108] (palmitoyl)

[0109] (naphthaleneylsulfonylamino group)

[0110]

[0111] (Diphenylcyclohexyl phosphate group),

[0112]

[0113] (4-(4-iodophenyl)butyrylamino group), or

[0114]

[0115] (6-(4-(4-iodophenyl)butyrylamino)hexanoate group).

[0116] In some embodiments, the protein-binding reactive group may comprise any peptide-fatty acid albumin-binding ligand disclosed in Zorzi et al., Nature Communications, vol. 8:16092, (2017) (the contents of which are incorporated herein by reference in their entirety). These peptide-fatty acid albumin-binding ligands comprise a fatty acid linked to a short peptide (e.g., a heptapeptide) via an amino acid side chain. The fatty acid may be linked to the short peptide via a carboxyl group on its lysine side chain. The fatty acid binds to albumin with an affinity in the micromolar range, while the short peptide enhances the affinity by forming additional contact with albumin. The peptide-fatty acid ligand may have the following general formula:

[0117]

[0118] Where X = any amino acid (such as Gly or Ser), K = Lys, and n = 12 (myristic acid), 14 (palmitic acid), or 16 (stearic acid).

[0119] In some embodiments, any albumin-binding functional group disclosed in US 9670482 (Bicycle Therapeutics) (the contents of which are incorporated herein by reference in their entirety) may be used as the protein-binding reactive group in this application. In some embodiments, the protein-binding reactive group comprises a fluorene ring and binds albumin non-covalently and / or reversibly. As a non-limiting example, the protein-binding reactive group comprises a fluorene methoxycarbonyl (FMOC). Optionally, the protein-binding reactive group comprises at least one amino acid, such as Lys, Trp, Gly, or Phe, linked to FMOC. For example, small molecules may comprise Fmoc-Lys-, Fmoc-Gly-, Fmoc-Phe-, Fmoc-GGSGD-, Fmoc-FGGGD-, Fmoc-FGSGD-, Fmoc-WGSGD-, Fmoc-WGGGA, or Fmoc-Trp-GGG.

[0120]

[0121] Non-limiting examples of conjugates

[0122] In some embodiments, the conjugate comprises at least one HSP90 targeting moiety connected via a linker to a chelating agent for the radioactive material. The HSP90 targeting moiety may be a ganesteib analog or derivative (e.g., TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, or TM14), an onaspiracil analog or derivative (e.g., TM6 or TM7), or TM15. The radioactive material may contain any radioisotope, such as lutetium-177 (Lu177 or...).177 Lu). Lutium isotope 177 in the conjugate ( 177 Lu) imparts 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, about 1000 Da to about 3000 Da, or about 1500 Da to 2500 Da.

[0123] In some implementations, the conjugate has the structure of formula X: Wherein TM refers to at least one target moiety binding 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 target moiety. In some embodiments, the conjugate has two target moieties. In some embodiments, the conjugate has three target moieties. In some embodiments, the conjugate has four target moieties.

[0124] In some embodiments, the conjugate has the structure of formula X10:

[0125] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7 or TM15.

[0126] In some embodiments, the conjugate has the structure of formula X1:

[0127] 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 covered by Formula X1 include CMP24, CMP26, and CMP27.

[0128] In some embodiments, the conjugate has the structure of formula X20:

[0129] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7 or TM15.

[0130] In some implementations, the conjugate has the structure of formula X2:

[0131] 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 covered by Formula X2 include CMP22, CMP8, CMP9, CMP10, CMP11, CMP12, CMP17, CMP18, CMP19, CMP20, CMP21, CMP25, CMP28, T25, T26, T30, T31, T40, and T43.

[0132] Non-limiting examples of conjugates of this disclosure covered by 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:

[0133] Table 1. Non-limiting examples of conjugates

[0134]

[0135]

[0136]

[0137]

[0138]

[0139]

[0140]

[0141]

[0142]

[0143]

[0144]

[0145]

[0146]

[0147]

[0148] The structures of conjugate analogs that do not contain radioactive materials are included in Table 1'. Conjugates may 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.

[0149] Table 1'. Non-limiting examples of conjugates (excluding radioactive materials)

[0150]

[0151]

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159]

[0160]

[0161]

[0162]

[0163] In some embodiments, the conjugate contains at least one TM1 as a ligand. In some embodiments, the conjugate contains at least one amino acid spacer group and contains one TM1 as a ligand. In some embodiments, the conjugate contains at least one amino acid spacer group and contains two TM1 as ligands. Non-limiting examples of conjugates include CMP11 and T18.

[0164] In some embodiments, the conjugate contains at least one TM2 as a ligand. In some embodiments, the conjugate contains at least one amino acid spacer group and contains one TM2 as a ligand. In some embodiments, the conjugate contains at least one amino acid spacer group and contains two TM2 as ligands. Non-limiting examples of conjugates include CMP12.

[0165] In some embodiments, the conjugate contains at least one TM3 as a ligand. In some embodiments, the conjugate contains at least one amino acid spacer group and contains one TM3 as a ligand. In some embodiments, the conjugate contains at least one amino acid spacer group and contains two TM3 as ligands. Non-limiting examples of conjugates include CMP10.

[0166] In some embodiments, the conjugate contains at least one TM5 as a ligand. In some embodiments, the conjugate contains at least one amino acid spacer group and contains one TM5 as a ligand, such as CMP22. In some embodiments, the conjugate contains at least one amino acid spacer group and contains 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 contains at least one amino acid spacer group and contains three TM5 as ligands. Non-limiting examples of conjugates include T40. In some embodiments, the conjugate contains at least one amino acid spacer group and contains four TM5 as ligands. Non-limiting examples of conjugates include CMP9.

[0167] In some embodiments, the conjugate contains at least one TM9 as a ligand. In some embodiments, the conjugate contains at least one amino acid spacer group and contains one TM9 as a ligand. In some embodiments, the conjugate contains at least one amino acid spacer group and contains two TM9 as ligands. Non-limiting examples of conjugates include CMP24, CMP25, CMP27, and CMP28.

[0168] In some embodiments, the conjugate contains at least one TM10 as a ligand. In some embodiments, the conjugate contains at least one amino acid spacer group and contains one TM10 as a ligand. In some embodiments, the conjugate contains at least one amino acid spacer group and contains two TM10 as ligands. Non-limiting examples of conjugates include CMP7 and T21.

[0169] In some embodiments, the conjugate contains at least one TM14 as a ligand. In some embodiments, the conjugate contains at least one amino acid spacer group and contains one TM14 as a ligand. In some embodiments, the conjugate contains at least one amino acid spacer group and contains two TM14 as ligands. Non-limiting examples of conjugates include T19.

[0170] amino acid spacer

[0171] In some embodiments, the conjugate comprises at least one HSP90 targeting moiety, which is linked to a chelating agent for a radioactive material via a linker. The HSP90 targeting moiety may be a ganesteib analog or derivative (e.g., TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, or TM14), an onaspiracil analog or derivative (e.g., TM6 or TM7), or TM15. The linker may comprise a spacer group made of at least one amino acid or its analogue, such as two amino acids or their analogues, three amino acids or their analogues, four amino acids or their analogues, or five amino acids or their analogues. The amino acid or its analogue may be a D-amino acid. The amino acid or its analogue may be anionic (e.g., DGlu), cationic (e.g., DLys), or uncharged (e.g., Sar, where Sar = N-methylglycine). The spacer group 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 being bound by any theory, the spacer group affects the biodistribution of the conjugate and can reduce hepatic uptake. HSP90 binding affinity is maintained regardless of the charge on the spacer group.

[0172] In some implementations, the conjugate has the structure of formula A:

[0173]

[0174] Wherein TM refers to at least one target moiety binding to HSP90. In some embodiments, the conjugate has two target moieties. In some embodiments, the conjugate has three target moieties. In some embodiments, the conjugate has four target moieties. Non-limiting examples of conjugates covered 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.

[0175] In some embodiments, the conjugate has the structure of formula A10:

[0176]

[0177] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7 or TM15.

[0178] In some embodiments, the conjugate has the structure of formula A1:

[0179]

[0180] 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 covered by Formula A1 include CMP23, CMP37, and CMP38.

[0181] In some embodiments, the conjugate has the structure of formula A20:

[0182]

[0183] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7 or TM15.

[0184] In some implementations, the conjugate has the structure of formula A2:

[0185]

[0186] 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 covered 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.

[0187] Non-limiting examples of the conjugates of this disclosure covered 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:

[0188] Table 2. Non-limiting examples of conjugates containing amino acid spacer groups

[0189]

[0190]

[0191]

[0192]

[0193]

[0194]

[0195]

[0196]

[0197]

[0198]

[0199]

[0200]

[0201]

[0202]

[0203]

[0204] Structures of conjugate analogs that do not contain radioactive materials are included in Table 2'. Conjugates may 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.

[0205] Table 2'. Non-limiting examples of conjugates containing amino acid spacer groups (excluding radioactive materials)

[0206]

[0207]

[0208]

[0209]

[0210]

[0211]

[0212]

[0213]

[0214]

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221] In some embodiments, the conjugate comprises at least one amino acid spacer group and at least one TM1 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer group and one TM1 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer group and two TM1 as ligands. Non-limiting examples include T10, T27, and T28.

[0222] In some embodiments, the conjugate comprises at least one amino acid spacer group and at least one TM2 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer group and one TM2 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer group and two TM2 as ligands. Non-limiting examples include CMP13 and CMP16.

[0223] In some embodiments, the conjugate comprises at least one amino acid spacer group and at least one TM5 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer group and one TM5 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer group 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.

[0224] In some embodiments, the conjugate comprises at least one amino acid spacer group and at least one TM10 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer group and one TM10 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer group and two TM10 as ligands. Non-limiting examples include T6, T29, CMP37, and CMP38.

[0225] In some embodiments, the conjugate comprises at least one amino acid spacer group and at least one TM15 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer group and one TM15 as a ligand. In some embodiments, the conjugate comprises at least one amino acid spacer group and two TM10 as ligands. Non-limiting examples include CMP23.

[0226] PEG spacer group

[0227] In some embodiments, the conjugate comprises an HSP90 targeting moiety of a chelating agent linked to a radioactive material via a linker. The HSP90 targeting moiety may be a ganesteib analog or derivative (e.g., TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, or TM14), an onaspirab analog or derivative (e.g., TM6 or TM7), or TM15. The linker may contain a spacer group comprising polyethylene glycol (PEG). The PEG spacer group may be made of (PEG). n The configuration is constructed, 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. HSP90 binding affinity is maintained regardless of the charge present on the spacer.

[0228] In some implementations, the conjugate has the structure of formula B:

[0229]

[0230] TM refers to at least one target moiety that binds to HSP90. In some embodiments, the conjugate has two target moieties. In some embodiments, the conjugate has three target moieties. In some embodiments, the conjugate has four target moieties.

[0231] In some embodiments, the conjugate has the structure of formula B10:

[0232]

[0233] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM6, TM7 or TM14.

[0234] In some embodiments, the conjugate has the structure of formula B1:

[0235]

[0236] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM6, TM7, or TM14. Non-limiting examples of conjugates covered by Formula B1 include CMP1, CMP29, CMP30, CMP31, CMP32, CMP33, CMP34, CMP35, CMP36, CMP39, CMP40, CMP41, and CMP42.

[0237] In some embodiments, the conjugate has the structure of formula B20:

[0238]

[0239] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM6, TM7 or TM14.

[0240] In some implementations, the conjugate has the structure of formula B2:

[0241]

[0242] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM6, TM7, or TM14. Non-limiting examples of conjugates covered by Formula B2 include CMP2.

[0243] Non-limiting examples of conjugates covered by Formula B include CMP1, CMP2, CMP29, CMP30, CMP31, CMP32, CMP33, CMP34, CMP35, CMP36, CMP39, CMP40, CMP41, CMP42, and CMP52.

[0244] Table 3. Non-limiting examples of conjugates containing one TM and a PEG spacer group

[0245]

[0246]

[0247]

[0248]

[0249]

[0250] The structures of conjugate analogs that do not contain radioactive materials are included in Table 3'. Conjugates may 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.

[0251] Table 3'. Non-limiting examples of conjugates (without radioactive materials) containing one TM and PEG spacer group.

[0252]

[0253]

[0254]

[0255]

[0256]

[0257] Table 4. Non-limiting examples of conjugates containing PEG spacer groups and more than one TM.

[0258]

[0259] The structures of conjugate analogs that do not contain radioactive materials are included in Table 4'. Conjugates may 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.

[0260] Table 4'. Non-limiting examples of conjugates (free of radioactive materials) containing PEG spacer groups and more than one TM.

[0261]

[0262] In some embodiments, the conjugate comprises at least one (PEG)4 spacer group. Non-limiting examples include CMP1, CMP2, CMP29, CMP30, CMP31, CMP39, CMP40, and CMP35.

[0263] In some embodiments, the conjugate comprises at least one (PEG)12 spacer group. Non-limiting examples include CMP32, CMP33, CMP34, CMP36, CPM41, and CMP42.

[0264] Albumin-binding pharmacokinetic regulatory unit

[0265] In some embodiments, the conjugate comprises an HSP90 targeting moiety linked via a linker to a chelating agent for radioactive materials 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 ganesteib analog or derivative (e.g., TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, or TM14), an onaspide analog or derivative (e.g., TM6 or TM7), or TM15. In some embodiments, the linker may comprise a spacer group comprising polyethylene glycol (PEG), such as (PEG)4 or (PEG)12. In some embodiments, the linker may comprise at least one amino acid. HSP90 binding affinity is maintained regardless of the charge present on the spacer group and / or the PMU.

[0266] In some implementations, the PMU contains functional groups that bind to albumin, such as (4-(4-iodophenyl)butyrylamino).

[0267] In some implementations, the conjugate has the structure of formula C:

[0268]

[0269] Wherein TM refers to at least one target moiety binding to HSP90. In some embodiments, the conjugate has two target moieties. In some embodiments, the conjugate has three target moieties. In some embodiments, the conjugate has four target moieties. In some embodiments, the linker comprises a PEG spacer group. In some embodiments, the linker comprises an amino acid spacer group.

[0270] In some embodiments, the conjugate has a structure of formula C10:

[0271]

[0272] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7 or TM15.

[0273] In some embodiments, the conjugate has the structure of formula C1:

[0274]

[0275] 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 covered by Formula C1 include CMP3, CMP4, CMP5, and CMP6.

[0276] In some implementations, the conjugate has the structure of formula C20:

[0277]

[0278] TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, TM14, TM6, TM7 or TM15.

[0279] In some implementations, the conjugate has the structure of formula C2:

[0280]

[0281] 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 covered 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.

[0282] Non-limiting examples of the conjugates of this disclosure covered 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:

[0283] Table 5. Non-limiting examples of conjugates containing albumin-binding PMU

[0284]

[0285]

[0286]

[0287]

[0288]

[0289]

[0290]

[0291]

[0292]

[0293]

[0294]

[0295]

[0296]

[0297]

[0298]

[0299]

[0300]

[0301]

[0302]

[0303]

[0304]

[0305]

[0306] The structures of conjugate analogs that do not contain radioactive materials are included in Table 5'. Conjugates may 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.

[0307] Table 5'. Non-limiting examples of conjugates containing albumin-bound PMU (free of radioactive materials)

[0308]

[0309]

[0310]

[0311]

[0312]

[0313]

[0314]

[0315]

[0316]

[0317]

[0318]

[0319]

[0320]

[0321]

[0322]

[0323]

[0324]

[0325]

[0326]

[0327]

[0328]

[0329]

[0330] In some embodiments, the conjugate contains at least one albumin-binding PMU and at least one TM1 as a ligand. In some embodiments, the conjugate contains at least one albumin-binding PMU and one TM1 as a ligand. In some embodiments, the conjugate contains at least one albumin-binding PMU and two TM1s as ligands, such as T17.

[0331] In some embodiments, the conjugate contains at least one albumin-binding PMU and at least one TM2 as a ligand. In some embodiments, the conjugate contains at least one albumin-binding PMU and one TM2 as a ligand. In some embodiments, the conjugate contains at least one albumin-binding PMU and two TM2s as ligands, such as T13.

[0332] In some embodiments, the conjugate contains at least one albumin-binding PMU and at least one TM5 as a ligand. In some embodiments, the conjugate contains at least one albumin-binding PMU and one TM5 as a ligand, such as T7. In some embodiments, the conjugate contains at least one albumin-binding PMU and two TM5s as ligands. Non-limiting examples include T3, T14, T15, T16, T22, T23, T32, T24, T33, T34, T35, T36, T37, T38, T41, and T42.

[0333] In some embodiments, the conjugate contains at least one albumin-binding PMU and at least one TM9 ligand. In some embodiments, the conjugate contains at least one albumin-binding PMU and one TM9 ligand, such as CMP3 and CMP4.

[0334] In some embodiments, the conjugate contains at least one albumin-binding PMU and at least one TM10 as a ligand. In some embodiments, the conjugate contains at least one albumin-binding PMU and one TM10 as a ligand, such as CMP5, CMP6, and T11. In some embodiments, the conjugate contains at least one amino acid spacer group and two TM10s as ligands, such as T12.

[0335] Conjugates T1, T2, T3, T4, T5, T6, T7, T8, T9, T10, T11, T12, T13, T14, T15, T16, T 17. T18, T19, T20, T21, T22, T23, T24, T25, T26, T27, T28, T29, T30, T31, T 32. T33, T34, T35, T36, T37, T38, T39, T40, T41, T42, T43, CMP1, CMP2, CMP3, CMP4, CMP5, CMP6, CMP7, CMP8, CMP9, CMP10, CMP11, CMP12, CMP13, CMP1 4. The lutetium (Lu) in CMP15, CMP16, CMP17, CMP18, CMP19, CMP20, CMP21, CMP22, CMP23, CMP24, CMP25, CMP26, CMP27, CMP28, CMP29, CMP30, CMP31, CMP32, CMP33, CMP34, CMP35, CMP36, CMP37, CMP38, CMP39, CMP40, CMP41, CMP42, CMP43, CMP44, CMP45, CMP46, CMP47, CMP48, CMP49, CMP50, or CMP52 can be replaced with Lu177 ( 177 Lu) or any other radioactive isotope (e.g., 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) to provide a radioactive analogue for the conjugate.

[0336] II. Preparations

[0337] In some embodiments, the composition is administered to a person, a human patient, or a subject. For the purposes of this disclosure, the phrase "active ingredient" generally refers to the conjugate as described herein.

[0338] Although the description of the pharmaceutical compositions provided herein primarily relates to pharmaceutical compositions suitable for administration to humans, those skilled in the art will understand that such compositions are generally suitable for administration to any other animal, such as non-human animals, including non-human mammals. It is fully understood that modifications to pharmaceutical compositions suitable for human administration are made to make the compositions suitable for administration to a variety of animals, and that such modifications can be designed and / or performed only by routine (if any) experiments by a veterinary pharmacologist of ordinary skill in the art. Subjects intended to administer the pharmaceutical compositions include, but are not limited to, humans and / or other primates; mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, cats, dogs, mice, and / or rats; and / or poultry, including commercially relevant poultry such as poultry, chickens, ducks, geese, and / or turkeys.

[0339] Formulations of the pharmaceutical compositions described herein can be prepared by any method known in or developed in the field of pharmacology. Generally, such preparation methods include the following steps: combining the active ingredient with an excipient and / or one or more other auxiliary ingredients, and then, if necessary and / or desired, dividing, shaping, and / or packaging the product into the desired single-dose or multi-dose units.

[0340] The pharmaceutical compositions of the present invention may be prepared, packaged, and / or sold in batches in single unit dose form and / or in multiple single unit dose forms. As used herein, a “unit dose” is an individual amount of a pharmaceutical composition containing a predetermined amount of active ingredient. The amount of active ingredient is generally equal to the dose of active ingredient to be administered to a subject, and / or an appropriate fraction of that dose, such as, for example, half or one-third of that dose.

[0341] The relative amounts of the active ingredient, pharmaceutically acceptable excipients, and / or any additional ingredients in the pharmaceutical compositions of the present invention will vary depending on the identity, size, and / or condition of the subject being treated, and further depending on the route of administration of the composition. For example, the composition may contain 0.1% to 100%, such as 0.5% to 50%, 1-30%, 5-80%, or at least 80% (w / w) of the active ingredient.

[0342] One or more excipients may be used to formulate the conjugates of the present invention to: (1) increase stability; (2) allow sustained or delayed release (e.g., from a reservoir of monomaleimide); (3) alter biodistribution (e.g., target the monomaleimide compound to a specific tissue or cell type); and (4) alter the release characteristics of the monomaleimide compound in vivo. Non-limiting examples of excipients include any and all solvents, dispersion media, diluents or other liquid solvents, dispersants or suspending agents, surfactants, isotonic agents, thickeners or emulsifiers, and preservatives. The excipients of the present invention may also include, but are not limited to, lipids, liposomes, lipid nanoparticles, polymers, lipid complexes, core-shell nanoparticles, peptides, proteins, hyaluronidase, nanoparticle mimics, and combinations thereof. Therefore, formulations of the present invention may include one or more excipients, each in a certain amount together, to increase the stability of the monomaleimide compound.

[0343] excipient

[0344] Pharmaceutical formulations may additionally contain pharmaceutically acceptable excipients suitable for the desired specific dosage form, as used herein, including any and all solvents, dispersion media, diluents or other liquid solvents, dispersants or suspending agents, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, etc. Remington's *The Science and Practice of Pharmacy*, 21st edition, ARGennaro (Lippincott, Williams & Wilkins, Baltimore, MD, 2006; incorporated herein by reference in its entirety) discloses various excipients for formulating pharmaceutical compositions and known techniques for their preparation. Unless any conventional excipient medium is incompatible with a substance or its derivatives due to any undesirable biological effect or otherwise harmful interaction with any other component of the pharmaceutical composition, its use is contemplated within the scope of this invention.

[0345] In some embodiments, the pharmaceutically acceptable purity of the excipient is at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%. 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 standards of the United States Pharmacopeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia, and / or the International Pharmacopoeia.

[0346] Pharmaceutically acceptable excipients for use in the manufacture of pharmaceutical compositions include, but are not limited to, inert diluents, dispersants and / or granulators, surfactants and / or emulsifiers, disintegrants, binders, preservatives, buffers, lubricants and / or oils. Such excipients may optionally be included in the pharmaceutical composition.

[0347] Examples of 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 / or combinations thereof.

[0348] Examples of granulating agents and / or dispersants include, but are not limited to, potato starch, corn starch, cassava starch, sodium starch glycolate, clay, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose and wood products, natural sponges, cation exchange resins, calcium carbonate, silicates, sodium carbonate, crosslinked poly(vinylpyrrolidone) (crosspovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethyl cellulose, crosslinked sodium carboxymethyl cellulose (crosslinked carboxymethyl cellulose), methyl cellulose, pregelatinized starch (starch 1500), microcrystalline starch, water-insoluble starch, calcium carboxymethyl cellulose, and magnesium aluminum silicate. Sodium lauryl sulfate, quaternary ammonium compounds, and / or combinations thereof.

[0349] Examples of surfactants and / or emulsifiers include, but are not limited to, natural emulsifiers (e.g., gum arabic, agar, alginic acid, sodium alginate, tragacanth gum, chondrux, cholesterol, xanthan gum, pectin, gelatin, egg yolk, casein, lanolin, cholesterol, waxes, and lecithin), colloidal clays (e.g., bentonite [aluminum silicate] and... Magnesium aluminum silicate), long-chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetyl glycerol monostearate, ethylene glycol distearate, glyceryl monostearate, propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxylated polymethyl methacrylate, polyacrylic acid, acrylic polymers, and carboxylated vinyl polymers), carrageenan, cellulose derivatives (e.g., sodium carboxymethyl cellulose, powdered cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose), and dehydrated sorbitol fatty acid esters (e.g., polyoxyethylene dehydrated sorbitol monolaurate). Polyoxyethylene dehydrated sorbitol Polyoxyethylene dehydrated sorbitan monooleate Sorbitol monopalmitate Sorbitol monostearate Sorbitol Tristearate Glyceryl monooleate, sorbitol monooleate ), polyoxyethylene esters (e.g., polyoxyethylene monostearate) Polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate and ), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g.) ), polyoxyethylene ethers (e.g., polyoxyethylene lauryl ether) Poly(vinylpyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl lauryl sulfate, sodium lauryl sulfate F 68、 Hexadecyltrimethylammonium bromide, hexadecylpyridine chloride, benzalkonium chloride, sodium docusate, and / or combinations thereof.

[0350] Examples of adhesives include, but are not limited to, starches (e.g., corn starch and starch paste); gelatin; sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol); natural and synthetic gums (e.g., gum arabic, sodium alginate, extracts of Irish moss, panwar gum, ghatti gum, mucilage of isapor husk, carboxymethyl cellulose, methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, microcrystalline cellulose, cellulose acetate, poly(vinylpyrrolidone), magnesium aluminum silicate). Larch arabinogalactan; alginate; polyethylene oxide; polyethylene glycol; inorganic calcium salts; silicic acid; polymethyl methacrylate; wax; water; alcohol; etc.; and combinations thereof.

[0351] Examples of preservatives may include, but are not limited to, antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, alcohol preservatives, acid preservatives, and / or other preservatives. Examples of antioxidants include, but are not limited to, α-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. Examples of chelating agents include ethylenediaminetetraacetic acid (EDTA), citrate monohydrate, disodium edetate, dipotassium edetate, edetate, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid, and / or trisodium edetate. Examples of antimicrobial preservatives include, but are not limited to, benzalkonium chloride, benzyl alcohol, bromonitrile glycol, hexadecyltrimethylammonium bromide, hexadecylpyridine chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethanol, glycerin, hexetidine, imidureurol, phenol, phenoxyethanol, phenylethanol, phenylmercuric nitrate, propylene glycol, and / or thimerosal. Examples of 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. Examples of alcoholic preservatives include, but are not limited to, ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, parabens, and / or phenylethanol. Examples of 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, tocopherol acetate, deteroxime mesylate, cetyltrimethylammonium bromide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, and glydant. Methylparaben, NEOLONE TM KATHON TM and / or

[0352] Examples of buffers include, but are not limited to, citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium gluconate, calcium gluconate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propionic acid, calcium levulinate, valeric acid, calcium hydrogen phosphate, phosphoric acid, calcium phosphate, calcium hydroxide, potassium acetate, potassium chloride, potassium gluconate, potassium mixture, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, potassium phosphate mixture, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium phosphate mixture, thiamethoxam, magnesium hydroxide, aluminum hydroxide, alginate, pyrogen-free water, isotonic saline, Ringer's solution, ethanol, and / or combinations thereof.

[0353] Examples of lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behenate, hydrogenated vegetable oil, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and combinations thereof.

[0354] Examples of oils include, but are not limited to, almond oil, apricot kernel oil, avocado oil, babassu oil, bergamot oil, blackcurrant seed oil, borage oil, juniper oil, chamomile oil, canola oil, coriander oil, Brazil palm oil, castor oil, cinnamon oil, cocoa butter, coconut oil, cod liver oil, coffee oil, corn oil, cottonseed oil, emu oil, eucalyptus oil, night-blooming jasmine oil, fish oil, flaxseed oil, geraniol, gourd oil, grapeseed oil, hazelnut oil, hyssop oil, isopropyl myristate, jojoba oil, macadamia nut oil, mixed lavender oil, lavender oil, lemon oil, litsea cubeba oil, and macadamia nut oil. Nut oil, mallow oil, mango seed oil, meadowfoam seed oil, mink oil, nutmeg oil, olive oil, orange oil, orange salmon 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, camellia oil, peppermint oil, sea buckthorn oil, sesame oil, shea butter, silicone oil, soybean oil, sunflower oil, tea tree oil, thistle oil, tsubaki oil, vetiver oil, walnut oil, and wheat germ oil. Example oils include, but are not limited to, butyl stearate, caprylic / capric triglyceride, caprylic / capric triglyceride, cyclomethicone, diethyl sebate, dimethyl silicone oil 360, isopropyl myristate, mineral oil, octyl dodecyl alcohol, oleyl alcohol, silicone oil, and / or combinations thereof.

[0355] Excipients such as cocoa butter and suppository wax, colorants, coating agents, sweeteners, flavoring agents and / or aromas may be present in the composition, depending on the formulator's judgment.

[0356] application

[0357] The conjugates of the present invention may be administered via any route that produces a therapeutically effective result. These routes include, but are not limited to, enteric, gastrointestinal, epidural, oral, percutaneous, epidural, intracerebral (to the brain), intraventricular (to the ventricles), epithelial (applied to the skin), intradermal (to the skin itself), subcutaneous (under the skin), nasal (through the nose), intravenous (to the vein), intraarterial (to the artery), intramuscular (to the muscle), intracardiac (to the heart), intraosseous infusion (to the bone marrow), intrathecal (to the spinal canal), intraperitoneal (infusion or injection into the peritoneum), intravesical infusion, intravitreal (through the eye), intracavernosal injection (to the base of the penis), intravaginal, intrauterine, extraamniotic, percutaneous (diffusion through intact skin to achieve systemic distribution), transmucosal (diffusion through mucous membranes), inhaled (nasal), sublingual, sublipal, enema, eye drops (to the conjunctiva), or ear drops. In certain embodiments, the composition may be administered in a manner that allows it to cross the blood-brain barrier, vascular barrier, or other epithelial barrier.

[0358] The formulations described herein contain an effective amount of the conjugate in a drug carrier suitable for administration to an individual in need. The formulations can be administered parenterally (e.g., by injection or infusion). The formulations or variations thereof can be administered in any manner, including enterally, topically (e.g., to the eyes), or through the lungs. In some embodiments, the formulations are administered topically.

[0359] Dosage

[0360] This invention provides a method of administering a conjugate as described herein to a subject in need. The conjugate as described herein can be administered to a subject in any amount and via any route of administration that is effective in preventing or treating or imaging diseases, conditions, and / or illnesses (e.g., diseases, conditions, and / or illnesses associated with working memory deficits). The precise amount required will vary depending on the subject's species, age and general condition, the severity of the disease, the specific composition, its administration method, its mode of action, etc.

[0361] The compositions of the present invention are typically formulated in dosage units to facilitate dosage application and uniformity. However, it should be understood that the total daily dosage of the compositions of the present invention will be determined by the attending physician within a reasonable medical judgment. The specific dosage level for any particular patient that is effective in treatment, prevention, or appropriate imaging will depend on a variety of factors, including the condition being treated and its severity; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health, sex, and diet; the timing, route of administration, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors well known in the medical field.

[0362] In some embodiments, the compositions of the present invention can be delivered once or more daily in doses sufficient to deliver approximately 0.0001 mg / kg to approximately 100 mg / kg, approximately 0.001 mg / kg to approximately 0.05 mg / kg, approximately 0.005 mg / kg to approximately 0.05 mg / kg, approximately 0.001 mg / kg to approximately 0.005 mg / kg, approximately 0.05 mg / kg to approximately 0.5 mg / kg, approximately 0.01 mg / kg to approximately 50 mg / kg, approximately 0.1 mg / kg to approximately 40 mg / kg, approximately 0.5 mg / kg to approximately 30 mg / kg, or approximately 0.01 mg / kg per day. The dose may be administered at dose levels of approximately 10 mg / kg, approximately 0.1 mg / kg to approximately 10 mg / kg, approximately 1 mg / kg to approximately 25 mg / kg, approximately 25 mg / kg to approximately 50 mg / kg, approximately 50 mg / kg to approximately 100 mg / kg, approximately 100 mg / kg to approximately 125 mg / kg, approximately 125 mg / kg to approximately 150 mg / kg, approximately 150 mg / kg to approximately 175 mg / kg, approximately 175 mg / kg to approximately 200 mg / kg, or approximately 200 mg / kg to approximately 250 mg / kg of the subject's body weight to achieve the desired therapeutic, diagnostic, preventative, or imaging effect. The required dose may be delivered three times a day, twice a day, once a day, every other day, every three days, weekly, every two weeks, every three weeks, or every four weeks. In some embodiments, the required dose may be delivered using multiple administrations (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, or more administrations). When multiple administrations are used, fractionated dosing regimens, such as those described herein, can be employed.

[0363] In the pharmaceutical composition, the concentration of the conjugate may be from about 0.01 mg / mL to about 50 mg / mL, from about 0.1 mg / mL to about 25 mg / mL, from about 0.5 mg / mL to about 10 mg / mL, or from about 1 mg / mL to about 5 mg / mL.

[0364] As used herein, “fragmented dosing” means dividing a single unit dose or total daily dose into two or more doses, such as administering a single unit dose in two or more separate doses. As used herein, “single unit dose” is the dose of any therapeutic agent administered in one dose / once / through a single route / at a single point of contact (i.e., a single administration event). As used herein, “total daily dose” is the amount given or prescribed over a 24-hour time period. It can be administered in single unit dose form.

[0365] Dosage form

[0366] The pharmaceutical compositions described herein can be formulated into the dosage forms described herein, such as topical, intranasal, intratracheal, or injectable (e.g., intravenous, intraocular, intravitreal, intramuscular, intracardiac, intraperitoneal, subcutaneous) dosage forms.

[0367] VI. Methods using conjugates

[0368] When appropriate, the conjugates described herein may be used to treat any proliferative disease, metabolic disease, infectious disease, or cancer. The preparations can be administered by injection, oral administration, or topically, typically to mucosal surfaces (lungs, nose, mouth, cheeks, sublingual, vaginal, rectal) or to the eyes (intraocular or ocular).

[0369] In various embodiments, methods for treating a subject suffering from cancer are provided, wherein the method comprises administering a therapeutically effective amount of the conjugate, or its salt form as described herein, to a subject who has cancer, is suspected of having cancer, or is susceptible to cancer. According to the invention, cancer includes any disease or ailment characterized by uncontrolled (e.g., excessive) cell proliferation. Cancer may be characterized by a tumor (e.g., a solid tumor) or any growth.

[0370] In some implementations, the cancer is a solid tumor. Large drug molecules have limited penetration into solid tumors. The penetration rate of large drug molecules is slow. On the other hand, small molecules, such as the conjugates of the present invention, can penetrate solid tumors rapidly and more deeply. Regarding the depth of drug penetration, large molecules penetrate less despite having more durable pharmacokinetics. Small molecules, such as the conjugates of the present invention, penetrate more deeply. Dreher et al. (Dreher et al., JNCI, vol. 98(5):335(2006), the contents of which are incorporated herein by reference in their entirety) investigated the penetration of dextran of different sizes into tumor xenografts.

[0371] In one embodiment, the conjugate of the present invention reaches a solid tumor at a distance of 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 from the vascular surface of the tumor. Zero distance is defined as the vascular surface of the tumor, and each distance greater than zero is defined as the distance to the nearest vascular surface measured in three dimensions.

[0372] In another embodiment, the conjugate of the present invention penetrates into the nucleus of a tumor. As used herein, the “nucleus” of a tumor refers to the central region of the tumor. The distance from any part of the nucleus region of the tumor to the vascular surface of the tumor is about 30% to about 50% of the length or width of the tumor. The distance from any part of the nucleus region of the tumor to the center point of the tumor is less than about 20% of the length or width of the tumor. The nucleus region of the tumor is approximately the central third of the tumor.

[0373] In another embodiment, the conjugate of the present invention penetrates into the center of a solid tumor. As mentioned herein, "center" of a tumor refers to the central region of the tumor. The distance from any portion of the central region of the tumor to the vascular surface of the tumor is about 15% to about 30% of the length or width of the tumor. The distance from any portion of the central region 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 central region of the tumor is generally located between the central third and the outer third of the tumor.

[0374] In some implementations, the subject may have no other indication for treatment with the conjugate. In some implementations, the method includes using cancer cells, including but not limited to mammalian cancer cells. In some cases, the mammalian cancer cells are human cancer cells.

[0375] In some embodiments, the conjugates taught in this invention have been found to inhibit cancer and / or tumor growth. They may also reduce cell proliferation, invasiveness, and / or metastasis, thereby making them suitable for the treatment of cancer.

[0376] In some embodiments, the conjugates taught in this invention can be used to prevent the growth of tumors or cancer, and / or to prevent the metastasis of tumors or cancer. In some embodiments, the compositions taught in this invention can be used to shrink or destroy cancer.

[0377] In some embodiments, the conjugates provided herein are suitable for inhibiting the proliferation of cancer cells. In some embodiments, the conjugates provided herein are suitable for inhibiting cell proliferation, such as inhibiting the rate of cell proliferation, preventing cell proliferation, and / or inducing cell death. Generally, the conjugates described herein can inhibit the cell proliferation of cancer cells, or inhibit the proliferation of cancer cells and / or induce cell death in cancer cells. In some embodiments, treatment with the conjugates of the present invention reduces cell proliferation by at least about 25%, about 50%, about 75%, or about 90% compared to untreated cells. In some embodiments, treatment with the conjugates of the present invention increases the cell cycle arrest marker phosphorylated histone H3 (PH3 or PHH3) by at least about 50%, about 75%, about 100%, about 200%, about 400%, or about 600% compared to untreated cells. In some embodiments, treatment with the conjugate of the present invention increased the apoptosis marker lysine caspase-3 (CC3) by at least 50%, about 75%, about 100%, about 200%, about 400%, or about 600% compared with untreated cells.

[0378] Furthermore, in some embodiments, the conjugates of the present invention effectively inhibit tumor growth in a variety of tumor types, whether measured by net size (weight, surface area, or volume) or by the rate of change over time.

[0379] In some embodiments, treatment with the conjugates of the present invention reduces the size of the tumor by about 60% or more. In some embodiments, the tumor size 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%, measured by weight and / or area and / or volume.

[0380] Cancers treatable by the methods taught in this invention typically occur in mammals. Mammals include, for example, humans, non-human primates, dogs, cats, rats, mice, rabbits, ferrets, guinea pigs, horses, pigs, sheep, goats, and cattle. In various embodiments, cancers include, but are not limited to, acoustic neuroma, acute leukemia, acute lymphoblastic leukemia, acute myeloid leukemia (monocyte, myeloblastic, adenocarcinoma, angiosarcoma, astrocytoma, myelomonocyte, and promyelocytic), acute T-cell leukemia, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, bronchial cancer, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid (granulocytic) leukemia, chronic myeloid leukemia, colon cancer, colorectal cancer, and colorectal cancer. Colorectal cancer, craniopharyngioma, cystadenocarcinoma, diffuse large B-cell lymphoma, Burkitt lymphoma, dysplastic changes (developmental abnormalities and metaplasia), embryonal carcinoma, endometrial cancer, endothelial sarcoma, ependymoma, epithelial carcinoma, erythroleukemia, esophageal cancer, estrogen receptor-positive breast cancer, essential thrombocytosis, Ewing's tumor, fibrosarcoma, follicular lymphoma, germ cell testicular cancer, glioma, heavy chain disease, angioblastoma, liver cancer, hepatocellular carcinoma, hormone-insensitive prostate cancer, leiomyosarcoma, liposarcoma, lung cancer, lymphangiocarcinoma. Lymphagiosarcoma, 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, T-cell or B-cell-derived malignant lymphoma cells, leukemia, lymphoma, medullary carcinoma, medulloblastoma, melanoma, meningioma, mesothelioma, multiple myeloma, myeloid leukemia, myeloma, myxosarcoma, neuroblastoma, Non-small cell lung cancer, oligodendroglioma, oral cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papillary gland carcinoma, papillary carcinoma, pineal tumor, 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, synovial tumor, sweat gland carcinoma, thyroid cancer, Waldenstrom's macroglobulinemia, testicular tumors, 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, urethral cancer, kidney cancer, urethral epithelial cancer, female reproductive tract cancer, uterine cancer, gestational trophoblastic disease, male reproductive tract cancer, seminal vesicle cancer, testicular cancer, germ cell tumors, endocrine gland tumors, thyroid cancer, adrenal cancer, pituitary cancer, hemangioma, bone and soft tissue sarcoma, Kaposi's sarcoma, nerve cancer, eye cancer, and meningeal cancer. Cancer), glioblastoma, neuroma, neuroblastoma, schwannoma, solid tumors caused by hematopoietic malignancies such as leukemia, metastatic melanoma, recurrent or persistent epithelial ovarian cancer, fallopian tube cancer, primary peritoneal cancer, gastrointestinal stromal tumors, colorectal cancer, gastric cancer, melanoma, glioblastoma multiforme, non-squamous non-small cell lung cancer, malignant glioma, epithelial ovarian cancer, primary peritoneal serous carcinoma, metastatic liver cancer, neuroendocrine carcinoma, refractory malignant tumors, triple-negative breast cancer, HER2-amplified breast cancer, nasopharyngeal carcinoma. Cancer, oral cancer, biliary tract cancer, hepatocellular carcinoma, squamous cell carcinoma of the head and neck (SCCHN), non-thyroid medullary carcinoma, recurrent glioblastoma multiforme, neurofibromatosis type 1, CNS cancer, liposarcoma, leiomyosarcoma, salivary gland cancer, mucosal melanoma, acral / lentigines melanoma, paraganglioma, pheochromocytoma, advanced metastatic cancer, solid tumors, triple-negative breast cancer, colorectal cancer, sarcoma, melanoma, renal cancer, endometrial cancer, thyroid cancer, rhabdomyosarcoma, multiple myeloma, ovarian cancer, glioblastoma, gastrointestinal stromal tumor, mantle cell lymphoma, and refractory malignancies.

[0381] In one embodiment, the conjugates described herein, or formulations containing the conjugates described herein, are used to treat small cell lung cancer. Approximately 12%–15% of lung cancer patients have small cell lung cancer. Survival rates for metastatic small cell lung cancer are very low. The five-year survival rate after diagnosis is less than 5%. In the United States, the incidence of small cell lung cancer is approximately 26,000–30,000.

[0382] In some embodiments, the conjugates described herein or formulations containing the conjugates described herein are used to treat tumor patients who express or overexpress HSP90.

[0383] The conjugates of the present invention are characterized by relatively low toxicity to organisms while maintaining inhibitory effects, such as slowing or stopping tumor growth. As used herein, "toxicity" refers to the ability of a substance or composition to be harmful or toxic to cells, tissues, or the cellular environment. Low toxicity means a reduced ability of a substance or composition to be harmful or toxic to cells, tissues, or the cellular environment. Such reduced toxicity or low toxicity can be relative to standard measures, relative to treatment, or relative to the absence of treatment. For example, the conjugates of the present invention may have lower toxicity than the active ingredient portion Z administered alone. For conjugates containing DM1, the toxicity is lower than that of DM1 administered alone.

[0384] Toxicity can be further measured relative to the subject's weight loss, where a weight loss exceeding 15%, 20%, or 30% of body weight indicates toxicity. Other toxicity measures, such as patient performance measures, including drowsiness and malaise, can also be measured. Neutropenia, thrombocytopenia, white blood cell (WBC) count, and complete blood cell (CBC) count may also be measures of toxicity. Pharmacological indicators of toxicity include elevated transaminase (AST / ALT) levels, neurotoxicity, kidney damage, GI damage, etc. In one embodiment, the conjugate of the present invention does not cause a significant change in the subject's weight. After treatment with the conjugate of the present invention, the subject's weight loss is less than about 30%, about 20%, about 15%, about 10%, or about 5%. In another embodiment, the conjugate of the present invention does not cause a significant increase in the subject's AST / ALT levels. After treatment with the conjugate of the present 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, treatment with the conjugate of the present invention does not cause a significant change in the CBC or WBC count of the subject. After treatment with the conjugate of the present invention, the CBC or WBC level of the subject decreases by less than about 30%, about 20%, about 15%, about 10%, or about 5%.

[0385] combination therapy

[0386] In some embodiments, the conjugate of the present invention is combined with at least one additional active substance. The active substance can be any suitable pharmaceutical agent. The conjugate and at least one additional active substance can be administered simultaneously, sequentially, or in any order. The conjugate and at least one additional active substance can be administered at different doses, at different frequencies of administration, or via different routes, as long as it is appropriate.

[0387] In some embodiments, additional active substances affect the biodistribution (i.e., tissue distribution) of the conjugates of the present invention. For example, radioactive substances can accumulate in the kidneys and may pose potential radiotoxicity problems to the kidneys and surrounding organs. Additional active substances can reduce renal accumulation or retention time. Preferably, renal uptake of the conjugate is reduced without affecting tumor uptake of the conjugate. The kidneys 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 its analogue. The amino acid or its analogue 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 substance that binds to HSP90 (such as an HSP90 inhibitor). Any ligand discussed in the "HSP90 Targeting Part" section, such as ganesespib or its derivatives / analytes, can be used. In another non-limiting example, the conjugates of the present invention can be administered in combination with monosodium glutamate (MSG) or glutamate. In yet another non-limiting example, the conjugate of the present invention can be administered in combination with ethyol (WR-2721), a solution of succinyl gelatin containing bovine gelatin (Gelofusine), or an albumin fragment. The molecular weight of the albumin fragment can be from 3 to 50 kDa.

[0388] The additional active substance may be selected from any active substance described herein, such as drugs used to treat cancer. It may also be a cancer symptom-relieving drug. Non-limiting examples of symptom-relieving drugs include: octreotide or lanreotide; interferon, cypoheptadine, or any other antihistamine. In some embodiments, the conjugate of the present invention does not have drug-drug interference with the additional active substance. 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 substance may be administered concurrently with the conjugate of the present invention.

[0389] In another instance, the conjugates of the present invention can be combined with moderate doses of chemotherapeutic agents such as mitomycin C, vincristine, and cisplatin (see Ellis et al., Br J Cancer, vol. 71(2):366–370 (1995), the contents of which are incorporated herein by reference in their entirety).

[0390] In yet another instance, a patient may first receive a pharmaceutically effective dose of the unconjugated active substance, followed by a pharmaceutically effective dose of a conjugate containing the same active substance.

[0391] In some embodiments, the 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 prior to the radioactive analog. In another instance, a subject can receive a mixture of the non-radioactive conjugate and its radioactive analog. In yet another instance, a subject can first receive treatment with the non-radioactive conjugate, and then receive a mixture of the non-radioactive conjugate and its radioactive analog.

[0392] In some embodiments, the conjugate of the present invention containing a radiolabel can be combined with at least one other conjugate of the present invention containing one or more different radiolabels. For example, a conjugate containing an imaging radiolabel can be combined with a conjugate containing a non-imaging radiolabel. In one embodiment, a conjugate containing lutetium (Lu) can be combined with a conjugate containing gallium (Ga).

[0393] The conjugates described herein, or formulations containing the conjugates described herein, can be used to deliver therapeutic, preventative, or diagnostic agents to selective tissues of individuals or patients in need. For example, the conjugates of the present invention are used to deliver radioactive materials to selective tissues. These tissues may be tumor tissue. Dosing regimens can be adjusted to provide an optimal desired response (e.g., a therapeutic or preventative response). For example, a single bolus injection can be administered, several fractionated doses can be administered over time, or the dose can be proportionally reduced or increased as indicated by the urgency of the treatment situation. As used herein, the form of dosage units refers to physically discrete units suitable as a unit dose for use in a mammalian subject to be treated; each unit contains a predetermined amount of active compound calculated to produce the desired treatment.

[0394] V. Medicine boxes and devices

[0395] This invention provides various kits and devices for conveniently and / or effectively performing the methods of the invention. Typically, the kit includes a sufficient quantity and / or number of components to allow the user to administer multiple treatments and / or conduct multiple experiments on a subject.

[0396] In one embodiment, the present invention provides a kit for inhibiting tumor cell growth in vitro or in vivo, comprising the conjugates of the present invention or combinations thereof, optionally in combination with any other active substance.

[0397] The kit may also include packaging and instructions for use and / or a delivery agent that forms the formulation composition. The delivery agent may include saline, buffer solutions, or any delivery agent disclosed herein. The amounts of the components may be varied to obtain consistent, reproducible, higher-concentration saline or buffer-only formulations. Components may also be varied to increase the stability of the conjugate over a period of time and / or under various conditions in buffer solutions.

[0398] This invention provides devices for incorporating the conjugates of this invention. These devices contain a stable formulation that can be immediately delivered to a subject in need, such as a human patient. In some embodiments, the subject has cancer.

[0399] Non-limiting examples of devices include pumps, catheters, needles, transdermal patches, pressurized olfactory delivery devices, iontophoresis devices, and multilayer microfluidic devices. The devices can be used to deliver the conjugates of the invention according to single, multiple, or fractionated dosing regimens. The devices can be used to deliver the conjugates of the invention through biological tissues, intradermis, subcutaneous tissue, or intramuscular tissue.

[0400] VI. Definition

[0401] As used herein, the term "compound" is intended to include all stereoisomers, geometric isomers, tautomers, and isotopes of the described structure. In this application, "compound" is used interchangeably with "conjugate." Therefore, as used herein, "conjugate" is also intended to include all stereoisomers, geometric isomers, tautomers, and isotopes of the described structure.

[0402] The compounds described herein may be asymmetric (e.g., having one or more stereocenters). Unless otherwise indicated, all stereoisomers, such as enantiomers and diastereomers, are intended. The compounds of this disclosure containing asymmetrically substituted carbon atoms can be isolated in optically active or racemic form. Methods for preparing optically active forms from optically active starting materials are known in the art, such as by resolving racemic mixtures or by stereoselective synthesis. Many geometric isomers, such as alkenes, C=N double bonds, etc., may also exist in the compounds described herein, and all such stable isomers are covered in this disclosure. Cis and trans geometric isomers of the compounds of this disclosure are described, and can be isolated in mixtures of isomers or in separate isomeric forms.

[0403] The disclosed compounds also include tautomeric forms. Tautomeric forms arise from the exchange of single bonds with adjacent double bonds and the accompanying proton migration. Tautomeric forms include proton-shift tautomers, which are isomeric protonated states having the same empirical formula and total charge. Examples of proton-shift tautomers include ketone-enol pairs, amide-imine pairs, lactam-lactamimide pairs, amide-imine pairs, enamine-imine pairs, and cyclic forms in which protons can occupy two or more positions in the heterocyclic system, such as 1H-imidazolium and 3H-imidazolium, 1H-1,2,4-triazole, 2H-1,2,4-triazole and 4H-1,2,4-triazole, 1H-isoindole and 2H-isoindole, and 1H-pyrazole and 2H-pyrazole. Tautomeric forms can be in equilibrium or spatially locked into one form by appropriate substitution.

[0404] The disclosed compounds also include all isotopes of the atoms present in the intermediates or final compounds. An "isotope" is an atom that has the same atomic number but different mass numbers due to differences in the number of neutrons in its nucleus. For example, isotopes of hydrogen include tritium and deuterium.

[0405] The compounds and salts of this disclosure can be prepared by conventional methods in combination with solvents or water molecules to form solvates and hydrates.

[0406] As used herein, the term "subject" or "patient" means any organism to which the conjugate may be administered, for example, for experimental, therapeutic, diagnostic, and / or preventative purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, guinea pigs, cattle, pigs, sheep, horses, dogs, cats, hamsters, llamas, non-human primates, and humans).

[0407] As used herein, the terms “treatment” or “prevention” may include preventing the occurrence of a disease, symptom, or condition in animals susceptible to said disease, symptom, and / or condition but not yet diagnosed with said disease, symptom, or condition; inhibiting said disease, symptom, or condition, such as preventing its progression; and alleviating said disease, symptom, or condition, such as causing the remission of said disease, symptom, and / or condition. Treating a disease, symptom, or condition may include improving at least one symptom of a particular disease, symptom, or condition, even if it does not affect the underlying pathophysiology, such as treating pain in a subject by administering an analgesic, even if the agent does not treat the cause of the pain.

[0408] As used herein, “target” refers to the site where the targeted construct binds. Targets can be in vivo or in vitro. In some embodiments, a target may be a cancer cell found in leukemia or tumors (e.g., tumors of the brain, lungs (small cell and non-small cell), ovaries, prostate, breast, and colon, as well as other carcinomas and sarcomas). In other embodiments, a target may refer to a molecular structure to which the targeting moiety or ligand binds, such as a hapten, epitope, receptor, dsDNA fragment, carbohydrate, or enzyme. Targets may be tissue types such as neuronal tissue, intestinal tissue, pancreatic tissue, liver, kidney, prostate, ovary, lung, bone marrow, or breast tissue.

[0409] "Target cells" that can serve as targets of the methods or conjugates are typically animal cells, such as mammalian cells. The methods of the present invention 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 otherwise exist within animal tissue). Therefore, target cells may include, for example, blood, lymphatic tissue, cells lining the digestive tract (such as the mucous membranes of the mouth and pharynx), cells forming 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 of the present invention), dermal / epidermal cells, cells of the vagina and rectum, cells of internal organs (including placental cells), and the so-called blood / brain barrier, etc. Generally, target cells express at least one type of HSP90. In some embodiments, target cells may be cells that express HSP90 and are targeted by the conjugates described herein, and are located near cells affected by the release of the active substance of the conjugate. For example, blood vessels near tumors that express HSP90 can be targets, and active substances released at that site will affect the tumor.

[0410] The term "therapeutic effect" is well known in the art and refers to the local or systemic effect caused by a pharmacologically active substance in animals, particularly mammals, and more particularly humans. Therefore, the term means any substance intended to enhance desired physical or mental development and condition in animals (e.g., humans) for the diagnosis, cure, alleviation, treatment, or prevention of disease, ailment, or illness.

[0411] The term "modulation" is well known in the art and refers to both upregulation (i.e., activation or stimulation), downregulation (i.e., inhibition or suppression), or a combination of both, of a response. Modulation is generally relative to a baseline or reference, which may be inside or outside the therapeutic entity.

[0412] As used herein, “parenteral administration” means administration by any means other than through the digestive tract (intestinal) or a non-invasive local route. For example, parenteral administration may include administration to the patient via intravenous, intradermal, intraperitoneal, intrapleural, intratracheal, intraosseous, intracranial, intrathecal, intramuscular, intramuscular, subcutaneous, subconjunctival, injection, and infusion.

[0413] As used herein, “topical application” refers to noninvasive application to the skin, orifices, or mucous membranes. Topical application can be delivered locally, meaning the therapeutic agent provides a local effect at the delivery area without or with minimal systemic exposure. Some topical preparations can provide systemic effects, for example, by absorption into the individual's bloodstream. Topical application can include, but is not limited to, dermal and transdermal application, buccal application, intranasal application, intravaginal application, intravesical application, ocular application, and rectal application.

[0414] As used in this article, "enteral administration" means administration via absorption through the gastrointestinal tract. Enteral administration can include oral and sublingual administration, gastric administration, or rectal administration.

[0415] As used herein, “lung administration” means administration to the lungs via inhalation or intratracheal administration. As used herein, the term “inhalation” refers to the intake of air into the alveoli of the lungs. Air intake can occur through the mouth or nose.

[0416] As used interchangeably herein, the terms “sufficient” and “effective” refer to the amount (e.g., mass, volume, dose, concentration, and / or time period) required to achieve one or more desired results. A “therapeutic effective amount” is the minimum concentration required to achieve a measurable improvement or prevention of at least one symptom or specific condition or ailment, a measurable increase in life expectancy, or a substantial improvement in a patient’s quality of life. Therefore, the therapeutic effective amount depends on the specific bioactive molecule and the specific condition or ailment to be treated. Therapeutic effective amounts of many active substances (such as antibodies) are known in the art. Therapeutic effective amounts of the compounds and compositions described herein, for example, for treating a specific condition, can be determined using techniques entirely within the skill of those skilled in the art, such as physicians.

[0417] As may be used interchangeably herein, the terms “bioactive substance” and “active substance” include, but are not limited to, physiologically or pharmacologically active substances that act locally or systemically in the body. A bioactive substance is a substance intended for the treatment (e.g., a therapeutic agent), prevention (e.g., a preventative agent), diagnosis (e.g., a diagnostic agent), cure or alleviate a disease or ailment; a substance that affects the structure or function of the body; or a prodrug that becomes bioactive or more active after being placed in a predetermined physiological environment.

[0418] The term "prodrug" refers to a substance that is converted into its biologically active form in vitro and / or in vivo, including small organic molecules, peptides, nucleic acids, or proteins. Prodrugs can be suitable because, in some cases, they can be more readily administered than the parent compound (the active compound). For example, a prodrug may be bioavailable through oral administration, while the parent compound may not. Prodrugs may also have improved solubility in pharmaceutical compositions compared to the parent drug. Prodrugs may also have lower toxicity compared to the parent drug. Prodrugs can be converted into the parent drug through various mechanisms, including enzymatic processes and metabolic hydrolysis. Harper, NJ (1962) Drug Latentiation, edited by Jucker, Progress in Drug Research, 4:221-294; Morozowich et al. (1977) Application of Physical Organic Principles to Prodrug Design, edited by EB Roche, Design of Biopharmaceutical Properties through Prodrug and Analogs, APhA; Acad.Pharm.Sci.; EB Roche (1977) Bioreversible Carriers in Drug Design, Theory and Application, APhA; H. Bundgaard (1985) Design of Prodrugs, Elsevier; Wang et al (1999) Prodrug approaches to the improved delivery of peptide peptide drug,Curr.Pharm.Design.5(4):265-287; Pauletti et al. (1997) Improvement in peptide bioavailability: Peptidomimetic and ProdrugStrategies, Adv.Drug.Delivery Rev.27:235-256; Mizen et al. (1998). The Use of Estersas Prodrugs for Oral Delivery of β-Lactam antibiotics, Pharm.Biotech.11:345-365; Gaignault et al. (1996) Designing Prodrug and Bioprecursors I.Carrier Prodrugs,Pract.Med.Chem. 671 - 696; M. Asgharnejad (2000). Improving Oral Drug Transport Via Prodrugs, G. L. Amidon, P. I. Lee and E. M. Topp eds., 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., 15(2): 143 - 53; Balimane and Sinko (1999). Involvement of multiple transporters in the oral absorption of nucleoside analogues, Adv. Drug Delivery Rev., 39(1 - 3): 183 - 209; Browne (1997). Fosphenytoin (Cerebyx), Clin. Neuropharmacol. 20(1): 1 - 12; Bundgaard (1979). Bioreversible derivatization of drugs--principle and applicability to improve the therapeutic effects of drugs, Arch. Pharm. Chemi. 86(1): 1 - 39; H. 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. 19(2): 115 - 130; Fleisher et al. (1985) Design of prodrugs for improved gastrointestinal absorption by intestinal enzyme targeting, Methods Enzymol.112:360-81; FarquharD et al. (1983) Biologically Reversible Phosphate-Protective Groups, J.Pharm.Sci., 72(3):324-325; Han, HK et al. (2000) Targeted prodrug design to optimize drug delivery, AAPS PharmSci., 2(1):E6; Sadzuka Y. (2000) Effective prodrug liposome and conversion to active metabolite, Curr. Drug Metab., 1(1): 31-48; DM Lambert (2000) Rationale and applications of lipids as prodrug carriers, Eur. J. Pharm. Sci., 11 Supplement 2: S15-27; Wang, W. et al. (1999) Prodrug approaches to the improved delivery of peptide drugs. Curr. Pharm. Des., 5(4):265-87. .

[0419] As used herein, "biocompatible" means that a substance and any of its metabolites or degradation products are generally non-toxic to the recipient and do not cause any significant adverse effects on the recipient. Generally, a biocompatible substance is one that does not elicit a significant inflammatory or immune response when administered to a patient.

[0420] As used in this article, "biodegradable" generally refers to a substance that will degrade or decay under physiological conditions into smaller units or chemical substances that can be metabolized, eliminated, or excreted by the subject. Degradation time varies depending on composition and form. Degradation time can range from hours to weeks.

[0421] As used herein, the term "pharmaceutically acceptable" means, according to guidelines from agencies such as the U.S. Food and Drug Administration, that a compound, substance, composition, and / or dosage form is suitable for use in human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, to the extent reasonably medically permissible, and in proportion to a reasonable benefit / risk ratio. As used herein, "pharmaceutically acceptable carrier" refers to all components of a pharmaceutical formulation that facilitate the 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.

[0422] As used herein, the term "molecular weight" generally refers to the mass or average mass of a substance. For polymers or oligomers, 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 various ways, including gel permeation chromatography (GPC) or capillary viscosity determination. This differs from the use of number-average molecular weight (M). n The weight-average molecular weight (M) w The molecular weight of GPC is reported using capillary viscosity determination, which provides an estimate of molecular weight in the form of specific logarithmic viscosity measured from a dilute polymer solution using a specific set of concentration, temperature, and solvent conditions.

[0423] As used in this article, the term "small molecule" generally refers to organic molecules with 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.

[0424] As used in this article, the term "hydrophilicity" refers to the presence of highly polar groups in a substance that readily interact with water.

[0425] As used in this article, the term "hydrophobicity" refers to a substance's lack of affinity for water; its tendency to repel and not absorb water, and its insolubility or miscibility with water.

[0426] As used in this article, the term "lipophilic" refers to the affinity of a compound for lipids.

[0427] As used herein, the term "amphiphilic" refers to the combination of hydrophilic and lipophilic (hydrophobic) properties of a molecular assembly. As used herein, "amphiphilic substance" refers to a substance containing both a hydrophobic or more hydrophobic oligomer or polymer (e.g., a biodegradable oligomer or polymer) and a hydrophilic or more hydrophilic oligomer or polymer.

[0428] As used herein, the term "targeted portion" refers to a portion that binds to or is located at a specific site. This portion may be, for example, a protein, nucleic acid, nucleic acid analogue, carbohydrate, or small molecule. The site may be a tissue, a specific cell type, or a subcellular compartment. In some embodiments, the targeted portion may specifically bind to selected molecules.

[0429] As used herein, the term "reactive coupling group" refers to any chemical functional group capable of reacting with a second functional group to form a covalent bond. The selection of reactive coupling groups is within the capabilities of those skilled in the art. Examples of reactive coupling groups may include primary amines (-NH2) and amine reactive linkers such as isothiocyanates, isocyanates, acyl azides, NHS esters, sulfonyl chlorides, aldehydes, glyoxal, epoxides, ethylene oxides, carbonates, aryl halides, imino esters, carbodiimides, anhydrides, and fluorophenyl esters. Most of these conjugates are conjugated to amines via acylation or alkylation. Examples of reactive coupling groups may include aldehydes (-COH) and aldehyde reactive linkers such as hydrazides, alkoxyamines, and primary amines. Examples of reactive coupling groups may include thiol groups (-SH) and mercapto reactive groups such as maleimides, haloacetyl groups, and pyridyl disulfides. Examples of reactive coupling groups may include photoreactive coupling groups, such as aryl azides or diaziridine. Coupling reactions may include the use of catalysts, heat, pH buffers, light, or combinations thereof.

[0430] As used herein, the term "protecting group" refers to a functional group that can be added to and / or substituted to another desired functional group to protect the desired functional group from certain reaction conditions, and can be selectively removed and / or replaced to deprotect or expose the desired functional group. Protecting groups are known to those skilled in the art. Suitable protecting groups may include those described in Greene and Wuts, Protective Groups in Organic Synthesis, (1991). Acid-sensitive protecting groups include dimethoxytriphenylmethyl (DMT), tert-butyl carbamate (tBoc), and trifluoroacetyl (tFA). Base-sensitive protecting groups include 9-fluorenylmethoxycarbonyl (Fmoc), isobutyryl (iBu), benzoyl (Bz), and phenoxyacetyl (pac). Other protecting groups include acetaminomethyl, acetyl, tert-amyloxycarbonyl, benzyl, benzyloxycarbonyl, 2-(4-biphenyl)-2-propyloxycarbonyl, 2-bromobenzyloxycarbonyl, tert-butyl, tert-butoxycarbonyl, 1-benzyloxycarbonylamide-2,2,2-trifluoroethyl, 2,6-dichlorobenzyl, 2-(3,5-dimethoxyphenyl)-2-propyloxycarbonyl, 2,4-dinitrophenyl, and dithiosuccinyl. Formyl, 4-methoxybenzenesulfonyl, 4-methoxybenzyl, 4-methylbenzyl, o-nitrophenylsulfinyl, 2-phenyl-2-propyloxycarbonyl, α-2,4,5-tetramethylbenzyloxycarbonyl, p-toluenesulfonyl, xanthonyl, benzyl ester, N-hydroxysuccinimide ester, p-nitrobenzyl ester, p-nitrophenyl ester, phenyl ester, p-nitrocarbonate, p-nitrobenzyl carbonate, trimethylsilyl and pentachlorophenyl ester.

[0431] As used herein, the term "activated ester" refers to an alkyl ester of a carboxylic acid, where the alkyl group is a favorable leaving group that makes the carbonyl group susceptible to nucleophilic attack by a molecule carrying an amino group. Therefore, activated esters are susceptible to ammonolysis and react with amines to form amides. Activated esters contain a carboxylic acid ester group -CO2R, where R is a leaving group.

[0432] The term "alkyl" refers to a saturated aliphatic group, including straight-chain alkyl, branched alkyl, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl, and cycloalkyl-substituted alkyl.

[0433] In some embodiments, the straight-chain or branched alkyl group has 30 or fewer carbon atoms (e.g., C1-C1) in its backbone. 30 (For straight chains), C3-C 30(For branched chains) 20 or fewer, 12 or fewer, or 7 or fewer carbon atoms. Similarly, in some embodiments, cycloalkyl groups have 3-10 carbon atoms in their ring structure, for example, 5, 6, or 7 carbon atoms in the ring structure. As used throughout the specification, examples, and claims, the term "alkyl" (or "lower alkyl") is intended to include both "unsubstituted alkyl" and "substituted alkyl," wherein the latter refers to an alkyl moiety having substituents on one or more carbons of a substituted hydrocarbon skeleton. Such substituents include, but are not limited to, halogens, hydroxyl groups, carbonyl groups (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl groups (such as thioesters, thioacetic esters, or thiocarbamates), alkoxy groups, phosphoryl groups, phosphate groups, phosphonates, hypophosphonates, amino groups, amide groups, amidine groups, imine groups, cyano groups, nitro groups, azide groups, mercapto groups, alkylthio groups, sulfate groups, sulfonates, aminosulfonyl groups, sulfonamide groups, sulfonyl groups, heterocyclic groups, aralkyl groups, or aromatic or heteroaromatic moieties.

[0434] Unless otherwise specified, “lower alkyl” as used herein means an alkyl group as defined above but having 1 to 10 carbon atoms or 1 to 6 carbon atoms in its skeletal structure. Similarly, “lower alkenyl” and “lower alkynyl” have similar chain lengths. In some embodiments, the alkyl group is a lower alkyl group. In some embodiments, the substituent designated as alkyl herein is a lower alkyl group.

[0435] Those skilled in the art will understand that the substituted portion on the hydrocarbon chain can be substituted itself where appropriate. For example, substituents of the substituted alkyl group may include halogens, hydroxyl groups, nitro groups, thiols, amino groups, azides, imino groups, amide groups, phosphoryl groups (including phosphonates and hypophosphonates), sulfonyl groups (including sulfate groups, sulfonamides, aminosulfonyl groups, and sulfonates), and silyl groups, as well as ethers, alkylthio groups, carbonyl groups (including ketones, aldehydes, carboxylic acid esters, and esters), -CF3, -CN, etc. Cycloalkyl groups can be substituted in the same manner.

[0436] As used herein, the term "heteroalkyl" refers to a straight-chain, branched, or cyclic carbonaceous group or combination thereof containing at least one heteroatom. Suitable heteroatoms include, but are not limited to, O, N, Si, P, Se, B, and S, wherein phosphorus and sulfur atoms are optionally oxidized, and nitrogen heteroatoms are optionally quaternized. Heteroalkyl groups may be substituted as defined above for alkyl groups.

[0437] The term "alkylthio" refers to an alkyl group having a sulfur group attached to it as defined above. In some embodiments, the "alkylthio" part is represented by one of -S-alkyl, -S-alkenyl, and -S-ynyl. Representative alkylthio groups include methylthio and ethylthio. The term "alkylthio" also covers cycloalkyl, alkenyl, and cycloalkenyl groups, as well as ynyl groups. "Arylthio" refers to an aryl or heteroaryl group. Alkylthio groups may be substituted as defined above for alkyl groups.

[0438] The terms “alkenyl” and “alkynyl” refer to alkyl groups that are similar in length and possible substitutions to those described above, but contain at least one double or triple bond, respectively, an unsaturated aliphatic group.

[0439] As used herein, the term "alkoxy" refers to an alkyl group having an oxygen group attached to it as defined above. Representative alkoxy groups include methoxy, ethoxy, propoxy, and tert-butoxy. An "ether" is two hydrocarbons covalently linked by oxygen. Therefore, the substituent that makes an alkyl group an ether is an alkoxy or similar alkoxy group, such as one of -O-alkyl, -O-alkenyl, and -O-ynyl. Aryloxy groups can be represented by –O-aryl or O-heteroaryl, where aryl and heteroaryl are defined below. Alkoxy and aryloxy groups can be substituted as described above for alkyl groups.

[0440] The terms "amine" and "amino" are well known in the art and refer to both unsubstituted and substituted amines, for example, portions that can be represented by the following general formula:

[0441]

[0442] Among them, R9, R 10 and R′ 10 Each can independently represent hydrogen, alkyl, alkenyl, or -(CH2). m -R8, or R9 and R 10 Together with the N atoms they are attached to, they form a heterocycle having 4 to 8 atoms in the ring structure; R8 represents aryl, cycloalkyl, cycloalkenyl, heterocyclic, or polycyclic; and m is 0 or an integer in the range of 1 to 8. In some embodiments, R9 or R 10 Only one of them can be a carbonyl group, such as R9, R 10 It does not form an imide with nitrogen. In other embodiments, the term "amine" does not cover amides, such as those where R9 and R... 10 One of them represents a carbonyl group. In another embodiment, R9 and R 10 (and optional R') 10Each of these groups independently represents hydrogen, alkyl or cycloalkyl, alkenyl or cycloalkenyl, or ynyl. Therefore, as used herein, the term "alkylamine" means an amine group as defined above, having a substituted (as described above for alkyl) or unsubstituted alkyl group attached thereto, namely R9 and R... 10 At least one of them is an alkyl group.

[0443] The term "amide" is known in the art to be an amino-substituted carbonyl group and includes a portion that can be represented by the following general formula:

[0444]

[0445] Among them, R9 and R 10 As defined above.

[0446] As used in this article, "aryl" refers to C5-C 10 Aromatic, heterocyclic, fused aromatic, fused heterocyclic, biaromatic, or biheterocyclic ring systems. In a broad sense, as used herein, "aryl" includes 5, 6, 7, 8, 9, and 10-membered monocyclic aromatic groups that may have 0 to 4 heteroatoms, such as benzene, pyrrole, furan, thiophene, imidazole, azole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine. Aromatic groups with heteroatoms in their ring structures may also be referred to as "aryl heterocycles" or "heteroaromatic compounds." The aromatic ring may be substituted at one or more ring positions by one or more substituents, including but not limited to halogens, azides, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxy, amino (or quaternized amino), nitro, mercapto, imino, amide, phosphonate, hypophosphonate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclic group, aromatic or heteroaromatic moiety, -CF3, -CN; and combinations thereof.

[0447] The term "aryl" also includes polycyclic systems having two or more cyclic rings, wherein two or more carbons are shared by two adjacent rings (i.e., "fused rings"), wherein at least one ring is aromatic, and for example, the other one or more cyclic rings may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, and / or heterocyclic. Examples of heterocyclic rings include, but are not limited to, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiopheneyl, benzozolinyl, benzothiazolinyl, benzotriazolyl, benzotetrazoleyl, benzoisoazolyl, benzoisothiazolinyl, benzimidazolyl, carbazolyl, 4aHcarbazolyl, carbazolinyl, chromanyl, chromenyl, cenylyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofurano[2,3b]tetrahydrofuran, furanyl, and furazolidyl. Imidazolyl, imidazolinyl, imidazolyl, 1H-indazole, pseudoindolenyl, dihydroindolenyl, indoleazinyl, indolenyl, 3H-indolenyl, indorubinyl, isobenzofuranyl, isochromyl, isoindazole, isodihydroindolenyl, isoindolenyl, isoquinolinyl, isothiazolyl, isozolyl, methylenedioxyphenyl, morpholinyl, naphthidyl, octahydroisoquinolinyl, diazolyl, 1,2,3-diazolyl, 1,2,4-diazolyl, 1,2 5-Diazolyl, 1,3,4-Diazolyl, Zolylalkyl, Azolyl, Hydroxyindolyl, Pyrimidinyl, Phenyridine, Phenanthrololinyl, Phenazinyl, Phenthiazinyl, Phenyrthiazinyl, Phenazinyl, Phthaloazinyl, Piperazinyl, Piperidinyl, Piperidinoneyl, 4-Piperidinoneyl, Piperidinyl, Pteridinyl, Puryl, Pyranyl, Pyrazinyl, Pyrazolylalkyl, Pyrazolinyl, Pyrazolyl, Pyridazinyl, Pyridobazole, Pyridobimazole, Pyridobthiazole, Pyridinyl, Pyridinyl, Pyrrolylalkyl, Pyrrololinyl 2H-pyrrole, pyrrole, quinazolinyl, quinolinyl, 4H-quinazinyl, quinoxalinyl, quininecycloyl, 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, thianyl, thiazolyl, thienyl, thienothiazolyl, thienozolyl, thienoimidazolyl, phenylthio, and xanthonyl. One or more rings may be substituted as defined above for "aryl".

[0448] As used herein, the term "aralkyl" refers to an alkyl group that has been substituted with an aryl group (e.g., an aromatic or heteroaromatic group).

[0449] As used in this article, the term "carbon ring" refers to an aromatic or non-aromatic ring in which every atom of the ring is carbon.

[0450] As used herein, "heterocyclic" or "heterocyclic" refers to a cyclic group connected by a monocyclic or bicyclic ring of carbon or nitrogen atoms, containing 3-10 ring atoms, for example 5-6 ring atoms, and optionally containing 1-3 double bonds, optionally substituted by one or more substituents, wherein the ring atoms consist of carbon and 1 to 4 heteroatoms, each selected from: non-peroxide oxygen, sulfur, and N(Y), wherein Y is absent or H, O, (C1-C) 10 Alkyl, phenyl, or benzyl. Examples of heterocycles include, but are not limited to, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiopheneyl, benzozolinyl, benzothiazolinyl, benzotriazolyl, benzotetrazoleyl, benzoisozolinyl, benzoisothiazolinyl, benzimidazolinyl, carbazoleyl, 4aH-carbazoleyl, carbolinyl, chromanyl, chromenyl, cenyl, decahydroquinolinyl, 2H,6H-1,5,2-diathiazinyl, dihydrofurano[2,3-b]tetrahydrofuran, furanyl, and furazolidyl. Imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazole, pseudoindole, dihydroindole, indoleazinyl, indole, 3H-indole, indorubinyl, isobenzofuranyl, isochoryl, isoindazole, isodihydroindole, isoindole, isoquinolinyl, isothiazolyl, isozolyl, methylenedioxyphenyl, morpholinyl, naphthidyl, octahydroisoquinolinyl, diazolyl, 1,2,3-diazolyl, 1,2,4-diazolyl, 1,2,5-diazolyl, 1,3,4-diazolyl Azolyl, azolealkyl, azole, oxetaneheptyl, oxetanebutyl, hydroxyindolyl, pyrimidinyl, phenanthidyl, phenanthrololinyl, phenazinyl, phenothiazinyl, phenanthiazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidoneyl, 4-piperidinoneyl, piperinyl, pteridinyl, purine, pyranyl, pyrazinyl, pyrazolylalkyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridozolyl, pyridozolyl, pyridozolyl, pyridozolyl, pyridozolyl, pyridozolyl, pyridozolyl, pyridozolyl, pyridinyl, pyridine, pyridine, pyrrolylalkyl, pyrrololinyl, 2H -Pyrroleyl, pyrroleyl, quinazolinyl, quinolinyl, 4H-quinazinyl, quinoxalinyl, quininecycloyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydropyranyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianyl, thiazolyl, thienyl, thienothiazolyl, thienozolyl, thienoimidazolyl, phenylthioyl, and xanthonyl. The heterocyclic group may optionally be substituted at one or more positions with one or more substituents as defined above for alkyl and aryl groups, such as halogens, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, mercapto, imino, amide, phosphate ester, phosphonate, hypophosphonate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclic group, aromatic or heteroaromatic moiety, -CF3 and -CN.

[0451] The term "carbonyl" is well known in the art and includes parts such as those that can be represented by the following general formula:

[0452]

[0453] Where X is a bond or represents oxygen or sulfur, and R 11 R′ represents hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, or ynyl. 11 This indicates hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, or alkynyl. When X is oxygen, and R... 11 or R' 11 When X is not hydrogen, the formula represents "ester". When X is oxygen, and R... 11 As defined above, the portion referred to herein is called a carboxyl group, and particularly when R 11 When X is hydrogen, the formula represents "carboxylic acid". When X is oxygen, and R′ 11 When X is hydrogen, the formula represents "formate ester". Generally, when the oxygen atom in the above formula is replaced by sulfur, the formula represents "thiocarbonyl". When X is sulfur, and R... 11 or R′ 11 When X is not hydrogen, the formula represents "thioester". When X is sulfur, and R... 11 When X is hydrogen, the formula represents "thiocarboxylic acid". When X is sulfur, and R' 11 When X is hydrogen, the formula represents "thiocarbamate". On the other hand, when X is a bond and R... 11 When X is not hydrogen, the above formula represents a "ketone" group. When X is a bond, and R... 11 When it is hydrogen, the above formula represents an "aldehyde" group.

[0454] As used herein, the term "monoester" refers to an analogue of a dicarboxylic acid, in which one carboxylic acid is functionalized into an ester, and the other carboxylic acid is a free carboxylic acid or a salt of a carboxylic acid. Examples of monoesters include, but are not limited to, monoesters of succinic acid, glutaric acid, adipic acid, octanoic acid, sebacic acid, azelaic acid, oxalic acid, and maleic acid.

[0455] As used herein, the term "heteroatom" refers to an atom of any element other than carbon or hydrogen. Examples of heteroatoms are boron, nitrogen, oxygen, phosphorus, sulfur, and selenium. Other applicable heteroatoms include silicon and arsenic.

[0456] As used herein, the term “nitro” means -NO2; the term “halogen” specifies -F, -Cl, -Br or -I; the term “thiol” means -SH; the term “hydroxyl” means -OH; and the term “sulfonyl” means -SO2-.

[0457] As used herein, the term "substituted" refers to all permissible substituents in the compounds described herein. In the broadest sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Illustrative substituents include, but are not limited to, halogens, hydroxyl groups, or any other organic group containing any number of carbon atoms, such as 1-14 carbon atoms, in the form of a straight-chain, branched, or cyclic structure, and optionally including one or more heteroatoms such as oxygen, sulfur, or nitrogen groups. Representative substituents include alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, phenyl, substituted phenyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halogen, hydroxyl, alkoxy, substituted alkoxy, phenoxy, substituted phenoxy, aryloxy, substituted aryloxy, alkylthio, substituted alkylthio, phenylthio, substituted phenylthio, arylthio, substituted arylthio, cyano, isocyano, substituted isocyano, carbonyl, substituted carbonyl, carboxyl, substituted carboxyl, amino, substituted amino, amide, substituted amide, sulfonyl, substituted sulfonyl, sulfonic acid, phosphoryl, substituted phosphoryl, phosphonyl, substituted phosphonyl, polyaryl, substituted polyaryl, C3-C 20 Cyclic groups, substituted C3-C 20 Cyclic groups, heterocyclic groups, substituted heterocyclic groups, amino acids, peptides, and polypeptide groups.

[0458] Heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents satisfying the heteroatom valence of the organic compounds described herein. It should be understood that “substitution” or “substituted” includes the implicit condition that the substitution conforms to the permissible valence of the substituted atom and the substituent, and that the substitution produces a stable compound, i.e., a compound that does not spontaneously undergo transformations, such as by rearrangement, cyclization, or elimination.

[0459] In a broad sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Illustrative substituents include, for example, those described herein. Permissible substituents may be one or more, and may be the same or different for a suitable organic compound. Heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of the organic compounds described herein that satisfy the heteroatom valence.

[0460] In various embodiments, the substituents are selected from alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamate, carboxyl, cyano, cycloalkyl, ester, ether, formyl, halogen, haloalkyl, heteroaryl, heterocyclic, hydroxyl, ketone, nitro, phosphate ester, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thione, each optionally substituted by one or more suitable substituents. In some embodiments, the substituents are selected from alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamate, carboxyl, cycloalkyl, ester, ether, formyl, haloalkyl, heteroaryl, heterocyclic, ketone, phosphate ester, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thion, wherein each of the alkoxy, aryloxy, alkyl, alkenyl, alkynyl, amide, amino, aryl, arylalkyl, carbamate, carboxyl, cycloalkyl, ester, ether, formyl, haloalkyl, heteroaryl, heterocyclic, ketone, phosphate ester, sulfide, sulfinyl, sulfonyl, sulfonic acid, sulfonamide, and thion may be further substituted with one or more suitable substituents.

[0461] Examples of substituents include, but are not limited to, halogens, azides, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxy, amino, nitro, mercapto, imino, acylamino, phosphonate, hypophosphonate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonylamino, ketone, aldehyde, thion, ester, heterocyclic, –CN, aryl, aryloxy, perhaloalkoxy, arylalkoxy, heteroaryl, heteroaryloxy, heteroarylalkyl, heteroarylalkoxy, azide, alkylthio, oxo, acylalkyl, carboxyl ester, carboxylamino, acyloxy, aminoalkyl, alkylaminoaryl, alkylaryl, alkylaminoalkyl, alkoxyaryl, arylamino, aralkylalkylamino, alkylsulfonyl, carboxylaminoalkylaryl, carboxylaminoaryl, hydroxyalkyl, haloalkyl, alkylaminoalkylcarboxyl, aminocarboxylaminoalkyl, cyano, alkoxyalkyl, perhaloalkyl, arylalkyloxyalkyl, etc. In some implementations, the substituents are selected from cyano, halogen, hydroxyl, and nitro groups.

[0462] As used herein, the term "copolymer" generally refers to a single polymeric substance comprising two or more different monomers. Copolymers can be in any form, such as random, block, or graft. Copolymers can have any end groups, including capped or acidic end groups.

[0463] The terms “polypeptide,” “peptide,” and “protein” generally refer to polymers of amino acid residues. As used herein, the terms also apply to amino acid polymers, where one or more amino acids are chemical analogs or modified derivatives of the corresponding naturally occurring amino acids, or to non-natural amino acids. The term “protein,” as generally used herein, refers to a polymer of amino acids linked together by peptide bonds to form a polypeptide with a chain length sufficient to produce tertiary and / or quaternary structures. By definition, the term “protein” excludes small peptides, which lack the necessary higher-order structures considered essential for proteins.

[0464] The terms “nucleic acid,” “polynucleotide,” and “oligonucleotide” are used interchangeably to refer to deoxyribonucleotide or ribonucleotide polymers in linear or cyclic conformations and in single- or double-stranded form. These terms should not be construed as limiting the length of the polymer. The terms may cover known analogs of natural nucleotides, as well as nucleotides modified in their base, sugar, and / or phosphate ester moieties (such as a thiophosphate backbone). Generally, and unless otherwise specified, analogs of a particular nucleotide have the same base-pairing specificity; that is, an analog of A will pair with a T base. The term “nucleic acid” is a term in the art that refers to a single unit of at least two base-sugar-phosphate ester monomers. A nucleotide is a monomeric unit of a nucleic acid polymer. The term includes deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) in the form of messenger RNA, antisense, plasmid DNA, portions of plasmid DNA, or viral-derived genetic material. Antisense nucleic acids are polynucleotides that interfere with the expression of DNA and / or RNA sequences. The term nucleic acid refers to a single unit of at least two base-sugar-phosphate ester combinations. Natural nucleic acids have a phosphate ester backbone. Artificial nucleic acids may contain other types of backbones, but they contain the same bases as natural nucleic acids. The term also includes PNA (peptide nucleic acid), phosphate thioesters, and other variants of the phosphate ester backbone of natural nucleic acids.

[0465] A “functional fragment” of a protein, polypeptide, or nucleic acid is a protein, polypeptide, or nucleic acid whose sequence differs from the full-length protein, polypeptide, or nucleic acid, but which retains at least one function similar to that of the full-length protein, polypeptide, or nucleic acid. The functional fragment may have more, fewer, or the same number of residues as the corresponding natural molecule, and / or may contain one or more amino acid or nucleotide substitutions. Methods for determining the function of nucleic acids (e.g., coding function, ability to hybridize with another nucleic acid) are well known in the art. Similarly, methods for determining the function of proteins are well known. For example, the DNA-binding function of a polypeptide can be determined, for example, by filter paper binding assay, electrophoretic mobility shift assay, or immunoprecipitation assay. DNA cleavage can be determined by gel electrophoresis. The ability of one protein to interact with another protein can be determined, for example, by co-immunoprecipitation, two-hybrid assay, or complementation (e.g., genetic complementation or biochemical complementation). See, for example, Fields et al. (1989) Nature 340:245-246; U.S. Patent No. 5,585,245; and PCT WO 98 / 44350.

[0466] As used herein, the term "linker" refers to a carbon chain that may contain heteroatoms (e.g., nitrogen, oxygen, sulfur, etc.) and may 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, or 50 atoms long. Linkers can be substituted with various substituents, including but not limited to hydrogen, alkyl, alkenyl, alkynyl, amino, alkylamino, dialkylamino, trialkylamino, hydroxyl, alkoxy, halogen, aryl, heterocyclic, aromatic heterocyclic, cyano, amide, carbamoyl, carboxylic acid, ester, thioether, alkyl thioether, thiol, and ureyl groups. Those skilled in the art will recognize that each of these groups can be further 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, photolytic linkers, thermally unstable linkers, enzyme-cleavable linkers (e.g., esterase-cleavable linkers), ultrasound-sensitive linkers, and X-ray-cleavable linkers.

[0467] The term "pharmaceutically acceptable counterion" refers to a pharmaceutically acceptable anion or cation. In various embodiments, a pharmaceutically acceptable counterion is a pharmaceutically acceptable ion. For example, a pharmaceutically acceptable counterion is selected from citrate, malate, acetate, oxalate, chloride, bromide, iodide, nitrate, sulfate, hydrogen sulfate, phosphate, acid phosphate, isonicotinic acid, acetate, lactate, salicylate, tartrate, oleate, tannic acid, pantothenate, hydrogen tartrate, ascorbate, succinate, maleate, gentianate, fumarate, gluconate, glucuronate, sucrose, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and dihydroxynaphthate (i.e., 1,1′-methylene-bis-(2-hydroxy-3-naphthate)). 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 specific embodiments, the pharmaceutically acceptable counterion is selected from chloride, bromide, iodide, nitrate, sulfate, bisulfate, and phosphate.

[0468] The term "pharmaceutically acceptable salt" refers to a salt containing an acidic or basic group present in a compound that can be used in the compositions of the present invention. Compounds included in the compositions of the present invention that are basic in nature are capable of forming a variety of salts with various inorganic and organic acids. The 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 sulfates, citrates, malates, acetates, oxalates, chlorides, bromides, iodides, nitrates, sulfates, hydrogen sulfates, phosphates, acid phosphates, isonicotinates, acetates, lactates, salicylates, citrates, tartrates, oleates, tannins, pantothenates, hydrogen tartrates, ascorbic acid salts, succinates, maleates, gentianates, fumarates, gluconates, glucurons, sucrose salts, formates, benzoates, glutamates, methanesulfonates, ethanesulfonates, benzenesulfonates, p-toluenesulfonates, and bis(hydroxynaphthyl)ates (i.e., 1,1′-methylene-bis(2-hydroxy-3-naphthylcarbamate)). In addition to the acids mentioned above, compounds comprising an amino moiety included in the compositions of the present invention can also form pharmaceutically acceptable salts with various amino acids. The acidic compounds included in the compositions of this invention are capable of forming alkaline salts with a variety of pharmacologically acceptable cations. Examples of such salts include alkali metal salts or alkaline earth metal salts, and in particular calcium, magnesium, sodium, lithium, zinc, potassium, and iron salts.

[0469] If the compound described herein is obtained as an acid addition salt, the free base can be obtained by alkalizing a solution of the acid salt. Conversely, if the product is a free base, the addition salt, particularly a pharmaceutically acceptable addition salt, can be produced according to conventional procedures for preparing acid addition salts from base compounds by dissolving the free base in a suitable organic solvent and treating the solution with acid. Those skilled in the art will recognize the various synthetic methodologies that can be used to prepare non-toxic, pharmaceutically acceptable addition salts.

[0470] Pharmaceutically acceptable salts are derived from acids selected from: 1-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-ketoglutaric acid, 4-acetaminobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, hexanoic acid, and caprylic acid. Acids, carbonic acid, cinnamic acid, citric acid, cyclohexane, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactosic acid, gentian acid, glucoheponic acid, gluconic acid, glucuronic acid, glutamic acid, glutamate, glycerophosphate, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, hydroxyethanesulfonic acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucilage, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, dihydroxynaphthalic acid, pantothenic acid, phosphoric acid, propionic acid, pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tartaric acid, thiocyanate, toluenesulfonic acid, trifluoroacetic acid, and undecanoic acid.

[0471] The term "bioavailable" is well known in the art and refers to a form of the present invention that allows a portion of the present invention or an amount thereof to be absorbed by, incorporated into, or otherwise physiologically available to the subject or patient to whom the present invention is administered.

[0472] It should be understood that the following embodiments are intended to illustrate, not limit, the invention. Various other embodiments and modifications to the foregoing description and embodiments will become apparent to those skilled in the art upon reading this disclosure without departing from the spirit and scope of the invention, and it is intended that all described embodiments or modifications be included within the scope of the appended claims. All publications and patents referenced herein are hereby incorporated by reference in their entirety.

[0473] It should be understood that in the following embodiments, some conjugates were prepared and characterized using a non-radioactive metal such as Lu-175. It will be apparent to those skilled in the art that corresponding radioactive Lu-177 analogs can be readily prepared using known methods, and that the distribution data of the Lu-175 conjugates can be representative of the distribution data of the Lu-177 analogs. Example

[0474] Example 1: Synthesis of Conjugates

[0475] The conjugates of the present invention can be prepared using any convenient method. In a reasonable approach, the conjugates are constructed from their respective components, targeting moieties, and in some cases, linkers and active material moieties. As is known in the art, the components can be covalently bonded to each other via functional groups, wherein these functional groups can be present on the components or introduced onto the components using one or more steps (e.g., oxidation, reduction, cleavage, etc.). Functional groups that can be used to covalently bind the components together to produce pharmaceutical conjugates include hydroxyl, thiol, amino, etc. Specific moieties of the different components will be selected to provide covalent bonding so as not to substantially adversely interfere with the desired binding activity of those components, such as the binding activity of the active material moieties. Regions that do not affect the target binding activity will be modified to maintain an adequate amount of the desired pharmaceutical activity. Where necessary and / or required, blocking groups can be used to protect certain moieties on the components, as is known in the art, see, for example, Green & Wuts, Protective Groups in Organic Synthesis (John Wiley & Sons) (1991).

[0476] Alternatively, known combinatorial methods can be used to generate a large library of potential conjugates, which can then be screened to identify bifunctional molecules with pharmacokinetic properties for conjugate production. Alternatively, conjugates can be produced using medicinal chemistry and known structure-activity relationships between the targeting and active moieties. In particular, this approach will determine where the two moieties are linked to the linker.

[0477] peptide synthesis

[0478] Peptide 1 was synthesized on a Liberty Blue peptide synthesizer starting with Fmoc-β-alanine (1.33 g, 0.75 mmol / g loading, 1.00 mmol) loaded onto 2-chlorotriphenylmethyl resin, 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 a 2% TFA solution in dichloromethane and purified by preparative HPLC to yield 624 mg (0.483 mmol, 48% yield) of peptide 1.

[0479]

[0480] Peptide 2 was synthesized on a Liberty Blue peptide synthesizer starting with Fmoc-β-alanine (4.00 g, 0.75 mmol / g loading, 3.00 mmol) loaded onto 2-chlorotriphenylmethyl resin, followed by the addition of Fmoc-D-Glu(OtBu)(x3) and Fmoc-AEEA. The crude peptide was cleaved by treating the resin with a dichloromethane solution of 2% TFA and purified by preparative HPLC to give 1.28 g (1.26 mmol, 42% yield) of peptide 2. Peptides 3 and 4 were synthesized in a similar manner.

[0481]

[0482] Peptide 5 was synthesized on a Liberty Blue peptide synthesizer starting with Fmoc-β-alanine (0.4 g, 0.5 mmol / g loading, 0.2 mmol) loaded onto 2-chlorotriphenylmethyl resin, followed by the addition of Fmoc-D-Glu(OtBu)(x3), Fmoc-AEEA, Dde-Lys(Fmoc), and (R)-tert-Bu4-DOTAGA, then treated with a 5% hydrazine DMF solution and 4-(p-iodophenyl)butyric acid. The crude peptide was cleaved by treating the resin with a 2% TFA solution in dichloromethane and purified by preparative HPLC to yield 50 mg (0.027 mmol, 13% yield) of peptide 5. Peptides 6 and 7 were synthesized in a similar manner.

[0483]

[0484] Peptide 8 was synthesized on a Liberty Blue peptide synthesizer starting with Fmoc-β-alanine (0.5 g, 0.75 mmol / g loading, 0.375 mmol) loaded onto 2-chlorotriphenylmethyl resin, followed by the addition of Fmoc-D-Glu(OtBu)(x3), Fmoc-AEEA, Fmoc-D-Lys(Dde), and 4-(p-iodophenyl)butyric acid, then treated with a 5% hydrazine DMF solution, followed by Fmoc-Gly, Fmoc-Val, Fmoc-Met, and Fmoc-Gly. The crude peptide was cleaved from the resin using a 2% TFA solution 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 to 70 °C for 1 h. All solvents were removed under vacuum, and the crude material 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 peptide 8 (124 mg, 55.9 mmol, 15% yield). Peptides 9-16 were prepared in a similar manner.

[0485]

[0486]

[0487]

[0488]

[0489] A solution of HATU (128 mg, 0.340 mmol) in DMF (3 mL) was added to a vial containing TM5 HCl salt (170 mg, 0.340 mmol) and Boc-glutamic acid (42 mg, 0.170 mmol), followed by the addition of diisopropylethylamine (0.60 mL). The solution was stirred at 50 °C for 1 h and then purified by preparative HPLC. The fraction containing the product was 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 substance was redissolved in a 1:1 acetonitrile:water mixture (10 mL), frozen, and lyophilized to give 17, trifluoroacetate salt (192 mg, 0.138 mmol, 81% yield). Compounds 18 and 19 were prepared in a similar manner.

[0490]

[0491] A vial was filled with T-1951 HCl salt (460 mg, 0.893 mmol) and Fmoc-DGlu-bAla (200 mg, 0.454 mmol). A solution of HATU (343 mg, 0.909 mmol) in DMF (5 mL) was added to the vial. 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 another 1 h, then acidified by the addition of 1 mL of TFA. The reaction mixture was then purified by preparative HPLC to give 20, trifluoroacetate (443 mg, 0.300 mmol, 65% yield). Compounds 21-32 were prepared in a similar manner.

[0492]

[0493]

[0494]

[0495] DMF (2 mL) and diisopropylethylamine (0.20 mL) were added to vials containing 20-trifluoroacetate (64.9 mg, 43.8 μmol), (R)-tert-Bu4-DOTAGA (74.9 mg, 109 μmol), and HATU (31.9 mg, 84.7 μmol). The solution was stirred at 50 °C for 1 h and then purified by preparative HPLC, giving compound 33 (43 mg, 23.6 μmol, 53% yield). Compounds 34-46 were prepared in a similar manner.

[0496]

[0497]

[0498]

[0499]

[0500]

[0501]

[0502] A vial was filled with 17-trifluoroacetate (120 mg, 86.8 μmol) and 1 (131 mg, 101 μmol). A solution of HATU (36.0 mg, 95.5 μmol) in DMF (4 mL) was added to the vial. 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.

[0503]

[0504]

[0505]

[0506]

[0507]

[0508] A vial was filled 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. A solution of (R)-tert-Bu4-DOTAGA NHS ester (18.6 mg, 23.3 μmol) in DMF (3 mL) was added to the remaining substance. Diisopropylethylamine (0.5 mL) was added, and the solution was stirred at 50 °C for 30 min, and 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.

[0509]

[0510]

[0511]

[0512]

[0513]

[0514] The vials were filled with 2 (41.0 mg, 40.5 μmol), 27 trifluoroacetate (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. Then, DBU (0.2 mL) was added, and the reaction was stirred at 50 °C for another 10 min. The reaction mixture was purified by preparative HPLC to give 61 trifluoroacetate (23.0 mg, 10.6 μmol, 33% yield). Compounds 62-67 were prepared in a similar manner.

[0515]

[0516]

[0517]

[0518]

[0519]

[0520] A vial was filled with 61 trifluoroacetate (29.0 mg, 14.5 μmol) and a 2 mL solution of (R)-tert-Bu4-DOTAGA NHS ester (16.3 mg, 20.4 μmol) in DMF was added. Diisopropylethylamine (0.2 mL) was added, and the reaction was stirred at room temperature for 18 h. The mixture was 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.

[0521]

[0522]

[0523]

[0524]

[0525]

[0526]

[0527]

[0528] The vials were filled with 5 (17.7 mg, 9.45 μmol) and 28 trifluoroacetate (17.2 mg, 11.6 μmol), and a 2 mL solution of HATU (4.6 mg, 12.3 μmol) in DMF was added. Then, 0.2 mL of diisopropylethylamine was added, and the reaction was stirred at 50 °C for 1 h. The mixture was then purified by preparative HPLC, giving 75 (10.7 mg, 3.57 μmol, 37% yield). Compounds 76-87 were prepared in a similar manner.

[0529]

[0530]

[0531]

[0532]

[0533]

[0534]

[0535]

[0536]

[0537]

[0538]

[0539]

[0540]

[0541]

[0542]

[0543]

[0544]

[0545] Typical procedure for using cold lutetium marking:

[0546] 33 (22.0 mg, 12.1 μmol) was dissolved in trifluoroacetic acid, and the solution was heated to 50 °C for 1 h. Toluene (5 mL) was added, and all solvent was removed under vacuum. A solution of lutetium(III) chloride (6.25 mg, 22.2 μmol) in pH 4.5 HCl / acetate buffer (1 mL) was added to the remaining residue. 0.2 N sodium acetate (1 mL) was added, and the solution was heated to 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).

[0547] The table below lists the starting materials, final products, and major ions found in mass spectrometry used in the above general lutetium conjugation process:

[0548]

[0549]

[0550]

[0551] Example 2: Biodistribution Study

[0552] lutetium accumulation was measured in tumors, plasma, and healthy tissues of mice with NCI-H460 tumors (lung cancer). Mice with NCI-H460 tumors were administered a lutetium conjugate at a dose of 0.5 mg / kg. Mice were sacrificed at specified time points, and tumors, liver, kidneys, and plasma were collected. Luteinium content in all tissues was analyzed by ICP-MS, and lutetium uptake was determined using the following equation, expressed as %ID / g:

[0553] %ID / g = ((tissue lutetium in ppb) / 175) * (molecular weight of the conjugate)) / (0.5 * (mouse weight in grams) * 10)

[0554] The table below shows the biological distribution at 24h and 72h.

[0555]

[0556] The table below shows the biodistribution and tumor / kidney ratio (T / K), tumor / liver ratio (T / L), and tumor / plasma ratio (T / P) at 24 h.

[0557]

[0558]

[0559] In another study using the NCI-H460 tumor model, the distribution of radioactive (thermal) analogues of CMP51 (177Lu) and the distribution of non-radioactive (cold) analogues of CMP51 (175Lu) were tested and compared at 24 and 72 hours. The radioactive analogues showed the same levels of tumor uptake and retention as the cold analogues. Figure 1 As shown, the level of thermal CMP51 in the kidneys was higher than that in the tumor. A high tumor / tissue ratio was observed in other tissues.

[0560] In another study using the NCI-H460 tumor model, the distribution of CMP6 cold analogues was measured at 24 and 72 hours. Figure 2 As shown, CMP6 exhibited significantly reduced renal uptake and retention. Hepatic uptake was also significantly reduced. High tumor-to-liver and tumor-to-kidney ratios were observed. These radioactive effects were enhanced at 72 hours.

[0561] Example 3: In vitro HSP90 binding study using conjugates

[0562] HSP90 binding was determined using purified N-terminal HSP90α via competitive fluorescence polarization assay. A series of dilutions of the test compound were prepared using assay buffer containing 10% DMSO, and 10 μl of the dilution was added to 100 μl of the reaction solution to ensure a final DMSO concentration of 1% in all reactions. The reaction was carried out at room temperature for 3 hours in a mixture containing assay buffer, 5 nM FITC-labeled geldamicin, 350 ng N-terminal HSP90α, and 100 μl of the test compound. Fluorescence intensity was measured using a TecanInfinite M1000 microplate reader at excitation at 485 nm and emission at 530 nm. Fluorescence intensity was converted to fluorescence polarization using TecanMagellan 6 software. The fluorescence polarization data were analyzed using Graphpad Prism software. The fluorescence polarization (FPt) value in each dataset without the compound was defined as 100% activity. The fluorescence polarization (FPb) value in each dataset without the protein and compound was defined as 0% activity. The percentage activity in the presence of the compound was calculated using the following equation: % activity = (FP - FPb) / (FPt - FPb) × 100%, where FP = fluorescence polarization in the presence of the compound. HSP90 binding data are as follows.

[0563] compound HSP90 Kd(nM) T4 0.45 T6 0.45 T8 0.48 T9 0.47 T10 0.77 T11 0.70 T15 0.43

[0564] Example 4: Determination of the permeability of the conjugate

[0565] To test the ability of the conjugate to enter cells, an artificial membrane permeability assay (“PAMPA”) was used. PAMPA is a useful tool for predicting the in vivo drug permeability of drugs entering cells via passive transport mechanisms. LC / MS was used in conjunction with the PAMPA assay to determine the ability of the conjugate to permeate cells.

[0566] Before adding the assay components, pre-coated PAMPA plates were heated to room temperature for at least 30 minutes.

[0567] Stock solutions were prepared using the conjugates to be tested. To prepare working solutions, 50 μL of 100 μM stock solution in DMSO + 950 μL of PBS or 50 μL of 200 μM stock solution was added to a 96-well plate to obtain a final concentration of 5 μM or 10 μM, respectively. 300 μL of working solution containing each conjugate to be tested was added to the appropriate wells of the donor PAMPA plate. 200 μL of PBS was added to the corresponding wells of the recipient PAMPA plate.

[0568] The acceptor plate was lowered onto the donor plate and incubated for 5 hours. After 5 hours, 50 μL aliquots were taken from each well of each plate and added to a new 96-well plate.

[0569] 100 μL of methanol containing a predetermined internal standard compound was added to each aliquot, and the mixture was analyzed by LC / MS. The permeability of each conjugate was calculated.

[0570] Example 5: Synthesis of complexes CMP1-CMP50

[0571] The conjugates of the present invention can be prepared using any convenient method. In a reasonable approach, the conjugates are constructed from their individual components, a targeting moiety, a linker in some cases, and an active substance moiety or a precursor thereof. In some embodiments, the linker or precursor is linked to the targeting moiety or precursor thereof, and subsequently coupled to the active substance moiety or precursor to provide the conjugate. In some embodiments, the linker or precursor is linked to the active substance moiety or precursor thereof, and subsequently coupled to the targeting moiety or precursor to provide the conjugate. Components can be covalently bonded to each other by functional groups as known in the art, wherein such functional groups may be present on the components or introduced onto the components using one or more steps, such as oxidation, reduction, cleavage, etc. Functional groups that can be used to covalently bond components together to produce pharmaceutical conjugates include hydroxyl, thiol, amino, etc. Specific portions of different components modified to provide covalent bonding are selected so as not to adversely interfere with the desired binding activity of the component. For example, for the active substance moiety, regions that do not affect target binding activity are modified to retain sufficient amounts of the desired pharmaceutical activity. When necessary and / or desired, blocking groups known in the art may be used to protect certain portions of the component, see, for example, Green & Wuts, Protective Groups in Organic Synthesis (John Wiley & Sons) (1991).

[0572] Alternatively, conjugates can be generated as follows: a large library of potential conjugates can be produced using known combinatorial methods, which can then be screened to identify bifunctional molecules with pharmacokinetic signatures. Alternatively, conjugates can be produced using medicinal chemistry and known structure-activity relationships between the target moiety and the active moiety. In particular, this method will provide insights into where the two moieties are linked to the linker. Conjugate CMP1-CMP50 can also be synthesized as shown in Example 1. The target moiety, linker, and active moiety of conjugate CMP1-CMP50 can also be synthesized as shown in Example 1.

[0573] The PEG spacer group disclosed herein may be commercially available or can be synthesized by those skilled in the art using conventional organic chemistry. In a reasonable approach, the PEG spacer group may be constructed from polyethylene glycol (PEG). n Construct a variable, where n is an integer between 1 and 20. (PEG) n It can be covalently bonded to functional groups as known in the art, wherein such functional groups may be present in (PEG). n The above or one or more steps are introduced into (PEG). n Above. When necessary and / or desired, protection with a blocking group (PEG) known in the art may be used. nFor certain parts thereof, see, for example, Green & Wuts, Protective Groups in Organic Synthesis (John Wiley & Sons) (1991). The PEG linkers of this disclosure can be homo- or hetero-bifunctional. For example, in some embodiments, the PEG linker may have a carboxylic acid group at one end and an amino group at the other end. PEG linkers can be synthesized using discrete, monodisperse, or polydisperse PEG reagents. The peptide / peptide linkers of this disclosure can be synthesized according to the method shown in Example 1.

[0574] CMP1-CMP50 can be synthesized similarly according to the method for synthesizing the conjugates shown in Example 1. As shown below, CMP2 is synthesized by binding the targeting portion TM10 to 100, then coupling it with (R)-tert-Bu4-DOTAGA and labeling it with cold lutetium. CMP1, CMP22, CMP29-CMP34, and CMP39-CMP42 are prepared in a similar manner.

[0575]

[0576] As shown below, CMP3 was synthesized by conjugating the linker with a suitable DOTA reagent to provide 101, then coupling it with the target moiety TMb, and labeling it with cold lutetium. CMP4-CMP6 and CMP23 were synthesized in a similar manner.

[0577]

[0578] As shown below, CMP10 was synthesized by reacting the targeted portion TM3 with 102, then coupling it with (R)-tert-Bu4-DOTAGA and labeling it with cold lutetium. CMP7-CMP9, CMP11, CMP12, CMP17-CMP21 and CMP24-CMP28 were synthesized in a similar manner.

[0579]

[0580] As shown below, CMP13 was synthesized by conjugating the targeting moiety TM2 with 104 to obtain 105. 105 was then conjugated with peptide 106 synthesized according to the method shown in Example 1, followed by conjugation with (R)-tert-Bu4-DOTAGA and cold lutetium labeling. CMP14-CMP16, CMP35-CMP38, and CMP43-CMP50 were prepared in a similar manner.

[0581]

Claims

1. A conjugate comprising an active substance coupled to at least one targeting moiety (TM) via a linker, wherein the TM binds HSP90, wherein the active substance comprises a radioactive substance or a chelating agent binding a radioactive substance, and wherein the conjugate has a structure of formula X10 or X20: (X10)， (X20)； in, In equations X10 and X20: (a) The linker comprises a spacer group, wherein the spacer group is made of (PEG). n Construct, where n is an integer between 1 and 20; the connector does not contain ; or (b) The active substance is coupled to at least two targeting moieties, wherein (1) the linker comprises a spacer group, wherein the spacer group is selected from 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; or (2) the conjugate comprises at least one pharmacokinetic regulating unit (PMU), wherein the PMU comprises the following structure: (4-(4-iodophenyl)butyrylamino group); Wherein, TM is selected from TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, or TM14, and TM1, TM2, TM3, TM4, TM5, TM8, TM9, TM10, TM11, TM12, TM13, or TM14 are represented by the following structures respectively: The chelating agent in X10 is selected from 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA); The chelating agent in X20 is selected from 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, α-(2-carboxyethyl) (DOTAGA); The radioactive materials include Y-90, Bi-212, Lu-177, Ac-225, or Bi-213. The condition is that the conjugate is not .

2. The conjugate according to claim 1, wherein the conjugate comprises a structure of formula A10 or formula A20: (A10)， (A20)。 3. The conjugate according to claim 1 or 2, wherein the TM is selected from TM1, TM5, TM9, TM10, TM11, TM12 or TM13.

4. A conjugate, which is T4, T5, T8, T9, T10, CMP44, CMP48, T4', T5', T8', T9', T10', CMP44', CMP48', or a pharmaceutically acceptable salt thereof, said conjugate having the following structure:

5. The conjugate according to claim 1, wherein the linker comprises a spacer group, wherein the spacer group comprises polyethylene glycol (PEG).

6. The conjugate according to claim 5, wherein the spacer group is (PEG)4 or (PEG)12.

7. The conjugate according to claim 5, wherein the conjugate comprises a structure of formula B10 or formula B20: (B10)， (B20)。 8. The conjugate according to any one of claims 5-7, wherein the TM is selected from TM1, TM5, TM9, TM10, TM11, TM12 or TM13.

9. The conjugate according to claim 5, wherein the conjugate 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', or a pharmaceutically acceptable salt thereof, having the following structural formula:

10. The conjugate according to claim 9, wherein the conjugate is CMP31': ， Or its pharmaceutically acceptable salt.

11. The conjugate according to claim 9, wherein the conjugate is CMP31: ， Or its pharmaceutically acceptable salt.

12. The conjugate according to claim 1, wherein the conjugate comprises at least one pharmacokinetic regulating unit (PMU), the PMU being... 。 13. The conjugate according to claim 12, wherein the conjugate comprises a structure of formula C10 or formula C20: (C10)， (C20)。 14. The conjugate according to any one of claims 12-13, wherein the TM is selected from TM1, TM5, TM9, TM10, TM11, TM12 or TM13.

15. The conjugate according to claim 12, wherein the conjugate is CMP3, CMP4, CMP5, CMP6, T3, T7, T11, T12, T15, CMP3', CMP4', CMP5', CMP6', T3', T7', T11', T12', T15', with the following structure: Or its pharmaceutically acceptable salt.

16. The conjugate according to claim 1, wherein the TM is selected from TM1, TM5, TM9, TM10, TM11, TM12 or TM13.

17. The conjugate according to claim 1, wherein the conjugate is CMP8, CMP24, CMP27, CMP28, CMP8', CMP24', CMP27', or CMP28'. Or its pharmaceutically acceptable salt.

18. A pharmaceutical composition comprising the conjugate of any one of claims 1-17 and at least one pharmaceutically acceptable excipient.

19. Use of the pharmaceutical composition of claim 18 in the preparation of a medicament for treating lung cancer.

20. The application according to claim 19, wherein the lung cancer is small cell lung cancer or non-small cell lung cancer.

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