Small zwitterionic multimeric targeted drugs and methods of use and manufacture thereof

Abstract

In accordance with at least one aspect of this disclosure, there is provided a small zwitterionic multimeric targeted anti-cancer drug which effectively addressed tumor cell heterogeneity and off-target / non-specific binding and uptake. The small zwitterionic multimeric targeted anti-cancer drug includes a nexus, three different targeting ligands conjugated to the nexus, a payload of a zwitterionic diagnostic or therapeutic agent, and zwitterionic flexible linkers connecting each of the three different targeting ligands and the pay load to the nexus.

Description

[0001] SMALL ZWITTERIONIC MULTIMERIC TARGETED DRUGS AND METHODS OF USE AND MANUFACTURE THEREOF

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U.S. Provisional Patent Application Serial No. 63 / 648,305 (Atty Doc. No. LL 1515138.111US0), filed on May 16, 2024; U.S. Provisional Patent Application Serial No. 63 / 658,121 (Atty Doc. No. LL 1515138.111US02), filed on June 10, 2024; U.S. Provisional Patent Application Serial No. 63 / 770,120 (Atty Doc. No. LL 1515138.111US03), filed on March 11, 2025. The disclosures of the applications are incorporated herein by reference.

[0004] This application also claims priority to U.S. Provisional Patent Application Serial No. 63 / 724, 707(Atty Doc. No. LL 1515138.116US02), filed on November 25, 2024, and U.S. Provisional Patent Application Serial No. 63 / 767, 817( Atty Doc. No. LL 1515138.121US0), filed on March 6, 2025. The disclosures of the applications are also incorporated herein by reference.

[0005] The disclosure of PCT applications PCT / US2024 / 045968 (Atty Doc. No. LL 1515138.105WO2), filed on September 10, 2024, PCT / US2024 / 045995 (Atty Doc. No. LL

[0006] 1515138.106WO2), filed on September 10, 2024, PCT / US2024 / 046072 (Atty Doc. No. LL

[0007] 1515138.107WO2), filed on September 11, 2024, PCT / US2024 / 046105 (Atty Doc. No. LL

[0008] 1515138.109WO2), filed on September 11, 2024, are hereby incorporated by references.

[0009] SEQUENCE LISTING

[0010] A Sequence Listing conforming to the rules of WIPO Standard ST.26 is hereby incorporated by reference. Said Sequence Listing has been filed as an electronic document via PatentCenter encoded as XML in UTF-8 text. The electronic document, created on April 28, 2025, is entitled “1515138_l l lUS2_SL.xml”, and is 32,908 bytes in size.

[0011] FIELD OF THE INVENTION The present invention relates to a small zwitterionic molecule drug having more than one targeting ligand binding to more than one disease-rel ted receptor on, e.g., receptors on tumor cells, and thus addresses diseases’ heterogeneity and background reduction more effectively than traditional drugs.

[0012] BACKGROUND

[0013] Many diseases involve the expression of abnormal receptors or antigens that are not present in healthy individuals. For example, tumor receptors on cancer cells and antigens on disease-induced pathogens are unique markers that distinguish diseased cells from normal cells. These abnormal receptors and antigens provide a critical target for diagnosis and therapy. Drugs designed to specifically recognize and bind to these molecules can selectively target and diagnose and / or treat the diseased cells while sparing healthy cells. For diagnosis, this approach improves the tumor-to-background ratio. For therapy, this approach enhances the efficacy of treatments and reduces side effects, offering a promising strategy for managing various diseases, including cancer and infectious diseases.

[0014] However, while targeting abnormal receptors or antigens on diseased cells or pathogens offers a promising therapeutic strategy, the heterogeneity of these receptors or antigens can significantly impede the effectiveness of such treatments. This heterogeneity means that within a single tumor or pathogens population, there can be variations in the receptors or antigens expressed. For instance, in cancer, different cells within the same tumor may express different levels or types of tumor receptors, leading to some cells being targeted by the drug while others evade treatment. Similarly, the human immunodeficiency virus (HIV) exhibits high variability in its antigens, which complicates the development of a universally effective vaccine. Additionally, bacterial pathogens such as Staphylococcus aureus can express different surface proteins that contribute to antibiotic resistance. This variability within and between pathogen populations and cancer cells underscores the challenge of developing drugs that can effectively target all disease-related receptors, highlighting the need for more sophisticated and adaptable diagnostic and therapeutic approaches.

[0015] Cancer, for example, is one of the diseases with the most severe consequences if treatment fails due to heterogeneity of the tumor cell. Cancer heterogeneity or tumor heterogeneity describes the observation that different tumor cells can show distinct morphological and phenotypic profiles, including cellular morphology, gene expression, metabolism, motility, proliferation, and metastatic potential.

[0016] Cancer / tumor heterogeneity is a common characteristic of cancer that can make it difficult for cancer imaging and treatment drugs to be effective. It can refer to differences between tumors in different patients, between cancer cells in a single tumor of one patient, or between primary and secondary tumors. Cancer heterogeneity can lead to drug resistance and therapeutic failure in several ways. First, cancer heterogeneity can directly affect therapeutic targets. Second, cancer heterogeneity can influence the tumor microenvironment (TME), which can then affect drug resistance. In addition, cancer heterogeneity can cause uneven distribution of genetic diversity within tumors, which can lead to drug resistance.

[0017] Moreover, cancer / tumor heterogeneity can occur over the life span of the tumor. For example, the cellular morphology and gene expression of the tumor may change over time through clonal evolution, and thus a drug which may be effective in treating the tumor at the beginning of treatment may lose its effectiveness after a treatment period. Cancer / tumor heterogeneity may also affect the correct and efficient diagnosis of the cancer / tumor due to a lower binding rate of the imaging diagnosing agents to the surfaces of cancer / tumor cells.

[0018] Another problem of paramount importance but unaddressed is off-target, aka nonspecific binding and uptake of anti-cancer drugs. Most anti-cancer drugs are hydrophobic and / or sticky, and make weak but significant interactions with normal, non-cancerous cells and tissues. Off-target binding and uptake leads to a high background thus a low tumor-to- background ratio (TBR) when imaging cancer cells and a low therapeutic window when treating cancer cells. This problem also exists for some other diseases involving the nonspecific binding and uptake of targeted drugs.

[0019] Accordingly, it is imperative to design drugs that can address the heterogeneity present in various diseases, as well as background reduction. By developing therapeutic agents that can adapt to or target multiple variations of abnormal receptors or antigens, one can improve the efficacy of treatments and reduce the likelihood of disease recurrence and progression. This approach is particularly crucial in the context of cancer, where the diverse nature of tumor cells can lead to treatment resistance and severe consequences if not effectively managed. Similarly, rendering targeted drugs less prone to non-specific uptake and binding, and ideally rapid elimination by renal filtration, will reduce background significantly. Addressing heterogeneity and background reduction in drug design not only enhances the potential for successful treatment outcomes but also paves the way for more personalized and precise medical interventions, ultimately improving patient prognosis and quality of life.

[0020] SUMMARY

[0021] The present invention is directed, at least in part, to a receptor binding drug comprising a nexus; multiple targeting ligands conjugated to the nexus through linkers; and a payload molecule conjugated to the nexus, wherein each of the multiple targeting ligands is conjugated to the nexus via a linker comprising one or more zwitterionic groups, which reduce non-specific binding and uptake, and wherein each of the multiple targeting ligands binds to at least one receptor or antigen on tumor cells.

[0022] In one embodiment, each of the multiple targeting ligands and the payload molecule is conjugated to the nexus via a linker comprising one or more zwitterionic groups, wherein the one or more zwitterionic groups are ammonium sulfobetaine groups or N-oxide groups or a combination thereof.

[0023] In one embodiment, the nexus having multiple attaching sites.

[0024] In one embodiment, the nexus is a trihedral core, a tetrahedral core, a pentahedral core, or a hexahedral core.

[0025] In one embodiment, the nexus has a rigid structure.

[0026] In one embodiment, the nexus is a branching point comprising two or more amino acids.

[0027] In one embodiment, each of the amino acids is either Lys or Glu.

[0028] In one embodiment, at least one of the multiple targeting ligands is different from the others of the multiple targeting ligands.

[0029] In one embodiment, the multiple targeting ligands comprise three targeting ligands, and wherein each of the multiple targeting ligands is different from the others of the multiple targeting ligands.

[0030] In one embodiment, each of the multiple targeting ligands is cRGD, dPSMA-617, MUC1, ATN-161, KUE, a FAP binding vector, octreotide, or bombesin.

[0031] In one embodiment, the payload molecule is a zwitterionic molecule. In one embodiment, the payload molecule comprises one or more of ZW700-Forte, ZW800-1 , ZW83O-1 , ZN-800-1 , ZN-830-1 , ZN- 800-1 -Forte, ZN-700- 1 -Forte, ZW-DOTA, ZW-PyC3A, ZW-Macropa, ZW-porphyrin, ZW-NOTA, and ZW-deferoxamine.

[0032] In one embodiment, the payload molecule comprises one or more of ZW-DOTA-Cu- 64, ZW-DOTA-Cu-67, ZW-DOTA-Gd, ZW Poryphyrin-Mn3+.

[0033] In one embodiment, the linker is a flexible zwitterionic linker.

[0034] In one embodiment, the linker comprises at least one azido derivative.

[0035] In one embodiment, the tumor cells demonstrate heterogeneity.

[0036] In one embodiment, at least one of the multiple targeting ligands is configured to bind to at least one cancer-independent target, and at least one of the multiple targeting ligands is configured to bind to at least one cancer-dependent target.

[0037] In one embodiment, the payload molecule comprises a base payload and a special payload; wherein the base payload comprises a zwitterionic molecule; and the special payload comprises a metal molecule.

[0038] In one embodiment, the at least one cancer -independent target comprises one or more of FAP, integrin a5pi, and integrin avP3; and the at least one cancer-dependent target comprises one or more of GRPR, MUC 1 , and PSMA.

[0039] In one embodiment, the nexus has a first attaching site conjugated to a first targeting ligand via a first flexible zwitterionic linker, a second attaching site conjugated to a second targeting ligand via a second flexible zwitterionic linker, and a third attaching site conjugated to a payload via a third flexible zwitterionic linker, wherein the payload comprises one of ZW700-Forte, ZW800-1, ZW830-1, ZN-800-1, ZN-830-1, ZN-800-1 -Forte, ZN-700- 1 -Forte, ZW-DOTA, ZW-PyC3A, ZW-Macropa, ZW-porphyrin, ZW-NOTA, ZW-deferoxamine, ZW-DOTP, ZW-PCTA, ZW-Amide-PCTA, and ZW-PTCP.

[0040] In another aspect of the invention, the disclosure provides a method of imaging tissue, cells, or lumen in a subject, the method comprising: (a) contacting the tissue, cells or lumen with a composition according to Claim 1 to the subject, (b) irradiating the tissue, cells, or lumen at a wavelength absorbed by the dye; (c) and detecting a signal from the imaging agent, thereby imaging the tissue, cells, or lumen.

[0041] In another aspect of the invention, the disclosure provides, a method of treating a tumor by administering an effective amount of a small zwitterionic trimeric targeted anticancer drug, wherein the method comprises a step to diagnose the cancer and a step of administering the therapeutic agent to a subject determined to be in need thereof; wherein the step to diagnose the cancer comprises: contacting cells, tissues or organs of a subject with an imaging agent, imaging the cells, tissues, or organs of the subject using positron emission tomography (PET), single-photon emission computerized tomography (SPECT), or magnetic resonance imaging (MRI), and diagnosing the cancer in the cells tissues, or organs of the subject based on imaging data collected; and wherein the anti-cancer drug comprises at least one zwitterionic molecule as the pay load wherein a metal atom complexed to the zwitterionic molecule is: a radioactive metal isotope known to emit ionizing radiation that results in the death of cells that take up the analogs; or a non-radioactive metal that is capable of releasing cytotoxic radiation upon irradiation with alpha emission, beta emission, neutron capture, or a combination thereof.

[0042] Other features and advantages of the invention will be apparent from the following detailed description, and from the claims.

[0043] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0044] BRIEF DESCRIPTION OF THE DRAWINGS

[0045] So that those skilled in the art to which the subject disclosure appertains will readily understand how to make and use the devices and methods of the subject disclosure without undue experimentation, other embodiments thereof will be described in detail herein below with reference to certain figures, wherein:

[0046] Figure 1 shows a schematic overview of the structure of a small zwitterionic multimeric targeted drug of the invention. Figure 2 shows a schematic overview of the structure of a small zwitterionic multimeric targeted drug with a “force field” of zwitterionic charge and water of hydration.

[0047] Figure 3 shows a schematic of how the expression of various binding receptors evolves over time in a solid tumor.

[0048] Figure 4 shows small zwitterionic multimeric targeted drug’s pharmacokinetic advantage over traditional antibody-drug conjugates.

[0049] Figure 5 shows a process of synthesizing two Tri-FABNO Derivatives [Fmoc (4) and Allyl (6)].

[0050] Figure 6 shows a process of synthesizing two Tri-FLNO Derivatives [Fmoc (4) and Allyl (6)].

[0051] Figure 7 shows a schematic overview of the structure of a small zwitterionic multimeric targeted drug with illustrations of detailed functional groups.

[0052] Figure 8 shows a schematic overview of the structure of targeting ligand 1 prepared for synthesis of a small zwitterionic multimeric targeted drug.

[0053] Figure 9 shows a synthesis process of a payload-less small zwitterionic multimeric targeted drug (CPI-000).

[0054] Figure 10 shows synthesis processes of small zwitterionic multimeric targeted drugs with different payloads from a payload-less small zwitterionic multimeric targeted drug (CPI- 000)

[0055] Figure 11 shows a schematic overview of the structure of a small zwitterionic multimeric targeted drug with four targeting ligands and one payload.

[0056] Figure 12 shows interactions between the small zwitterionic multimeric targeted drug and the cancer cells I its supporting tissues.

[0057] DETAILED DESCRIPTION

[0058] It has now been found that compounds comprising a small zwitterionic, multimeric receptor-binding targeted drug with multiple different targeting ligands which addresses both disease heterogeneity and background reduction, in both diagnostic and therapeutic applications.

[0059] Definitions

[0060] The following definitions will be useful in understanding the instant invention.

[0061] As used herein, the term “comprising” is intended to mean that the compositions and methods include the recited elements, but do not exclude other elements. “Consisting essentially of”, when used to define compositions and methods, shall mean excluding other elements of any essential significance to the combination. Thus, a composition consisting essentially of the elements as defined herein would not exclude trace contaminants from the isolation and purification method and pharmaceutically acceptable carriers, such as phosphate buffered saline, preservatives, and the like. “Consisting of” shall mean excluding more than trace elements of other ingredients and substantial method steps for administering the compositions of this invention. Embodiments defined by each of these transition terms are within the scope of this invention.

[0062] As used in the specification and claims, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise.

[0063] Ranges provided herein are understood to be shorthand for all of the values within the range. For example, a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting of 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, and 50.

[0064] Unless specifically stated or obvious from context, as used herein, the term "or" is understood to be inclusive.

[0065] The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable or aspect herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof. As used herein, the term “subject” or “patient” encompasses mammals and nonmammals. Examples of mammals include, but are not limited to, humans, chimpanzees, apes monkeys, cattle, horses, sheep, goats, swine; rabbits, dogs, cats, rats, mice, guinea pigs, and the like. Examples of non-mammals include, but are not limited to, birds, fish, parasites, microbes, and the like.

[0066] As used herein, the term “administration” or “administering” of the subject compound refers to providing a combination composition of the invention and / or prodrugs thereof to a subject in need of diagnosis or treatment.

[0067] As used herein, the term “ligand”, “targeting vector”, or “targeting ligand” refers to a moiety which is bound to or coordinated to the therapeutic / imaging agents or zwitterionic metal chelators of the composition of the invention to provide enhanced binding to particular cell types or an increased concentration in the presence of particular cell types. In certain embodiments, the targeting vector can be bound to the therapeutic / imaging agents or zwitterionic metal chelators of the compositions in addition to the zwitterionic groups thereon. In still other embodiments, the targeting vector can be bound to the zwitterionic metal chelator in place of one or more zwitterionic groups provided that the zwitterionic metal chelator retains at least one zwitterionic group.

[0068] As used herein, the term “therapeutic window” or “therapeutic index” refers to the relationship between the therapeutic and toxic dose of a given drug and is calculated using the ED50 and TD50 (Therapeutic Index = TD50 / ED50). In certain embodiments of the invention, the zwitterionic metal chelators of the invention have a higher therapeutic index relative to other metal chelators. In certain other embodiments, the therapeutic window refers to a certainty safety factor (CSF) which is defined herein as the ratio of [TD1 / ED99]. A CSF > 1 indicates that the dose effective in 99% of the population is less than the dose that would be toxic in 1% of the population. In certain embodiments of the invention, the combination compositions of the invention have a higher CSF relative individual active agents.

[0069] As used herein, the term “carrier” refers to chemical compounds or agents that facilitate the incorporation of a compound described herein into cells or tissues.

[0070] As used herein, the term “acceptable” with respect to a formulation, composition or ingredient, as used herein, means having no persistent detrimental effect on the general health of the subject being treated.

[0071] As used herein, the term “azido derivative(s)” refers to chemical compounds that contain one or more azido groups (-Ns) attached to their molecular structure. In certain embodiments, the azido group is a functional group consisting of three nitrogen atoms (N) connected in a linear arrangement, typically represented as -Ns. In certain embodiments, the azido derivatives include one or more of Fmoc-Azidoalanine, Fmoc-Azidohomoalanine, Fmoc-Azidophenylalanine, Fmoc-Azidoglycine, Fmoc-Azidoomithine, Fmoc-Azidolysine, Fmoc-Azidoaspartic Acid.

[0072] As used herein, the term “diluent” refers to chemical compounds that are used to dilute a compound described herein prior to delivery. Diluents can also be used to stabilize compounds described herein.

[0073] As used herein, the term “contacting” refers to the bringing together of substances in physical contact such that the substances can interact with each other. For example, when an agent is “contacted” with tissue or cells, the tissue or cells can interact with the agent, for example, allowing the possibility of binding interactions between the agent and molecular components of the tissue or cells. “Contacting” is meant to include the administration of a substance such as an agent of the invention to an organism. Administration can be, for example, oral or parenteral.

[0074] As used herein, the term “ionic group” refers to a moiety comprising one or more charged substituents. The “charged substituent” is a functional group that is generally anionic or cationic when in substantially neutral aqueous conditions (e.g. a pH of about 6.5 to 8.0 or about physiological pH (7.4)). As recited above, examples of charged anionic substituents include anions of inorganic and organic acids such as sulfonate (-SO31), oxide, sulfinate, carboxylate, phosphinate, phosphonate, phosphate, and esters (such as alkyl esters) thereof. In some embodiments, the charged substituent is sulfonate or oxide. Examples of charged cationic substituents include quaternary ammonium ions (-NR3+) and phosphonium ions (-PR3+), where R is independently selected from C1-6 linear alkyl, C4-6 branched alkyl, C3-6 cycloalkyl, aryl, heteroaryl and arylalkyl or heteroarylalkyl. Other charged cationic substituents include protonated primary, secondary, and tertiary amines, and as well as guanidinium or amidinium or pyridinium or other protonated, alkylated or oxygenated nitrogen heterocycles. In some embodiments, the charged substituent is -N(CH3)3+.

[0075] As used herein, the phrase “non-ionic multimeric or polymeric solubilizing groups” refers to soluble polymers such as, for example, polyethylene glycol, polypropylene glycol, polyethylene oxide and propylene oxide copolymer, a carbohydrate, a dextran, polyacrylamide, a peptide and the like. The solubilizing group can be attached by any desired mode. The point of attachment can be, e.g., a carbon-carbon bond, a carbon-oxy gen bond, or a nitrogen-carbon bond. The attachment group can be, e.g., an ester group, a carbonate group, an urea group, an alcohol group, an ether group, a sulfide group, an amino group, an alkylene group, an alkyne group, an azide group, a tetrazine, an amide group, a carbonyl group, or a phosphate group.

[0076] Some examples of solubilizing groups include polyethylene glycols, such as -(CH2CH2O)a-H, -OC(=O)O(CH2CH2O)aH, -OC(=O)O(CH2CH2O)aCH3, - O(CH2CH2O)aCH3, and -S(CH2CH2O)2CH3, “a” being an integer between about 2 and about 250. In some embodiments, “a” is 4 to 12 or 5 to 10. In further embodiments, “a” is 6, 7, or 8. Other examples of solubilizing groups include dextrans such as -OC(=O)O(dextran).

[0077] The solubilizing moiety can have an absolute molecular weight of from about 500 amu to about 100,000 amu, e.g., from about 1,000 amu to about 50,000 amu or from about 1,500 to about 25,000 amu.

[0078] Further examples of solubilizing groups include: -(CH2)c-(OCH2CH2)d-ORa, wherein “c” is 0 to 6, “d” is 1 to 200, and Rais H or Ci-6 alkyl. In some embodiments, “c” is 1 to 4, “d” is 1 to 10, and Rais H. In some embodiments, “d” is 6 or 7.

[0079] See WO 2008 / 017074, U.S. Ser. No. 12 / 376,243 (filed February 3, 2009), and U.S. Ser. No. 12 / 376,225 (filed February 3, 2009), each of which is incorporated herein by reference in its entirety, for a further description of suitable non-ionic oligomeric or polymeric solubilizing groups, and method for incorporating them into dyes. It is further appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, can also be provided in combination in a single embodiment. Conversely, various features of the invention which are, for brevity, described in the context of a single embodiment, can also be provided separately or in any suitable subcombination.

[0080] Compounds of the invention can also include all isotopes of atoms occurring in the intermediates or final compounds. Isotopes include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium.

[0081] The chemical substances represented herein by name, chemical formula, or structure are meant to include all stereoisomers, geometric isomers, tautomers, resonance structures, and isotopes of the same, unless otherwise specified.

[0082] The chemical substances described herein may be charged or include substituents with formal charges. When such chemical substances are represented as charged, it is understood that, unless otherwise specified, the charges are generally countered with an appropriate counterion. For example, chemical substances or functional groups having a charge of -I are understood to be countered with an ion have a +1 charge. Suitable counterions with +1 charge include Na+, K+, tetraalkylammonium ions, and the like. Conversely chemical substances or functional groups having a charge of + 1 are understood to be countered with an ion having a -1 charge. Suitable counterions with -1 charge include F-, C1-, Br-, I-, perchlorate, acetate, trifluoroacetate, and the like.

[0083] As used herein, the term “zwitterionic group,” “zwitterionic ligand,” or “zwitterion” refer to one or more charged moieties with balanced (electrically neutral) polyionicity. The zwitterionic feature could be part of the linker and / or payload and / or targeting ligand. The zwitterionic multimeric drugs of the claimed invention are decorated by one or more zwitterionic groups. In the case of a payload that is a metal chelator, these zwitterionic groups are distinct from metal chelator cores, which typically has negative charges to chelate positively-charged metals. For example, a zwitterionic metal chelator with a chelator core of - 4 that binds a +4 metal would have a total net charge of zero. Although a total net charge of zero is considered ideal, a zwitterionic metal chelator with a chelator core of -4 that binds a +2 metal, resulting in an overall charge of -2, would still he expected to exhibit improved properties in vivo because of shielding of the chelator core / metal complex by one or more zwitterionic groups. In the absence of zwitterionic groups, the molecule would have no such shielding or expanded water of hydration and would be more likely to bind non-specifically.

[0084] A particular active agent molecule may have several attached "zwitterionic groups” or charge pairs. In general, the anion portion and the cation portion of the zwitterionic group (charge pair) will be part of the same moiety, though it is possible for two ionic groups to be used as separate moieties to form a zwitterionic group. In particular embodiments, the zwitterionic group is covalently bound to the base structure via a carbon-carbon bond, a carbon-oxygen bond, or a nitrogen-carbon bond. Examples of zwitterionic groups (charge pairs) that can be included in the compounds and complexes of the claimed invention include, but are not limited to, ammoniophosphates, ammoniophosphonates, ammoniophosphinates, ammoniosulfonates, ammoniosulfates, ammoniocarboxylates, ammoniosulfonamides, ammonio-sulfon-imides, guanidiniocarboxylates, pyridiniocarboxylates, pyridiniosulfonates, ammonio(alkoxy)dicyanoethenolates, ammonioboronates, sulfoniocarboxylates, phophoniosulfonates, and phosphoniocarboxylates. The charged groups in these zwitterions can be separated by suitable spacer groups like linear or branched alkyl chains, aryl or heteroaryl moieties. In certain embodiments, the zwitterionic groups can be derivatives of amino acids, such as amino carboxylic acids, amino phosphonic acids, amino phosphinic acids or amino sulfonic acids, furthermore, aminoalkyl substituted sulfates or phosphates. Zwitterions can also be derivatives of betaines, such as carboxybetaines, sulfobetaines, sulfabetaines, phosphobetaines or phosphabetaines or N-oxides or derivatives of sulfamic acid. Particular examples of zwitterionic groups include ammonium sulfobetaines or N- oxides. A simple example of a zwitterionic group at physiological pH is the charge pair of a carboxylic acid (deprotonated at physiological pH) and an amine (protonated at physiological pH).

[0085] In some embodiments, the targeting ligands of the zwitterionic multimer drugs of the invention can also comprise a targeting vector for an agricultural process, chemical process, disease, or tissue-specific epitope, such as the cyclic peptide cRGDyK (aka cRGD). cRGD is a cyclic derivative of the tripeptide Arg-Gly-Asp which can be conjugated to one or more arms of the metal chelators of the invention. In still other embodiments, the targeting vector is octreotide or bombesin. Tn other embodiments, the targeting vector is KUE or dPSMA- 617, a small molecule capable of targeting Fibroblast Activation Protein (FAP) also called FAP-inhibitor or FAPI, an amino acid or combination of amino acids, or derivatives thereof. In such embodiments, the targeting vector-conjugates can be formed in place of one or more zwitterionic groups. In certain embodiments, the targeting ligand includes one or more of LyP-1 peptide having a sequence of CGQKRTRGC (SEQ ID NO: 1) and binding to P32 for diagnosing / treating melanoma; K237 peptide having a sequence of HTMYYHHYQHHL(SEQ ID NO: 2) and binding to VEGFR-2 for diagnosing / treating breast tumor; IL4RPep-l peptide having a sequence of CRKRLDRNC(SEQ ID NO: 3) and binding to IL4R for diagnosing / treating lung tumor, breast tumor, colon tumor; mUNO peptide having a sequence of CSPGAK (SEQ ID NO: 4) and binding to CD206 for diagnosing / treating breast tumor; folate receptors for diagnosing / treating ovarian and lung cancer; GE11, a dodecapeptide, binding to epidermal growth factor receptor (EGFR or ErbB l) for diagnosing / treating tumors of epithelial origin.

[0086] An ideal active agent conjugated to a targeting vector would adopt the total net charge of the targeting vector, which is purposeful because in most cases the charges on the targeting vector are crucial for the ability to bind its target. Targeted zwitterionic trimer drugs thus retain the major advantage of minimizing non-specific binding while maximizing specific binding. It should be apparent to those skilled in the art that additional charges can be added to the zwitterionic multimer drugs, if needed, to balance overall surface charge to zero.

[0087] In certain embodiments, the zwitterionic multimeric drugs comprise a reactive conjugation group. Such reactive conjugation groups are typically an activated derivative of a carboxylic acid, such as an n-hydroxysuccinimide (NHS) ester, a sulfo-NHS ester, a pentafluorophenyl (PFP) ester, a hydroxybenzotriazole (HOBt) ester, a hydroxyazabenzotriazole (HO At) ester, a tetrafluorophenyl (TFP) ester, an acid anhydride, an acid azide or an acid halide. Such reactive conjugation groups can be bound or substituted onto the chelator at any suitable structural location as would be understood by one of ordinary skill in the synthesis of such compounds. Reactive conjugation groups also include, but are not limited to, alkynes, azides, maleimides, thiols, amines, alkohols, phenols, carbonyls, phosphanes, alkenes and tetrazines. As used herein, the phrase “core”, “central core”, “branching point” or “nexus” refers to a chemical structure which is located at the center of the drug with four or more substituents for binding zwitterionic linkers and / or targeting ligands. The nexus or “branching” may also be distributed in a more polymer- like configuration.

[0088] As used herein, the phrase “linker” refers to a chemical structure connects two or more functional groups.

[0089] As used herein, the phrase “payload” refers to a diagnosis or a treatment functional group. When being used as a treatment functional group, the payload refers to, for example, a potent cytotoxin drug that kills cancer cells. Another example would be a payload that radiosensitizes the malignant cell to radiation. When being used as a diagnosis functional group, the payload refers to an imaging agent that provides a signal facilitating the identification of cancer cells.

[0090] Other definitions appear in context throughout the disclosure.

[0091] Diseases, Related Receptors, and Zwitterionic Multimeric Targeted Drugs

[0092] The present invention is directed to a small zwitterionic multimeric receptor / antigen- binding drug with multiple targeting ligands, which addresses diseases’ heterogeneity efficiently. That is, each of the multiple targeting ligands of the small zwitterionic multimeric drug binds to one type of receptor or antigen expressed in diseases related cells or tissues. As such, by having multiple different targeting ligands, the small zwitterionic multimeric drug can bind to more than one type of receptors / antigens expressed in diseases related cells and tissues, and thus addresses the heterogeneity demonstrated by the diseases.

[0093] In certain embodiments, the diseases can be treated by the small zwitterionic multimeric receptor-binding drug of the present invention include infectious diseases, cancerous diseases, genetic and congenital disorders, nutritional and metabolic diseases, cardiovascular diseases, respiratory diseases, neurological disorders, endocrine disorders, gastrointestinal diseases, musculoskeletal disorders, autoimmune diseases, dermatological diseases, psychiatric disorders, hematologic diseases, renal and urological diseases, and reproductive system diseases. The targeting ligands of the present invention can be one or more of the following: antibiotics, anti-fungal ligands, anti-viral ligands, immune suppressors, angiogenesis inhibitors; anti-pain medicines (i.e., analgesics); anesthetics; hypertensive medications (blood pressure meds); anticoagulants; cardiac stimulants; cardiac suppressors (blockers); insulin- derived drugs; and muscle relaxants.

[0094] By identifying disease related receptors, and choosing proper targeting ligands binding to these receptors, the small zwitterionic multimeric receptor- binding drug of the present invention can be used as analgesics; anesthetics; anti- addiction agents; antibacterials; anticonvulsants; antidementia agents; antidepressants; antiemetics; antifungals; antigout agents; antimigraine agents; antimy asthenic agents; antimycobacterials; antineoplastics; antiparasitics; antiparkinson agents; antipsychotics; antispasticity agents; antivirals; anxiolytics; bipolar agents; blood glucose regulators; blood products; cardiovascular agents; central nervous system agents; dental and oral agents; dermatological agents; electrolytes, minerals, metals, vitamins; gastrointestinal agents; genetic / enzyme / protein disorder agents; genitourinary agents; hormonal agents including adrenal, pituitary, prostaglandins, sex hormones, thyroid; hormone suppressant including adrenal, pituitary, thyroid; immunological agents; inflammatory bowel disease agents; metabolic bone disease agents; ophthalmic agents; otic agents; respiratory tract agents; skeletal muscle relaxants; sleep disorder agents.

[0095] It should be noted that cancer is used as an example below to illustrate the structure, function, and working principles of the small zwitterionic multimeric receptor-binding drug of the present invention, as well as its manufacturing process. A skilled artisan can modify this drug by replacing the multiple targeting ligands that bind to cancer-related receptors with targeting ligands that bind to receptors associated with other diseases, such as those on pathogens. This modification would yield a small zwitterionic multimeric receptor-binding drug applicable to other diseases.

[0096] General Structure of Anti-Cancer Drugs It has now been found that certain small zwitterionic multimeric targeted anti-cancer drugs are effective in diagnosing and treating cancers / tumors demonstrating tumor heterogeneity.

[0097] Any compositions or methods provided herein can be combined with one or more of any of the other compositions and methods provided herein.

[0098] In one aspect, the present invention is directed to a small zwitterionic heteromultimeric receptor-binding drug with multiple targeting ligands, which addresses tumor heterogeneity efficiently.

[0099] In another aspect, the present invention is directed to a zwitterionic hetero-multimeric small molecule drug with multiple targeting ligands, which effectively addresses receptor- involved diseases having more than one type of receptors involved.

[0100] In certain embodiments, the structure of the small zwitterionic multimeric targeted anti-cancer drug includes a nexus, more than one targeting ligands, linkers, a payload for imaging or treating the tumors, and optionally spacing groups. Figure 1 shows a generalized structure of the small zwitterionic multimeric targeted anti-cancer drug, particularly a small zwitterionic trimeric targeted drug with three targeting ligands. Figure 11 shows a generalized structure of the small zwitterionic multimeric targeted anti-cancer drug of a different embodiment, particularly a small zwitterionic trimeric targeted drug with four targeting ligands, and a payload having a zwitterionic chelator (base payload) and isotope (special payload). A forcefield of hydration and balanced charge are formed around the nexus and linkers conjugating the targeting ligands to the nexus, while the targeting ligands are exposed out of the forcefield such that they can bind to targets on cancer cells or cancer- associated tissue. In this embodiment showed in Figure 1 1 , the targeting ligands include MUC1 binding receptor, FAP, Bombesin binding receptor, and

[0101] In certain embodiments, a small zwitterionic multimeric targeted drug includes a nexus having multiple attaching sites, multiple targeting ligands conjugated to the nexus via the multiple attaching sites, one or more payloads of a zwitterionic diagnostic or therapeutic agent conjugated to the nexus via one of the multiple attaching sites, and zwitterionic flexible linkers connecting each of the multiple different targeting ligands and the payload to the nexus. In certain embodiments, in addition to the zwitterionic flexible linkers, one or more spacing groups e.g. Ahx locate between a targeting ligand and the nexus.

[0102] In certain embodiments, each of the multiple targeting ligands is conjugated to one of the attaching sites of the nexus independent of other multiple targeting ligands. That is, each of the multiple targeting ligands is conjugated to the nexus rather than being conjugated to other multiple targeting ligands. For example, when a nexus has n attaching sites, n-1 targeting ligands are conjugated to the nexus. In particular, one attaching site is conjugated to the payload, with each of n-1 targeting ligands conjugated to one of the n-1 attaching sites. In another example, when a nexus has m attaching sites, m-2 targeting ligands are conjugates to the nexus. In particular, two attaching sites are conjugated to two payloads, with each of the m-2 targeting ligands conjugated to one of the m-2 attaching sites.

[0103] In certain embodiments, a small zwitterionic multimeric targeted anti-cancer drug includes a nexus having three attaching sites, two targeting ligands conjugated to the nexus via two of the three attaching sites, a payload of a zwitterionic diagnostic or therapeutic agent conjugated to the nexus via the third attaching site, and zwitterionic flexible linkers connecting each of the two targeting ligands and the payload to the nexus.

[0104] In certain embodiments, a small zwitterionic multimeric targeted anti-cancer drug includes a nexus having four attaching sites, three targeting ligands conjugated to the nexus via three of the four attaching sites, a pay load of a zwitterionic diagnostic or therapeutic agent conjugated to the nexus via the fourth attaching site, and zwitterionic flexible linkers connecting each of the three targeting ligands and the payload to the nexus.

[0105] In certain embodiments, a small zwitterionic multimeric targeted drug includes a nexus having five attaching sites, four targeting ligands conjugated to the nexus via four of the five attaching sites, a payload of a zwitterionic diagnostic or therapeutic agent conjugated to the nexus via the fifth attaching site, and zwitterionic flexible linkers connecting each of the four targeting ligands and the payload to the nexus.

[0106] In certain embodiments, a small zwitterionic multimeric targeted drug includes a nexus having five attaching sites, three targeting ligands conjugated to the nexus via three of the five attaching sites, and two payloads of a zwitterionic diagnostic or therapeutic agent conjugated to the nexus via the fourth and fifth attaching sites, and zwitterionic flexible linkers connecting each of the three targeting ligands and two payloads to the nexus. In this configuration, one of the payloads may not be zwitterionic.

[0107] In certain embodiments, a small zwitterionic multimeric targeted drug includes a nexus having six attaching sites, five targeting ligands conjugated to the nexus via five of the six attaching sites, a payload of a zwitterionic diagnostic or therapeutic agent conjugated to the nexus via the sixth attaching site, and zwitterionic flexible linkers connecting each of the five targeting ligands and the payload to the nexus.

[0108] In certain embodiments, a small zwitterionic multimeric targeted drug includes a nexus having six attaching sites, four targeting ligands conjugated to the nexus via four of the six attaching sites, and two pay loads of a zwitterionic diagnostic or therapeutic agent conjugated to the nexus via the fifth and sixth attaching sites, and zwitterionic flexible linkers connecting each of the four targeting ligands and two pay loads to the nexus. In this configuration, one of the payloads may not be zwitterionic.

[0109] In certain embodiments, a small zwitterionic multimeric targeted drug includes a nexus having seven attaching sites, six targeting ligands conjugated to the nexus via six of the seven attaching sites, a payload of a zwitterionic diagnostic or therapeutic agent conjugated to the nexus via the seventh attaching site, and zwitterionic flexible linkers connecting each of the six targeting ligands and the payload to the nexus.

[0110] In certain embodiments, a small zwitterionic multimeric targeted drug includes a nexus having seven attaching sites, six targeting ligands conjugated to the nexus via six of the seven attaching sites, a payload of a zwitterionic diagnostic or therapeutic agent conjugated to the nexus via the seventh attaching site, and zwitterionic flexible linkers connecting each of the six targeting ligands and the payload to the nexus.

[0111] In certain embodiments, a small zwitterionic multimeric targeted drug includes a nexus having eight attaching sites, seven targeting ligands conjugated to the nexus via seven of the eight attaching sites, a payload of a zwitterionic diagnostic or therapeutic agent conjugated to the nexus via the eighth attaching site, and zwitterionic flexible linkers connecting each of the seven targeting ligands and the payload to the nexus. In certain embodiments, a small zwitterionic multimeric targeted drug includes a nexus having nine to twelve attaching sites, a payload of a zwitterionic diagnostic or therapeutic agent conjugated to the nexus via the one of the attaching sites, eight to eleven targeting ligands conjugated to the nexus via the rest of the twelve attaching sites, and zwitterionic flexible linkers connecting each of the seven targeting ligands and the payload to the nexus.

[0112] In certain embodiments, two or more of the multiple targeting ligands are attached to each other forming a linear chain, and thus the multiple targeting ligands form one or more linear chains. Each of the one or more linear chains is conjugated to one of the attaching sites of the nexus. Therefore, the number of the multiple targeting ligands conjugated to the nexus is not limited by the attaching sites on the nexus. For example, when a nexus has n attaching sites, one attaching site is conjugated to the payload, and more than n-1 targeting ligands can be conjugated to the n-1 attaching sites.

[0113] In certain embodiments, the nexus, the payload of the zwitterionic diagnostic or therapeutic agent, and zwitterionic flexible linkers of the small zwitterionic multimeric targeted anti-cancer drug form a “force field” of charge-balanced polyionicity and water of hydration such that only the targeting ligands are exposed out of the force field, as shown in Figure 2.

[0114] In certain embodiments, the multiple targeting ligands conjugated to the nexus are different from each other, such that each of the multiple targeting ligands binds to a particular receptor on the tumor cells. Therefore, by having multiple different targeting ligands, the small zwitterionic multimeric targeted anti-cancer drug is capable of binding to multiple different receptors on the tumor cells, thus significantly improves the small zwitterionic multimeric targeted anti-cancer drug’s capability of diagnosing or treating the tumors demonstrating heterogeneity. That is, at least one of the multiple possible receptors should be present on every malignant cell.

[0115] In certain embodiments, two or more of the multiple targeting ligands conjugated to the nexus are identical, with at least one targeting ligand different from these identical targeting ligands. In certain embodiments, all of the multiple targeting ligands conjugated to the nexus are the same.

[0116] The small zwitterionic multimeric targeted anti-cancer drugs of the present invention have significant advantages over traditional Antibody-Drug Conjugates (ADCs). First, with respect to the pharmacokinetic profile, the small zwitterionic multimeric targeted anti-cancer drugs of the present invention have a rapid effect and can be applied in a form of multiple retreatment to prevent resistance to the drugs, as shown in Figure 4. Moreover, the small zwitterionic multimeric targeted anti-cancer drugs of the present invention are heteromultimeric monovalent molecules which addresses the tumor heterogeneity more efficiently. In addition, the small zwitterionic multimeric targeted anti-cancer drugs of the present invention can be synthesized more easily and at a lower cost than ADCs.

[0117] Nexus

[0118] Central Core

[0119] In certain embodiments, the nexus is a central core has a geometry structure comprising multiple vertexes, in which each of the vertexes serves as an attaching site for being conjugated to the payload or the targeting ligands, such that the multiple targeting ligands and the payload are maximally spaced apart from each other in 3D space, so as to reduce steric interference among the multiple targeting ligands and the payload.

[0120] In certain embodiments, for example, the central core has a pentahedral geometry such that four targeting ligands (tetramer of targeting ligands) and one payload are attached to the pentahedral core and are maximally spaced apart from each other in 3D space, so as to reduce steric interference among the four targeting ligands and the payload.

[0121] In certain embodiments, the pentahedral core is attached to three targeting ligands and two payloads.

[0122] In one preferred embodiment for breast cancer, the central core of pentahedral geometry comprises ligands specific for FAP, integrin alpha5betal, MUC1, and GRPR while the payload comprises an N-oxide-based zwitterionic DOTA. In one preferred embodiment for prostate cancer, the central core of pentahedral geometry comprises ligands specific for FAP, integrin alpha5betal , PSMA, and GRPR while the payload is an N-oxide-based zwitterionic DOTA.

[0123] In certain embodiments, for example, the central core has a tetrahedral geometry such that the three targeting ligands and the payload are maximally spaced apart from each other in 3D space, so as to reduce steric interference among the three targeting ligands and the payload.

[0124] In one preferred embodiment, the central core comprises adamantane which is a rigid, symmetrical, and stable organic compound with the chemical formula (CH CFL In yet another preferred embodiment, the central core comprises orthocarbonic acid.

[0125] In certain embodiments, the central core comprises one or more of the following: 1,3,5,7-tetrasila-adamantane; 1,3, 5, 7 tetrabora- adamantane; 1,3,5,7-tetraaza-adamantane; 2,4,6,8,9,10-hexamethyl-2,4,6,8,9,10-hexabora-adamantane; and etc.

[0126] In certain embodiments, the central core comprises diglutamate, which provides three carboxylic acids for conjugation of zwitterionic linkers and individual targeting ligands and a single primary amine for conjugation of zwitterionic linkers and the payload.

[0127] In certain embodiments, instead of being attached to the central core having a tetrahedral geometry, the multiple targeting ligands and the pay load are conjugated to one or more molecule to form other arrangements such as a linear arrangement. In certain embodiments, while one or more of the multiple targeting ligands are attached to the central core directly, at least one of the targeting ligands is attached to another targeting ligand, such that it is conjugated to the central core indirectly. In certain embodiments, a first group of multiple targeting ligands are conjugated to the central core directly, a second group of multiple targeting ligands are conjugated to the first group of multiple targeting ligands.

[0128] In these embodiments, the arrangement for the multiple targeting ligands and the payload effectively maximizes the space between the three targeting ligands and the payload in 3D space, so as to prevent the interactions among the other chemical groups. Branching Point

[0129] In certain embodiments, the nexus is a branching point which connects to the multiple targeting ligands via the one or more zwitterionic linkers and / or one or more spacing groups, e.g. Ahx. In certain embodiments, the branching point comprises one or more amino acids, each of which have multiple attaching sites for being conjugated to the multiple targeting ligands / payload. In addition to the pay load and targeting ligands, the attaching sites on one amino acid can be conjugated to another amino acids, either directly or through linkers and / or spacing groups, as shown in Figure 7.

[0130] In certain embodiments, the branching point comprises two amino acids, and each amino acid comprises at least one attaching site. In certain embodiments, the branching point comprises three amino acids, and each amino acid comprises at least one attaching site. In certain embodiments, the branching point comprises four amino acids, and each amino acid comprises at least one attaching site. In certain embodiments, the branching point comprises five amino acids, and each amino acid comprises at least one attaching site. In certain embodiments, the branching point comprises six amino acids, and each amino acid comprises at least one attaching site. In certain embodiments, the branching point comprises six amino acids, and each amino acid comprises at least one attaching site. In certain embodiments, the branching point comprises seven amino acids, and each amino acid comprises at least one attaching site. In certain embodiments, the branching point comprises eight amino acids, and each amino acid comprises at least one attaching site.

[0131] In certain embodiments, the branching point comprises two amino acids, and each amino acid comprises three attaching sites. While one of these three attaching sites is attached to the other amino acid, two of these attaching sites are attached to either targeting ligands or payload. As such, a branching point of two amino acids can attach to three targeting ligands and one payload. Alternatively, the branching point of two amino acids can attach to two targeting ligands and two payloads.

[0132] In certain embodiments, the branching point comprises three amino acids, and each amino acid comprises three attaching sites. While one of these three attaching sites is attached to another amino acids, two of these attaching sites are attached to either targeting ligands or payload. As such, a branching point of three amino acids can attach to four targeting ligands and one payload. Alternatively, the branching point of three amino acids can attach to three targeting ligands and two payloads.

[0133] In certain embodiments, the branching point comprises four amino acids, and each amino acid comprises three attaching sites. While one of these three attaching sites is attached to another amino acids, two of these three attaching sites are attached to either targeting ligands or payload. As such, a branching point of four amino acids can attach to five targeting ligands and one payload. Alternatively, the branching point of four amino acids can attach to four targeting ligands and two payloads.

[0134] As shown in Figures 7-9, in certain embodiments, the branching point comprises one or more Lys. In certain embodiments, the branching point comprises one or more Glu. In certain embodiments, the branching point comprises at least one Lys and at least one Glu. In certain embodiments, each of the Lys and Glu includes three attaching sites. In certain embodiments, one attaching site on Lys is connected to one attaching site on Glu, and the other two attaching sites of each amino acids are conjugated to either the targeting ligands or the pay load via the linkers and / or spacing groups.

[0135] It should be noted that other amino acids can be used instead of Glu and / or Lys, or other non-amino acid chemical groups can be used. In certain embodiments, the branching point can be one or more of polyethylene glycol (PEG), polysaccharides, polylactic-co- glycolic acid / poly(e-caprolactone) (PCL), polyLactic Acid, poly(3- hydroxybutyrate-co-3- hydroxyvalerate) (PHBV), chitosan, and cellulose..

[0136] In certain embodiments, the one or more amino acids are modified or decorated before and / or during the synthesis of the zwitterionic multimeric targeted anti-cancer drug. In certain embodiments, the modifications and / or decorations on the one or more amino acids protect the side chain groups of the amino acids. In certain embodiments, the modifications and / or decorations on the one or more amino acids include Fmoc (9- fluorenylmethoxycarbonyl) group, alloc (CH2=CH-CH2-O-CO-) group, allyl (CH2=CH- CH2-) group, etc. It should be noted that some or all of these modifications and / or decorations are for group / side chain protection purpose and only exists on the branching point before and / or during the synthesis of the zwitterionic multimeric targeted anti-cancer drug. Once the synthesis is accomplished, some or all of these modifications and / or decorations are removed.

[0137] In certain embodiments, some or all of these modifications and / or decorations are converted to other chemical / functional groups during or at the last step of the synthesis.

[0138] In certain embodiments, the branching point comprises one or more polymers, each of which comprises multiple monomers. In certain embodiments, the monomers are connected end-to-end in single chains such that the polymer is a linear polymer form without branches. In certain embodiments, monomers form side chains which attach to a main chain such that the polymer has branches. In certain embodiments, monomers form chains which are interconnected in a network.

[0139] In certain embodiments, some or all of the monomers serve as attaching sites which are conjugated to either targeting ligands or payloads via linkers and / or spacing groups.

[0140] In certain embodiments, the polymer comprises one or more of polyethylene glycol (PEG), polysaccharides, polylactic-co-glycolic acid / poly(s-caprolactone) (PCL), polyLactic Acid, poly(3- hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), chitosan, cellulose, etc.

[0141] Linkers and Spacing Groups

[0142] In certain embodiments, the linkers attaching the targeting ligands and the payload to the nexus comprises linear polymers of alternating quaternary amines and sulphonates. In certain embodiment, the linkers attaching the targeting ligands and the payload to the nexus comprise linear polymers of nitric oxides or similar zwitterionic groups.

[0143] In certain embodiments, the linkers are zwitterionic flexible linkers. In certain embodiments, the zwitterionic flexible linkers reduce the targeting ligands’ non-specific or off-target binding. In certain embodiments, the zwitterionic flexible linkers provide spacing of the targeting ligands and the payload from the nexus, and from each other. Therefore, the zwitterionic flexible linkers reduce the risk of steric hindrance or interference between the different functional groups of the drug. In certain embodiments, the zwitterionic flexible linkers create the “force field’’ of charge-balanced polyionicity and water of hydration around the nexus, such that the targeting ligands are exposed for binding to receptors on the tumor cells, thus reducing non-specific and off-target interactions with normal cells and tissues.

[0144] In certain embodiments, the zwitterionic flexible linkers have desirable properties that maximize solubility in aqueous environments, minimize non-specific interactions, and retain the ability to target thus resulting in an improved performance in a variety of medical, agricultural, and chemical processes. In in vivo and medical applications, zwitterionic flexible linkers improve the signal-to-background ratio and therapeutic window as compared to other metal chelators while retaining high stability.

[0145] In certain embodiments, the zwitterionic flexible linkers demonstrate same properties described for the zwitterionic payloads below.

[0146] In certain embodiments, the linker comprises Fmoc- Azidoalanine, Fmoc- Azidohomoalanine or other azido derivatives, which can be converted to ZW groups using click chemistry. In certain embodiments, the Fmoc- Azidoalanine has CAS number 684270- 46-0. In certain embodiments, the Fmoc- Azidohomoalanine has a CAS number 942518-20-9.

[0147] In certain embodiments, the Fmoc- Azidoalanine and Fmoc-Azidohomoalanine include an azido group (-N3) attached to the side chain of an amino acid, e.g., alanine, and the amino group of the amino acid is protected by a 9-fluorenylmethoxycarbonyl (Fmoc) group. This compound is typically used in solid-phase peptide synthesis (SPPS) to introduce azido functionalities into peptides, which can then be used for further chemical modifications, such as click chemistry reactions.

[0148] In certain embodiments, the azido derivatives includes one or more of Fmoc- Azidophenylalanine (Fmoc-AzidoPhe), Fmoc-Azidoglycine (Fmoc-AzidoGly), Fmoc- Azidoornithine (Fmoc-AzidoOrn), Fmoc-Azidolysine (Fmoc-AzidoLys), Fmoc- Azidoaspartic Acid (Fmoc-AzidoAsp).

[0149] In certain embodiment, the zwitterionic flexible linker comprises Fmoc- Aminobutanoic acid N-Oxide (FABNO). The FABNO has a formula as follow:

[0150]

[0151] In certain embodiments, the FABNO is made from Fmoc-2-amino-4- (dimethylamino)butanoic acid by treating the acid with 10 eq. of H2O2 or organic peroxide.

[0152] The FABNO is synthesized through the steps shown in Figure 5.

[0153] Fmoc-Tri-FABNO 4 in Figure 5 has a formula

[0154] Tri-FABNO-Allyl 6 in Figure 5 has a formula

[0155] In certain embodiment, the zwitterionic flexible linker comprises Fmoc-Lysine N- Oxide (FLNO). The FLNO has a formula as follow:

[0156]

[0157] In certain embodiments, the FLNO is made from Fmoc-Dimethyl-Lysine by treating it with 10 eq. of H2O2 or organic peroxide.

[0158] The FLNO is synthesized through the steps shown in Figure 6.

[0159] Fmoc-Tri-FLNO 4 in Figure 6 has a formula

[0160] Tri-FLNO-Allyl 6 in Figure 6 has a formula In certain embodiments, in addition to the linkers, one or more spacing groups, e.g. Ahx, locate between one targeting ligand and the nexus / branching point. For example, as shown in Figure 7, one Ahx group connects the zwitterionic flexible linker and the targeting ligand.

[0161] Targeting Ligands

[0162] In certain embodiments, the targeting ligand comprises one or more of small molecules, peptides / antibodies, DNAs, RNAs or other molecules which bind to one or more receptors on the plasma membrane of tumor cells.

[0163] In certain embodiments, a multimeric targeted anti-cancer drug has at least one targeting ligand binding to a cancer-independent target, and at least one targeting ligand binding to a cancer-dependent target.

[0164] Cancer-independent targets are typically normal cells and / or molecule structures within the body that have been co-opted and / or influenced by the tumor to enhance its growth and / or evade the immune system. These targets are not inherently cancerous but play a crucial role in the tumor microenvironment. By targeting these cells or structures, therapies can disrupt the supportive conditions that the tumor relies on, potentially leading to more effective cancer treatments. Examples include fibroblast activation protein (FAP) expressed by cancer-associated fibroblasts (CAFs) and certain integrins overexpressed in the neo vasculature of tumors.

[0165] Importantly, normal cells are much less prone to genetic mutations and clonal evolution compared to cancer cells. Therefore, targeting these cancer-independent cells can have profound effects on tumor growth and destruction. By focusing on these stable targets, therapies can potentially reduce the likelihood of the tumor developing resistance, leading to more effective and durable cancer treatments by the small zwitterionic multimeric targeted anti-cancer drug having targeting ligands binding to these cancer-independent cells / molecules. Cancer-dependent targets are specific molecules or structures found on the surface of malignant cells. These targets are integral to the cancer cells' survival, proliferation, and ability to evade the immune system. The present invention focuses on the cancer-dependent targets characterized by high surface expression level, known as BMax (the number of binding sites per cell), which allows for the delivery of a substantial therapeutic payload to each cancer cell. By focusing on these targets via ligands, the multimeric targeted anti-cancer drug can directly attack and destroy cancer cells while minimizing damage to normal cells. Examples of cancer-dependent targets include receptors and antigens such as the Gastrin- Releasing Peptide Receptor (GRPR), Prostate-Specific Membrane Antigen (PSMA), and MUC1. These targets are selected for their high expression levels on cancer cells, making them effective points of intervention in cancer treatment strategies.

[0166] Figure 12 illustrates a cancer tissued developed into cancer cells and cancer- associated tissue, and a process of how the multimeric targeted anti-cancer drug including both cancer-independent and cancer dependent target binding ligands treats the cancer tissues. As shown in the upper panel, the cancer issues include cancer cells with cancerdependent targets, as such MUC1, bombesin, etc., and cancer-associated tissues, e.g. cancer- associated fibroblasts and tumor neovasculature. The cancer-associated tissues include one or more cancer-independent targets. For example, the cancer-associated fibroblasts include FAP, and the tumor neovasculature includes integrin 0.5( 1. Therefore, a multimeric targeted anticancer drug efficient for treating the cancer includes one or more cancer-dependent targets binding ligands and one or more cancer-independent targets binding ligands. For example, in this scenario, the cancer-dependent targets binding ligands include a MUC1 binding ligand and a GRPR binding ligand; the cancer-independent targets binding ligands include a FAP binding ligand and an integrin a5pi binding ligand.

[0167] During the treatment process, the FAP binding ligand binds to the FAP on the cancer- associated fibroblasts such that the immune blocking for the cancer cells are removed, while the integrin a501 binding ligand binds to the integrin cx5(31 on the tumor neovasculature such that the blood supply to the cancer cells is cut off. Meanwhile, the cancer-dependent binding ligands, e.g., MUC1 binding ligand and bombesin binding ligand, binds to the cancer cell targets directly, targeting them for destruction. The cancer-dependent and cancer-independent targets shown in Figure 12 are examples for illustration. Targeting ligands binding to other cancer-dependent and cancerindependent targets can be selected according to the cancer conditions to be treated.

[0168] Examples of cancer-independent targets include:

[0169] Fibroblast Activation Protein (FAP): Expressed by cancer-associated fibroblasts in the stroma of both primary and metastatic tumors, FAP contributes to immune suppression. Using a targeting ligands binding to FAP in the multimeric drug can eradicate CAFs, and thus could unleash the immune system against malignant cells.

[0170] Integrin cc5|31 : This integrin is overexpressed in the neovasculature of most human solid tumors. It serves as an ideal target for therapies aimed at destroying the tumor's blood supply. By targeting integrin a501, treatments can disrupt the vasculature system that provides essential nutrients to the tumor, thereby inhibiting its growth.

[0171] Integrin av|33: Similar to integrin a5[31, integrin avP3 is also overexpressed in the neovasculature of most human solid tumors. Targeting this integrin can similarly disrupt the tumor's blood supply, depriving it of the nutrients necessary for its growth and survival.

[0172] Other cancer-independent targets can be targeted by the multimeric targeted anticancer drug as well, depending on the specific cancer conditions being treated.

[0173] Examples of cancer-dependent targets include:

[0174] Gastrin- Releasing Peptide Receptor (GRPR; also known as bombesin receptor): This receptor is expressed on over 80% of luminal breast cancer cells, prostate cancer cells with a basal phenotype, glioblastoma multiforme, and various other common human cancers. In cells that express GRPR, the BMax is typically around 105binding sites per cell.

[0175] Prostate-Specific Membrane Antigen (PSMA): PSMA is highly expressed in the majority of early-stage prostate cancers and also in the neovasculature of some other human cancers. In cells expressing PSMA, the BMax is typically around 105binding sites per cell.

[0176] MUC1: This target is highly expressed in most epithelial malignancies in various forms, including the full-length form with variable numbers of tandem repeats (VNTRs), a truncated form without the VNTRs, and various truncated forms with aberrant glycosylation. The BMax for MUC1 is highly variable depending on its form but can be as high as 10A6 binding sites per cell. Unlike GRPR, which may be limited to certain histologic types of breast cancer, MUC1 is more universally expressed across different types of epithelial cancers.

[0177] These examples of cancer-independent and cancer-dependent targets are not exhaustive but rather representative of the general strategy of combining targeting ligands binding to cancer-independent targets and cancer-dependent ligands in a single therapeutic approach to maximize the therapeutic benefit. This strategy aims to enhance the efficacy of cancer treatments by targeting both the tumor cells directly and the supportive cells in the tumor microenvironment.

[0178] Each small zwitterionic multimeric targeted anti-cancer drug includes an optimal combination of both targeting ligands binding to cancer-independent targets and targeting ligands binding to cancer-dependent targets. This strategic approach is designed to ensure that the tumor cannot develop resistance through its usual mechanism of clonal evolution. By addressing multiple pathways and mechanisms simultaneously, these drugs aim to outmaneuver the tumor's ability to adapt and survive, thereby enhancing the effectiveness of the treatment and potentially leading to better therapeutic effects. This multifaceted targeting strategy is crucial in the ongoing battle against cancer, as it seeks to prevent the tumor from exploiting its genetic diversity to evade therapeutic interventions.

[0179] In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a nexus, one or more targeting ligands binding to cancer-independent targets, one or more targeting ligands binding to career-dependent targets, a payload, and zwitterionic linkers conjugate the targeting ligands and the payload to the nexus.

[0180] In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes two targeting ligands binding to cancer-independent targets, two targeting ligands binding to career-dependent targets, a payload which include a base payload and a specific payload. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes one targeting ligands binding to cancer-independent targets, three targeting ligands binding to career-dependent targets, a pay load which include a base pay load and a specific pay load. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes three targeting ligands binding to cancer-independent targets, one targeting ligands binding to career-dependent targets, a payload which include a base payload and a specific payload.

[0181] In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes one targeting ligands binding to cancer- independent targets, four targeting ligands binding to career-dependent targets, a payload which include a base payload and a specific payload. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes two targeting ligands binding to cancer-independent targets, three targeting ligands binding to career-dependent targets, a payload which include a base payload and a specific payload. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes three targeting ligands binding to cancer- independent targets, two targeting ligands binding to career-dependent targets, a pay load which include a base pay load and a specific pay load. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes four targeting ligands binding to cancer-independent targets, one targeting ligands binding to career-dependent targets, a pay load which include a base pay load and a specific pay load.

[0182] In certain preferred embodiments, the targeting ligand is a hydrophilic and relatively charge-balanced molecule.

[0183] Table 1. List of examples of cancer-independent and dependent targets, their binding ligands.

[0184]

[0185] In certain preferred embodiments, the three targeting ligands of a small zwitterionic trimeric targeted anti-cancer drug comprises KUE (a PSMA-binding small molecule), FAPI- 74 (a Fibroblast Activation Protein binding small molecule) and JMV594 (a GRPR, bombesin-like peptide). This combination of three targeting ligands binds to over 90% of all solid tumors and thus effectively addresses tumor heterogeneity and / or reduces drug resistance. As shown in Figure 3, the prostate tumors (top panel) and most solid tumors (bottom panel) evolve over the time with receptors on the tumor cells changes. By being able to bind by any one of the three targeting ligands of the small zwitterionic trimeric targeted anti-cancer drug, all tumor cells within a heterogeneous tumor are killed during the early stage of the tumor development, and thus avoiding drug resistance.

[0186] In certain embodiments, the targeting ligand comprises one or more of the followings. cRGD

[0187] In certain embodiments, the targeting ligand includes cyclic-RGD (cRGD). In one embodiment, the cRGD peptide as the targeting ligand has the formula:

[0188]

[0189] Integrins are a family of cell adhesion receptors that provide for cell motility and invasion. The integrin family in humans comprise 18 a and subunits which assemble into different functional heterodimers. Some integrins, act as mediators of angiogenesis in solid tumors. Integrin expression is important for tumor progression and metastasis by promoting tumor cell migration, invasion, proliferation, and survival. In addition to cancer, integrins are also highly expressed during many normal and abnormal processes, such as fibrosis, wound healing, and inflammation. Integrin av 3 expression has been established as a surrogate marker of angiogenic activity. Similarly, Integrin v 6 is a subtype of the integrin family that is expressed exclusively on epithelial cells. av 6 is usually expressed at low or undetectable levels in normal adult tissues but can be highly upregulated during pathological and physiological processes such as wound healing, fibrosis, inflammation, and cancer. RGD mimetics have been used in some studies as a targeting moiety to deliver integrin antagonists to a variety of cell types. However, continuous infusion of RGD peptides have been found to stimulate tumor growth (See, Reynolds, L. E. et al. Enhanced pathological angiogenesis in mice lacking 3 integrin or and 5 integrins. Nature Med. 8, 27-34 (2002)). In addition, many RGD bound agents have expensive, complex, time-consuming and low-yield synthetic procedures. Similarly, with regard to imaging, degradation of agents accumulation of RGD bound agents in the bladder have been found to impair some imaging methods. Given their stability and clearance rates, the conjugates of the invention have the potential to provide a long-lasting visual identification and imaging of tumors which overexpress integrins while avoiding the setbacks of prolonged systemic administration of RGD peptide ligands. In certain embodiments, the cRGD peptide (e.g. cRGDyK) binds to Integrin av|k

[0190] In certain embodiments, the cRGD peptide as targeting ligand includes a dimer or a multimer of cRGD peptide, or a derivative thereof.

[0191] Integrin Alpha5-Betal

[0192] Integrin alpha5-betal is overexpressed on the neovasculature of a variety of malignant tumors. Multiple small molecule and peptide targeting ligands specific for integrin alpha5- betal have been reported, including the acetylated peptide ATN-161. In general, integrin alpha5-betal is more highly ovexpressed in tumor neo vasculature than integrin alphaV-beta3 and therefore may be a superior target.

[0193] In certain embodiments, the targeting ligand includes a5pi binding ligands. In certain embodiments, the a5pi binding ligand includes one or more of Peptide ATN-161 (SEQ ID No. 7), ATN-345, peptide CRRETAWAC (SEQ ID No: 8) and its cyclic form, PR_b peptide (SEQ ID No: 9), Fibronectin Cell-Adhesion Module Type III- 10 (FNIII10), or a derivative thereof. Additional information of ATN-161, ATN-345 and their derivatives can be found in - Livant, Donna L., et al., Cancer research 60.2 (2000): 309-320, and Khalili, Parisa, et al., Molecular cancer therapeutics 5.9 (2006): 2271-2280 and - which are incorporated herein by reference.

[0194] In certain embodiments, the alpha5-betal binding ligand includes a dimer or a multimer of one or more of Peptide ATN-161, peptide CRRETAWAC, PR_b, FNIII10, or a derivative thereof.

[0195] MUC1 binding ligands

[0196] MUC1(SEQ ID No. 10) is highly overexpressed in a large number of malignant carcinomas, especially breast cancer. Multiple peptide and peptidomimetic targeting ligands specific for MUC1 have been reported, most of which either bind or complement the core peptide sequence PDTRP. Because of the extremely high levels of expression of MUC1 on malignant cells, and lack of expression on most normal cells, MUC1 could be an ideal target. In certain embodiments, the targeting ligand includes MUC1 binding ligands. In certain embodiments, the MUC1 binding ligand includes one or more of Galectin-3(SEQ ID No. 11), Intercellular Adhesion Molecule l(ICAM-l) (SEQ ID No. 12), Epidermal Growth Factor Receptor(EGFR)(SEQ ID No. 13), P-catenin(SEQ ID No. 14), Sialic acid-binding Ig- like lectin 9 (Siglec-9) (SEQ ID No. 15), Nucleoside diphosphate kinase A (NME1) (SEQ ID No. 27-28), Nucleoside diphosphate kinase homolog 7 (NME7) (SEQ ID No: 29-30), etc., or a derivative thereof.

[0197] In certain embodiments, the MUC1 binding ligand includes a dimer or a multimer of one or more of Galectin-3, ICAM-1, EGFR, P-catenin, Siglec-9, NME1, NME7, etc., or a derivative thereof.

[0198] PSMA and KUE

[0199] In certain embodiments, the targeting ligand includes PSMA, e.g., KUE or dPSMA- 617. In one embodiment, PSMA includes a targeting ligand based on (((S)-5-((S)-2-(( lr,4S)- 4-(aminomethyl)cyclohexane-l-carboxamido)-3-(naphthalen-2-yl)propanamido)-l- carbo xypentyl)carbamoyl)-L-glutamic acid, which is a derivative of PSMA-617 or Vipivotide tetraxetan. This PSMA-targeting ligand is called dPSMA-617 (for “derivative of PSMA-617) in this disclosure.

[0200] An alternative PSMA-targeting ligand is (((S)-5-amino-l-carboxypentyl)carbamoyl)- L-glutamic acid (KUE).

[0201] In particular embodiment, the KUE or PSMA-617-conjugated zwitterionic metal chelators can be complexed with Ga3+, Cu2+, Lu3+, Zr4+, Mn2+, Mn3+, Tb3+, Gd3+, or similar metal cations. In certain embodiments of the invention, the targeting ligand according to the invention can be dPSMA-617 having the following structure:

[0202]

[0203] In certain embodiments of the invention, the targeting ligand according to the invention can be KUE having the following structure: In certain embodiments, the targeting ligand includes a dimer or a multimer of one or more of KUE and dPSMA-617, or a derivative thereof.

[0204] FAP

[0205] In certain embodiments, the targeting ligand includes FAP targeting binding ligand. In certain embodiments, the FAPI-targeting ligand includes NH2-FAPI-74. Specific details regarding NH2-FAPI-74 are described in: Linder et al., “Radioligands Targeting Fibroblast Activation Protein (FAP),’’ the entirety of which is incorporated herein by reference. In general, NH2-FAPL74 has the following structure: the FAPI-targeting ligand includes NH2- FAPI-74

[0206] NH2-FAPl-74

[0207] In certain embodiments, the targeting ligand includes a dimer or a multimer of one or more of FAPI-targeting binding ligand, or a derivative thereof.

[0208] Bombesin receptor binding ligand

[0209] In certain embodiments, the targeting ligand includes bombesin receptor binding ligand. In general, the structure of bombesin (SEQ ID No: 6) is:

[0210]

[0211] , or a derivative thereof.

[0212] In certain embodiments, the targeting ligand includes one or more derivatives of bombesin receptor binding ligands, including, but not limited to, BIM26226 (SEQ ID No. 16; {D-F5 Phe}-Gln-Trp-{D-Ala}-Val-Ala-His-Leu-OMe), IMV594 (SEQ ID No. 17), JMV641(SEQ ID No. 18), JMV736, JMV717, JMV845, and JMV542. Additional bombesin derivatives, including bombesin agonists, bombesin antagonists, and bombesin pseudopeptides can be found in Azay et al., Peptides, Vol 19, No. 1, pp 57-63, 1988 - which is incorporated herein by reference.

[0213] In certain embodiments, the bombesin receptor binding ligand are configured to bind to Gastrin-releasing peptide receptor (GRPR).

[0214] In certain embodiments, the targeting ligand includes a dimer or a multimer of one or more of BIM26226, IMV594, JMV641, IMV736, JMV717, IMV845, JMV542, other bombesin receptor binding ligands, or a derivative thereof.

[0215] Octreotide receptor binding ligand

[0216] In certain embodiments, the targeting ligand includes octreotide receptor binding ligand. In general, the structure of octreotide (SEQ ID No: 5) is:

[0217] , or a derivative thereof.

[0218] In certain embodiments, the targeting ligand includes a dimer or a multimer of one or more of octreotide receptor binding ligands, or a derivative thereof.

[0219] EpCAM binding ligands

[0220] In certain embodiments, the targeting ligand includes one or more EpCAM binding ligands. In certain embodiments, a EpCAM binding ligand includes one or more of Pep 10 having SEQ ID No. 19, Edrecolomab, Catumaxomab, Solitomab (MT110), YQ-S2- MPA(SEQ ID No. 20) ((Ser-Asn-Phe-Tyr-Met-Pro-Leu-Lys)cycie), or a derivative thereof.

[0221] In certain embodiments, the EpCAM binding ligand includes a dimer or a multimer of PeplO, Edrecolomab, Catumaxomab, MT110 (Solitomab), or YQ-S2-MPA.

[0222] Integrin avp6 binding ligands

[0223] In certain embodiments, the targeting ligand includes avP6 binding ligands. In certain embodiments, the avP6 binding ligand includes one or more of A20FMDV2 Peptide (SEQ ID No. 21), A2OFMDV1(SEQ ID No. 22), A20LAP (SEQ ID No. 23), or a derivative thereof. Additional information of A20FMDV2 Peptide and its derivatives can be found in - Hausner, Sven H., et al. Cancer research 67.16 (2007): 7833-7840 - which is incorporated herein by reference.

[0224] In certain embodiments, the avP6 binding ligand includes a dimer or a multimer of one or more of A20FMDV2 Peptide, A20FMDV1, A20LAP, or a derivative thereof.

[0225] Integrin av 3 binding ligands

[0226] Integrin avP3 is prominently overexpressed in the neovasculature of most human solid tumors. This characteristic makes integrin avp3 a compelling target for therapeutic strategies aimed at disrupting the tumor's blood supply. The neovasculature is crucial for tumor growth and survival as it supplies essential nutrients and oxygen. By targeting integrin avP3, treatments can effectively compromise the vascular system that sustains the tumor, potentially inhibiting its growth and progression. This approach is part of a broader strategy in oncology to starve tumors by cutting off their nutrient supply, thereby enhancing the efficacy of cancer treatments. Integrin avP3's role in angiogenesis and its prevalence in tumor vasculature underscore its importance as a target in cancer therapy, offering a pathway to potentially improve patient outcomes through targeted interventions.

[0227] In certain embodiments, the targeting ligand includes avP3 binding ligands. In certain embodiments, the avf>3 binding ligand includes one or more of cilengitide(SEQ ID No. 24 cyclofRGDFV]), RGD Peptides, vitronectin(SEQ ID No. 25), osteopontin(SEQ ID No. 26), or a derivative thereof. In certain embodiments, the EpCAM binding ligand includes a dimer or a multimer of one or more of cilengitide, RGD Peptides, vitronectin, osteopontin, or a derivative thereof.

[0228] Other Anti-Cancer Targeting Ligands

[0229] In certain embodiments, the targeting ligand includes one or more of LyP-1 peptide having a sequence of CGQKRTRGC (SEQ ID NO: 1) and binding to P32 for diagnosing / treating melanoma; K237 peptide having a sequence of HTMYYHHYQHHL(SEQ ID NO: 2) and binding to VEGFR-2 for diagnosing / treating breast tumor; IL4RPep-l peptide having a sequence of CRKRLDRNC(SEQ ID NO: 3) and binding to IL4R for diagnosing / treating lung tumor, breast tumor, colon tumor; mUNO peptide having a sequence of CSPGAK (SEQ ID NO: 4) and binding to CD206 for diagnosing / treating breast tumor; folate receptors for diagnosing / treating ovarian and lung cancer; GEII, a dodecapeptide, binding to epidermal growth factor receptor (EGFR or ErbBI) for diagnosing / treating tumors of epithelial origin; a-M2 (EPPT / dTPPE) or PDTRP peptides binding to MUCI; [F7,P34]-NPY peptide binding to NPYIR; and Met-Pepl or equivalent binding to c-Met.

[0230] In certain embodiments, the targeting ligands are agonists or antagonists of the target, including, but not limited to, agonists or antagonists of integrins, agonists or antagonists of FAP, agonists or antagonists of PSMA, agonists or antagonists of somatostatin receptor, and agonists or antagonists of GRPR. In still other embodiments, the targeting ligand may be a derivative of the targeting ligand, including, but not limited to, derivatives of cRGD, derivatives of FAP, derivatives of PSMA-binding vectors, derivatives of octreotide, and derivatives of bombesin.

[0231] In certain embodiments, the targeting ligand can further include a molecular scaffold moiety to which the binding moiety and other groups can attach. For example, the molecule scaffold can bear one or more of the following: (1) a moiety designed to react with the reactive linking group of the dye to form a covalent bond, (2) a charge balancing moiety, such as any of the ionic groups described herein, and (3) a moiety that binds to the biological target. An example of a molecular scaffold is an adamantane derivative, such as described in U.S. Pat. App. Pub. No. 2006 / 0063834, which is incorporated herein by reference in its entirety, and illustrates the preparation of a targeting ligand that incorporates an adamantane scaffold. Specifically, the adamantane core holds (1) an amino group capable of reacting with the dye compounds, (2) a charge-balancing moiety that will neutralize a negative charge on the dye molecule, and (3) two moieties that bind to the biological target PSMA. For a description of moieties that bind to PSMA, see, Humblet, V. et al. Mol. Imaging, 2005, 4: 448-62; Misra P. et al. J. Nucl. Med. 2007, 48: 1379-89; Chen, Y„ et al. J. Med. Chem, 2008, 51: 7933-43; Chandran, S.S., et al. Cancer Biol. Ther., 2008, 7:974-82; Banerjee, S.R., J. Med. Chem. 2008, 51: 4504-17; Mease, R.C., et al. Clin. Cancer Res., 2008, 14:3036-43; Foss, C.A. et al. Clin. Cancer. Res., 2005, 11:4022-8, each of which is incorporated herein by reference in its entirety.

[0232] Payload In certain embodiments, the payload comprises one or more imaging agents. In certain embodiments, the payload comprises one or more therapeutic / treatment agents.

[0233] In certain preferred embodiments, the pay load comprises one or more zwitterionic groups, which extends the force field of hydration to the whole drug molecule except for the targeting ligands. In certain embodiments, the zwitterionic group is a zwitterionic metal chelator complex.

[0234] In certain embodiments, a zwitterionic metal chelator complex has one or more of the zwitterionic metal chelators. These zwitterionic metal chelators have desirable properties that maximize solubility in aqueous environments, minimize non-specific interactions, and retain the ability to target thus resulting in an improved performance in a variety of medical, agricultural, and chemical processes. In in vivo and medical applications, zwitterionic metal chelators improve the signal-to-background ratio and therapeutic window as compared to other metal chelators while retaining high stability.

[0235] The zwitterionic groups described herein are useful in, for example, the detection of abnormal or diseased biological tissues and cells. In some aspects, the zwitterionic metal chelators are particularly useful for imaging whole organisms as they have improved in vivo behavior, such as low non-specific binding to non-targeted tissues and high stability, resulting in an improved signal-to-background ratio in connection with the detected signal and / or improved therapeutic window. Similarly, in an agricultural or chemical process that now uses a metal chelator, zwitterionic metal chelators will improve solubility and minimize nonspecific interactions. It is believed that these improved properties result from the balancing of formal charges on the metal chelator, rendering a polyionic yet “charge-balanced” molecule having a net charge that is neutral or close to neutral, with an extended sphere of hydration and better isolation of the metal.

[0236] In certain embodiments, the payload includes functional molecule listed in Table 2.

[0237] Table 2. Payload functional group of small zwitterionic multimeric targeted anti-cancer drugs

[0238] In certain embodiments, the payload includes at least one base payload and at least one specific payload. In certain embodiments, one particular base payload can be combined with different specific payloads.

[0239] In certain embodiments, the payload carried by zwitterionic multimeric drugs is tailored to the specific clinical application for which they are intended. One of the notable efficiencies of these drugs lies in their versatility. As the base payload, a single zwitterionic chelator can be combined with various types of metals as the specific payload, allowing for the creation of different subsets of payloads. This adaptability enables the development of customized therapeutic agents that can be fine-tuned to target specific diseases or conditions, thereby maximizing their efficacy and minimizing potential side effects. By leveraging this modular approach, it is possible to design a wide range of treatments that can address diverse medical needs with precision and effectiveness. Table 3 below provides a list of payload examples and their clinical application.

[0240] Table 3. Payload examples and their clinical application.

[0241] In certain embodiments, the zwitterionic groups are introduced into the payload and / or linkers by using Fmoc-Azidoalanine, Fmoc-Azidohomoalanine, or other azido derivatives, which can be converted to ZW groups using click chemistry. In certain embodiments, the Fmoc-Azidoalanine has CAS number 684270-46-0. In certain embodiments, the Fmoc-Azidohomoalanine has a CAS number 942518-20-9.

[0242] In certain embodiments, the payload includes a combination of one or more radioisotope and one or more radiosensitizer. In certain embodiments, the payload includes a combination of one or more radioisotope and one or more chemotherapeutic agent. In certain embodiments, the payload includes a combination of one or more radioisotope and one or more external cyberknife radiotherapeutics.

[0243] ZW700-1 Forte

[0244] In certain embodiments, the payload includes a ZW700-1 Forte, which is a 700-nm zwitterionic pentamethine indocyanine near-infrared fluorophore which permits dual-channel image-guided surgery. In certain embodiments, the ZW700-1 Forte as the payload having the formula: ZW700-1C can be prepared using the following Scheme 1:

[0245] Scheme 1:

[0246] In certain embodiments, the zwitterionic groups are introduced into the ZW700-1 Forte using one or more azido-zwitterion derivatives via click chemistry.

[0247] ZW800-1

[0248] In certain embodiments, the pay load includes ZW800-1, which is a 800-nm zwitterionic near-infrared fluorophore. In certain embodiments, the ZW800-1 having the formula: In certain embodiments, the zwitterionic groups are introduced into the ZW800-1 using one or more azido-zwitterion derivatives via click chemistry.

[0249] ZW830-1

[0250] In certain embodiments, the pay load includes ZW83O-1, which is a 830-nm zwitterionic near- infrared fluorophore. In certain embodiments, the ZW830-1 having the formula:

[0251] In certain embodiments, the zwitterionic groups are introduced into the ZW830-1 using one or more azido-zwitterion derivatives via click chemistry.

[0252] ZN-800-1

[0253] In certain embodiments, the pay load includes ZN-800-1, which is an 800-nm zwitterionic near- infrared fluorophore comprising N-oxide-based zwitterionic groups. In certain embodiments, the ZN-800-1 has the formula:

[0254]

[0255] In certain embodiments, the zwitterionic groups are introduced into the ZN 800-1 using one or more azido-zwitterion derivatives via click chemistry.

[0256] ZN-830-1

[0257] In certain embodiments, the payload includes ZN-830-1, which is an 830-nm zwitterionic near-infrared fluorophore comprising N-oxide-based zwitterionic groups. In certain embodiments, the ZN-830-1 has the formula:

[0258] In certain embodiments, the zwitterionic groups are introduced into the ZN83O-1 using one or more azido-zwitterion derivatives via click chemistry.

[0259] ZN-800-l-Forte In certain embodiments, the payload includes ZN-800-l-Forte, which is an 800-nm zwitterionic near-infrared fluorophore comprising N-oxide-based zwitterionic groups. In certain embodiments, the ZN-800-l-Forte has the formula:

[0260] In certain embodiments, the zwitterionic groups are introduced into the ZN800-1 Forte using one or more azido-zwitterion derivatives via click chemistry.

[0261] ZN-700-l-Forte

[0262] In certain embodiments, the payload includes ZN-700-1, which is an 700-nm zwitterionic near-infrared fluorophore comprising N-oxide-based zwitterionic groups. In certain embodiments, the ZN-800-1 has the formula: In certain embodiments, other zwitterionic NIR fluorophores are used as payload in applications such as image-guided surgery where optical imaging of the malignant cells is required.

[0263] In certain embodiments, the zwitterionic groups are introduced into the ZN700-1 Forte using one or more azido-zwitterion derivatives via click chemistry.

[0264] ZW-DOTA

[0265] In certain embodiments, the payload includes one or more ZW-DOTA. In certain embodiments, the ZW-DOTA includes the one of the following formulas: in which ZWI represents a zwitterionic group; each instance of W and Y independently represent a linking group, and each instance of X represents a reactive group. In certain embodiments, ZWI represents an ammonium sulfobetaine group.

[0266] In such embodiments, a ZW-DOTA contains stereogenic centers, any of these might have R or S -configuration. The ZW-DOTA might be a single stereoisomer or might be a mixture of stereoisomers. The ZW-DOTA includes 1, 2, 3, or 4 zwitterionic groups and 1, 2 or 3 reactive groups X for conjugation to the nexus or other molecules or materials. This reactive group can be a carboxylic acid, an activated derivative of a carboxylic acid, such as an NHS ester, a sulfo-NHS ester, a PFP ester, a HOBt ester, a HOAt ester, a TFP ester, an acid anhydride, an acid azide or an acid halide. The reactive group X can also be an amine, azide, alkyne, alkene, ketone, aldehyde, alcohol, phenol, maleimide, thiol, phosphane or a tetrazine. The zwitterionic groups ZWI and the reactive groups X can be separated from the chelator core structure by appropriate spacer moieties W and Y including alkyl, aryl or heteroaryl groups. In particular embodiments, the ZW-DOTA includes 1, 2, 3, or 4 zwitterionic groups and 4 carboxylate groups for complexation to the metal or metal isotope. In still other embodiments, the ZW-DOTA includes 1, 2, 3, or 4 zwitterionic groups, 4 carboxylate groups for complexation to the metal or metal isotope, and one or more targeting vectors. These targeting vectors can be bound to the chelator at any suitable structural location (e.g. reactive group X) as would be understood by one of ordinary skill in the synthesis of such compounds.

[0267] Such ZW- DOTA can be synthesized using the protocol described in Scheme 2.

[0268] Scheme 2

[0269] In a particular embodiment, the ZW-DOTAs can be complexed with Zr, Cu, Ga, In, Y, Gd, Lu, Tb or other metals.

[0270] In certain embodiments, the DOTA-based zwitterionic payload is not ZWI3-DOTA- Ahx-cRGD.

[0271]

[0272] In certain embodiments, the DOTA-based zwitterionic payload is not:

[0273] In certain embodiments, the DOTA-based zwitterionic payload is not:

[0274]

[0275] In certain embodiments, other zwitterionic DOTA derivatives are used as payload in applications such as radiodiagnosis or radiotherapeutics where chelation of a radiometal is required.

[0276] In certain embodiments, the zwitterionic groups are introduced into the ZW-DOTA using one or more azido-zwitterion derivatives via click chemistry.

[0277] ZW-PyC3A

[0278] In certain embodiments, the payload includes one or more ZW-PyC3A. Tn certain embodiments, the ZW-PyC3A includes the one of the following formulas: wherein each of W and Y may independently be absent. In certain embodiments, ZWI represents an ammonium sulfobetaine group.

[0279] In such embodiments, a ZW-PyC3A includes 1, 2 3 or 4, zwitterionic groups and 1, 2 or 3 reactive groups X for conjugation to the nexus or other molecules or materials. This reactive group can be a carboxylic acid, an activated derivative of a carboxylic acid, such as an NHS ester, a sulfo-NHS ester, a PFP ester, a HOBt ester, a HO At ester, a TFP ester, an acid anhydride, an acid azide or an acid halide. The reactive group X can also be an amine, azide, alkyne, alkene, ketone, aldehyde, alcohol, phenol, maleimide, thiol, phosphane or a tetrazine. The zwitterionic groups ZWI and the reactive groups X can be separated from the chelator core structure by appropriate spacer moieties W and Y including alkyl, aryl or heteroaryl groups. In certain embodiments, each of W and Y may independently be absent. In certain embodiments, ZWI represents an ammonium sulfobetaine group.

[0280] Such ZW-PyC3A can be synthesized using the protocol described in Scheme 3. Scheme 3

[0281] In particular embodiment, the ZW-PyC3A can be complexed with Mn or similar metals.

[0282] In certain embodiments, the agent further comprises a PEG-moiety to alter the circulation time in blood. Such moiety can be bound to the conjugate at any suitable structural location as would be understood by one of ordinary skill in the synthesis of such compounds.

[0283] In certain embodiments, the zwitterionic groups are introduced into the ZW-PyC3A using one or more azido-zwitterion derivatives via click chemistry.

[0284] ZW-Macropa

[0285] In certain embodiments, the payload includes one or more ZW-Macropa. In certain embodiments, the ZW-Macropa includes the one of the following formulas:

[0286] wherein ZW represents a zwitterionic group.

[0287] In such embodiments, a ZW-Macropa includes 1 or 2 zwitterionic groups as defined in Fig. 3 and 1 or 2 reactive groups X for conjugation to the nexus or other molecules or materials. This reactive group can be a carboxylic acid, an activated derivative of a carboxylic acid, such as an NHS ester, a sulfo-NHS ester, a PFP ester, a HOBt ester, a HO At ester, a TFP ester, an acid anhydride, an acid azide or an acid halide. The reactive group X can also be an amine, azide, alkyne, alkene, ketone, aldehyde, alcohol, phenol, maleimide, thiol, phosphane or a tetrazine. The zwitterionic groups ZWI and the reactive groups X can be separated from the chelator core structure by appropriate spacer moieties W and Y including alkyl, aryl or heteroaryl groups. In certain embodiments, each of W and Y may independently be absent. Such ZW-Macropa can be synthesized using the protocol described in Scheme 4.

[0288] -THIS AREA INTENTIONALLY LEFT BLANK- Scheme 4

[0289] In a particular embodiment, the ZW-Macropa can he complexed with Ac3+, Bi3+, or similar metal cations.

[0290] In certain embodiments, the agent further comprises a PEG-moiety to alter the circulation time in blood. Such moiety can be bound to the conjugate at any suitable structural location as would be understood by one of ordinary skill in the synthesis of such compounds.

[0291] In certain embodiments, the zwitterionic groups are introduced into the ZW-Macropa using one or more azido-zwitterion derivatives via click chemistry.

[0292] ZW-porphyrin

[0293] In certain embodiments, the payload includes one or more ZW-porphyrin. In certain embodiments, the ZW-porphyrin includes the one of the following formulas:

[0294]

[0295] (ZWI)2-porphyrin-X2V2 ZWI-porphyrin-X3wherein ZW represents a zwitterionic group. In certain embodiments, ZW represents an ammonium sulfobetaine group.

[0296] In such embodiments, a ZW-Porphyrin includes 1, 2, 3, or 4 zwitterionic groups and 1 or 2 reactive groups X for conjugation to the nexus or other molecules or materials. This reactive group can be a carboxylic acid, an activated derivative of a carboxylic acid, such as an NHS ester, a sulfo-NHS ester, a PFP ester, a HOBt ester, a HO At ester, a TFP ester, an acid anhydride, an acid azide or an acid halide. The reactive group X can also be an amine, azide, alkyne, alkene, ketone, aldehyde, alcohol, phenol, maleimide, thiol, phosphane or a tetrazine. The zwitterionic groups ZWI and the reactive groups X can be separated from the chelator core structure by appropriate spacer moieties W and Y including alkyl, aryl or heteroaryl groups. In certain embodiments, each of W and Y may independently be absent.In particular embodiments, the ZW-Porphyrin includes 4 zwitterionic groups. In certain embodiments, ZWI represents an ammonium sulfobetaine group. In certain embodiments, the chelator comprises a reactive linking group, typically an activated derivative of a carboxylic acid, such as an NHS ester, a sulfo-NHS ester, a PFP ester, a HOBt ester, a HO At ester, a TFP ester, an acid anhydride, an acid azide or an acid halide. Such reactive linking groups can be bound or substituted onto the chelator at any suitable structural location as would be understood by one of ordinary skill in the synthesis of such compounds.

[0297] In particular embodiments, the ZW-Porphyrin has the formula: wherein the zwitterion is a pyridine-N-oxide (as drawn above) or an aromatic N-oxide derived from imidazole, pyrimidine or a similar heteroaromatic residue. X is a reactive group for conjugation to the nexus or other molecules or materials. This reactive group can be a carboxylic acid, an activated derivative of a carboxylic acid, such as an NHS ester, a sulfo- NHS ester, a PFP ester, a HOBt ester, a HOAt ester, a TFP ester, an acid anhydride, an acid azide or an acid halide. The reactive group X can also be an amine, azide, alkyne, alkene, ketone, aldehyde, alcohol, phenol, maleimide, thiol, phosphane or a tetrazine. The reactive group X can be separated from the chelator core structure by appropriate spacer moieties W including alkyl, aryl or heteroaryl groups. In certain embodiments, each of W and Y may independently be absent.

[0298] ZW-Porphyrins can be synthesized using the protocol described in Scheme 5. Scheme 5

[0299] In a particular embodiment, the porphyrin -based zwitterionic metal chelators can be complexed with Mn2+, Mn3+, Fe2+, Fe3+, Gd3+, Ac3+, Bi3+, or similar metal cations.

[0300] In certain embodiments, the zwitterionic groups are introduced into the ZW-porphyrin using one or more azido-zwitterion derivatives via click chemistry.

[0301] ZW-NOTA

[0302] In certain embodiments, the payload includes one or more ZW-NOTA. In certain embodiments, the ZW-NOTA includes one of the following formulas:

[0303]

[0304] (ZWI)3-NOTA (ZWI)2-NOTA-X ZWI-NOTA-X2

[0305] In such embodiments, a NOTA-based zwitterionic metal chelator includes 1, 2 or 3 zwitterionic groups. In particular embodiments, the NOTA-based zwitterionic metal chelator includes 2 zwitterionic groups and 1 or 2 reactive groups X for conjugation to the nexus or other molecules or materials. This reactive group can be a carboxylic acid, an activated derivative of a carboxylic acid, such as an NHS ester, a sulfo-NHS ester, a PFP ester, a HOBt ester, a HO At ester, a TFP ester, an acid anhydride, an acid azide or an acid halide. The reactive group X can also be an amine, azide, alkyne, alkene, ketone, aldehyde, alcohol, phenol, maleimide, thiol, phosphane or a tetrazine. The zwitterionic groups ZWI and the reactive groups X can be separated from the chelator core structure by appropriate spacer moieties W and Y including alkyl, aryl or heteroaryl groups. In certain embodiments, each of W and Y may independently be absent.

[0306] In particular embodiments, the NOTA -based zwitterionic metal chelator has the formula: protected (ZWI)2-NOTA-CO2H protected (ZWI)2-NOTA-NH2protected (ZWI)2-NOTA-N3 Such NOTA -based zwitterionic metal chelators can be synthesized using the protocol described in Scheme 6.

[0307] Scheme 6

[0308] In a particular embodiment, the NOTA-based zwitterionic metal chelators can be complexed with Ga3+, Cu2+, Gd3+, Ac3+, Bi3+, or similar metal cations.

[0309] In certain embodiments, the zwitterionic groups are introduced into the ZW-NOTA using one or more azido-zwitterion derivatives via click chemistry.

[0310] ZW-deferoxamine

[0311] In certain embodiments, the payload includes one or more ZW-deferoxamine. In certain embodiments, the ZW-deferoxamine has the formula:

[0312] -THIS AREA INTENTIONALLY LEFT BLANK-

[0313]

[0314] In such embodiments, a deferoxamine-based zwitterionic metal chelator with four hydroxamate groups is derived from DFO* and one with three hydroxamate groups is derived from DFO. A deferoxamine-based zwitterionic metal chelator includes 1, 2, 3 or 4 zwitterionic groups as defined in Fig. 3 and 1 or 2 reactive groups X for conjugation to the nexus or other molecules or materials. This reactive group can be a carboxylic acid, an activated derivative of a carboxylic acid, such as an NHS ester, a sulfo-NHS ester, a PFP ester, a HOBt ester, a HO At ester, a TFP ester, an acid anhydride, an acid azide or an acid halide. The reactive group X can also be an amine, azide, alkyne, alkene, ketone, aldehyde, alcohol, phenol, maleimide, thiol, phosphane or a tetrazine. The zwitterionic groups ZWI and the reactive groups X can be separated from the chelator core structure by appropriate spacer moieties W and Y including alkyl, aryl or heteroaryl groups. In certain embodiments, each of W and Y may independently be absent. In still other embodiments, W-X can represent a hydrogen or an alkyl group.

[0315] In certain embodiments, the chelator comprises a reactive linking group, typically an activated derivative of a carboxylic acid, such as an NHS ester, a sulfo-NHS ester, a PFP ester, a HOBt ester, a HO At ester, a TFP ester, an acid anhydride, an acid azide or an acid halide. Such reactive linking groups can be bound or substituted onto the chelator at any suitable structural location as would be understood by one of ordinary skill in the synthesis of such compounds.

[0316] Such deferoxamine -based zwitterionic metal chelators can be synthesized using the protocol described in Scheme 7below:

[0317] In certain embodiments, the zwitterionic groups are introduced into the ZW- deferoxamine using one or more azido-zwitterion derivatives via click chemistry. ZW-DOTAM

[0318] In one aspect, the ZW-DOTAM payload includes one or more of 1,4,7,10- Tetraazacyclododecane-l,4,7,10-tetraacetic amide, also known as DOTAM. DOT AM is a derivative of DOTA in which -COOH groups are replaced with -CONR2 groups. Such DOTAM has a formula of the following: in which ZWI represents a zwitterionic group; each instance of W and Y independently represent a linking group, and each instance of X represents a reactive group. In certain embodiments, ZWI represents an ammonium sulfobetaine group. In other embodiments ZWI represents an N-oxide group. Substituents R are hydrogen or alkyl groups or a combination of both.

[0319] In such embodiments, a DOT AM-based zwitterionic payload contains stereogenic centers, any of these might have R or S -configuration. The DOTAM-based zwitterionic pay load might be a single stereoisomer or might be a mixture of stereoisomers. The DOTAM-based zwitterionic payload includes 1, 2, 3, or 4 zwitterionic groups and 1, 2 or 3 reactive groups X for conjugation to targeting vectors or other molecules or materials. This reactive group can be a carboxylic acid, an activated derivative of a carboxylic acid, such as an NHS ester, a sulfo-NHS ester, a PFP ester, a HOBt ester, a HOAt ester, a TFP ester, an acid anhydride, an acid azide or an acid halide. The reactive group X can also be an amine, azide, alkyne, alkene, ketone, aldehyde, alcohol, phenol, maleimide, thiol, phosphane or a tetrazine. The zwitterionic groups ZWI and the reactive groups X can be separated from the chelator core structure by appropriate spacer moieties W and Y including alkyl, aryl or heteroaryl groups. In particular embodiments, the DOTAM-based zwitterionic payload includes 1, 2, 3, or 4 zwitterionic groups and 4 carboxylate groups for complexation to the metal or metal isotope. In still other embodiments, the DOTAM-based zwitterionic payload includes 1, 2, 3, or 4 zwitterionic groups, 4 carboxylate groups for complexation to the metal or metal isotope, and one or more targeting vectors. These targeting vectors can be bound to the chelator at any suitable structural location (e.g. reactive group X) as would be understood by one of ordinary skill in the synthesis of such compounds.

[0320] DOTAM-based zwitterionic payload can be synthesized using the protocol described in Scheme 8.

[0321] Scheme 8

[0322] In a particular embodiment, the DOTAM-based zwitterionic payload can be complexed with Pb, Zr, Cu, Ga, In, Y, Gd, Lu, Ac, Tb, Pb, Pd or other metals.

[0323] In certain embodiments, the agent further comprises a PEG-moiety to alter the circulation time in blood. Such moiety can be bound to the conjugate at any suitable structural location as would be understood by one of ordinary skill in the synthesis of such compounds.

[0324] In a particular embodiment, the deferoxamine -based zwitterionic metal chelators can be complexed with Zr4+, Fe3+, Mn2+, Mn3+, or similar metal cations.

[0325] In certain embodiments, the payload has zwitterionic metal chelators coordinated to or labeled with a metal or metal isotope for detection, imaging, and treatment in a subject or in a biological sample. Non-limiting examples of metals that may be complexed with the chelators of the invention include Gd, Mn, Cu, Co, Y, In, Ga, Zr, Tc, Eu, Tb, Ac, Lu and other lanthanide or actinide metals. In some embodiments, the metals can be radioactive metal isotopes. Non-limiting examples of radioactive metal isotopes that may be used include Ga-66, Ga-67, Ga-68, Cu-64, Cu-67, Y-86, Co-55, Zr-89, Sr-83, Mn-52, As-72, Sc- 44, Gd-153, Co-57, In-111, Zr-89, Ac-225, Tc-99m, or F-18 FDG.

[0326] For the disclosed methods of therapeutically treating malignant tumors, any radioactive metal isotope known to emit ionizing radiation in a form that would result in the death of cells that take up the analogs labeled with the radioactive metal isotope can be incorporated by chelation in the payload of the invention. So, too, can non-radioactive metals that are capable of capturing neutrons and releasing cytotoxic radiation (neutron capture therapy). In some embodiments, the radioactive metal isotope emits its ionizing radiation in a form that minimizes damage to tissue outside of the cells that take up the labeled analogs.

[0327] In certain embodiments, the payload of the invention comprise zwitterionic metal chelators, which comprise at least one radioactive isotope of technetium, rhenium, gallium, indium, copper, yttrium, actinium, bismuth, samarium, dysprosium, holmium, or lutetium, including radioactive isotopes selected from Tc-99m, Tc-94m, Re- 186, Re- 188, Ga-68, Cu- 64, Cu-67, Y-90, Y-86, Ac-225, Bi-213, In-111, Sm-153, Ho-166, Lu-177, Sc-43, Sc-44, Sc- 47, Tb-149, Tb-152, Tb-155, Tb-161, and Dy-166. In other embodiments, non-limiting examples of metals used in neutron capture therapy include Zr-88 or Gd-157.

[0328] In certain embodiments, the payload is a conventional chemotherapeutic and / or radiosensitizer such as SN-38, doxorubicin, daunorubicin, maitansine, monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), mertansine, calicheamicin, and exatecan. In certain embodiments, the zwitterionic groups are introduced into the ZW-DOTAM using one or more azido-zwitterion derivatives via click chemistry.

[0329] ZW-DOTP

[0330] In certain embodiments, the payload includes one or more ZW-DOTP. In certain embodiments, the ZW-DOTP includes the one of the following formulas: a zwitterionic metal chelator based on l,4,7,10-tetraazacyclododecane-l,4,7,10-(tetra(methylene phosphonic acid, also known as DOTP. DOTP is a derivative of DOTA in which -COOH groups are replaced with -PO(OH)2 groups. Such zwitterionic metal chelators include the following: in which ZWI represents a zwitterionic group; each instance of W and Y independently represent a linking group, and each instance of X represents a reactive group. In certain embodiments, ZWI represents an ammonium sulfobetaine group. In other embodiments ZWI represents an N-oxide group. Substituents R are hydrogen or alkyl groups or a combination of both.

[0331] In such embodiments, a DOTP -based zwitterionic metal chelator contains stereogenic centers, any of these might have R or S -configuration. The DOTP -based zwitterionic metal chelator might be a single stereoisomer or might be a mixture of stereoisomers. The DOTP - based zwitterionic metal chelator includes 1, 2, 3, or 4 zwitterionic groups and 1, 2 or 3 reactive groups X for conjugation to targeting vectors or other molecules or materials. This reactive group can be a carboxylic acid, an activated derivative of a carboxylic acid, such as an NHS ester, a sulfo-NHS ester, a PFP ester, a HOBt ester, a HOAt ester, a TFP ester, an acid anhydride, an acid azide or an acid halide. The reactive group X can also be an amine, azide, alkyne, alkene, ketone, aldehyde, alcohol, phenol, maleimide, thiol, phosphane or a tetrazine. The zwitterionic groups ZWI and the reactive groups X can be separated from the chelator core structure by appropriate spacer moieties W and Y including alkyl, aryl or heteroaryl groups. In particular embodiments, the DOTP -based zwitterionic metal chelator includes 1, 2, 3, or 4 zwitterionic groups and 4 carboxylate groups for complexation to the metal or metal isotope. In still other embodiments, the DOTP -based zwitterionic metal chelator includes 1, 2, 3, or 4 zwitterionic groups, 4 carboxylate groups for complexation to the metal or metal isotope, and one or more targeting vectors. These targeting vectors can be bound to the chelator at any suitable structural location (e.g. reactive group X) as would be understood by one of ordinary skill in the synthesis of such compounds.

[0332] DOTP-based zwitterionic metal chelators can be synthesized using the protocol described in Scheme 9.

[0333] -THIS AREA INTENTIONALLY LEFT BLANK- Scheme 9

[0334] DOTP-based zwitterionic metal chelators can also be synthesized using the protocol described in Scheme 10.

[0335] Scheme 10

[0336] In a particular embodiment, the DOTP-based zwitterionic payload (zwitterionic metal chelators), as a base payload, can be complexed with Bi, Pb, Zr, Cu, Ga, In, Y, Gd, Lu, Ac, Tb, Pb, Pd or other metals as a special payload. In particular embodiments, the DOTP-based zwitterionic metal chelators chelate Bi3+ more tightly than commercially available metal chelators. This has the effect of minimizing the ejection of Pb-212 / Bi-3+ from the chelated complex.

[0337] In certain embodiments, the agent further comprises a PEG-moiety to alter the circulation time in blood. Such moiety can be bound to the conjugate at any suitable structural location as would be understood by one of ordinary skill in the synthesis of such compounds.

[0338] In certain embodiments, the conjugate of the DOTP -based zwitterionic metal chelator and the targeting vector has the following formula with KUE, dPSMA-617, cRGD, FAPI, octreotide, bombesin, or as the targeting vector (tv):

[0339] (ZWI)4-DOTP-tv (ZWI)3-DOTP-tv

[0340] In certain embodiments, the zwitterionic groups are introduced into the ZW-DOTP using one or more azido-zwitterion derivatives via click chemistry.

[0341] ZW-PCTA

[0342] In certain embodiments, the payload includes one or more ZW-PCTA. In certain embodiments, the ZW-PCTA includes the one of the following formulas: a zwitterionic metal chelator based on 2-[3,9-bis[l-carboxylato-4-(2,3-dihydroxypropylamino)-4-oxobutyl]- 3,6,9, 15-tetrazabicyclo[9.3.1]pentadeca- 1(15), 1 l,13-trien-6-yl]-5-(2,3- dihydroxypropylamino)-5-oxopentanoate;gadolinium(3+), also known as gadopiclenol. In particular, the metal chelator is a derivative of 3,6,9, 15 -Tetraazabicyclo [9.3.1] pentadeca- l(15),l l,13-triene-3,6,9-triacetic acid (PCTA). Such zwitterionic metal chelators include the following:

[0343] ZW-PCTA Targeted- ZW-PCTA in which ZW represents a zwitterionic group. In certain embodiments, ZW represents an ammonium sulfobetaine group.

[0344] In such embodiments, a PCTA-based zwitterionic metal chelator contains stereogenic centers, any of these might have R or S -configuration. The PCTA-based zwitterionic metal chelator might be a single stereoisomer or might be a mixture of stereoisomers. The PCTA- based zwitterionic metal chelator includes 1, 2, 3, or 4 zwitterionic groups and 1, 2 or 3 reactive groups X for conjugation to targeting vectors or other molecules or materials. This reactive group can be a carboxylic acid, an activated derivative of a carboxylic acid, such as an NHS ester, a sulfo-NHS ester, a PFP ester, a HOBt ester, a HOAt ester, a TFP ester, an acid anhydride, an acid azide or an acid halide. The reactive group X can also be an amine, azide, alkyne, alkene, ketone, aldehyde, alcohol, phenol, maleimide, thiol, phosphane or a tetrazine. The zwitterionic groups ZWI and the reactive groups X can be separated from the chelator core structure by appropriate spacer moieties W and Y including alkyl, aryl or heteroaryl groups. In particular embodiments, the PCTA-based zwitterionic metal chelator includes 1, 2, 3, or 4 zwitterionic groups and 4 carboxylate groups for complexation to the metal or metal isotope. In still other embodiments, the PCTA-based zwitterionic metal chelator includes 1, 2, 3, or 4 zwitterionic groups, 4 carboxylate groups for complexation to the metal or metal isotope, and one or more targeting vectors. These targeting vectors can be bound to the chelator at any suitable structural location (e.g. reactive group X) as would be understood by one of ordinary skill in the synthesis of such compounds.

[0345] PCTA-based zwitterionic metal chelators can be synthesized using the protocols described in Scheme 11.

[0346] Scheme 11

[0347] The protocols shown in the scheme provide an example for ZW to be an / V-oxide group or a sulphobetaine group.

[0348] Alternatively, A-oxide functionalities may be synthesized by nucleophilic displacement of a suitable leaving group (such as a halogenide or a sulfonate) instead of the group R in the scheme above. The nucleophile in these conversions is an A,A-dialkylated hydroxy] amine, which can be protected at the hydroxyl group.

[0349] In a particular embodiment, the PCTA-based zwitterionic metal chelators, as a base payload, can be complexed with Zr, Cu, Ga, In, Y, Gd, Lu, Ac, Pb, Mn, Tb or other metals as a special payload.

[0350] In certain embodiments, the agent further comprises a PEG-moiety to alter the circulation time in blood. Such moiety can be bound to the conjugate at any suitable structural location as would be understood by one of ordinary skill in the synthesis of such compounds.

[0351] In certain embodiments, the zwitterionic groups are introduced into the ZW-PCTA using one or more azido-zwitterion derivatives via click chemistry.

[0352] ZW-Amide-PCTA

[0353] In certain embodiments, the payload includes one or more ZW- Amide-PCTA. In certain embodiments, the ZW- Amide-PCTA includes the one of the following formulas: Amide-PCTA. Amide-PCTA is a derivative of PCTA in which -COOH groups are replaced with -CONR2 groups. Such zwitterionic metal chelators include the following:

[0354] ZW-amide-PCTA Targeted ZW-amide-PCTA wherein ZW represents a zwitterionic group. Substituents R can be targeting vectors or reactive groups. Substituents R1and R2can be the same or different and are H, alkyl or aryl groups. In certain embodiments, substituents R1and R2are each H.

[0355] In certain embodiments, ZW represents an ammonium sulfobetaine group. In other embodiments ZW represents an A-oxide group.

[0356] In such embodiments, an amide-PCTA-based zwitterionic metal chelator contains stereogenic centers, any of these might have R or S-configuration. The amide-PCTA-based - based zwitterionic metal chelator might be a single stereoisomer or might be a mixture of stereoisomers. The amide-PCTA-based zwitterionic metal chelator includes 1 , 2, 3, or 4 zwitterionic groups and 1, 2 or 3 reactive groups for conjugation to targeting vectors or other molecules or materials. This reactive group can be a carboxylic acid, an activated derivative of a carboxylic acid, such as an NHS ester, a sulfo-NHS ester, a PFP ester, a HOBt ester, a HO At ester, a TFP ester, an acid anhydride, an acid azide or an acid halide. The reactive group can also be an amine, azide, alkyne, alkene, ketone, aldehyde, alcohol, phenol, maleimide, thiol, phosphane or a tetrazine. The zwitterionic groups ZW and the reactive groups can be separated from the chelator core structure by appropriate spacer moieties including alkyl, aryl or heteroaryl groups. In particular embodiments, the amide-PCTA- based zwitterionic metal chelator includes 1, 2, 3, or 4 zwitterionic groups and 4 carboxylate groups for complexation to the metal or metal isotope. In still other embodiments, the amide-PCTA-based zwitterionic metal chelator includes 1, 2, 3, or 4 zwitterionic groups, 4 carboxylate groups for complexation to the metal or metal isotope, and one or more targeting vectors. These targeting vectors can be bound to the chelator at any suitable structural location (e.g. at a reactive group) as would be understood by one of ordinary skill in the synthesis of such compounds.

[0357] Amide-PCTA-based zwitterionic metal chelators can be synthesized using the protocol described in Scheme 12.

[0358] -THIS AREA INTENTIONALLY LEFT BLANK-

[0359] Scheme 12

[0360] The protocols shown in the scheme provide an example for ZW to be an / V-oxide group or a sulphobetaine group.

[0361] Alternatively, A-oxide functionalities may be synthesized by nucleophilic displacement of a suitable leaving group (such as a halogenide or a sulfonate) instead of the group R in the scheme above. The nucleophile in these conversions is a N, A-dialkylated hydroxylamine, which can be protected at the hydroxyl group.

[0362] In certain embodiments, the amide-PCTA-based zwitterionic metal chelators, as a base payload, can be complexed with Pb, Mn, Zr, Cu, Ga, In, Y, Gd, Lu, Ac, Tb, Pb, Pd or other metals, as a special payload.

[0363] In certain embodiments, the zwitterionic groups are introduced into the ZW-Amide- PCTA using one or more azido-zwitterion derivatives via click chemistry.

[0364] ZW-PCTP

[0365] In certain embodiments, the payload includes one or more ZW-PCTP. In certain embodiments, the ZW-PCTP includes the one of the following formulas: a zwitterionic metal chelator based on 2-[3,9-bis[l-carboxylato-4-(2,3-dihydroxypropylamino)-4-oxobutyl]- 3,6,9,15-tetrazabicyclo[9.3.1]pentadeca-l(15),l l,13-trien-6-yl]-5-(2,3- dihydroxypropylamino)-5-oxopentanoate;gadolinium(3+), also known as gadopiclenol. In particular, the metal chelator is a derivative of 3,6,9, 15-Tetraazabicyclo[9.3.1] pentadeca- l(15),l l,13-triene-3,6,9-triacetic acid (PCTA) in which -COOH groups are replaced with - P0(0H)2 groups. Such zwitterionic metal chelators include the following:

[0366] Targeted-ZW-PCTP in which ZW represents a zwitterionic group. In certain embodiments, ZW represents an ammonium sulfobetaine group. In such embodiments, a PCTA-based zwitterionic metal chelator contains stereogenic centers, any of these might have R or S-configuration. The PCTA-based zwitterionic metal chelator might be a single stereoisomer or might be a mixture of stereoisomers. The PCTA- based zwitterionic metal chelator includes 1, 2, 3, or 4 zwitterionic groups and 1, 2 or 3 reactive groups X for conjugation to targeting vectors or other molecules or materials. This reactive group can be a carboxylic acid, an activated derivative of a carboxylic acid, such as an NHS ester, a sulfo-NHS ester, a PFP ester, a HOBt ester, a HOAt ester, a TFP ester, an acid anhydride, an acid azide or an acid halide. The reactive group X can also be an amine, azide, alkyne, alkene, ketone, aldehyde, alcohol, phenol, maleimide, thiol, phosphane or a tetrazine. The zwitterionic groups ZWI and the reactive groups X can be separated from the chelator core structure by appropriate spacer moieties W and Y including alkyl, aryl or heteroaryl groups. In particular embodiments, the PCTA-based zwitterionic metal chelator includes 1, 2, 3, or 4 zwitterionic groups and 4 carboxylate groups for complexation to the metal or metal isotope. In still other embodiments, the PCTA-based zwitterionic metal chelator includes 1 , 2, 3, or 4 zwitterionic groups, 4 carboxylate groups for complexation to the metal or metal isotope, and one or more targeting vectors. These targeting vectors can be bound to the chelator at any suitable structural location (e.g. reactive group X) as would be understood by one of ordinary skill in the synthesis of such compounds.

[0367] PCTP-based zwitterionic metal chelators can be synthesized using the protocols described in Scheme 13.

[0368] -THIS AREA INTENTIONALLY LEFT BLANK-

[0369] Scheme 13 The protocols shown in the scheme can be modified to provide an example for ZW to be an A-oxide group or a sulphobetaine group.

[0370] Alternatively, A-oxide functionalities may be synthesized by nucleophilic displacement of a suitable leaving group (such as a halogenide or a sulfonate) instead of the group R in the scheme above. The nucleophile in these conversions is an N, A-di alkylated hydroxylamine, which can be protected at the hydroxyl group.

[0371] In a particular embodiment, the PCTA-based zwitterionic metal chelators, as a base payload, can be complexed with Zr, Cu, Ga, In, Y, Gd, Lu, Ac, Pb, Mn, Tb or other metals, as a special payload.

[0372] In certain embodiments, the zwitterionic groups are introduced into the ZW-PCTP using one or more azido-zwitterion derivatives via click chemistry.

[0373] Imaging Methods

[0374] In one aspect, the invention provides for methods of biomedical imaging of tissues or cells in a biological sample. In particular embodiment, the invention encompasses a method for detecting or imaging one or more cancer cells in a biological sample.

[0375] In some embodiments, the method is suitable for imaging of abnormal, but not malignant, tissue, such as defects of the musculoskeletal system using FAP as a targeting ligand, vascular system using cRGD as a targeting ligand, melanoma using LyP- 1 peptide as a targeting ligand, breast cancer using K237 peptide and / or mUNO as a targeting ligand, Lung tumor, breast tumor, colon tumor using IL4RPep-l as a targeting ligand.

[0376] In some embodiments, the method is suitable for imaging of cancer, tumor or neoplasm. In a further embodiment, the cancer is selected from eye or ocular cancer, rectal cancer, colon cancer, cervical cancer, prostate cancer, breast cancer and bladder cancer, oral cancer, benign and malignant tumors, stomach cancer, liver cancer, pancreatic cancer, lung cancer, corpus uteri, ovary cancer, prostate cancer, testicular cancer, renal cancer, brain / cns cancer (e.g., gliomas), throat cancer, skin melanoma, acute lymphocytic leukemia, acute myelogenous leukemia, Ewing's Sarcoma, Kaposi's Sarcoma, basal cell carinoma and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms Tumor, neuroblastoma, mouth / pharynx cancer, esophageal cancer, larynx cancer, lymphoma, neurofibromatosis, tuberous sclerosis, hemangiomas, and lymphangiogenesis.

[0377] In some embodiments, the cancer cells are adult solid tumor cells or pediatric solid tumor cells. Non-limiting examples of such cells include melanoma cells, neuroblastoma cells, lung cancer cells, adrenal cancer cells, colon cancer cells, colorectal cancer cells, ovarian cancer cells, prostate cancer cells, liver cancer cells, subcutaneous cancer cells, squamous cell cancer cells, intestinal cancer cells, retinoblastoma cells, cervical cancer cells, glioma cells, breast cancer cells, pancreatic cancer cells, Ewings sarcoma cells, rhabdomyosarcoma cells, osteosarcoma cells, retinoblastoma cells, Wilms' tumor cells, and pediatric brain tumor cells.

[0378] In particular embodiments, the cancer cells are prostate cancer cells.

[0379] In some embodiments, the biological sample is part or all of a subject. In some embodiments, the biological sample is obtained from a subject.

[0380] In particular embodiments, the method includes the steps of (a) contacting the biological sample with a combination of the imaging agent conjugates described above, wherein each of the imaging agent conjugates in the combination comprises a targeting ligand linked to an imaging agent. The imaging agent is capable of being detected by one or more conventional scanning methods.

[0381] In some embodiments, the compound is administered by parenteral, intranasal, sublingual, rectal, or transdermal delivery. In some such embodiments, the compound is administered intravenously. In some embodiments, the compound is administered intratumorally.

[0382] In some embodiments, the tumor or cell is found in a subject. The subject treated by the presently disclosed methods in their many embodiments is desirably a human subject, although it is to be understood that the methods described herein are effective with respect to all vertebrate species, which are intended to be included in the term “subject.” Accordingly, a “subject” can include a human subject for medical purposes, such as for the treatment of an existing condition or disease or the prophylactic treatment for preventing the onset of a condition or disease, or an animal (non-human) subject for medical, veterinary purposes, or developmental purposes. Suitable animal subjects include mammals including, but not limited to, primates, e.g., humans, monkeys, apes, and the like; bovines, e.g., cattle, oxen, and the like; ovines, e.g., sheep and the like; caprines, e.g., goats and the like; porcines, e.g., pigs, hogs, and the like; equines, e.g., horses, donkeys, zebras, and the like; felines, including wild and domestic cats; canines, including dogs; lagomorphs, including rabbits, hares, and the like; and rodents, including mice, rats, and the like. An animal may be a transgenic animal. In some embodiments, the subject is a human including, but not limited to, fetal, neonatal, infant, juvenile, and adult subjects. Further, a “subject” can include a patient afflicted with or suspected of being afflicted with a condition or disease. Thus, the terms “subject” and “patient” are used interchangeably herein. In some embodiments, the subject is human. In other embodiments, the subject is non-human.

[0383] In particular embodiments, the subject is a human.

[0384] The methods of imaging tissue or cells include the following basic steps:

[0385] (a) contacting the tissue or cells with a combination of imaging agent conjugates; and

[0386] (b) imaging the tissue or cells using positron emission tomography (PET), single-photon emission computerized tomography (SPECT), or magnetic resonance imaging (MRI).

[0387] The imaging agent conjugate described herein is a substance to that can be used to image tissues or cells, such as those of a living organism, for purposes of diagnosis, therapy, image-guided surgery, and the like. In some embodiments, the organism is a mammal, such as a human.

[0388] The imaging agent conjugates described herein generally has improved “signal-to- background ratio” (SBR) compared to presently known imaging agents. The improvement in SBR is believed to be a result of improved in vivo properties due to “charge-balancing.” SBR is a measure of the intensity of the signal obtained from a target (peak signal) over the measure of the intensity of the signal obtained nearby the target (background signal), the target being the tissues or cells targeted by the imaging agent. SBR measurements can be readily obtained through routine measurement procedures. Higher SBR values are more desirable, resulting in greater resolution of the imaged tissues. In some embodiments, the imaging agents achieve an SBR of at least about 1.1 (i.e., peak signal is at least 10% over background). In further embodiments, the imaging agents achieve an SBR of at least about 1.2, at least about 1.3, at least about 1.4, at least about 1.5, at least about 1.6, at least about 1.7, at least about 1.8, at least about 1.9, or at least about 2.0. In yet further embodiments, the imaging agents achieve an SBR of about 1.1 to about 50, about 1.5 to about 30, about 2.0 to about 20, about 2.0 to about 5.0, or about 5.0 to about 10.

[0389] In another aspect, the disclosure encompasses a method of measuring and / or monitoring the effectiveness of various biological functions of a subject. In particular embodiments, the method provides for measuring the effectiveness of hepatic function, renal function, or blood pooling in a subject. In such embodiments, the zwitterionic metal chelators may be used with or without the addition of a targeting ligand. The method includes the step of administering to a subject in need of measurement of a biological function with a quantifiable amount of one or more of the zwitterionic metal chelators described above; imaging the subject using positron emission tomography (PET), singlephoton emission computerized tomography (SPECT), or magnetic resonance imaging (MRI); and determining the amount of the zwitterionic metal chelator present in the biological function being observed.

[0390] Treatment, Diagnosis, and Monitoring Methods

[0391] Cancerous Conditions

[0392] In another aspect, the disclosure encompasses a method for inhibiting the proliferation or growth of malignant or non-malignant cells. The method includes the step of contacting one or more cells with an effective amount of a combination of imaging agent conjugates described above.

[0393] In some embodiments, the method is performed in vivo, ex vivo, or in vitro.

[0394] In some embodiments, the malignant cells are adult solid tumor cells or pediatric solid tumor cells. Non-limiting examples of such cells include melanoma cells, neuroblastoma cells, lung cancer cells, adrenal cancer cells, colon cancer cells, colorectal cancer cells, ovarian cancer cells, prostate cancer cells, liver cancer cells, subcutaneous cancer cells, squamous cell cancer cells, intestinal cancer cells, retinoblastoma cells, cervical cancer cells, glioma cells, breast cancer cells, pancreatic cancer cells, Ewings sarcoma cells, rhabdomyosarcoma cells, osteosarcoma cells, retinoblastoma cells, Wilms' tumor cells, and pediatric brain tumor cells.

[0395] In another aspect, the disclosure encompasses a method of diagnosing cancer in a subject. The method includes one or more of the imaging / detection steps outlined above. Tn the method, the biological sample is obtained from, part of, or all of a subject. If cancer cells are detected or imaged in the method steps, the subject is diagnosed with cancer. In certain embodiments, the method of diagnosing cancer in a subject is followed by a step of treating the subject diagnosed with cancer with a cancer therapy. In some embodiments, the cancer therapy is, surgery, chemotherapy or radiotherapy.

[0396] In some embodiments, the cancer that is diagnosed is an adult solid tumor or a pediatric solid tumor. Non-limiting examples of such cancer include melanoma, neuroblastoma, lung cancer, adrenal cancer, colon cancer, colorectal cancer, ovarian cancer, prostate cancer, liver cancer, subcutaneous cancer, squamous cell cancer, intestinal cancer, retinoblastoma, cervical cancer, glioma, breast cancer, pancreatic cancer, Ewings sarcoma, rhabdomyosarcoma, osteosarcoma, retinoblastoma, Wilms’ tumor, and pediatric brain tumors.

[0397] In still another aspect, the disclosure encompasses a method of monitoring the efficacy of a cancer therapy in a human subject. The method includes performing one or more of the imaging / detection steps outlined above at two or more different times on the biological sample, wherein the biological sample is obtained from, part of, or all of a subject. The change in strength of the signals characteristic of the metal isotope between the two or more different times is correlated with the efficacy of the cancer therapy.

[0398] In another aspect, the disclosure encompasses a method of treatment of abnormal, but not malignant, tissue, such as defects of the musculoskeletal system using FAP as a targeting ligand, vascular system using cRGD as a targeting ligand, melanoma using LyP- 1 peptide as a targeting ligand, breast cancer using K237 peptide and / or mUNO as a targeting ligand, Lung tumor, breast tumor, colon tumor using IL4RPep-l as a targeting ligand.

[0399] The method includes the step of contacting one or more abnormal cells with an effective amount of a small zwitterionic multimeric targeted anti-cancer drugs described above, wherein the metal atom is a radioactive metal isotope known to emit ionizing radiation in a form that would result in a therapeutic effect on the cells that take up the analogs.

[0400] In another aspect, the disclosure encompasses a method of treating a cancer by administering an effective amount of a therapeutic agent comprising a small zwitterionic multimeric targeted anti-cancer drugs and a pharmaceutically acceptable carrier or excipient. The method comprises a step to diagnose the cancer and a step of administering the therapeutic agent to a subject determined to be in need thereof; wherein the step to diagnose the cancer comprises: contacting cells, tissues or organs of a subject with a combination of the imaging agent conjugates, imaging the cells, tissues, or organs of the subject using positron emission tomography (PET), single-photon emission computerized tomography (SPECT), or magnetic resonance tomography (MRT), and diagnosing the cancer in the cells tissues, or organs of the subject based on imaging data collected; and wherein the metal atom complexed to the imaging agent dyes of the conjugates is: a radioactive metal isotope known to emit ionizing radiation that results in the death of cells that take up the analogs; or a non-radioactive metal that is capable of releasing cytotoxic radiation upon irradiation with alpha emission, beta emission, neutron capture, or a combination thereof.

[0401] In still another aspect, the disclosure encompasses a method of treating a rare or childhood cancer by administering an effective amount of a therapeutic agent comprising a combination of conjugates described above and a pharmaceutically acceptable carrier or excipient. In such aspects, the therapeutic agent may further comprise one or more targeting ligands selected from the group consisting of cRGD, PS MA, FAP, bombesin, octreotide, octreotide or a dimer formed from their combination.

[0402] In particular embodiments, the rare or childhood cancer is: Acinic cell carcinoma, ACTH-secreting tumor, Actinic keratosis, Adamantinoma, Adenoid cystic carcinoma, Alveolar soft part sarcoma, Ampullary cancer, Angiosarcoma, Appendix (appendiceal) neuroendocrine (carcinoid) tumor, Askin tumor - a type of Ewing tumor (Ewing sarcoma), Bartholin gland cancer, Basaloid squamous cell carcinoma of the anus, Bowen disease, Bronchioloalveolar carcinoma, Carcinoid tumor, Carcinoma of the ampulla of Vater, Cardiac angiosarcoma, Castleman disease, Cholangiocarcinoma, Choriocarcinoma, Choroid plexus tumor, Chondrosarcoma, Chordoma, Chromophobe renal cell carcinoma, Clear cell sarcoma, Craniopharyngioma, Dermatofibrosarcoma protuberans, Desmoid tumor, Desmoplastic small round cell tumor, Dysgerminoma, Embryonal carcinoma, Endodermal sinus tumor, Endometrial stromal sarcoma, Ependymoma, Epithelial appendix (appendiceal) cancer, Epithelial-myoepithelial carcinoma, Epithelioid hemangioendothelioma (EHE), Epithelioid sarcoma, Essential thrombocythemia, Esthesioneuroblastoma (olfactory neuroblastoma), Extra-cranial malignant rhabdoid tumor (MRT), Extranodal NK / T-cell lymphoma - nasal type, Fallopian tube cancer, Fibrolamellar carcinoma, Fibromatosis, Fibromyxoid sarcoma (Evans’ tumor), Fibrosarcoma Folliculotropic mycosis fungoides, Ganglioglioma, Ganglioneuroblastoma, Gastric Adenocarcinoma and Proximal Polyposis of the Stomach (GAPPS), Gastrinoma, Gastroesophageal junction (GEJ) cancer, Gestational trophoblastic disease (GTD) (hydatidiform mole; gestational trophoblastic neoplasia), Germ cell tumor, Giant cell tumor of bone, Glucagonoma, Granulomatous slack skin, Heart cancer (cardiac angiosarcoma), Hemangioendothelioma, Hemangiosarcoma, Hepatobiliary cancer, Hepatoblastoma, Hepatocellular carcinoma, Hepatoma, Hereditary diffuse gastric cancer (HDGC), Hurthle cell cancer (oxyphil cell carcinoma), Insulinoma, Islet cell tumor, Keratoacanthoma, Klatskin tumor, Large cell neuroendocrine carcinoma, Leiomyosarcoma, Leydig cell tumor, Lip cancer, Liposarcoma, Lymphomatoid papulosis, Lymphoplasmacytic lymphoma, Malignant mesenchymoma, Malignant mixed mullerian tumor, Malignant peripheral nerve sheath tumor (MPNST), Malignant rhabdoid tumor of the kidney, Medulloepithelioma, Meningioma, Mesoblastic nephroma, Metaplastic cancer of the breast, Monoclonal gammopathy of undetermined significance (MGUS), Mouth cancer, Mucinous cystic neoplasm, Mucoepidermoid carcinoma, Muscle cancer (myosarcoma), Myoepithelial carcinoma, Mycosis fungoides, Myelofibrosis, Myxofibrosarcoma, Nephroblastoma, Neuroendocrine carcinoma of the skin, NUT carcinoma, Oat cell cancer, Occult primary cancer, Ocular or intraocular melanoma, Olfactory neuroblastoma (esthesioneuroblastoma), Oligodendroglioma, Oncocytic carcinoma, Ovarian small cell cancer, Paget disease, Pagetoid reticulosis, Paraganglioma, Parathyroid cancer, Periosteal osteosarcoma, Peripheral primitive neuroectodermal tumor (PPNET), Pheochromocytoma, Phyllodes tumor, Pineoblastoma, Plasmacytoma, Polycythemia vera, Polymorphous low-grade adenocarcinoma, Primary cutaneous lymphoma, Primary peritoneal carcinoma, Prolactinoma (lactotroph adenoma), Renal cell carcinoma, Sarcomatoid carcinoma (carcinosarcoma), Schwannoma, Sclerosing epithelioid fibrosarcoma, Sebaceous carcinoma, Seminoma , Sertoli cell tumor, Sezary syndrome, Sinus cancer, Skin adnexal tumors, Solid pseudopapillary neoplasm, Solitary fibrous tumor, Solitary plasmacytoma, Octreotideoma, Spermatocytic seminoma, Spindle cell neoplasm, spindle cell tumor, spindle cell carcinoma, spindle cell sarcoma, Subcutaneous panniculitis -like T-cell lymphoma, Synovial sarcoma, T-cell lymphoma, Teratoma, Throat cancer, Thymoma, Tongue cancer, Tonsil cancer, Trabecular cancer, Translocation renal cell carcinoma, Transitional cell carcinoma (urothelial carcinoma), Undifferentiated pleomorphic sarcoma, Urachal cancer, Urethral cancer, Urothelial carcinoma (transitional cell carcinoma), Uterine cancer, Verrucous carcinoma, VIPoma, Vocal cord / voice box cancer, Womb cancer, or Yolk sac tumor.

[0403] Non-cancerous Conditions

[0404] In another aspect, the invention provides a method of treating a non-cancerous condition in a subject in need thereof, the method comprising: administering to the subject an effective amount of a small zwitterionic multimeric receptor binding targeted drugs described above and a pharmaceutically acceptable carrier or excipient.

[0405] In certain embodiments, the non-cancerous condition involves in cells or pathogens expressing one or more receptors which are targeted by the targeting ligands of the small zwitterionic multimeric targeted drugs.

[0406] In certain embodiments, the non-cancerous condition includes infectious diseases, genetic and congenital disorders, nutritional and metabolic diseases, cardiovascular diseases, respiratory diseases, neurological disorders, endocrine disorders, gastrointestinal diseases, musculoskeletal disorders, autoimmune diseases, dermatological diseases, psychiatric disorders, hematologic diseases, renal and urological diseases, and reproductive system diseases.

[0407] In still other embodiments, the subject is a human.

[0408] Efficacy Measurements of Biological Systems.

[0409] In another aspect, the disclosure encompasses a method of measuring the efficacy of a biological system of a subject. In particular embodiments, the biological system to be measured or monitored is the renal system, the hepatic system, or the blood pool.

[0410] In particular embodiments, the method of measuring the efficacy of a biological system includes:

[0411] (a) administering a quantifiable amount of a diagnostic agent comprising a combination of imaging agent conjugates and a pharmaceutically acceptable carrier or excipient to the subject;

[0412] (b) imaging the subject with positron emission tomography (PET), single-photon emission computerized tomography (SPECT), or magnetic resonance imaging (MRI); and

[0413] (c) determining the amount of therapeutic agent present in the biological system being observed in the subject.

[0414] In a particular aspect, the disclosure encompasses a method of measuring the efficacy of renal function a subject. In particular embodiments, the method comprises:

[0415] (a) administering a quantifiable amount of a diagnostic agent comprising a combination of imaging agent conjugates and a pharmaceutically acceptable carrier or excipient to the subject;

[0416] (b) imaging the subject with positron emission tomography (PET), single-photon emission computerized tomography (SPECT), or magnetic resonance imaging (MRI); and (c) determining the amount of therapeutic agent present in the biological system being observed in the subject.

[0417] In another aspect, the invention provides a method of quantifying the glomerual filtration rate of a subject. In particular embodiments, the method comprises:

[0418] (a) administering a quantifiable amount of a diagnostic agent comprising a combination of imaging agent conjugates and a pharmaceutically acceptable carrier or excipient to the subject; and

[0419] (b) determining the amount of diagnostic agent present in the blood and urine of the subject as a function of time using either measurements of each bodily fluid or imaging the subject with positron emission tomography (PET), single-photon emission computerized tomography (SPECT), or magnetic resonance imaging (MRI).

[0420] Radiosurgery

[0421] Radiosurgery is a known method of treating targets in the body. During radiosurgery, the target is bombarded with a series of X-ray beams fired from various different positions and orientations by using a radiation delivery system, to affect the tumor biology using the cumulative radiation dose at the target. The radiation can be delivered invasively in conjunction with traditional scalpel surgery, or through a percutaneous catheter. CyberKnife™ (Accuray Inc.) and Trilogy™ (Varian Medical Systems) are two such radiation delivery systems. Advances in stereotactic surgery have provided increased accuracy in registering the position of tissue targeted for treatment and a radiation source. For example, see U.S. Pat. Nos. 6,351,662 and 6,402,762. Stereotactic radiosurgery systems may be commercially available from ACCURAY, INC. of Sunnyvale, Calif., and BRAINLAB. The Accuray Cyberknife™ stereotactic radiosurgery system has reportedly been used to provide targeted, painless, and fast treatment of tumors.

[0422] In one aspect, the invention provides a radiosurgical method for treating a patient body, the method comprising: receiving a desired lesion pattern and planned radiation distribution; administering an effective amount of a diagnostic agent comprising a combination of imaging agent conjugates and a pharmaceutically acceptable carrier or excipient to the subject to effectively image the desired lesion pattern; and performing surgery on the desired lesion pattern to treat the patient body.

[0423] In certain embodiments, the surgery can be using scalpel surgery or a stereotactic radiosurgery system to administer a cumulative radiation dose.

[0424] In other embodiments, the imaging agent conjugates of the diagnostic agent further comprises one or more targeting ligands wherein the one or more targeting ligands are cRGD, PSMA, FAP, bombesin, octreotide, octreotide, or a dimer formed from their combination.

[0425] In still other embodiments, the desired lesion pattern is received from a user interface of a treatment planning module. Such planning modules can be pre-programmed with specifications for various disease states and cancerous conditions. Alternatively, specifications for desired lesion patterns can be identified and produced using artificial intelligence data.

[0426] Examples

[0427] Example 1

[0428] Table 4 discloses examples of zwitterionic multimeric targeted anti-cancer drug, their respective targeting ligands and payload, clinical application, and diseases to be diagnosed or treated.

[0429] Table 4. Examples of zwitterionic multimeric targeted anti-cancer drug

[0430] As shown in the example above, a zwitterionic multimeric targeted anti-cancer drug includes one or more targeting ligands binding to cancer- independent targets, one or more targeting ligands binding to career-dependent targets, a payload which include a base payload and a specific payload.

[0431] In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a nexus having five attaching sites, which two of them conjugate to two targeting ligands binding to cancer-independent targets, two of them conjugate to two targeting ligands binding to career-dependent targets, and one of them conjugates to a payload which include a base payload and a specific payload. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a targeting ligand binding to FAP, a targeting ligand binding to cx5p I , a targeting ligand binding to GDPR, and a targeting ligand binding to MUC1.

[0432] In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a targeting ligand binding to FAP, a targeting ligand binding to ot5 1 , a targeting ligand binding to GDPR, and a targeting ligand binding to PSMA.

[0433] In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a payload having a base payload of zwitterionic DOTA and a special payload of Gd3+. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a payload having a base payload of zwitterionic Gadopiclenol and a special payload of Gd3+. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a pay load having a base payload of zwitterionic porphyrin and a special payload of Mn3+. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a pay load having a base payload of zwitterionic DOTA and a special payload of Cu-64. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a payload having a base payload of zwitterionic DOTA and a special payload of Pb-212. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a payload having a base payload of zwitterionic Fluorophore ZN800-1 and a special payload of Pb-212. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a payload having a base payload of zwitterionic Fluorophore ZN700-land a special payload of Pb-212. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a pay load having a base payload of zwitterionic DOTA and a special payload of Gd3+. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a payload having a base payload of zwitterionic Gadopiclenol and a special payload of Gd3+. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a payload having a base payload of zwitterionic porphyrin and a special payload of Mn3+. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a pay load having a base payload of zwitterionic DOTA and a special payload of Cu-64. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a payload having a base payload of zwitterionic DOTA and a special payload of Pb-212. In certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a payload having a base payload of zwitterionic Fluorophore ZN800-1 and a special payload of Pb-212. Tn certain embodiments, a zwitterionic multimeric targeted anti-cancer drug includes a payload having a base payload of zwitterionic Fluorophore ZN700-1 and a special pay load of Pb-212.

[0434] Example 2

[0435] In one embodiment, a small zwitterionic multimeric targeted anti-cancer drug comprises an adamantane as a nexus, a KUE, a FAPI, and a Bombesin as its three different targeting ligands.

[0436] The synthesis process of this small zwitterionic multimeric targeted anti-cancer drug is described in Scheme 8 as follow:

[0437] -THIS AREA 1NTENTJOANLLY LEFT BLANK-

[0438] Scheme 8:

[0439] In the Scheme 8 above, the zwitterionic linkers attaching the core to the targeting ligands and payload are not shown. However, it is envisioned that the carboxylic acids shown is adjacent to the end of a zwitterionic linker.

[0440] In certain embodiments, the Scheme 8 is accomplished in solid-phase synthesis. The synthesis process in Scheme 8 can be used for synthesis of other zwitterionic multimeric targeted anti-cancer drugs with different nexuses, targeting ligands, and / or pay loads.

[0441] Example 3

[0442] Figures 9-10 shows an embodiment of synthesis process of the small zwitterionic multimeric targeted drug.

[0443] In certain embodiments, the synthesis of the small zwitterionic multimer targeted drug performed using solid-phase synthesis (SPS), which provides maximal flexibility, costeffectiveness, and drug purity, especially with complex drugs.

[0444] In certain embodiments, the synthetic strategy for small zwitterionic multimer targeted drug is modular. That is, the targeting ligands and payloads can be replaced at will, provided certain chemical requirements are met. A list of these requirements and their rationale are disclosed as follows.

[0445] In certain embodiments, all targeting ligands and payloads are compatible and stable during standard Fmoc SPS and final resin cleavage with high concentrations of TFA. This means that reactive chemical groups need to be protected with acid-labile protection groups until final deprotection and resin cleavage. In addition, the targeting ligands maintain their affinities when conjugated to other domains.

[0446] In certain embodiments, zwitterionic linkers and zwitterionic payloads are formed by nucleophilic substitution of hydroxyl groups with N-oxides at the last step before deprotection and cleavage of the multimer from the resin. Because of this, amino acids or other molecules used at any time during SPS cannot use trityl groups for side chain protection. As shown in Figures 7-9, trityl is reserved for the linker precursor Fmoc- Homoserine (Trt) and ZN-DOTA precursors, with the former being selectively de-protected using 1% TFA and triisopropylsilane (TIS) after payload conjugation. Acceptable alternatives to trityl for side chain protection include: Asn (N): DMCP; His (H): Boc; Statine: t-Butyl;Cys (C): Mob; Ser (S): t-Butyl; Gin (Q): DMCP; and Thr (T): t-Butyl.

[0447] In certain embodiments, different from Figures 7-9, trityl-protected homoserine is no longer being used for zwitterionic linkers and OH derivatives are no longer being used for payloads. Instead, the zwitterionic linkers and zwitterionic payloads are formed by using Fmoc- Azidoalanine, Fmoc- Azidohomoalanine or other azido derivatives, which can be converted to ZW groups using click chemistry.

[0448] In certain embodiments, alloc (i.e., allyl) side chain protection is reserved for the two branching point residues. Because of this, other amino acids or molecules used at anytime during solid-phase synthesis (SPS) cannot use alloc (allyl) groups for side chain protection.

[0449] As shown in Figure 8, the synthesis of the small zwitterionic multimer targeted drug starts with targeting ligand 1. In certain embodiments, the targeting ligand 1 can either be synthesized via standard Fmoc solid-phase synthesis (SPS) on resin (i.e., if a peptide or peptide derivative) or can he attached to the resin via a free primary amine or carboxylic acid without losing its binding affinity. If using the former method, targeting ligand 1 Fmoc-SPS proceeds from the resin as shown in the top panel of Figure 8. If using the latter method, a Fmoc-Lysine-Amide or Fmoc-Glutamate-Amide resin (or similar) is used and the targeting ligand 1 is conjugated as shown in the lower panel of Figure 8 depending on orientation of conjugation. In certain embodiments, when conjugating pre-formed targeting ligand 1 to the resin, Ahx or similar spacing may or may not be needed depending on targeting ligand l’s properties. Cleavage from the resin should create the amide of either Lys or Glu.

[0450] As shown in Figure 9, in certain embodiments, the small zwitterionic multimer targeted drug is synthesized using a branched chain approach so that targeting ligand 1, the multiple zwitterionic linkers, any Ahx (or similar) spacing groups, and ideally, other targeting ligands can all be added in an uninterrupted fashion. This is made possible by the fact that N- oxide addition to the linkers and payload occurs after payload addition and immediately before deprotection and cleavage.

[0451] In certain embodiments, the synthesis of the small zwitterionic multimer targeted drug uses Lys(Alloc) as the branch point between the targeting ligand 1 and targeting ligand 2 and either Fmoc-Lys(Alloc) or Fmoc-Glu(Allyl), depending on the targeting ligand conjugation groups, between targeting ligand 2 and targeting ligand 3. As shown in Figure 9, HS stands for homoserine, the precursor for zwitterionic linkers.

[0452] Schematically, synthesis proceeds as shown in Figure 9. The Fmoc Lys(Alloc) is deprotected and the homoserine linker precursors and spacing elements for targeting ligand 2 are added by simply continuing SPS. Whenever possible, the targeting ligand 2 is also synthesized by SPS.

[0453] The Alloc of the first branch (Lys) is then deprotected with Pd, and either Fmoc- Lys(Alloc) or Fmoc-Glu(Allyl) is added as the second branch. The side chain (NH2 for Lys and COOH for Glu) is chosen based on the appropriate attachment to the payload. If the payload uses an NH2 group for conjugation, the Fmoc-Glu (Allyl) should be chosen. If the payload uses a COOH group for conjugation, the Fmoc-Lys(Alloc) should be chosen. After addition of the second branch residue, the Fmoc is deprotected and the homoserine linker precursors and spacing elements for targeting ligand 3 are added by simply continuing SPS. Whenever possible, the targeting ligand 3 is also synthesized by SPS. The final product is stored on-resin until payload addition through the Allyl (COOH) is accomplished. The following example assumes a payload that uses NH2 for conjugation.

[0454] In certain embodiments, if multiple branching is needed, e.g., to create tetramers and pentamers, additional Fmoc-Lys-E- Allocs can be added after growing the targeting ligand 2. New targeting ligands are then grown off the epsilon amines using standard Fmoc chemistry.

[0455] In certain embodiment, to add the payload, Alloc / Allyl is deprotected, the carboxylate (if Glu) is activated, and free NH2 (if Glu) or COOH (if Lys) of payload is conjugated to the synthesized complex. In certain embodiments, the payload has 3-4 hydroxyl (OH) groups that will be substituted later with N-oxides.

[0456] In certain embodiments, the synthesis process includes a step of N-oxide formation by nucleophilic substitution. Fisr, Trt groups of homoserine is deprotect. All exposed hydroxyl groups (on linkers and payload) are mesylated and convert to N-oxides. The entire molecule is deprotect and cleaved from resin with TFA.

[0457] As shown in Figure 10, this process creates CPL001 (for radiometal chelation to create either CPL002 [Cu-64] or CPL003 [Pb-212]), or CPL004 (for optical imaging). As shown in Figure 7, below with two ZW linkers per ligand there are a total of 9 N-oxides on CPI-001 / -002 / -003 and 10 N-oxides on CPI-004.

[0458] OTHER EMBODIMENTS It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

[0459] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein.

[0460] Such equivalents are intended to be encompassed by the following claims.

Claims

1. CLAIMS1. A receptor-binding or antigen-binding drug comprising: a nexus; multiple targeting ligands conjugated to the nexus; and a pay load molecule conjugated to the nexus, wherein each of the multiple targeting ligands is conjugated to the nexus via a linker comprising one or more zwitterionic groups, and wherein each of the multiple targeting ligands binds to at least one receptor on disease-related cells.

2. The receptor-binding or antigen-binding drug according to claim 1 , wherein each of the multiple targeting ligands and the payload molecule is conjugated to the nexus via a linker comprising one or more zwitterionic groups, wherein the one or more zwitterionic groups are ammonium sulfobetaine groups or N-oxide groups or a combination thereof.

3. The receptor-binding or antigen-binding drug according to claim 1, wherein the nexus having multiple attaching sites.

4. The receptor-binding or antigen-binding drug according to claim 3, wherein the nexus is a trihedral core, a tetrahedral core, a pentahedral core, or a hexahedral core.

5. The receptor-binding or antigen-binding drug according to claim 3, wherein the nexus is a branching point comprises two or more monomers.

6. The receptor-binding or antigen-binding drug according to claim 5, wherein each of the monomer is an amino acid.

7. The receptor-binding or antigen-binding drug according to claim 6, wherein the amino acid is either Lys or Glu.

8. The receptor-binding or antigen-binding drug according to claim 1, wherein at least one of the multiple targeting ligands is different from the others of the multiple targeting ligands.

9. The receptor-binding or antigen-binding drug according to claim 8, wherein each of the multiple targeting ligands is different from the others of the multiple targeting ligands10. The receptor-binding or antigen-binding drug according to claim 8, wherein each of the multiple targeting ligands is cRGD, dPSMA-617, KUE, a FAP binding vector, octreotide, or bombesin.

11. The receptor-binding or antigen-binding drug according to claim 8, wherein the payload molecule is a zwitterionic molecule.

12. The receptor-binding or antigen-binding drug according to claim 11 , wherein the payload molecule comprises one or more of ZW700-Forte, ZW800-1, ZW830-1, ZN-800-1, ZN-830-1, ZN-800-1 -Forte, ZN-700-1 -Forte, ZW-DOTA, ZW-PyC3A, ZW-Macropa, ZW- porphyrin, ZW-NOTA, ZW-deferoxamine, ZW-DOTP, ZW-PCTA, ZW-Amide-PCTA, and ZW-PTCP.

13. The receptor-binding or antigen-binding drug according to claim 11, wherein the payload molecule comprises one or more of ZW-DOTA-Cu-64, ZW-DOTA-Cu-67, ZW- DOTA-Gd, ZW Poryphyrin-Mn3+.

14. The receptor-binding or antigen-binding drug according to claim 2, wherein the linker is a flexible zwitterionic linker.

15. The receptor-binding or antigen-binding drug according to claim 14, wherein the linker comprises at least one azido derivative.

16. The receptor-binding or antigen-binding drug according to claim 1, wherein the tumor cells demonstrate heterogeneity.

17. The receptor-binding or antigen-binding drug according to claim 1 , wherein at least one of the multiple targeting ligands is configured to bind to at least one cancer-independent target, and at least one of the multiple targeting ligands is configured to bind to at least one cancer-dependent target.

18. The receptor-binding or antigen-binding drug according to claim 17, wherein the payload molecule comprises a base payload and a special payload; wherein the base payload comprises a zwitterionic molecule; and the special payload comprises a metal molecule.

19. The receptor-binding or antigen-binding drug according to claim 18, wherein the at least one cancer -independent target comprises one or more of FAP, integrin a5pi, and integrin avP3; and the at least one cancer-dependent target comprises one or more of GRPR, MUC1, and PS MA.

20. The receptor-binding or antigen binding drug according to claim 3, wherein the nexus has a first attaching site conjugated to a first targeting ligand via a first flexible zwitterionic linker, a second attaching site conjugated to a second targeting ligand via a second flexible zwitterionic linker, and a third attaching site conjugated to a payload via a third flexible zwitterionic linker, wherein the payload comprises one of ZW700-Forte, ZW800-1, ZW830- 1, ZN-800-1, ZN-830-1, ZN- 800-1 -Forte, ZN-700-1 -Forte, ZW-DOTA, ZW-PyC3A, ZW- Macropa, ZW-porphyrin, ZW-NOTA, ZW-deferoxamine, ZW-DOTP, ZW-PCTA, ZW- Amide-PCTA, and ZW-PTCP.

21. A method of imaging tissue, cells, or lumen in a subject, the method comprising:(a) contacting the tissue, cells or lumen with a composition according to Claim 1 to the subject,(b) irradiating the tissue, cells, or lumen at a wavelength absorbed by the dye;(c) and detecting a signal from the imaging agent, thereby imaging the tissue, cells, or lumen.

22. A method of treating a tumor by administering an effective amount of an anti-cancer drug according to Claim 1, wherein the method comprises a step to diagnose the cancer and a step of administering the therapeutic agent to a subject determined to be in need thereof; wherein the step to diagnose the cancer comprises: contacting cells, tissues or organs of a subject with an imaging agent, imaging the cells, tissues, or organs of the subject using positron emission tomography (PET), single-photon emission computerized tomography (SPECT), or magnetic resonance imaging (MRI), and diagnosing the cancer in the cells tissues, or organs of the subject based on imaging data collected; and wherein the anti-cancer drug according to Claim 1 comprises at least one zwitterionic molecule as the payload wherein a metal atom complexed to the zwitterionic molecule is: a radioactive metal isotope known to emit ionizing radiation that results in the death of cells that take up the analogs; or a non-radioactive metal that is capable of releasing cytotoxic radiation upon irradiation with alpha emission, beta emission, neutron capture, or a combination thereof.

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