Programmable polymeric drugs

Abstract

To provide a potent, targeted drug conjugate having a high therapeutic index.SOLUTION: The present invention provides a compound having the following structure, or its stereoisomer, pharmaceutically acceptable salt or tautomer. (M: NSAID, kinase inhibitor or the like. L: amide bond, ester bond or the like. L1, L2, L3: alkylene, alkenylene or the like. R1: H, alkyl, alkoxy. R2, R3: H, OH, SH or the like. n: an integer of 1 or greater.)SELECTED DRAWING: None

Description

[Technical field]

[0001] FIELD OF THE DISCLOSURE Embodiments of the present invention relate generally to dimeric and polymeric bioactive compounds having spacing groups, and methods for their preparation and use in various therapeutic methods. [Background technology]

[0002] Unlike chemotherapy, targeted drug conjugates are intended to target only diseased cells and spare healthy cells.Typically, conjugates are composed of a targeting molecule linked to a bioactive payload or drug.By combining the therapeutic effectiveness of bioactive drugs with the unique targeting ability, conjugates can deliver drugs only to the intended target and minimize potential side effects. Antibody-drug conjugates (ADCs) are a class of targeted drug conjugates that are particularly important for cancer treatment. ADCs combine the targeting features of monoclonal antibodies with the cancer-killing capabilities of cytotoxic agents to provide therapeutic agents with several advantages over other chemotherapeutic agents. However, challenges associated with the complexity of the ADC construct, specifically the chemical linker between the antibody and the drug, have presented significant difficulties for the development of novel and effective therapeutic agents. The first ADC was approved in 2001, but it took almost a decade for the next ADC to be approved. As of today, only Adcetris® and Kadcyla® are commercially available on a global scale (Zevalin® is only approved in China). Pioneer Pfizer / Wyeth withdrew Mylotarg® in 2010 after safety issues were noted during controlled clinical trials. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] US Patent Application Publication No. 2010 / 0092386 [Patent Document 2] US Patent Application Publication No. 2016 / 0039850 [Patent Document 3] US Patent Application Publication No. 2013 / 0137755 [Patent Document 4] US Patent Application Publication No. 2016 / 0176903 Summary of the Invention [Problem to be solved by the invention]

[0004] Thus, there is a need in the art for potent targeted drug conjugates with high therapeutic index. Ideally, such drug conjugates should sensitively discriminate between healthy and diseased tissues (e.g., tumor cells). The present invention fulfills this need and further provides related advantages. [Means for solving the problem]

[0005] In summary, embodiments of the present invention relate to compounds that are generally useful as targeted drug conjugates, which may include fluorescent and / or colored dyes that allow selective delivery to targets such as tumor cells, as well as reagents for their preparation. Methods for the preparation of such molecules and their use to provide therapeutic treatment to patients in need thereof are also described. The compound embodiments disclosed herein include one or more biologically active moieties covalently linked by a linker ("L"). Advantageously, the present invention provides compounds that can be combined during polymer synthesis or attached post-synthetically. Furthermore, the embodiments described herein allow for the incorporation of multiple biologically active moieties and the optional inclusion of a targeting moiety within the same compound. In one embodiment, there is provided a compound having the following structure (I): or a stereoisomer, tautomer, or salt thereof:

[0006] [ka] (I) (In the formula, R 1 , R 2 , R 3 , L 1 , L 2 , L 3 , M, and n are as defined herein. The compounds of structure (I) are useful in a number of applications, including use as therapeutic agents in a variety of methods of treatment.

[0007] In another embodiment, a method of treating a disease is provided comprising administering to a subject in need thereof a therapeutically effective amount of a compound of structure (I) or a composition comprising a compound of structure (I), wherein each M is independently a biologically active moiety effective for treating the disease. These and other aspects of the present invention will become evident upon reference to the following detailed description. In the drawings, identical reference numbers indicate similar elements. The sizes and relative positions of elements in the drawings are not necessarily drawn to scale, and some of these elements have been enlarged and positioned to facilitate drawing legibility. Furthermore, the particular shapes of the elements as shown are not intended to convey any information regarding the actual shape of the particular elements, but have only been selected for ease of recognition in the drawings. [Brief description of the drawings]

[0008] [Figure 1] FIG. 1 provides a SEC spectrum showing compound 7 conjugated to UCHT-1 antibody. [Diagram 2] FIG. 1 shows the PAGE results of the synthesis of compound 17. [Diagram 3] FIG. 13 shows the results of a potency assay of HCC1954 cells as described in Example 13. [Figure 4] FIG. 13 shows the results of a potency assay of HS578 cells as described in Example 13. [Diagram 5]FIG. 13 shows the results of a potency assay of SKO3 cells as described in Example 13. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] In the following description, certain specific details are set forth in order to provide a thorough understanding of various embodiments of the invention. However, it will be understood by those skilled in the art that the invention may be practiced without these details. Unless the context requires otherwise, throughout this specification and claims, the word "comprise" and variations thereof, such as "comprises" and "comprising," are to be interpreted in an open and inclusive sense, i.e., as "including but not limited to." Throughout this specification, a reference to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with that embodiment is included in at least one embodiment of the invention. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0010] "Amino" refers to the group -NH2. "Carboxy" refers to the group -CO2H. "Cyano" refers to the radical -CN. "Formyl" refers to the group -C(=O)H. "Hydroxy" or "hydroxyl" refers to the group --OH. "Imino" refers to the group ═NH. "Nitro" refers to the -NO2 group. "Oxo" refers to the =O substituent. "Sulfhydryl" refers to the -SH group. "Thioxo" refers to the group ═S. "Alkyl" means an alkyl group containing no unsaturation and having 1 to 12 carbon atoms (C1-C 12 "C1-C8 alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms having 1 to 8 carbon atoms (C1-C8 alkyl) or 1 to 6 carbon atoms (C1-C6 alkyl) and attached to the remainder of the molecule by a single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl (iso-propyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl, etc. Unless otherwise specified in the specification, an alkyl group may be optionally substituted.

[0011] "Alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain containing no unsaturation and consisting only of carbon and hydrogen having 1 to 12 carbon atoms, linking the remainder of the molecule to a radical group, e.g., methylene, ethylene, propylene, n-butylene, ethenylene, propenylene, n-butenylene, propynylene, n-butynylene, etc. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless otherwise specified in the specification, alkylene can be optionally substituted. "Alkenylene" or "alkenylene chain" refers to a straight or branched divalent hydrocarbon chain consisting only of carbon and hydrogen, containing at least one carbon-carbon double bond and having 2 to 12 carbon atoms, linking the rest of the molecule to a radical group, e.g., ethenylene, propenylene, n-butenylene, etc. The alkenylene chain is attached to the rest of the molecule through a single bond and to the radical group through a double bond or a single bond. The points of attachment of the alkenylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless otherwise specified in the specification, alkenylene may be optionally substituted.

[0012] "Alkynylene" or "alkynylene chain" refers to a straight or branched divalent hydrocarbon chain consisting only of carbon and hydrogen, containing at least one carbon-carbon triple bond and having 2 to 12 carbon atoms, linking the rest of the molecule to a radical group, e.g., ethenylene, propenylene, n-butenylene, and the like. The alkynylene chain is attached to the rest of the molecule through a single bond and to the radical group through a double bond or a single bond. The points of attachment of the alkynylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. Unless specified otherwise in the specification, alkynylene may be optionally substituted.

[0013] "Alkyl ether" refers to any alkyl group, as defined above, in which at least one carbon-carbon bond has been replaced with a carbon-oxygen bond. The carbon-oxygen bond may be terminal (as in an alkoxy group) or the carbon-oxygen bond may be internal (i.e., COC). An alkyl ether contains at least one carbon-oxygen bond, but may contain more than one. For example, polyethylene glycol (PEG) is included within the meaning of alkyl ether. Unless otherwise specified specifically in the specification, an alkyl ether group may be substituted. For example, in some embodiments, an alkyl ether is an alkyl group selected from the group consisting of an alcohol or -OP(=R a )(R b )R c (In the formula, R a , R b and R c Each of is substituted with as defined for compounds of structure (I).

[0014] "Alkoxy" means a group of the formula -OR a (wherein R a is an alkyl group, as defined above, containing 1 to 12 carbon atoms. Unless specified otherwise specifically in the specification, an alkoxy group may be optionally substituted. An "alkoxyalkyl ether" is a group represented by the formula -OR a R b(wherein R a is an alkylene group, as defined above, containing 1 to 12 carbon atoms, and Rb is an alkyl ether group, as defined herein. Unless otherwise specifically specified herein, an alkoxyalkyl ether group may be optionally substituted, for example, with an alcohol or -OP(=Ra)(Rb)Rc, where each of Ra, Rb, and Rc is as defined for the compounds of structure (I). "Heteroalkyl" refers to an alkyl group, as defined above, that contains at least one heteroatom (e.g., Si, N, O, P, or S) within or at the terminus of the alkyl group. In some embodiments, the heteroatom is within the alkyl group (i.e., a heteroalkyl has at least one carbon-[heteroatom] x -carbon bond (where x is 1, 2, or 3). In other embodiments, the heteroatom is at the end of the alkyl group, thus serving to join the alkyl group to the remainder of the molecule (e.g., M1-HA, where M1 is a portion of the molecule, H is a heteroatom, and A is an alkyl group). Unless otherwise specifically stated herein, heteroalkyl groups may be substituted. Exemplary heteroalkyl groups include ethylene oxide (e.g., polyethylene oxide), and may include a phosphorus-oxygen bond, such as a phosphodiester bond.

[0015] "Heteroalkoxy" refers to a group of the formula -OR a (wherein R a is a heteroalkyl group, as defined above, containing 1 to 12 carbon atoms. Unless specified otherwise specifically in the specification, a heteroalkoxy group may be optionally substituted. "Heteroalkylene" refers to an alkylene group, as defined above, containing at least one heteroatom (e.g., Si, N, O, P, or S) within the alkylene chain or at the end of the alkylene chain. In some embodiments, the heteroatom is within the alkylene chain (i.e., the heteroalkylene contains at least one carbon-[heteroatom]-carbon bond, where x is 1, 2, or 3). In other embodiments, the heteroatom is at the end of the alkylene, thus serving to join the alkylene to the remainder of the molecule (e.g., M1-HA-M2, where M1 and M2 are part of the molecule, H is a heteroatom, and A is an alkylene). Unless otherwise specified in the specification, a heteroalkylene group may be substituted. Exemplary heteroalkylene groups include ethylene oxide (e.g., polyethylene oxide) and the "C", "HEG", and "PEG 1K" linking groups illustrated below.

[0016] [ka] Various embodiments of the heteroalkylene linker include multimers of the above C-linker, HEG linker and / or PEG 1K linker. In some embodiments of the PEG 1K linker, n ranges from 20 to 25, e.g., n is 20, 21, 22, 23, 24, or 25. Multimers can be, for example, of the following structure:

[0017] [ka] (wherein x is 0 or an integer greater than 0, for example, x ranges from 0 to 100 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10)).

[0018] "Heteroalkenylene" is a heteroalkylene, as defined above, containing at least one carbon-carbon double bond. Unless otherwise specified in the specification, a heteroalkenylene group may be optionally substituted. "Heteroalkynylene" is a heteroalkylene containing at least one carbon-carbon triple bond. Unless stated otherwise specifically in the specification, a heteroalkynylene group may be optionally substituted. "Heteroatom" with respect to a "heteroatom linker" refers to a linker group consisting of one or more heteroatoms. Exemplary heteroatom linkers include single atoms selected from the group consisting of Si, O, N, P, and S, as well as multiple heteroatoms, such as those of the formula -P(O - )(=O)O- or -OP(O - )(=O)O-- linkers and polymers and combinations thereof. "Phosphate" is -OP(=O)(R a )R b group (in the formula, R a OH, O - OR c and R b OH, O - , OR c , a thiophosphate group or a further phosphate group, where R c refers to the counter ion (e.g., Na+, etc.).

[0019] "Phosphoalkyl" is -OP(=O)(R a )R b group (in the formula, R a OH, O - OR c and R b is -Oalkyl, where R c refers to a counter ion (e.g., Na+, etc.). Unless otherwise specifically stated herein, a phosphoalkyl group may be substituted. For example, in certain embodiments, the -Oalkyl portion in a phosphoalkyl group is hydroxyl, amino, sulfhydryl, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkylether, thiophosphoalkylether, or -OP(=R a )(R b )R c (In the formula, R a , R b and Rc Each of may be substituted with one or more of, as defined for compounds of structure (I). "Phosphoalkyl ether" is -OP(=O)(R a )R b group (in the formula, R a OH, O - or R c and R b is an -O alkyl ether, where R c is the counter ion (e.g., Na + Unless otherwise specified specifically in the specification, the phosphoalkyl ether group may be substituted. For example, in certain embodiments, the -O alkyl ether moiety in the phosphoalkyl ether group is hydroxyl, amino, sulfhydryl, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkyl ether, thiophosphoalkyl ether, or -OP(=R a )(R b )R c (In the formula, R a , R b and R c Each of may be substituted with one or more of, as defined for compounds of structure (I).

[0020] "Thiophosphate" is -OP(=R a )(R b )R c group (in the formula, R a is O or S, R b OH, O - , S - , OR d or S.R. d and R c OH, SH, O - , S - , OR d , S.R. d , a phosphate group or a further thiophosphate group, where R d is the counter ion (e.g., Na + etc.), where i) R ais S, and ii) R b is S - or S.R. d and iii) R c , SH, S - or S.R. d or iv) a combination of i), ii) and / or iii). "Thiophosphoalkyl" is -OP(=R a )(R b )R c group (in the formula, R a is O or S, R b OH, O - , S - , OR d or S.R. d and R c is -Oalkyl, where R d is the counter ion (e.g., Na + etc.), where i) R a is S, and ii) R b is S - or S.R. d or iii) R a is S and R b is S - or S.R. d Unless otherwise specifically stated in the specification, a thiophosphoalkyl group may be optionally substituted. For example, in certain embodiments, the -Oalkyl portion in a thiophosphoalkyl group is hydroxyl, amino, sulfhydryl, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkylether, thiophosphoalkylether, or -OP(=R a )(R b )R c (In the formula, R a , R b and R c Each of may be substituted with one or more of, as defined for compounds of structure (I).

[0021] "Thiophosphoalkyl ether" is -OP(=R a )(Rb )R c group (in the formula, R a is O or S, R b OH, O - , S - , OR d or S.R. d and R c is an -O alkyl ether, where R d is the counter ion (e.g., Na + etc.), where i) R a is S, and ii) R b is S - or S.R. d or iii) R a is S and R b is S - or S.R. d Unless otherwise specifically stated herein, a thiophosphoalkyl ether group may be optionally substituted. For example, in certain embodiments, the -O alkyl ether moiety in a thiophosphoalkyl group is hydroxyl, amino, sulfhydryl, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkyl ether, thiophosphoalkyl ether, or -OP(=R a )(R b )R c (In the formula, R a , R b and R c Each of may be substituted with one or more of, as defined for compounds of structure (I).

[0022] "Carbocyclic" refers to a stable 3- to 18-membered aromatic or non-aromatic ring containing 3 to 18 carbon atoms. Unless otherwise specified in the specification, a carbocyclic ring may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system which may include fused or bridged ring systems and may be partially or fully saturated. Non-aromatic carbocyclyl radicals include cycloalkyl, whereas aromatic carbocyclyl radicals include aryl. Unless otherwise specified in the specification, a carbocyclic group may be optionally substituted. "Cycloalkyl" refers to a stable non-aromatic monocyclic or polycyclic carbocyclic ring, which may include fused or bridged ring systems having 3 to 15 carbon atoms, preferably 3 to 10 carbon atoms, is saturated or unsaturated, and is attached to the remainder of the molecule by a single bond. Monocyclic cyclocalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptly, and cyclooctyl. Polycyclic cycloalkyls include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo-[2.2.1]heptanyl, and the like. Unless otherwise specified in the specification, cycloalkyl groups may be optionally substituted. "Aryl" refers to a ring system containing at least one carbocyclic aromatic ring. In some embodiments, the aryl contains 6 to 18 carbon atoms. The aryl ring may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems. Aryl includes, but is not limited to, aryl derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, pleiadene, pyrene and triphenylene. Unless otherwise specified in the specification, the aryl group may be substituted.

[0023] "Heterocyclic" refers to a stable 3- to 18-membered aromatic or non-aromatic ring containing 1-12 carbon atoms and 1-6 heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur. Unless otherwise specified herein, a heterocyclic ring may be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems, the nitrogen, carbon, or sulfur atoms in the heterocyclic ring may be oxidized, the nitrogen atom may be quaternized, and the heterocyclic ring may be partially or fully saturated. Examples of aromatic heterocyclic rings are listed below in the definition of heteroaryl (i.e., heteroaryl is a subset of heterocyclic). Examples of non-aromatic heterocyclic rings include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, pyrazolopyrimidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trioxanyl, trithianyl, triazinanyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in the specification, a heterocyclic group may be optionally substituted.

[0024] "Heteroaryl" refers to a 5-14 membered ring system containing 1-13 carbon atoms, 1-6 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, and at least one aromatic ring. For purposes of certain embodiments of the present invention, the heteroaryl radical may be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems, and the nitrogen, carbon or sulfur atoms in the heteroaryl radical may be optionally oxidized and the nitrogen atom may be optionally quaternized.Examples are azepinyl, acridinyl, benzimidazolyl, benzthiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzo Thienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, benzoxazolinonyl, benzimidazolethionyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolinyl, indolizinyl, isoxazolyl n-phenyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl, 1-oxidopyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, pteridinonyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyridinonyl, pyrazinyl, pyrimidinyl, pyrimidino Heteroaryl groups include, but are not limited to, aryl, pyridazinyl, pyrrolyl, pyrido[2,3-d]pyrimidinonyl, quinazolinyl, quinazolinonyl, quinoxalinyl, quinoxalinonyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, thieno[3,2-d]pyrimidin-4-onyl, thieno[2,3-d]pyrimidin-4-onyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e., thienyl). Unless stated otherwise specifically in the specification, a heteroaryl group may be optionally substituted.

[0025] "Fused" refers to a ring system that contains at least two rings, the two rings sharing at least one common ring atom, for example, two common ring atoms. When the fused ring is a heterocyclyl or heteroaryl ring, the common ring atom can be carbon or nitrogen. Fused rings include bicyclic, tricyclic, tertracyclic, etc.

[0026] As used herein, the term "substituted" refers to an alkyl group in which at least one hydrogen atom (e.g., one, two, three or all hydrogen atoms) has been replaced with a halogen atom, such as F, Cl, Br, and I; an oxygen atom in groups such as hydroxyl, alkoxy, and ester groups; a sulfur atom in groups such as thiol, thioalkyl, sulfone, sulfonyl, and sulfoxide groups; a nitrogen atom in groups such as amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, imides, and enamines; a trialkylsilyl group, a dialkylarylsilyl group, an alkyldiarylsilyl group, and an alkyldiarylsilyl group. "Substituted" refers to any of the above groups (e.g., alkyl, alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, alkoxy, alkyl ether, alkoxyalkyl ether, heteroalkyl, heteroalkoxy, phosphoalkyl, phosphoalkyl ether, thiophosphoalkyl, thiophosphoalkyl ether, carbocyclic, cycloalkyl, aryl, heterocyclic, and / or heteroaryl) in which one or more hydrogen atoms have been replaced by a bond to a non-hydrogen atom, such as, but not limited to, a silicon atom in groups such as, for example, alkyl, alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, alkoxy, alkyl ether, alkoxyalkyl ether, heteroalkyl, heteroalkoxy, phosphoalkyl, phosphoalkyl ether, thiophosphoalkyl, thiophosphoalkyl ether, carbocyclic, cycloalkyl, aryl, heterocyclic, and / or heteroaryl. "Substituted" also refers to any of the above groups in which one or more hydrogen atoms have been replaced by a heteroatom, such as oxygen in oxo, carbonyl, carboxyl, and ester groups; and a higher order bond (e.g., double or triple bond) to a nitrogen in groups such as imine, oxime, hydrazone, and nitrile. For example, "substituted" refers to any of the above groups in which one or more hydrogen atoms have been replaced by a bond to a non-hydrogen atom, such as, for example, a silicon atom in groups such as, for example, -NR g R h , -NR g C(=O)R h, -NR g C(=O)NR g R h , -NR g C(=O)OR h , -NR g SO2R h , -OC(=O)NR g R h , -OR g , -SR g , -SOR g , -SO2R g , -OSO2R g , -SO2OR g , =NSO2R g and -SO2NR g R h "Substituted" includes any of the above groups, wherein one or more hydrogen atoms have been replaced with -C(=O)R. g , -C(=O)OR g , -C(=O)NR g R h , -CH2SO2R g , -CH2SO2NR g R h In the above, R also means any of the above groups replaced by g and R h are the same or different and are independently hydrogen, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl, and / or heteroarylalkyl. "Substituted" further refers to any of the above groups in which one or more hydrogen atoms are replaced by a bond to an amino, cyano, hydroxyl, imino, nitro, oxo, thioxo, halo, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl, and / or heteroarylalkyl group. In some embodiments, the optional substituents are -OP(=R a)(R b )R c (In the formula, R a , R b and R c each of which is as defined for compounds of structure (I). In addition, each of the foregoing substituents may be substituted with one or more of the substituents described above.

[0027] "Conjugation" or "bioconjugation" refers to a chemical strategy for forming a stable covalent bond between two molecules. The term "bioconjugation" is generally used when one of the molecules is a biomolecule (e.g., an antibody). The product or compound resulting from such a strategy is a conjugate, conjugated, or grammatical equivalents.

[0028] "Fluorescent" refers to a molecule that can absorb light of a particular frequency and emit light of a different frequency. Fluorescence is well known to those of skill in the art. "Colored" refers to molecules that absorb light within the color spectrum (i.e., red, yellow, blue, etc.).

[0029] "Linker" refers to a continuous chain of at least one atom, such as carbon, oxygen, nitrogen, sulfur, phosphorus, and combinations thereof, that connects one portion of a molecule to another portion of the same molecule, or to a different molecule, moiety, or solid support (e.g., a microparticle). Linkers may connect molecules through covalent bonds or other means, such as ionic or hydrogen bonding interactions. The term "biomolecule" refers to any of a variety of biological materials, including nucleic acids, carbohydrates, amino acids, polypeptides, glycoproteins, hormones, aptamers, and mixtures thereof. More specifically, the term is intended to include, but is not limited to, RNA, DNA, oligonucleotides, modified or derivatized nucleotides, enzymes, receptors, prions, receptor ligands (including hormones), antibodies, antigens, and toxins, as well as bacteria, viruses, blood cells, and tissue cells. The visually detectable biomolecules of the present invention (e.g., compounds of structure (I) having a biomolecule linked thereto) are prepared by contacting a biomolecule with a compound having a reactive group or a functional group such as amino, hydroxyl, carboxyl, or sulfhydryl group on the biomolecule, which allows the binding of the biomolecule to the compound via any available atom, as further described herein.

[0030] A "reactive group" is a moiety that can react with a second reactive group (e.g., a "complementary reactive group") to form one or more covalent bonds, for example, by substitution, oxidation, reduction, addition, or cycloaddition reactions. Exemplary reactive groups are provided in Table 1 and include, for example, nucleophiles, electrophiles, dienes, dienophiles, aldehydes, oximes, hydrazones, alkynes, amines, azides, acyl azides, acyl halides, nitriles, nitrones, sulfhydryls, disulfides, sulfonyl halides, isothiocyanates, imidoesters, activated esters, ketones, α,β-unsaturated carbonyls, alkenes, maleimides, α-haloimides, epoxides, aziridines, tetrazines, tetrazoles, phosphines, biotins, thiiranes, and the like. "Solid support" refers to any solid substrate known in the art as a solid phase support for molecules, for example, "microparticle" refers to any of a number of small particles useful for binding to the compounds of the invention, including, but not limited to, glass beads, magnetic beads, polymeric beads, non-polymeric beads, etc. In certain embodiments, the microparticle comprises a polystyrene bead. "Solid support residue" refers to a functional group that remains attached to a molecule when the molecule is cleaved from the solid support. Solid support residues are known in the art and can be easily derived based on the structure of the solid support and the group linking the molecule thereto.

[0031] A "targeting moiety" is a moiety that selectively binds or associates with a particular target, such as a tumor cell antigen. "Selectively" binding or association means that the targeting moiety preferentially associates or binds with a desired target over other targets. In some embodiments, the compounds disclosed herein include a linkage to a targeting moiety for the purpose of selectively binding or associating the compound with a tumor cell antigen (i.e., the target of the targeting moiety), thereby enabling delivery of a biologically active moiety to the tumor cell. Exemplary targeting moieties include, but are not limited to, antibodies, antigens, nucleic acid sequences, enzymes, proteins, cell surface receptor antagonists, and the like. In some embodiments, a targeting moiety is a moiety, such as an antibody, that selectively binds or associates with a target feature on or in a cell, such as a target feature on a cell membrane or other cellular structure, thereby enabling delivery of a biologically active moiety to or into the cell of interest. Small molecules that selectively bind or associate with a desired biological target are also contemplated as targeting moieties in certain embodiments. Those skilled in the art will recognize other biological targets and corresponding targeting moieties that will be useful in various embodiments.

[0032] A "physiologically cleavable linker" refers to a molecular linkage that can be split or separated in a predetermined manner while in the presence of an in vivo or in vitro environment of an organism or cell system to produce two or more separate molecules. In general, physiological conditions that induce such a cleavage or scission event can include temperatures ranging from about 20 to 40° C., atmospheric pressure of about 1 atmosphere (101 kPa or 14.7 psi), pH of about 6 to 8, glucose concentration of about 1 to 20 mM, atmospheric oxygen concentration, and gravity of the Earth. In some embodiments, physiological conditions include enzymatic conditions (i.e., enzymatic cleavage). Bond cleavage or scission can be isotropic or anisotropic.

[0033] The embodiments of the invention disclosed herein are also intended to encompass all compounds of structure (I) that are isotopically labeled by replacing one or more atoms with atoms having a different atomic mass or mass number. Examples of isotopes that may be incorporated into the disclosed compounds are: 2 H, 3 H, 11 C. 13 C. 14 C. 13 N, 15 N, 15 O. 17 O. 18 O. 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I and 125 Includes isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, such as I. Isotopically labeled compounds of structure (I) can generally be prepared by conventional techniques known to those of skill in the art, or by processes analogous to those described below and in the examples below, substituting an appropriate isotopically labeled reagent for the non-labeled reagent used above. "Stable compound" and "stable structure" are meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent. "Optional" or "may" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances in which the event or circumstance occurs and instances in which the event or circumstance does not occur. For example, "optionally substituted alkyl" means that the alkyl group may or may not be substituted, and that the description includes both substituted and unsubstituted alkyl groups. "Salt" includes both acid and base addition salts.

[0034] "Acid addition salts" include salts of inorganic acids, such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, as well as salts of acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxoglucose ... "N" refers to salts formed with organic acids such as, but not limited to, rutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenic acid. "Base addition salt" refers to a salt prepared by the addition of an inorganic or organic base to a free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine.

[0035] Crystallization may produce solvates of the compounds described herein. The embodiments of the present invention include all solvates of the compounds described. As used herein, the term "solvate" refers to an aggregate that includes one or more molecules of the compound of the present invention together with one or more molecules of a solvent. The solvent may be water, in which case the solvate may be a hydrate. Alternatively, the solvent may be an organic solvent. Thus, the compounds of the present invention may exist as hydrates, including monohydrates, dihydrates, hemihydrates, sesquihydrates, trihydrates, tetrahydrates, etc., and corresponding solvate forms. The compounds of the present invention may be true solvates, whereas in other cases, the compounds of the present invention may simply retain adventitious water or another solvent, or may be a mixture of water plus some adventitious solvent.

[0036] The compounds of the present invention (e.g., compounds of structure I), or salts, tautomers, or solvates thereof, may contain one or more stereocenters and thus give rise to enantiomers, diastereomers, and other stereoisomers that can be defined in terms of absolute stereochemistry relative to the amino acid as (R)- or (S)- or (D)- or (L)-. The embodiments of the present invention are meant to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, e.g., chromatography and fractional crystallization. Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from suitable optically pure precursors, or resolution of the racemates (or racemates of salts or derivatives), e.g., using chiral high pressure liquid chromatography (HPLC). When the compounds described herein contain olefinic double bonds or other features which result in geometric asymmetry, unless otherwise specified, the compounds are intended to include both the E and Z geometric isomers, likewise, all tautomeric forms are intended to be included. "Stereoisomer" refers to a compound made up of the same atoms linked by the same bonds but with different three-dimensional structures that are not interchangeable. The present invention contemplates various stereoisomers and mixtures thereof, and includes "enantiomers," which refers to two stereoisomers whose molecules are non-superimposable mirror images of one another.

[0037] "Tautomer" refers to a proton transfer from one atom of a molecule to another atom of the same molecule. The present invention includes tautomers of any of the above compounds. Various tautomeric forms of the compounds can be readily derived by one skilled in the art. The chemical naming protocol and structure diagrams used herein are a modification of the IUPAC naming system using the ACD / Names version 9.07 software program and / or the ChemDraw Ultra version 11.0 software naming program (CambridgeSoft). Common names familiar to those skilled in the art are also used.

[0038] As noted above, in one embodiment of the present invention, compounds are provided that are useful as covalent linkers between biologically active moieties and targeting moieties. In other embodiments, compounds are provided that are useful as synthetic intermediates for the preparation of compounds that include one or more biologically active moieties. In general terms, embodiments of the present invention relate to polymers with pendant biologically active moieties. The biologically active moieties are linked by a linking moiety. Without wishing to be bound by a particular theory, it is believed that the linker helps to protect or suspend biological activity until the drug is released at the desired target. In another aspect, the linker provides a link between the biologically active moiety and the targeting moiety and serves to increase the accumulation of the biologically active moiety at the desired target. That is, biological activity is only effective or increased as a result of accumulation at the intended target, thus minimizing potential side effects (e.g., cytotoxicity) of the therapeutic agent. In other embodiments, there is provided a compound having the following structure (I): or a stereoisomer, pharma- ceutically acceptable salt, or tautomer thereof:

[0039] [ka] (I) (In the formula, M, at each occurrence, is independently a biologically active moiety or fragment thereof, a prodrug of a biologically active moiety or fragment thereof, a fluorescent dye, an imaging agent, or a radioisotope binding moiety, with the proviso that at least one occurrence of M is not a fluorescent dye; L is a physiologically cleavable linker; L1 , L 2 and L 3 is independently at each occurrence an optional alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, or a heteroatom linker; R 1 is independently, at each occurrence, H, alkyl, or alkoxy; R 2 and R 3 are each independently H, OH, SH, alkyl, alkoxy, alkyl ether, heteroalkyl, -OP(=R a )(R b )R c , Q or a protected form thereof, or L′; R a is O or S, R b OH, SH, O - , S - , OR d or S.R. d and R c OH, SH, O - , S - , OR d , O.L.', S.R. d , alkyl, alkoxy, heteroalkyl, heteroalkoxy, alkyl ether, alkoxyalkyl ether, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkyl ether, or thiophosphoalkyl ether; R d is the counter ion, Q, in each occurrence, is independently a moiety that contains a reactive group or a protected form thereof that is capable of forming a covalent bond with a complementary reactive group Q' of a targeting moiety; L', in each occurrence, is independently a linker that includes a covalent bond to a targeting moiety, Q, a linker that includes a covalent bond to a targeting moiety, a linker that includes a covalent bond to a solid support, a linker that includes a covalent bond to a solid support residue, a linker that includes a covalent bond to a nucleoside, or a linker that includes a covalent bond to an additional compound of structure (I); n is an integer equal to or greater than 1. The various linkers and substituents (e.g., M, Q, R 1 , R 2 , R 3 , R c , L, L 1 , L 2 , L 3 and L') may be substituted with another substituent. For example, in some embodiments, the optional substituents are selected to optimize the water solubility, permeability, retention, or other properties of the compound of structure (I). In certain embodiments, each alkyl, alkoxy, alkyl ether, alkoxyalkyl ether, phosphoalkyl, thiophosphoalkyl, phosphoalkyl ether, and thiophosphoalkyl ether in the compound of structure (I) may be substituted with another substituent selected from the group consisting of hydroxyl, alkoxy, alkyl ether, alkoxyalkyl ether, sulfhydryl, amino, alkylamino, carboxyl, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkyl ether, and thiophosphoalkyl ether. In certain embodiments, the optional substituent is -OP(=R a )(R b )R c (In the formula, R a , R b and R c is as defined for compounds of structure (I). In certain embodiments, the substituents are selected to increase cell or tissue permeability. In related embodiments, the substituents are selected to increase cell or tissue retention.

[0040] In some embodiments, L 1is, at each occurrence, independently an optional alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, or heteroatom linker. 1 is a heteroalkylene linker. For example, in some embodiments, the compound has the following structure (IA):

[0041] [ka] (IA) (In the formula, L 4 is independently at each occurrence an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, or heteroalkynylene linker; R 4 is, in each occurrence, independently, OH, SH, O - , S - , OR d or S.R. d and R 5 is independently, at each occurrence, oxo, or thioxo, or absent; m, in each occurrence, is independently an integer greater than or equal to 0. In some embodiments, L 4 is, at each occurrence, independently a heteroalkylene linker. 4 is, at each occurrence, independently an alkylene oxide linker. For example, in some embodiments, L 4 is polyethylene oxide, and the compound has the following structure (IB):

[0042] [ka] (IB) (wherein z is an integer from 1 to 100). In some embodiments of (IB), z is an integer from 3 to 6, such as about 6. In some embodiments, z is 3. In some embodiments, z is 4. In some embodiments, z is 5. In some embodiments, z is 6.

[0043] The linker L can be used as a point of attachment of the M moiety to the remainder of the compound. For example, in some embodiments, a synthetic precursor of the compound of structure (I) is prepared, and the M moiety is attached to the synthetic precursor using several facile methods known in the art, such as the method known as "click chemistry." To this end, any fast and substantially irreversible reaction can be used to attach M to the synthetic precursor to form the compound of structure (I). Exemplary reactions include the copper-catalyzed reaction of azides with alkynes to form triazoles (Husgen 1,3-dipolar cycloaddition), the reaction of dienes with parent dienes (Diels-Alder), strain-promoted alkyne-nitrone cycloaddition, the reaction of strained alkenes with azides, tetrazines or tetrazoles, the [3+2] cycloaddition of alkenes with azides, the reverse demand Diels-Alder of alkenes with tetrazines, the photoreaction of alkenes with tetrazoles, and various substitution reactions such as the replacement of leaving groups by nucleophilic attack on an electrophilic atom. Exemplary substitution reactions include reactions of amines with activated esters, N-hydroxysuccinimide esters, isocyanates, isothioscyanates, etc. In some embodiments, the reaction to form L can be carried out in an aqueous environment.

[0044] Thus, in some embodiments, L is a linker that, in each occurrence, contains a functional group capable of being formed by reaction of two complementary reactive groups, e.g., a functional group that is the product of one of the aforementioned "click" reactions. In various embodiments, in at least one occurrence of L, the functional group can be formed by reaction of an aldehyde, oxime, hydrazone, alkyne, amine, azide, acyl azide, acyl halide, nitrile, nitrone, sulfhydryl, disulfide, sulfonyl halide, isothiocyanate, imidoester, activated ester (e.g., N-hydroxysuccinimide ester), ketone, α,β-unsaturated carbonyl, alkene, maleimide, α-haloimide, epoxide, aziridine, tetrazine, tetrazole, phosphine, biotin, or thiirane functional group with a complementary reactive group, e.g., reaction of an amine with an N-hydroxysuccinimide ester or an isothiocyanate.

[0045] In other embodiments, in at least one occurrence of L, the functional group can be formed by reaction of an alkyne with an azide. In other embodiments, in at least one occurrence of L, the functional group can be formed by reaction of an amine (e.g., a primary amine) with an N-hydroxysuccinimide ester or an isothiocyanate. In further embodiments, at least one occurrence of L comprises a functional group that is an alkene, ester, amide, thioester, disulfide, carbocyclic, heterocyclic or heteroaryl group. In further embodiments, at least one occurrence of L comprises a functional group that is an alkene, ester, amide, thioester, thiourea, disulfide, carbocyclic, heterocyclic or heteroaryl group. In other embodiments, the functional group comprises an amide or thiourea. In some more specific embodiments, at least one occurrence of L comprises a linker that is a triazolyl functional group. In other embodiments, at least one occurrence of L comprises a linker that is a amide or thiourea functional group.

[0046] Some embodiments provide L that can be cleaved under appropriate conditions (e.g., physiological conditions). Thus, in some embodiments, L comprises an amide bond, an ester bond, a disulfide bond, a hydrazone, a phosphotriester, a diester, a β-glucuronide, a double bond, a triple bond, an ether bond, a ketone, a diol, a cyano, a nitro, or a combination thereof. In some embodiments, L comprises tert-butyloxycarbonyl, paramethoxybenzyl, dialkyl or diaryldialkoxysilane, orthoester, acetal, β-thiopropionate, ketal, phosphoramidate, hydrazone, vinyl ether, imine, aconityl, trityl, polyketal, bisarylhydrazone, diazobenzene, vicinal diol, pyrophosphate diester, or valine citrulline.

[0047] In certain embodiments, L, at each occurrence, is independently a linker that is cleavable at a pH ranging from 6 to 8. For example, in some embodiments, L is a linker that is cleavable at pH 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, or 8.0.

[0048] In certain embodiments, L, at each occurrence, is independently a linker that is cleavable at a temperature ranging from 20° C. to 40° C., 25° C. to 35° C., 30° C. to 35° C., 30° C. to 37° C., 35° C. to 37° C., 35° C. to 40° C., or 32° C. to 38° C. In certain embodiments, L, at each occurrence, is independently a linker that is cleavable at a temperature ranging from about 20° C., about 21° C., about 22° C., about 23° C., about 24° C., about 25° C., about 26° C., about 27° C., about 28° C., about 29° C., about 30° C., about 31° C., about 32° C., about 33° C., about 34° C., about 35° C., about 36° C., about 37° C., about 38° C., about 39° C., or about 40° C. In some embodiments, L, at each occurrence, is independently an enzyme-cleavable linker. For example, in some embodiments, the enzyme is a hydrolase, oxidoreductase, or lyase. In certain embodiments, the enzyme is an EC4.1 (e.g., EC4.1.1, EC4.1.2, EC4.1.3, or EC4.1.99), EC4.2, EC4.3, EC4.4, EC4.5, EC4.6, or EC4.99 enzyme. In certain embodiments, L has one of the following structures:

[0049] [ka] wherein R is H, methyl, ethyl, isopropyl, tert-butyl, or phenyl; X is O or CH2; n is an integer greater than 0. In yet another embodiment, in at least one occurrence of L, LM has the structure:

[0050] [ka] (In the formula, L 1a and L 1b are each independently an optional linker. In a different embodiment, in at least one occurrence of L, LM has the structure

[0051] [ka] (In the formula, L 1a and L 1b are each independently an optional linker. In various embodiments of the above, L 1a Or L 1b In another embodiment, L 1a Or L 1b , or both are present. In some embodiments, L 1a and L 1b When present, each independently is alkylene or heteroalkylene. For example, in some embodiments, L 1a and L 1b When present, independently has one of the following structures:

[0052] [ka] In yet another embodiment of structure (I), L, at each occurrence, is independently an optional alkylene or heteroalkylene linker. In certain embodiments, L1 has one of the following structures:

[0053] [ka] In further embodiments, L2 and L3, in each occurrence, are independently C1-C6 alkylene, C2-C6 alkenylene, or C2-C6 alkynylene. For example, in some embodiments, the compound has the following structure (IC):

[0054] [ka] (I C) (In the formula, x 1 , x 2 , x 3 and x 4 is, in each occurrence, independently an integer from 0 to 6; and z is an integer from 1 to 100.

[0055] In certain embodiments of the compound of structure (IC), x 1 , x 2 , x 3 or x 4 At least one occurrence of x is 1. 1 , x 2 , x3 and x 4 In another embodiment, x 1 and x 3 In some embodiments, x 2 and x 4 In still other embodiments, x 1 and x 3 is 0 in each occurrence, and x 2 and x 4 are 1 for each occurrence. In some more specific embodiments of the compounds of structure (IC), L comprises a triazolyl functional group. In some other specific embodiments of the compounds of structure (IC), L comprises an amide or thiourea functional group. In other embodiments of the compounds of structure (IB), L comprises an optional alkylene or heteroalkylene linker. In some embodiments, L 4 is, at each occurrence, independently, C1-C6 alkylene or C2-C6 alkynylene. For example, in some embodiments, the compound has the structure (ID):

[0056] [ka] (ID) (In the formula, x 1 , x 2 , x 3 and x 4 is independently, in each occurrence, an integer from 0 to 6; In some embodiments, y is 1. In some embodiments, y is 2. In some embodiments, y is 3, 4, 5, or 6. In certain embodiments, x 1 , x 2 , x 3 or x 4At least one occurrence of x is 1. 1 , x 2 , x 3 and x 4 are 1 for each occurrence. In still other embodiments of any of the compounds of structure (I), R 4 is, in each occurrence, independently, OH, O - OR d "OR d " and "SR d " is an O bonded to a cation - and S - For example, the disodium salt of the phosphate group is

[0057] [ka] (In the formula, R d is sodium (Na + ) can be expressed as In other embodiments of any of the compounds of structure (I), R 5 is oxo in each occurrence. In some different embodiments of any of the foregoing compounds, R 1 is H in each occurrence.

[0058] In various other embodiments, R 2 and R 3 are each independently OH or -OP(=R a )(R b )R c In some different embodiments, R 2 or R 3 is OH or -OP(=R a )(R b )R c and R 2 or R 3 The other is Q or a linker comprising a covalent bond to Q. In yet another embodiment of any of the compounds of structure (I) above, R 2 and R 3 are each independently -OP(=R a )(R b )R c In some of these embodiments, R c is OL'. In some more specific embodiments, L' is a targeting moiety or a linker to a targeting moiety. In related embodiments, L' is a linker to a targeting moiety, which linker comprises an alkylene oxide or a phosphodiester moiety, or a combination thereof. In other embodiments, R 2 and R 3 are each independently -OP(=R a )(R b ) OL', where L' is an alkylene or heteroalkylene linker to Q, a targeting moiety, an analyte (e.g., an analyte molecule), a solid support, a solid support moiety, a nucleoside, or an additional compound of structure (I).

[0059] The linker L' can be any linker suitable for linking Q, a targeting moiety, an analyte (e.g., an analyte molecule), a solid support, a solid support residue, a nucleoside, or an additional compound of structure (I) to the compound of structure (I). Advantageously, certain embodiments include the use of an L' moiety selected to increase or optimize the water solubility of the compound. In certain embodiments, L' is a heteroalkylene moiety. In some other certain embodiments, L' includes an alkylene oxide or a phosphodiester moiety, or a combination thereof. In certain embodiments, L' has the structure:

[0060] [ka] (In the formula, m" and n" are independently an integer from 1 to 10; R eis H, an electron pair or a counterion, L″ is a targeting moiety or a linkage to a targeting moiety.

[0061] In some embodiments, m" is an integer from 4 to 10, such as 4, 6, or 10. In other embodiments, n" is an integer from 3 to 6, such as 3, 4, 5, or 6. In certain embodiments, the targeting moiety is an antibody or a cell surface receptor antagonist. In related embodiments, the antibody or cell surface receptor antagonist is an epidermal growth factor receptor (EGFR) inhibitor, a hepatocyte growth factor receptor (HGFR) inhibitor, an insulin-like growth factor receptor (IGFR) inhibitor, a folate, or a MET inhibitor. In certain embodiments, the antibody or cell surface receptor antagonist is a tyrosine kinase inhibitor (e.g., gefitinib, erlotinib), lapatinib, vandetanib, neratinib, osimertinib, tivantinib (ARQ197), crizotinib, cabozantinib, tyrphostin (e.g., AG538, AG1024), pyrrolo(2,3-d)-pyrimidine derivative (e.g., NVP -AEW541), monoclonal antibodies (e.g., figitumumab, cetuximab, panitumumab, necitumumab, ganitumumab, cizutumumab, dalotuzumab, lobatumumab, onartuzumab, K1, labetuzumab, milatuzumab, lorvotuzumab, inotuzumab), BMS-777607, PF-02341066, PF-04217903, AMG-458, MK-2461, JNJ-38877605, GSK 1363089 (foretinib), XL880, XL 184, ARQ197, E7050, or INCB28060.

[0062] In certain embodiments, the antibody or cell surface receptor antagonist targets EGFR (e.g., EGFRvIII), HER2, phorate receptor, CD19, CD20, CD22, CD27L, CD30, CD33, CD37, CD56, CD66e, CD70, CD74, CD79b, CA6, CD138, CA6, mesothelin, nectin 4, STEAP1, MUC16, MaPi2b, GCC, Trop-2, AGS-5, ENPP3, carbonic anhydrase IX, GPNMB, PDMA. In some other embodiments, L" is an alkylene or heteroalkylene moiety. In some other certain embodiments, L" comprises an alkylene oxide, a phosphodiester moiety, a sulfhydryl, a disulfide, or a maleimide moiety, or a combination thereof. In other more specific embodiments of any of the compounds of structure (I) above, R 2 or R 3 has one of the following structures:

[0063] [ka] JPEG2025000634000017.jpg43170 JPEG2025000634000018.jpg38118 JPEG2025000634000019.jpg43170 JPEG2025000634000020.jpg27150 Certain embodiments of the compound of structure (I) can be prepared according to solid phase synthesis methods similar to those known in the art for the preparation of oligonucleotides. Thus, in some embodiments, L' is a linkage to a solid support, a solid support residue, or a nucleoside. Solid supports containing activated deoxythymidine (dT) groups are readily available and can be used, in some embodiments, as starting materials for the preparation of compounds of structure (I). Thus, in some embodiments, R 2or R 3 has the following structure:

[0064] [ka] Those skilled in the art will appreciate that the dT group shown above is included merely for ease of synthesis and economic efficiency, and is not required. Other solid supports can be used, which would result in different nucleosides or solid support moieties being present on L', or the nucleosides or solid support moieties can be removed or modified after synthesis.

[0065] In yet other embodiments, Q, at each occurrence, is independently a moiety that includes a reactive group that can form a covalent bond with an analyte molecule or a solid support. In other embodiments, Q, at each occurrence, is independently a moiety that includes a reactive group that can form a covalent bond with a complementary reactive group, Q'. For example, in some embodiments, Q' is a moiety that includes a further compound of structure (I) (e.g., R 2 or R 3 and Q and Q' contain complementary reactive groups such that reaction of a compound of structure (I) with another compound of structure (I) results in a covalently linked dimer of a compound of structure (I). Multimeric compounds of structure (I) can also be prepared in an analogous manner and are included within the scope of embodiments of the present invention.

[0066] The type of Q group and its connectivity to the remainder of the compound of structure (I) are not limited, provided that Q contains a moiety with appropriate reactivity to form the desired bond. In certain embodiments, Q is a moiety that is not susceptible to hydrolysis under aqueous conditions, but is sufficiently reactive to form with a corresponding group (eg, an amine, azide, or alkyne) on an analyte molecule or solid support.

[0067] Certain embodiments of the compound of structure (I) include Q groups commonly used in the field of bioconjugation. For example, in some embodiments, Q includes a nucleophilic reactive group, an electrophilic reactive group, or a cycloaddition reactive group. In some more specific embodiments, Q includes a sulfhydryl, a disulfide, an activated ester, an isothiocyanate, an azide, an alkyne, an alkene, a diene, a dienophile, an acid halide, a sulfonyl halide, a phosphine, an α-haloamide, a biotin, an amino, or a maleimide functional group. In some embodiments, the activated ester is an N-succinimide ester, an imido ester, or a polyflourophenyl ester. In other embodiments, the alkyne is an alkyl azide or an acyl azide.

[0068] The Q group can be conveniently provided in a state to enhance storage stability or other desired properties, and the protecting group can then be removed at the appropriate time, for example, for coupling with a targeting moiety or analyte. Thus, the Q group includes "protected forms" of reactive groups, including any of the reactive groups described above and in Table 1 below. A "protected form" of Q refers to a moiety that is less reactive than Q under a given reaction condition, but can preferably be converted to Q under conditions that do not degrade or react with other moieties of the compound of structure (I). Those skilled in the art can derive a suitable protected form of Q based on the particular Q and the desired end use and storage conditions. For example, when Q is SH, the protected form of Q includes a disulfide, which can be reduced to reveal the SH moiety using commonly known techniques and reagents. Exemplary Q moieties are set forth in Table 1 below.

[0069] [Table 1] JPEG2025000634000024.jpg250139 It should be noted that in some embodiments where Q is SH, the SH moiety tends to form a disulfide bond with another sulfhydryl group of another compound, for example of structure (I). Thus, some embodiments include compounds of structure (I) in the form of a disulfide dimer in which the disulfide bond is derived from the Q group of SH.

[0070] Within certain embodiments, R 2 and R 3 Also included are compounds of structure (I) in which one or both of R 2 and R 3 Either or both of the following are -OP(=R a )(R b )R c and R c Also included are compounds in which L' is OL' and L' is a linker comprising a covalent bond to an additional compound of structure (I). Such compounds can be prepared, for example, by preparing a first compound of structure (I) having about 10 "M" moieties (i.e., n=9) and a "Q" suitable for reaction with a complementary Q' group of a second compound of structure (I). In this manner, compounds of structure (I) having multiple "M" moieties, for example 100 or more, can be prepared without the need to sequentially couple each monomer. An exemplary embodiment of such a compound of structure (I) is shown below in structure (I'):

[0071] [ka] (I') (In the formula, R 1 , R 2 , R 3 , R 4 , R 5 , L, L 2 , L 3 , L 4 each occurrence of M, m, and n is independently as defined for compounds of structure (I); L″ is a linker containing a functional group resulting from reaction of a Q moiety with the corresponding Q′ moiety; and α is an integer greater than 1, for example, an integer from 1 to 100 or from 1 to 10. Other compounds of structure (I') are compounds that one of skill in the art can derive, for example, by dimerizing or polymerizing the compounds of structure (I) described herein. In other embodiments, the Q moiety is conveniently masked (e.g., protected) as a disulfide moiety, which can be subsequently reduced to provide an activated Q moiety for attachment to a desired targeting moiety. For example, the Q moiety can be represented by the following structure:

[0072] [ka] where R is an optionally substituted alkyl group. For example, in some embodiments, Q is provided as a disulfide moiety having the structure:

[0073] [ka] (wherein n' is an integer of 1 to 10, for example 6). In some other embodiments, R 2 or R 3 One of the groups is OH or -OP(=R a )(R b )R c and R 2 or R 3 The other is a linker that includes a covalent bond to the targeting moiety or a linker that includes a covalent bond to the solid support. For example, in some embodiments, the targeting moiety is an antibody or a cell surface receptor antagonist. In yet another embodiment, the solid support is a polymeric bead or a non-polymeric bead.

[0074] The value of m is another variable that can be selected based on the desired solubility, permeation effect, or therapeutic application. In some embodiments, m, in each occurrence, is independently an integer from 1 to 10. In other embodiments, m, in each occurrence, is independently an integer from 1 to 5, e.g., 1, 2, 3, 4, or 5. In other embodiments, m, in each occurrence, is independently an integer greater than 2 and z is an integer from 3 to 10; for example, in some embodiments, m, in each occurrence, is independently an integer greater than 2, such as 3, 4, 5, or 6, and z is an integer from 3 to 6.

[0075] Solubility, permeability or retention can also be adjusted by selection of different values ​​of n. In certain embodiments, n, at each occurrence, is an integer from 1 to 100. In other embodiments, n is an integer from 1 to 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10.

[0076] Tumor cell antigens include tumor-specific antigens and tumor-associated antigens, such as EGFR, HER 2, phorate receptor, CD20, CD33, carcinoembryonic antigens (e.g., alpha-fetoprotein, carcinoembryonic antigen, immature laminin receptor, TAG-72), CA-125, MUC-1, epithelial tumor antigens, tyrosinase, melanoma-associated antigens (MAGE), and aberrant products of RAS or p53. Tumor cell antigens can also include antigens considered to be oncofetal, oncoviral (e.g., HPV E6, E7), overexpressed / accumulated (e.g., BING-4, calcium activated chloride channel 2, 9D7, Ep-CAM, EphA3, HER2, telomerase, mesothelin, SAP-1, survivin), cancer-tetis (e.g., BAGE family, CAGE family, GAGE ​​family, MAGE family, SAGE family, XAGE family), lineage restricted, mutated, post-translationally modified, idiotypic, CT9 or CT10 (e.g., NY-ESO-1 / LAGE-1, PRAME). In certain embodiments, M, at each occurrence, is independently an NSAID, a kinase inhibitor, an anthracycline, and an EGFR inhibitor or an alkylating agent.

[0077] In some embodiments, the biologically active moiety is an anti-cancer drug. In certain specific embodiments, M, at each occurrence, is independently an anti-cancer drug, and the targeting moiety is an antibody specific to a tumor cell antigen. As used herein, an anti-cancer drug includes derivatives. That is, an anti-cancer drug that has been modified or derivatized (e.g., to include a Q moiety) so that the anti-cancer drug can be conjugated or attached to another molecule. For example, maytansine is an anti-cancer drug, and maytansinoids are anti-cancer drug derivatives. In certain embodiments, the anticancer drug is an epidermal growth factor receptor (EGFR) inhibitor, a phosphatidylinositol kinase (PI3K) inhibitor, an insulin-like growth factor receptor (IGF1R) inhibitor, a Janus kinase (JAK) inhibitor, a Met kinase inhibitor, an SRC family kinase inhibitor, a mitogen-activated protein kinase (MEK) inhibitor, an extracellular signal-regulated kinase (ERK) inhibitor, a topoisomerase inhibitor (such as irinotecan, or such as etoposide, or such as doxorubicin), a taxane (such as an anti-microtubule agent, including paclitaxel and docetaxel), an antimetabolite (such as 5-FU or such as gemcitabine), an alkylating agent (such as cisplatin or such as cyclophosphamide), or a taxane. Anti-cancer drugs that can be modified and incorporated into the compound embodiments of the present disclosure include, for example, auristatin F; auristatin E; maytansine; calicheamicin; paclitaxel; doxorubicin; cryptophycin; erlotinib; CC-1065; carzelesin; SJG-136; DSB-120; afatinib; Iressa; methotrexate; DNA methylating agents (e.g., procarbazine, temozolomide, dacarbazine, N-methyl-N-nitrourea, N-methyl-N'-nitro-N-nitroguanine, etc.).

[0078] Other non-limiting examples of anti-cancer drugs include Gleevec® (imatinib mesylate), Velcade® (bortezomib), Casodex (bicalutamide), Iressa® (gefitinib), and adriamycin; alkylating agents such as thiotepa and cyclosphosphamide (CYTOXAN®); alkylsulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; Ethyleneimines and methylamelamines, including lutretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphaoramide, and trimethylolmelamine; chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nobuenbiquine, phenesterine, prednisone nitrogen mustards such as uracil mustard, trofosfamide, and uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine; aclacinomycin, actinomycin, anthramycin, azaserine, bleomycin, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, Casodex®, chromomycin, dactinomycin, and daunorubicin. Antibiotics such as detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcelomycin, mitomycin, mycophenolic acid, nogalamycin, olivomycin, peplomycin, porfiromycin, puromycin, queramycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, and zorubicin; metabolic antagonists such as methotrexate and 5-fluorouracil (5-FU);folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; antiadrenal agents such as aminoglutethimide, mitotane, trilostane; folinic acid acid);aceglatone;aldophosphamide glycosides;aminolevulinic acid;amsacrine;bestrabulil;bisantrene;edatrexate;defofamine;demecolcine;diaziquone;eflornithine;elfomithine;elliptinium acetate;etoglucide;gallium nitrate;hydroxyurea;lentinan;lonidamine;mitoguazone;mitoxantrone;mopidamol;nitracrine;pentostatin;phenamet;pirarubicin syn; podophyllic acid; 2-ethylhydrazide; procarbazine; PSK.RTM.; razoxane; schizophyllan; spirogermanium; tenuazonic acid; triazicon; 2,2',2"-trichlorotriethylamine; urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxanes such as paclitaxel (TAXOL™, Bristol-Myers Squibb Oncology, Princeton, NJ) and docetaxel (TAXOTERE™, Rhone-Poulenc Rorer, Antony, France); retinoic acid; esperamycin or capecitabine. Suitable anti-cancer drugs include, for example, antiestrogens, including tamoxifen (Nolvadex™), raloxifene, aromatase-inhibiting 4(5)-imidazole, 4-hydroxytamoxifen, trioxyphene, keoxyphene, LY 117018, onapristone, and toremifene (Fareston);and antihormonal agents that act to control or inhibit hormone action on tumors, such as antiandrogens, such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs, such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; camptothecin-11 (CPT-11); the topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO);

[0079] If desired, embodiments of the compounds or compositions of the present disclosure can be administered in combination with other agents such as Herceptin®, Avastin®, Erbitux®, Rituxan®, Taxol®, Arimidex®, Taxotere®, ABVD, AVICINE, abagovomab, acridine carboxamide, adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, alpharadin, alvocidib, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone, amonafide, anthracenedione, anti-CD22 immunotoxins, antineoplastic agents, antitumor herbs, apaziquone, atiprimod, azathioprine, belotecan, bendamustine, BIBW 2992, biricoderm, brostallicin, bryostatin, buthionine sulfoximine, CBV (chemotherapy), calyculin, cell cycle nonspecific antineoplastic agents, dichloroacetic acid, discodermolide, elsamitrucin, enocitabine, epothilone, eribulin, everolimus, exatecan, exisulind, ferruginol, forodesine, fosfestrol, ICE chemotherapy regimen, IT-101, imexon, imiquimod, indolocarbazole, irofulven, laniquidar, larotaxel, lenalidomide, lucantone, lurtotecan, mafosfamide, mitozolomide, nafoxid It can be used in combination with commonly prescribed anticancer drugs such as cyclophosphamide, nedaplatin, olaparib, ortataxel, PAC-1, pawpaw, pixantrone, proteasome inhibitors, rebeccamycin, resiquimod, rubitecan, SN-38, salinosporamide A, sapacitabine, Stanford V, swainsonine, talaporfin, tariquidar, tegafur-uracil, temodar, tesetaxel, triplatin tetranitrate, tris(2-chloroethyl)amine, troxacitabine, uramustine, vadimezan, vinflunine, ZD6126 or zosuquidar.

[0080] M is selected based on the desired therapeutic and / or optical properties, for example, the treatment of a specific disease or condition (e.g., cancer) or the generation of a particular color and / or fluorescent emission wavelength. In some embodiments, M is the same in each occurrence. However, it is important to note that each occurrence of M does not have to be the same M, and certain embodiments include compounds in which M is not the same in each occurrence. For example, in some embodiments, M is not the same in each occurrence, and different M moieties are selected to have different therapeutic properties (e.g., cytotoxicity and anti-inflammatory). In some embodiments, M is not the same in each occurrence, and different M moieties are selected to have the same or similar therapeutic properties (e.g., cytotoxicity). In certain embodiments, M is independently selected from the following:

[0081] [ka] For ease of illustration, the specific points of attachment to the remainder of the molecule (i.e.,

[0082] [ka] Although the formula is shown having a linker group, the M moiety can be attached via any available point. One of skill in the art can determine the appropriate point of attachment. In certain embodiments, LM has one of the following structures:

[0083] [ka] JPEG2025000634000033.jpg166170 In yet more embodiments of any of the foregoing, M are the same. In other embodiments, M are each different. In yet more embodiments, one or more M are the same and one or more M are different.

[0084] In some embodiments, the selected occurrences of M are not the same, and the different M moieties are selected to have absorbance and / or emission for use in fluorescence resonance energy transfer (FRET) methods. For example, in such embodiments, the different M moieties are selected so that radiation absorption at one wavelength causes radiation emission at a different wavelength by FRET mechanism. Exemplary M moieties can be appropriately selected by those skilled in the art based on the desired end use. Exemplary M moieties for FRET methods include fluorescein and 5-TAMRA (5-carboxytetramethylrhodamine, succinimidyl ester) dyes. M can be attached to the remainder of the molecule from any position (i.e., atom) of M. Those of skill in the art will recognize means for attaching M to the remainder of the molecule. Exemplary methods include the "click" reaction described herein.

[0085] In some embodiments, M is a fluorescent or colored moiety. Any fluorescent and / or colored moiety can be used, such as those known in the art and typically used in colorimetric, UV, and / or fluorescent assays. Examples of M moieties that are useful in various embodiments of the invention include xanthene derivatives (e.g., fluorescein, rhodamine, Oregon Green, eosin, or Texas Red); cyanine derivatives (e.g., cyanine, indocarbocyanine, oxacarbocyanine, thiacarbocyanine, or merocyanine); squaraine derivatives and ring-substituted squaraines, including Seta, SeTau, and Square dyes; naphthalene derivatives (e.g., dansyl and prodan derivatives); coumarin derivatives; oxadiazole derivatives (e.g., pyridyloxazole, nitrobenzoxadiazole, or benzoxadiazole); anthracene derivatives (e.g., DRAQ5, DRAQ7, and CyTRAK). Examples of M moieties include, but are not limited to, anthraquinones, including Orange; pyrene derivatives such as Cascade Blue; oxazine derivatives (e.g., Nile Red, Nile Blue, Cresyl Violet, Oxazine 170); acridine derivatives (e.g., Proflavine, Acridine Orange, Acridine Yellow); arylmethine derivatives: Auramine, Crystal Violet, Malachite Green; and tetrapyrrole derivatives (e.g., Porphine, Phthalocyanine or Bilirubin). Other exemplary M moieties include cyanine dyes, xanthate dyes (e.g., Hex, Vic, Nedd, Joe or Tet); Yakima Yellow; Redmond Red; tamra; Texas Red and Alexa Fluor® dyes.

[0086] In yet another embodiment of any of the above, M also includes three or more aryl or heteroaryl rings or combinations thereof, such as four or more aryl or heteroaryl rings or combinations thereof, or five or more aryl or heteroaryl rings or combinations thereof. In some embodiments, M also includes six aryl or heteroaryl rings or combinations thereof. In other embodiments, the rings are fused. For example, in some embodiments, M also includes three or more fused rings, four or more fused rings, five or more fused rings, or six or more fused rings.

[0087] In some embodiments, M is cyclic. For example, in some embodiments, M is carbocyclic. In other embodiments, M is heterocyclic. In yet other embodiments of the above, M independently comprises an aryl moiety at each occurrence. In some of these embodiments, the aryl moiety is polycyclic. In other more specific examples, the aryl moiety is a fused polycyclic aryl moiety, and can comprise, for example, at least three, at least four, or even more than four aryl rings. In other embodiments of any of the foregoing compounds of structure (I), (IA), (IB), (IC), (ID) or (I'), M comprises at least one heteroatom. For example, in some embodiments, the heteroatom is nitrogen, oxygen or sulfur. In further embodiments of any of the foregoing, M includes at least one substituent. For example, in some embodiments, the substituent is a fluoro, chloro, bromo, iodo, amino, alkylamino, arylamino, hydroxy, sulfhydryl, alkoxy, aryloxy, phenyl, aryl, methyl, ethyl, propyl, butyl, isopropyl, t-butyl, carboxy, sulfonate, amido, or formyl group.

[0088] In some more specific embodiments of the foregoing, M is a dimethylaminostilbene, quinacridone, fluorophenyl-dimethyl-BODIPY, bis-fluorophenyl-BODIPY, acridine, terylene, sexiphenyl, porphyrin, benzopyrene, (fluorophenyl-dimethyl-difluorobora-diaza-indacene)phenyl, (bis-fluorophenyl-difluorobora-diaza-indacene)phenyl, quaterphenyl, bi-benzothiazole, ter-benzothiazole, bi-naphthyl, bi-anthracyl, squaraine, squarylium, 9,10-ethynylanthracene, or ter-naphthyl moiety. In other embodiments, M is p-terphenyl, perylene, azobenzene, phenazine, phenanthroline, acridine, thioxanthrene, chrysene, rubrene, coronene, cyanine, perylene imide, or perylene amide, or a derivative thereof. In still other embodiments, M at each occurrence is independently a coumarin dye, a resorufin dye, a dipyrrometheneboron difluoride dye, a ruthenium bipyridyl dye, an energy transfer dye, a thiazole orange dye, a polymethine, or an N-aryl-1,8-naphthalimide dye. In some embodiments, M is pyrene, perylene, perylene monoimide or 6-FAM or a derivative thereof. In some other embodiments, M has one of the following structures:

[0089] [ka]

[0090] The M moiety containing the carboxylic acid group is in the anionic form (CO2 - ), but one of skill in the art will appreciate that this varies with pH and that the protonated form (COH) is included in various embodiments. In some specific embodiments, the compound is a compound selected from Table 2. In some embodiments, the compound is a compound selected from Table 3. In some embodiments, the compound is a compound selected from Table 4. The compounds in Table 2 were prepared according to the procedures described in the Examples, and their identities were confirmed by mass spectrometry.

[0091] [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8] [Table 2-9]

[0092] [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4]

Table 3-5

Table 3-6

Table 3-7

Table 3-8

Table 3-9

Table 3-10

Table 3-11

Table 3-12

Table 3-13

Table 3-14

Table 3-15

Table 3-16

[0093]

Table 4-1

Table 4-2

Table 4-3

Table 4-4

Table 4-5

[0094] As used in Tables 2, 3, and 4, and throughout this specification, R 2 , R 3 , M, m, n, and L' have the definitions given for the compounds of structure (I) unless otherwise indicated. In some embodiments, M is F, F', F", N', I', D', or D". F, F', and F" each refer to a fluorescein moiety having the following structure:

[0095] [ka] "N'" refers to the following structure:

[0096] [ka] "I'" refers to the following structure:

[0097] [ka] "D'" refers to the following structure:

[0098] [ka] "D" refers to the following structure:

[0099] [ka] "dT" has the following structure

[0100] [ka] (In the formula, R is H or a direct bond. "AF" refers to the following structure:

[0101] [ka] Thus, in some embodiments, at least one occurrence of M has one of the following structures:

[0102] [ka] In some more specific embodiments, each occurrence of M has the structure:

[0103] [ka] In certain embodiments, the treatment comprises reducing or alleviating pain or inflammation. In certain embodiments, the treatment comprises pain control or pain management. In some specific embodiments, at least one occurrence of M has one of the following structures:

[0104] [ka]

[0105] In some more specific embodiments, each occurrence of M has one of the following structures:

[0106] [ka]

[0107] Some embodiments include any of the aforementioned compounds, including the specific compounds provided in Table 2, conjugated to a targeting moiety, such as an antibody. In some embodiments, one compound of structure (I) is conjugated to the antibody. In some embodiments, one to two compounds of structure (I) are conjugated to the antibody. In some embodiments, two compounds of structure (I) are conjugated to the antibody. In some embodiments, three compounds of structure (I) are conjugated to the antibody. In some embodiments, four compounds of structure (I) are conjugated to the antibody. In some embodiments, five compounds of structure (I) are conjugated to the antibody. In some embodiments, five or fewer compounds of structure (I) are conjugated to the antibody.

[0108] In various embodiments, reactive polymers can be used to prepare compounds of structure (I). In certain embodiments, these reactive polymers are synthetic intermediates that contain moieties that are useful for reacting with complementary moieties by some synthetic methods (e.g., the "click" reaction described above) to form a covalent bond between M and the reactive polymer, thereby forming compounds of structure (I). Thus, in various embodiments, compounds of structure (I), or stereoisomers, salts, or tautomers thereof, are formed using reactive polymers having the following structure (V):

[0109] [ka] (V) (In the formula, G, in each occurrence, is independently a moiety that contains a reactive group or a protected analogue thereof that is capable of forming a covalent bond with a complementary reactive group; L 1a , L 2 and L 3 is independently at each occurrence an optional alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, or a heteroatom linker; L 4 is, independently at each occurrence, a heteroalkylene, heteroalkenylene, or heteroalkynylene linker greater than 3 atoms in length, and the heteroatoms in the heteroalkylene, heteroalkenylene, and heteroalkynylene linkers are selected from O, N, and S; R 1 is independently, at each occurrence, H, alkyl, or alkoxy; R 2 and R 3 are each independently H, OH, SH, alkyl, alkoxy, alkyl ether, heteroalkyl, -OP(=R a )(R b )R c , Q or L′, R 4 is, in each occurrence, independently, OH, SH, O - , S - , OR d or S.R. d and R 5 is independently, at each occurrence, oxo, or thioxo, or absent; R a is O or S, R b OH, SH, O - , S - , OR d or S.R. d and R c OH, SH, O - , S - , OR d , O.L.', S.R. d , alkyl, alkoxy, alkyl ether, alkoxyalkyl ether, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkyl ether or thiophosphoalkyl ether; R d is the counter ion, Q, at each occurrence, is independently a moiety that contains a reactive group or a protected analogue thereof that is capable of forming a covalent bond with an analyte molecule, a targeting moiety, a solid support, or a complementary reactive group Q'; L', in each occurrence, is independently a linker that includes a covalent bond to Q, a linker that includes a covalent bond to a targeting moiety, a linker that includes a covalent bond to an analyte molecule, a linker that includes a covalent bond to a solid support, a linker that includes a covalent bond to a solid support residue, a linker that includes a covalent bond to a nucleoside, or a linker that includes a covalent bond to an additional compound of structure (V); m, in each occurrence, is independently an integer greater than or equal to 0, provided that at least one occurrence of m is an integer greater than or equal to 1; (n is an integer greater than or equal to 1) In some embodiments, the reactive polymer is selected from Table 5 below.

[0110] [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4] [Table 5-5] [Table 5-6] [Table 5-7] [Table 5-8] [Table 5-9] In various embodiments, G in the compounds of Table 5 is alkynyl, such as ethynyl. In other embodiments, G in the compounds of Table 5 is azide. In other embodiments, G in the compounds of Table 5 is amino (NH2). In other embodiments, G in the compounds of Table 5 is isothiocyanate. In other embodiments, G in the compounds of Table 5 is an activated ester, such as an ester of N-hydroxysuccinimide.

[0111] Certain embodiments relate to therapeutically effective fluorescent compounds, with the proviso that at least one occurrence of M is not a fluorescent dye and at least one occurrence of M is a fluorescent dye. Therapeutically effective fluorescent compounds include compounds that contain at least one biologically active moiety or fragment thereof, or a prodrug of a biologically active moiety or fragment thereof, which emit a fluorescent signal upon excitation with light, such as ultraviolet light. In further embodiments, the compound of structure (I) includes at least one ethylene oxide moiety. Also provided is a composition comprising any one of the claimed compounds and a targeting moiety.

[0112] The embodiments of the compounds disclosed herein are "tunable," meaning that by appropriate selection of the variables in any of the aforementioned compounds, one of skill in the art can arrive at a compound with a desired and / or predetermined molar fluorescence (molar brightness). The "tunability" of certain embodiments of the compounds allows the user to easily arrive at a compound with the desired fluorescence and / or color for use in a particular assay. While all variables can affect the molar fluorescence of certain embodiments of the compounds disclosed herein, the M, L, and 4 It is believed that the appropriate selection of m, n and n plays an important role in the molar fluorescence of the disclosed compound embodiments. Thus, in one embodiment, a method for obtaining a compound with a desired molar fluorescence comprises selecting an M moiety with known fluorescence, preparing a compound of structure (I) containing the M moiety, and reacting L 4, m and n to arrive at a desired molar fluorescence.

[0113] For ease of illustration, various compounds containing phosphorus moieties (e.g., phosphate, etc.) are shown in their anionic state (e.g., -OPO(OH)O - , -OPO3 2- ) as a charge. One of skill in the art will readily appreciate that charge is pH dependent, and that uncharged (e.g., protonated or salts such as sodium or other cations) forms are also within the scope of embodiments of the present invention. Compositions comprising any of the aforementioned compounds and one or more targeting moieties (e.g., antibodies or cell surface receptor antagonists) are provided in various other embodiments. In some embodiments, the use of such compositions in a method for treating a disease is also provided, the method comprising administering to a subject in need of treatment of the disease a therapeutically effective amount of a compound of structure (I) or a composition comprising a compound of structure (I), wherein each M is independently a biologically active moiety effective for treating the disease.

[0114] Pharmaceutical Compositions Certain embodiments provide a composition comprising a compound according to any one of the embodiments disclosed herein and a pharma- ceutically acceptable carrier. Another embodiment provides a composition comprising a plurality of conjugates, the conjugates comprising an antibody covalently bound to two or more biologically active moieties via a single linkage, the plurality of conjugates having at least 90% structural uniformity. In a more specific embodiment, the plurality of conjugates has at least 95% structural uniformity. In a related embodiment, the plurality of conjugates has greater than 99% structural uniformity. In certain embodiments, the single linkage is to a polymer backbone, the polymer backbone comprising two or more biologically active moieties covalently bound thereto. In certain embodiments, the conjugates each independently comprise a compound according to an embodiment disclosed herein, 2and R 3 One of them is -OP(=R a )(R b )OL' or L', where L' is an antibody or a linker comprising a covalent bond to an antibody.

[0115] Other embodiments relate to pharmaceutical compositions. The pharmaceutical composition comprises any one (or more) of the aforementioned compounds and a pharma- ceutically acceptable carrier. In some embodiments, the pharmaceutical composition is formulated for oral administration. In other embodiments, the pharmaceutical composition is formulated for injection. In still other embodiments, the pharmaceutical composition comprises a compound disclosed herein and an additional therapeutic agent (e.g., an anti-cancer agent). Non-limiting examples of such therapeutic agents are described herein below. Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ocular, pulmonary, mucosal, transdermal, intravaginal, otic, nasal, and topical administration.Further, by way of example only, parenteral delivery includes intramuscular, subcutaneous, intravenous, intramedullary injection, as well as intrathecal, direct intraventricular, intraperitoneal, intralymphatic, and intranasal injection. In certain embodiments, the compounds described herein are administered locally, rather than systemically, for example, by injecting the compounds directly into an organ, often in a depot preparation or sustained release formulation. In specific embodiments, long-acting formulations are administered by implantation (e.g., subcutaneous or intramuscular) or intramuscular injection. Furthermore, in other embodiments, the drug is delivered in a targeted drug delivery system, for example, liposomes coated with organ-specific antibodies. In such embodiments, the liposomes target and are selectively taken up by the organ. In still other embodiments, the compounds described herein are provided in the form of a fast-release formulation, a sustained-release formulation, or an intermediate-release formulation. In still other embodiments, the compounds described herein are administered locally.

[0116] The compounds according to the invention are effective over a wide range of doses. For example, in treating adults, doses of 0.01-1000 mg, 0.5-100 mg, 1-50 mg, and 5-40 mg per day are examples of doses used in some embodiments. An exemplary dose is 10-30 mg per day. The exact dose depends on the route of administration, the form in which the compound is administered, the subject being treated, the weight of the subject being treated, and the preferences and experience of the attending physician. In some embodiments, the compound of the present invention is administered in a single dose.Typically, such administration is by injection, for example intravenous injection, in order to rapidly introduce the drug.However, other routes can be used as appropriate.A single dose of the compound of the present invention can also be used to treat acute conditions. In some embodiments, the compound of the present invention is administered in multiple doses. In some embodiments, administration is about once, twice, three times, four times, five times, six times, or more than six times per day. In other embodiments, administration is about once a month, once every other week, once a week, or once every other day. In another embodiment, the compound of the present invention and another agent are administered together about once a day to about six times a day. In another embodiment, administration of the compound of the present invention and the agent continues for less than about 7 days. In yet another embodiment, administration continues for more than about 6, 10, 14, 28 days, 2 months, 6 months, or 1 year. In some cases, continuous administration is achieved and maintained as long as necessary.

[0117] The administration of the compound of the present invention can be continued as long as necessary.In some embodiments, the compound of the present invention is administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days.In some embodiments, the compound of the present invention is administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day.In some embodiments, the compound of the present invention is administered chronically for a long period of time, for example, to treat chronic effects. In some embodiments, the compound of the present invention is administered in multiple doses.It is known in the art that due to subject-to-subject variability of compound pharmacokinetics, individualization of dosing regimen is necessary for optimal treatment.The administration of the compound of the present invention can be found by routine experimentation in view of the present disclosure.

[0118] In some embodiments, the compounds described herein are formulated into pharmaceutical compositions.In specific embodiments, the pharmaceutical compositions are formulated in a conventional manner using one or more physiologically acceptable carriers, including excipients and auxiliary agents that facilitate the processing of active compounds into pharma-ceutically usable preparations.The appropriate formulation depends on the route of administration selected.Any pharma-ceutically acceptable technique, carrier, and excipient can be used as suitable for formulating the pharmaceutical compositions described herein.Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, HA and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999).

[0119] Described herein is a pharmaceutical composition comprising a compound of structure (I) and a pharma- ceutically acceptable diluent, excipient, or carrier.In certain embodiments, the described compound is administered as a pharmaceutical composition in which the compound of structure (I) is mixed with other active ingredients, as in combination therapy.All combinations of actives described in the combination therapy section below and throughout this disclosure are encompassed herein.In a specific embodiment, the pharmaceutical composition comprises one or more compounds of structure (I). As used herein, a pharmaceutical composition refers to a mixture of a compound of structure (I) and other chemical components, such as carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents, and / or excipients. In certain embodiments, the pharmaceutical composition facilitates administration of the compound to an organism. In some embodiments, the methods of treatment or use described herein are practiced by administering a therapeutically effective amount of a compound of structure (I) described herein as a pharmaceutical composition to a mammal having a disease, disorder, or medical condition to be treated. In specific embodiments, the mammal is a human. In certain embodiments, the therapeutically effective amount will vary depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors. The compounds described herein are used alone or in combination with one or more therapeutic agents as components of a mixture.

[0120] In one embodiment, one or more compounds of structure (I) are formulated as an aqueous solution. In a specific embodiment, the aqueous solution is selected from physiologically compatible buffers, such as Hank's solution, Ringer's solution, or physiological saline buffer, merely by way of example. In another embodiment, one or more compounds of structure (I) are formulated for transmucosal administration. In a specific embodiment, the transmucosal formulation comprises a permeabilizing agent appropriate for the barrier to be permeated. In yet another embodiment, in which the compounds described herein are formulated for other parenteral injections, suitable formulations comprise aqueous or non-aqueous solutions. In a specific embodiment, such solutions comprise physiologically compatible buffers and / or excipients. In another embodiment, the compound described herein is formulated for oral administration.The compound described herein is formulated by combining active compound with, for example, pharmaceutically acceptable carrier or excipient.In various embodiments, the compound described herein is formulated as oral dosage form, which includes, by way of example only, tablet, powder, pill, dragee, capsule, liquid, gel, syrup, elixir, slurry, suspension and the like.

[0121] In certain embodiments, pharmaceutical preparations for oral use can be obtained by mixing one or more solid excipients with one or more of the compounds described herein, grinding the resulting mixture, and processing the mixture of granules to obtain tablets or dragee cores, after adding suitable auxiliary agents if desired.Suitable excipients are fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol, among others; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, crystalline cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or others such as polyvinylpyrrolidone (PVP or povidone) or calcium phosphate.In specific embodiments, disintegrants can be added.Disintegrants include, by way of example only, cross-linked croscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or its salts, such as sodium alginate.

[0122] In one embodiment, dosage forms such as dragee cores and tablets are coated with one or more suitable coatings. In a specific embodiment, sugar concentrates are used to coat the dosage forms. The sugar concentrates may contain additional components such as gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures, merely by way of example. Dyes and / or pigments may also be added to the coating for identification purposes. Furthermore, dyes and / or pigments may be utilized to characterize different combinations of active compound doses. In certain embodiments, at least one of the therapeutically effective amount of the compounds described herein is formulated into other oral dosage forms. Oral dosage forms include push-fit capsules made of gelatin, and sealed soft capsules made of gelatin and plasticizers such as glycerol or sorbitol. In specific embodiments, the push-fit capsules contain active ingredient mixed with one or more fillers. Fillers include, by way of example only, binders such as lactose, starch, and / or lubricants such as talc or magnesium stearate, and may contain stabilizers. In other embodiments, the soft capsules contain one or more active compounds dissolved or suspended in a suitable liquid. Suitable liquids include, by way of example only, one or more fatty oils, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers may be added. In other embodiments, a therapeutically effective amount of at least one of the compounds described herein is formulated for buccal or sublingual administration. Formulations suitable for buccal or sublingual administration include, by way of example only, tablets, lozenges, or gels. In yet other embodiments, the compounds described herein are formulated for parental injection, including formulations suitable for bolus injection or continuous infusion. In specific embodiments, the formulations for injection are provided in unit dosage form (e.g., ampoules) or in multi-dose containers. Preservatives may be added to the injection formulation. In still other embodiments, the pharmaceutical composition is formulated in a form suitable for parenteral injection as a sterile suspension, solution, or emulsion in an oily or aqueous vehicle. Parenteral injection formulations may include formulating agents such as suspending agents, stabilizing agents, and / or dispersing agents. In specific embodiments, pharmaceutical formulations for parenteral administration include an aqueous solution of the active compound in water-soluble form. In additional embodiments, a suspension of the active compound (e.g., a compound of structure (I)) is prepared as a suitable oily injection suspension. Lipophilic solvents or vehicles suitable for use in the pharmaceutical compositions described herein include, by way of example only, fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes.In certain specific embodiments, aqueous suspension injections contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran.Suspensions may also contain suitable stabilizers or agents that increase the solubility of compounds, allowing the preparation of highly concentrated solutions.Alternatively, in other embodiments, the active ingredient is in powder form for constitution with a suitable vehicle, such as sterile pyrogen-free water, before use.

[0123] In yet another embodiment, the compound of structure (I) is administered topically.The compounds described herein are formulated into various topically administrable compositions, such as solutions, suspensions, lotions, gels, pastes, medicated sticks, balms, creams or ointments.Such pharmaceutical compositions may include solubilizers, stabilizers, tonicity enhancers, buffers and preservatives.

[0124] In yet other embodiments, the compounds of structure (I) are formulated for transdermal administration. In specific embodiments, the transdermal formulations can be lipophilic emulsions or buffered aqueous solutions dissolved and / or dispersed in polymers or adhesives using transdermal delivery devices and transdermal delivery patches. In various embodiments, such patches are constructed for continuous, pulsatile, or on-demand delivery of the drug. In additional embodiments, transdermal delivery of the compounds of structure (I) is accomplished by iontophoretic patches and the like. In certain embodiments, the transdermal patch provides controlled delivery of the compounds of structure (I). In specific embodiments, the absorption rate is slowed by using rate-controlling membranes or by entrapment of the compound within a polymer matrix or gel. In alternative embodiments, absorption enhancers are used to enhance absorption. Absorption enhancers or carriers include pharma- ceutically acceptable absorbent solvents that aid in passage through the skin. For example, in one embodiment, the transdermal device takes the form of a bandage comprising a backing member, a reservoir containing the compound, optionally with a carrier, and optionally a rate-controlling barrier that delivers the compound to the host's skin at a controlled, predetermined rate over an extended period of time, and a means for securing the device to the skin.

[0125] In other embodiments, the compound of structure (I) is formulated for inhalation administration. Various forms suitable for inhalation administration include, but are not limited to, aerosols, mists or powders. Any pharmaceutical composition of the compound of structure (I) is conveniently delivered in the form of an aerosol spray presentation from a pressurized pack or nebulizer using a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas). In a specific embodiment, the dosage unit of the pressurized aerosol is determined by providing a valve that delivers a metered amount. In certain embodiments, capsules and cartridges of gelatin, etc., for use in inhalers or insufflators, are formulated containing a powder mix of the compound and a suitable powder base, such as lactose or starch, by way of example only. In yet other embodiments, the compounds of structure (I) are formulated in rectal compositions such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, jelly suppositories, or retention enemas containing conventional suppository bases such as cocoa butter or other glycerides and synthetic polymers such as polyvinylpyrrolidone, PEG, and the like. In suppository forms of the composition, a low melting point wax, such as, but not limited to, a mixture of fatty acid glycerides, optionally in combination with cocoa butter, is melted first.

[0126] In certain embodiments, pharmaceutical compositions are formulated in any conventional manner using one or more physiologically acceptable carriers, including excipients and auxiliaries that facilitate the processing of active compounds into medicament-usable preparations.The appropriate formulation depends on the route of administration selected.Any pharma-ceutically acceptable technique, carrier, and excipient may be used as appropriate.The pharmaceutical composition containing the compound of structure (I) is prepared in a conventional manner, such as by ordinary mixing, dissolving, granulating, sugar-making, sieving, emulsifying, encapsulating, encapsulating, or compressing processes, merely for illustrative purposes. The pharmaceutical composition comprises at least one pharma- ceutically acceptable carrier, diluent or excipient, and at least one compound of structure (I) as described herein as an active ingredient. The active ingredient is in the form of a free acid or free base, or in the form of a pharma- ceutically acceptable salt. In addition, the methods and pharmaceutical compositions described herein include the use of N-oxides, crystalline forms (also known as polymorphs), and active metabolites of these compounds having the same type of activity. All tautomers of the compounds described herein are included within the scope of the compounds presented herein. In addition, the compounds described herein include unsolvated and solvated forms with pharma- ceutically acceptable solvents such as water, ethanol, and the like. The solvated forms of the compounds presented herein are also considered to be disclosed herein. In addition, the pharmaceutical composition may include other medicinal or pharmaceutical agents, carriers, adjuvants such as preservatives, stabilizers, wetting agents or emulsifiers, dissolution enhancers, salts for adjusting osmotic pressure, buffers, and / or other therapeutically useful substances.

[0127] Methods for preparing compositions containing the compounds described herein include formulating the compounds with one or more inert pharma- ceutically acceptable excipients or carriers to form solids, semi-solids, or liquids. Solid compositions include, but are not limited to, powders, tablets, dispersible granules, capsules, cachets, and suppositories. Liquid compositions include solutions in which the compounds are dissolved, emulsions containing the compounds, or solutions containing liposomes, micelles, or nanoparticles containing the compounds disclosed herein. Semi-solid compositions include, but are not limited to, gels, suspensions, and creams. Forms of the pharmaceutical compositions described herein include liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to use, or as emulsions. These compositions may also contain small amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, and the like.

[0128] In some embodiments, the pharmaceutical composition comprising at least one compound of structure (I) is in the form of a liquid, illustratively, the drug is present as a solution, a suspension, or both.Typically, when the composition is administered as a solution or a suspension, a first portion of the drug is present as a solution, and a second portion of the drug is present as a suspension in the liquid matrix, in the form of particulates.In some embodiments, the liquid composition comprises a gel formulation.In other embodiments, the liquid composition is aqueous.

[0129] In certain embodiments, useful aqueous suspensions comprise one or more polymers as suspending agents.Useful polymers include water-soluble polymers such as cellulose-based polymers, for example hydroxypropylmethylcellulose, and water-insoluble polymers such as cross-linked carboxyl-containing polymers.Certain pharmaceutical compositions described herein comprise mucoadhesive polymers selected from, for example, carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methyl methacrylate), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate and dextran. Useful pharmaceutical compositions may also include solubilizing agents that aid in the dissolution of the compound of structure (I). The term "solubilizing agent" generally includes agents that result in the formation of a micellar or true solution of the drug. Certain acceptable non-ionic surfactants, such as polysorbate 80, are useful as solubilizing agents, as can ophthalmically acceptable glycols, polyglycols, such as polyethylene glycol 400, and glycol ethers.

[0130] Additionally, useful pharmaceutical compositions may contain one or more pH adjusting or buffering agents, including acids such as acetic acid, boric acid, citric acid, lactic acid, phosphoric acid, and hydrochloric acid; bases such as sodium hydroxide, sodium phosphate, borax, sodium citrate, sodium acetate, sodium lactate, and tris-hydroxymethylaminomethane; and buffers such as citrate / dextrose, sodium bicarbonate, and ammonium chloride. Such acids, bases, and buffers are included in the amounts required to maintain the pH of the composition in an acceptable range. In addition, useful compositions may contain one or more salts in an amount required to bring the osmolality of the composition into an acceptable range. Such salts include those having sodium, potassium or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate or bisulfite anions. Suitable salts include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite and ammonium sulfate.

[0131] Other useful pharmaceutical compositions may contain one or more preservatives to inhibit microbial activity. Suitable preservatives include mercury-containing substances such as merfen and thiomersal; stabilized chlorine dioxide; and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride. Still other useful compositions contain one or more surfactants to improve physical stability or for other purposes. Suitable nonionic surfactants include polyoxyethylene fatty acid glycerides and vegetable oils, such as polyoxyethylene (60) hydrogenated castor oil; and polyoxyethylene alkyl ethers and alkyl phenyl ethers, such as octoxynol 10, octoxynol 40. Still other useful compositions optionally contain one or more antioxidants to enhance chemical stability. Suitable antioxidants include, by way of example only, ascorbic acid and sodium metabisulfite.

[0132] In certain embodiments, aqueous suspension compositions are packaged in non-resealable single-dose containers. Alternatively, resealable multi-dose containers are used, in which case it is typical to include a preservative in the composition.

[0133] In alternative embodiments, other delivery systems for hydrophobic pharmaceutical compounds are used. Liposomes and emulsions are examples of delivery vehicles or carriers useful herein. In certain embodiments, organic solvents such as N-methylpyrrolidone are also used. In additional embodiments, the compounds described herein are delivered using sustained release systems, such as semipermeable matrices of solid hydrophobic polymers containing therapeutic agents. A variety of sustained release materials are useful herein. In some embodiments, sustained release capsules release compounds for several weeks, up to 100 days. Depending on the chemical nature and biological stability of the therapeutic reagent, additional strategies for protein stabilization are used. In certain embodiments, the formulations described herein include one or more antioxidants, metal chelators, thiol-containing compounds, and / or other general stabilizers. Examples of such stabilizers include, but are not limited to, (a) about 0.5% to about 2% glycerol by weight / volume, (b) about 0.1% to about 1% methionine by weight / volume, (c) about 0.1% to about 2% monothioglycerol by weight / volume, (d) about 1 mM to about 10 mM EDTA, (e) about 0.01% to about 2% ascorbic acid by weight / volume, (f) 0.003% to about 0.02% polysorbate 80 by weight / volume, (g) 0.001% to about 0.05% polysorbate 20 by weight / volume, (h) arginine, (i) heparin, (j) dextran sulfate, (k) cyclodextrin, (l) pentosan polysulfate and other heparinoids, (m) divalent cations such as magnesium and zinc; or (n) combinations thereof.

[0134] In some embodiments, the concentration of one or more compounds used in the pharmaceutical composition may be 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.0 ... lower than .09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001%.

[0135] In some embodiments, the concentration of one or more compounds is 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19.75%, 19.50%, 19.25% 19%, 18.75%, 18.50%, 18.25% 18%, 17.75%, 17.50%, 17.25% 17%, 16.75%, 16.50%, 16.25% 16%, 15.75%, 15.50%, 15.25% 15%, 14.75%, 14.50%, 14.25% 14%, 13.75%, 13.50%, 13.25% by mass / mass, mass / volume, or volume / volume. 13%, 12.75%, 12.50%, 12.25% 12%, 11.75%, 11.50%, 11.25% 11%, 10.75%, 10.50%, 10.25% 10%, 9.75%, 9.50%, 9.25% 9%, 8.75%, 8.50%, 8.25% 8%, 7.75%, 7.50%, 7.25% 7%, 6.75%, 6.50%, 6.25% 6%, 5.75%, 5.50%, 5.25% 5%, 4.75%, 4.50%, 4.25%, 4%, 3.75%, 3.50%, 3.25%, 3%, 2.75%, 2.50%, 2.25%, 2%, 1.75%, 1.50%, 125% , 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or higher than 0.0001%.

[0136] In some embodiments, the concentration of the one or more compounds is from about 0.0001% to about 50%, from about 0.001% to about 40%, from about 0.01% to about 30%, from about 0.02% to about 29%, from about 0.03% to about 28%, from about 0.04% to about 27%, from about 0.05% to about 26%, from about 0.06% to about 25%, by mass / mass, mass / volume, or volume / volume. , about 0.07% to about 24%, about 0.08% to about 23%, about 0.09% to about 22%, about 0.1% to about 21%, about 0.2% to about 20%, about 0.3% to about 19%, about 0.4% to about 18%, about 0.5% to about 17%, about 0.6% to about 16%, about 0.7% to about 15%, about 0.8% to about 14%, about 0.9% to about 12%, and about 1% to about 10%. In some embodiments, the concentration of the one or more compounds ranges from about 0.001% to about 10%, from about 0.01% to about 5%, from about 0.02% to about 4.5%, from about 0.03% to about 4%, from about 0.04% to about 3.5%, from about 0.05% to about 3%, from about 0.06% to about 2.5%, from about 0.07% to about 2%, from about 0.08% to about 1.5%, from about 0.09% to about 1%, or from about 0.1% to about 0.9%, by mass / mass, mass / volume, or volume / volume.

[0137] In some embodiments, the amount of one or more compounds is 10 g, 9.5 g, 9.0 g, 8.5 g, 8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g, 3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g, 0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g, 0.4 g, 0.35 g, 0.3 g, 0.25 g, 0.2 g , 0.15g, 0.1g, 0.09g, 0.08g, 0.07g, 0.06g, 0.05g, 0.04g, 0.03g, 0.02g, 0.01g, 0.009g, 0.008g, 0.007g, 0.006g, 0.005g, 0.004g, 0.003g, 0.002g, 0.001g, 0.0009g, 0.0008g, 0.0007g, 0.0006g, 0.0005g, 0.0004g, 0.0003g, 0.0002g, or 0.0001g.

[0138] In some embodiments, the amount of one or more compounds is 0.0001g, 0.0002g, 0.0003g, 0.0004g, 0.0005g, 0.0006g, 0.0007g, 0.0008g, 0.0009g, 0.001g, 0.0015g, 0.002g, 0.0025g, 0.0 03g, 0.0035g, 0.004g, 0.0045g, 0.005g, 0.0055g, 0.006g, 0.0065g, 0.007g, 0. 0075g, 0.008g, 0.0085g, 0.009g, 0.0095g, 0.01g, 0.015g, 0.02g, 0.025g, 0.03g , 0.035g, 0.04g, 0.045g, 0.05g, 0.055g, 0.06g, 0.065g, 0.07g, 0.075g, 0.08g, 0.085g, 0.09g, 0.095g, 0.1g, 0.15g, 0.2g, 0.25g, 0.3g, 0.35g, 0.4g, 0.45g, 0.5 g, 0.55g, 0.6g, 0.65g, 0.7g, 0.75g, 0.8g, 0.85g, 0.9g, 0.95g, 1g, 1.5g, 2g, 2.5, 3g, 3.5, 4g, 4.5g, 5g, 5.5g, 6g, 6.5g, 7g, 7.5g, 8g, 8.5g, 9g, 9.5g, or more than 10g. In some embodiments, the amount of one or more compounds ranges from 0.0001-10 g, 0.0005-9 g, 0.001-8 g, 0.005-7 g, 0.01-6 g, 0.05-5 g, 0.1-4 g, 0.5-4 g, or 1-3 g.

[0139] Treatment In yet another embodiment, the compound is useful in various methods of treating disease or condition.Thus, one embodiment provides a method of treating disease, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of the embodiments disclosed herein or a composition according to any one of the embodiments disclosed herein, wherein each M is independently a biologically active moiety effective for treating disease.In some embodiments, the disease is cancer, and each M is independently an anti-cancer drug.In some embodiments, at least one occurrence of M has the following structure:

[0140] [ka] In some embodiments, each occurrence of M has the structure:

[0141] [ka] For example, in some embodiments, the disclosure provides methods of treating solid tumors, multiple myeloma, glioma, renal clear cell carcinoma, prostate cancer, ovarian cancer, non-small cell lung cancer, GI malignancies, acute lymphoblastic leukemia, acute myeloid leukemia, renal cell carcinoma, colorectal cancer, epithelial cancer, pancreatic and gastric cancer, renal cell carcinoma, non-Hodgkin's lymphoma, metastatic renal cell carcinoma, malignant mesothelioma, pancreatic, ovarian and / or lung adenocarcinoma, B-cell malignancies, breast cancer, melanoma, recurrent multiple myeloma, small cell lung cancer, CD22-positive B-cell malignancies, Hodgkin's lymphoma / anaplastic large cell lymphoma, or HER2-positive breast cancer.

[0142] Certain embodiments also relate to a method of treating a hyperproliferative disorder in a mammal (e.g., a human), comprising administering to the mammal a therapeutically effective amount of a compound, or a pharma- ceutically acceptable salt, ester, prodrug, solvate, hydrate, or derivative thereof.In some embodiments, the method is directed to treating acute myeloid leukemia, cancer in adolescents, adrenocortical carcinoma in childhood, AIDS-related cancers (e.g., lymphoma and Kaposi's sarcoma), anal cancer, appendix cancer, astrocytoma, atypical teratoid, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain stem glioma, brain tumor, breast cancer, bronchial tumor, Burkitt's lymphoma, carcinoid tumor, atypical teratoid, embryonal tumor, germ cell tumor, primary lymphoma, cervical cancer, childhood cancer, chordoma, cardiac tumor, chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), chronic myeloproliferative (MM)-associated cancer (CLL), chronic myelogenous leukemia (CML), chronic myeloproliferative (MM)-associated cancer (CML), chronic myeloproliferative (MM)-associated cancer (CML), chronic myeloproliferative (MM)-associated cancer (CML), chronic myelogenous leukemia ... yleoproliferative disorders, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic ductal carcinoma in situ (DCIS), germinoma, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, esthesioneuroblastoma, Ewing's sarcoma, extracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, osteofibrous histiocytoma, gallbladder cancer, gastric (stomach) cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic tumor, hairy cell leukemia, head and neck cancer, heart cancer, liver cancer, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumor, pancreatic Neuroendocrine tumors, Renal cancer, Laryngeal cancer, Lip and oral cavity cancer, Liver cancer, Lobular carcinoma in situ (LCIS), Lung cancer, Lymphoma, Metastatic squamous neck cancer of unknown primary site, Midline carcinoma, Oral cancer, Multiple endocrine neoplasia syndrome, Multiple myeloma / plasma cell neoplasm, Mycosis fungoides, Myelodysplastic syndrome, Myelodysplastic / myeloproliferative neoplasm, Multiple myeloma, Merkel cell carcinoma, Malignant mesothelioma, Malignant fibrous histiocytoma of bone and osteosarcoma, Nasal cavity and paranasal sinus cancer, Nasopharyngeal cancer, Neuroblastoma, Non-Hodgkin's lymphoma, Non-small cell lung cancer (NSCLC), Oral cancer, Lip and oral cavity cancer, Oropharyngeal cancer, Ovarian cancer for the treatment of cancers such as pancreatic cancer, papillomatosis, paraganglioma, paranasal sinus and nasal cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pleuropulmonary blastoma, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, transitional cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer, gastric (stomach) cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, trophoblastic tumor, rare cancers of childhood, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer, or virus-induced cancer.In some embodiments, the methods relate to the treatment of a non-cancerous hyperproliferative disorder, such as benign hyperplasia of the skin (eg, psoriasis), benign hyperplasia of restenosis, or benign hyperplasia of the prostate (eg, benign prostatic hyperplasia (BPH)).

[0143] Certain embodiments provide a method for treating lung cancer, comprising administering to a subject in need thereof an effective amount of any of the above compounds (or pharmaceutical compositions containing them). In some embodiments, the lung cancer is non-small cell lung cancer (NSCLC), such as adenocarcinoma, squamous cell lung cancer or large cell lung cancer. In other embodiments, the lung cancer is small cell lung cancer. Other lung cancers treatable with the disclosed compounds include, but are not limited to, ductal tumors, carcinoid tumors and undifferentiated carcinomas.

[0144] Thus, in some embodiments of the aforementioned method, R 2 is a linker that includes a covalent bond to a targeting moiety, such as an antibody or a cell surface receptor antagonist, for example, an epidermal growth factor receptor (EGFR) inhibitor, a hepatocyte growth factor receptor (HGFR) inhibitor, an insulin-like growth factor receptor (IGFR) inhibitor, a phorate, or a MET inhibitor. In many more embodiments, the method further comprises the step of inducing apoptosis. In some embodiments, the method of treating a disease comprises: (a) a compound of structure (I), e.g., 2 or R 3 is a linker that contains a covalent bond to the analyte molecule, R 2 or R 3 The other is H, OH, alkyl, alkoxy, alkyl ether or -OP(=R a )(R b )R c and (b) detecting the compound by its visual properties; Further includes: In some embodiments, the analyte molecule is a nucleic acid, an amino acid, or a polymer thereof (e.g., a polynucleotide or a polypeptide). In many embodiments, the analyte molecule is an enzyme, a receptor, a receptor ligand, an antibody, a glycoprotein, an aptamer, or a prion.

[0145] In certain embodiments, a method is provided that further comprises the step of combining a compound of structure (I) with an analyte molecule. Thus, embodiments of the compounds are useful in a number of ways, including, but not limited to, drug delivery; quantification of apoptosis; qualification of therapeutic drug delivery; quantification of apoptosis; and diagnosis and treatment of diseases such as hematological cancers. In addition to the methods described above, embodiments of compounds of structure (I) are useful in a variety of fields and methods, including, but not limited to, cancer treatment and imaging by including targeting moieties in the compounds of structure (I), such as antibodies or sugars or other moieties that preferentially bind cancer cells; and / or drug delivery. In some embodiments, the method of treatment includes treating a tumor having tumor cells that contain tumor cell receptors. In some embodiments, the tumor cells have between 1,000 and 100,000, between 1,000 and 50,000, between 1,000 and 25,000, between 1,000 and 10,000 receptors per cell. For example, in some embodiments, the tumor cells have less than about 1,000, about 10,000, or 100,000 receptors per cell.

[0146] The embodiments of the present disclosure are not limited to the treatment of cancer.In fact, the present disclosure is not limited in terms of the types of diseases, symptoms, conditions, indications, and treatments that the compounds and methods of the present disclosure can be adapted to.That is, the present disclosure provides compounds, compositions, and methods for treating or preventing a wide variety of diseases.For example, the compounds and compositions disclosed herein can be modified by selecting the appropriate biological moiety or combination of biological moieties to treat a particular disease as desired.Based on the present disclosure, it will be readily apparent to those skilled in the art how to modify the compounds and compositions of the present disclosure to treat, prevent, or target a disease, symptom, or clinical indication.

[0147] Thus, methods of the disclosure include methods of administering a compound of the disclosure to treat a disease, condition, or a symptom of a disease or condition, methods of preventing a disease or a symptom of a disease or condition, methods of prophylactically treating a disease, condition, or a symptom of a disease or condition, methods of identifying at-risk subjects and treating a disease, condition, or a symptom of a disease or condition, methods of slowing or halting the progression of a disease, condition, or a symptom of a disease or condition, methods of increasing the survival of a subject having a disease, condition, or a symptom of a disease or condition, methods of ameliorating a symptom of a disease, condition, or a symptom of a disease or condition, and the like. Additionally, disorders, conditions, symptoms, illnesses, side effects, diseases, syndromes, biological occurrences, biological abnormalities, medical conditions, ailments, pathological conditions, pathologies, and the like are intended to be encompassed within the present disclosure, again without limitation. Examples include, but are not limited to, cancer, inflammation, pain, pain control, inflammatory diseases, infectious diseases, viral infections, genetic disorders, bacterial infections, fungal infections, skin conditions, endocrine conditions, eye disorders, intestinal diseases, neurological disorders, liver disorders, pulmonary infections, cardiac conditions and disorders, psychiatric disorders (e.g., eating disorders, mood disorders, personality disorders), norovirus infections, blood-borne pathogens, protozoal infections, viral hepatitis, HIV / AIDS, diabetes, sclerosis, colonic Crohn's disease, lupus, arthritis, allergies and asthma, celiac disease, polychondritis, scleroderma, liver disease, cardiac disease, acquired diseases, acute diseases, chronic conditions or diseases, congenital diseases or disorders, genetic diseases or disorders, iatrogenic diseases, idiopathic diseases, primary diseases, secondary diseases, terminal diseases, or the like. The foregoing may be acute, chronic, clinical, relapsing, progressive, refractory, subclinical, localized, diffuse, systemic, and the like. Any of the above examples include airborne, foodborne infections, or occurring as a result of lifestyle events.

[0148] Preparation method Any of the embodiments of the compound of structure (I) above, as well as the variable R in the compound of structure (I) above, 1 , R 2 , R 3 , L, L 1 , L 2 , L 3 It is understood that any specific choice described herein for R, M, and / or n can be independently combined with other embodiments and / or variables of the compounds of structure (I) to form embodiments of the invention not specifically described above. Additionally, any specific variable R in a particular embodiment and / or claim can be independently combined with other embodiments and / or variables of the compounds of structure (I) to form embodiments of the invention not specifically described above. 1 , R 2 , R 3 , L, L 1 , L 2 , L 3, M and / or n are listed, it is understood that each individual selection can be deleted from a particular embodiment and / or claim, and the remainder of the list of selections is considered to be within the scope of the invention. It is understood that combinations of substituents and / or variables of the depicted formulae herein are permissible only if such contributions result in stable compounds.

[0149] It will also be appreciated by those skilled in the art that in the methods described herein, it may be necessary to protect the functional groups of intermediate compounds with suitable protecting groups. Such functional groups include hydroxy, amino, mercapto, and carboxylic acid. Suitable protecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl (e.g., t-butyldimethylsilyl, t-butyldiphenylsilyl, or trimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitable protecting groups for amino, amidino, and guanidino include t-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable protecting groups for mercapto include -C(O)-R" (wherein R" is alkyl, aryl, or arylalkyl), p-methoxybenzyl, trityl, and the like. Suitable protecting groups for carboxylic acid include alkyl, aryl, or arylalkyl esters. Protecting groups can be added or removed according to standard techniques known to those skilled in the art and described herein. The use of protecting groups is described in detail in Green, TW and PGM Wutz, Protective Groups in Organic Synthesis (1999), 3rd Edition, Wiley. As one skilled in the art will recognize, the protecting group can also be a polymeric resin, such as a Wang resin, a Rink resin, or a 2-chlorotrityl-chloride resin.

[0150] Additionally, all compounds of the invention that exist in free base or acid form can be converted to their salts by treatment with an appropriate inorganic or organic base or acid by methods known to those skilled in the art. Salts of compounds of the invention can be converted to their free base or acid form by standard techniques.

[0151] The following reaction schemes show exemplary methods for making the compounds of the present invention. It is understood that these compounds can be made by those skilled in the art through similar methods or combinations of other methods known to those skilled in the art. It is understood that those skilled in the art can make other compounds of structure (I) that are not specifically shown below in the following similar manner by using appropriate starting components and adjusting the parameters of synthesis as necessary. In general, starting components can be obtained from suppliers such as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI, and Fluorochem USA, or can be synthesized according to suppliers known to those skilled in the art (see, for example, Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5th Edition (Wiley, December 2000)), or can be prepared as described in the present invention.

[0152] Reaction Scheme I [ka] Reaction Scheme I illustrates an exemplary method for preparing intermediates useful for the preparation of compounds of structure (I). In the scheme, R 1 , L 2 , L 3 and M is as defined above, R 2 and R 3is as defined above or a protected version thereof, and L is an optional linker. With reference to Reaction Scheme 1, compounds of structure a can be purchased or prepared by methods known to those skilled in the art. Reaction of a with MX under Suzuki coupling conditions known in the art (wherein x is a halogen, such as bromo) produces compounds of structure b. Compounds of structure b can be used to prepare compounds of structure (I) below.

[0153] Reaction Scheme II [ka] Reaction Scheme II illustrates an alternative method for preparing intermediates useful for the preparation of compounds of structure (I). 1 , L, L 2 , L 3 and M is as defined above, R 2 and R 3 With reference to Reaction Scheme II, where L is as defined above or a protected version thereof, a compound of structure c, which can be purchased or prepared by known techniques, is reacted with M-G' to give a compound of structure d, where G and G' represent functional groups with complementary reactivity (i.e., functional groups that react to form a covalent bond). G' can be pendant to M or can be part of the structural backbone of M. La can be prepared by the reaction shown above to form L a is the intermediate selected to be converted to L. G and G' can be several functional groups described herein, such as an alkyne and azide, respectively, an amine and activated ester, respectively, or an amine and isothiocyanate, respectively.

[0154] Compounds of structure (I) can be prepared by synthesis of the following structure (e) from one of structures b or d under well-known automated DNA synthesis conditions: [ka] wherein A is as defined herein and each L is independently an optional linker.

[0155] Methods for DNA synthesis are well known in the art. Briefly, the two alcohol groups of intermediates b or d above, e.g., R 2 and R 3 are functionalized with a dimethoxytrityl (DMT) group and a 2-cyanoethyl-N,N-diisopropylamino phosphoramidite group, respectively. The phosphoramidite group is typically coupled to an alcohol group in the presence of an activating agent such as tetrazole, followed by oxidation of the phosphorus atom with iodine. The dimethoxytrityl group can be removed with an acid (e.g., chloroacetic acid) to expose a free alcohol, which can react with the phosphoramidite group. The 2-cyanoethyl group can be removed by treatment with aqueous ammonia after oligomerization.

[0156] The preparation of phosphoramidites used in oligomerization methods is also well known in the art. For example, primary alcohols (e.g., R 3 ) can be protected as a DMT group by reaction with DMT-Cl. A secondary alcohol (e.g., R 2 ) is functionalized as a phosphoramidite by reaction with a suitable reagent such as 2-cyanoethyl N,N-diisopropylchlorophosphoramidite. Methods for the preparation of phosphoramidites and their oligomerization are well known in the art. Compounds of structure (I) are prepared by oligomerization of intermediates b or d and e according to the well-known phosphoramidite chemistry described above. The desired number of repeat units n is incorporated into the molecule by repeating the phosphoramidite coupling a desired number of times.

[0157] In exemplary embodiments, the G moiety can be selected from any of the Q moieties described herein, including the specific examples set forth in Table 1. In some embodiments, G, at each occurrence, independently comprises a moiety suitable for reactions, including copper-catalyzed reaction of azides with alkynes to form triazoles (Husgen 1,3-dipolar cycloaddition), reaction of dienes with parent dienes (Diels-Alder), strain-promoted alkyne-nitrone cycloaddition, reaction of strained alkenes with azides, tetrazines, or tetrazoles, [3+2] cycloaddition of alkenes with azides, reverse demand Diels-Alder of alkenes with tetrazines, photoreaction of alkenes with tetrazoles, and various substitution reactions, such as replacement of leaving groups by nucleophilic attack on an electrophilic atom. In some embodiments, G, at each occurrence, is independently a moiety that includes an aldehyde, oxime, hydrazone, alkyne, amine, azide, acyl azide, acyl halide, nitrile, nitrone, sulfhydryl, disulfide, sulfonyl halide, isothiocyanate, imidoester, activated ester, ketone, α,β-unsaturated carbonyl, alkene, maleimide, α-haloimide, epoxide, aziridine, tetrazine, tetrazole, phosphine, biotin, or thiirane functional group.

[0158] In other embodiments, G, at each occurrence, independently comprises an alkyne or azide group. In other embodiments, G, at each occurrence, independently comprises an amino, isothiocyanate, or activated ester group. In different embodiments, G, at each occurrence, independently comprises a reactive group that can form a functional group comprising an alkene, ester, amide, thioester, disulfide, carbocyclic, heterocyclic, or heteroaryl group when reacted with a complementary reactive group. For example, in some embodiments, heteroaryl is triazolyl. Methods of preparing compounds of structure (II) and methods of using automated DNA synthesis techniques to prepare compounds are described in U.S. Patent Application No. 15 / 481,378, WO 2015 / 027176, WO 2016 / 138461, and WO 2016 / 183185, all of which are incorporated by reference herein in their entireties. The following examples are offered by way of illustration and not by way of limitation. EXAMPLES

[0159] General method Mass spectral analysis was performed using MassLynx 4.1 acquisition software on a Waters / Micromass Quattro micro MS / MS system (MS-only mode). The mobile phase used for LC / MS for the dyes was 100 mM 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP), 8.6 mM triethylamine (TEA), pH 8. The phosphoramidite and precursor molecules were also analyzed using a Waters Acquity UHPLC system equipped with a 2.1 mm x 50 mm Acquity BEH-C18 column held at 45°C, using an acetonitrile / water mobile phase gradient. Molecular weights of monomeric intermediates were obtained using tropylium cation injection-enhanced ionization on a Waters / Micromass Quattro micro MS / MS system (MS-only mode). Excitation and emission profile experiments were recorded on a Cary Eclipse spectrophotometer. All reactions were carried out in oven-dried glassware under a nitrogen atmosphere unless otherwise noted. Commercial DNA synthesis reagents were purchased from Glen Research (Sterling, VA). Anhydrous pyridine, toluene, dichloromethane, diisopropylethylamine, triethylamine, acetic acid, pyridine, and THF were purchased from Aldrich. All other chemicals were purchased from Aldrich or TCI and used as received without further purification.

[0160] Example 1 Synthesis of Ibuprofen-NHS and Naproxen-NHS [ka]

[0161] Ibuprofen-NHS and naproxen-NHS were synthesized using standard coupling conditions: ibuprofen and naproxen were dissolved separately in dichloromethane, and N,N'-dicyclohexylcarbodiimide (DCC) and N-hydroxysuccinimide (NHS) were added to the mixture. The products were then purified as necessary and used in the next synthetic step. Ibuprofen-NHS is easily synthesized in one step on a multigram scale.

[0162] Example 2 Synthesis of ibuprofen phosphoramidite [ka]

[0163] Expandable derivatives of ibuprofen were synthesized according to the reactions shown above as representative derivatives that can be incorporated into the compound embodiments described herein. Ibuprofen-NHS was reacted with 2-1 (2-amino-1,3-propanediol) and then a trityl protecting group was added to obtain intermediate 2-3. The protected product 2-3 was then reacted with 2-4 to obtain the ibuprofen-phosphoramidite of the final product 2-5 (79%). The ibuprofen-phosphoramidite was then used in automated DNA synthesis to incorporate ibuprofen as a representative biologically active moiety into the compound embodiments described herein. Alternatively, ibuprofen-NHS was reacted with 4-(aminomethoxy)butane-1,2-diol and subsequently derivatized according to the reaction sequence shown above, with the final product resulting in an alternative ibuprofen phosphoramidite with an extended linker compared to the ibuprofen-phosphoramidite shown above.

[0164] Example 3 Synthesis of naproxen phosphoramidite [ka]

[0165] Naproxen-phosphoramidite (3-5) was synthesized using similar reaction conditions and reagents described in the synthesis of ibuprofen-phosphoramidite shown in Example 2. The desired product was obtained in good yield (79%) and was incorporated into compound embodiments of the present disclosure as a representative biologically active moiety. Alternatively, naproxen-NHS was reacted with 4-(aminomethoxy)butane-1,2-diol and subsequently derivatized according to the reaction sequence shown above, with the final product resulting in an alternative naproxen phosphoramidite with an extended linker compared to the naproxen phosphoramidite shown above.

[0166] Example 4 Exemplary compounds prepared using automated DNA synthesis methods Representative sequences were prepared using ibuprofen-phosphoramidites and / or naproxen-phosphoramidites, as described in Examples 2 and 3, respectively. Sequences were successfully prepared using automated DNA synthesis techniques known in the art. Among the 10 representative constructs prepared, some were homogeneous (i.e., containing only one biologically active moiety) and some were heterogeneous (i.e., containing a combination of the two).

[0167] Representative embodiments synthesized according to this method include other advantageous structural features. For example, fluorescein was easily and conveniently included as part of the construct. Four of the synthesized compounds contained a protected thiol moiety (C6SS) and were then used for conjugation to a targeting moiety (e.g., an antibody) as described in Example 5. Representative compounds are summarized in Table 6 below. [Table 6-1] [Table 6-2] [Table 6-3] [Table 6-4]

[0168] [ka]

[0169] Example 5 Activation and antibody conjugation of compound 7 [ka]

[0170] The thiol protecting group of compound 7 was removed using standard reduction conditions (i.e., TCEP) and the deprotected thiol was functionalized with 5-1 (bis-maleimidoethane; "BMOE") to give 5-2. Concurrently, the UCHT-1 antibody was treated with BMOE to reduce the disulfide bond. The reduced antibody was reacted with 5-2 (1.5 g) at a polymer to antibody molar ratio of 5:1. The reaction gave the final product 5-3 with a polymer to antibody ratio of 1:1 as detected by size exclusion chromatography (Figure 1).

[0171] Example 6 Synthesis of compound 11 [ka]

[0172] Polymer A, bearing three pendant amine functional groups, was coupled with ibuprofen-NHS as shown in the reaction sequence above (Note - for clarity, not all structural features of polymer A are shown). The reaction was carried out using a borate-buffered HO / DMSO mixture (1:3) and magnesium chloride. The reaction successfully appended an ibuprofen moiety to each of the three amine functional groups to give compound 11, which was identified by LC-MS (observed MW=4523.6). Two additional representative polymers were reacted under similar reaction conditions to give compounds 12 and 13, respectively. The structures of polymers A, B and C are shown below in Table 7.

[0173] [Table 7]

[0174] Example 8 Synthesis of compound 14 [ka] Polymer A was coupled to naproxen-NHS in the same manner as ibuprofen-NHS described in Example 7. Naproxen-NHS was less reactive than ibuprofen-NHS, possibly due to differences in solubility. However, the fully reacted polymer was successfully identified by LC-MS. Reactions with naproxen-NHS and reactive polymers B and C also showed the desired products.

[0175] Example 9 Synthesis of doxorubicin-NHS [ka] Doxorubicin was reacted with dihydrofuran-2,5-dione to give a carboxylic acid-containing intermediate, which was activated using TFA-NHS and pyridine to give doxorubicin-NHS.

[0176] Example 10 Synthesis of doxorubicin polymer (compound 17) [ka] Polymer D (structure shown below), which has five pendant amine functional groups, was coupled to doxorubicin-NHS. Because the solubility of the doxorubicin derivative is limited, experimentation was required to determine the reaction conditions. Different reaction conditions were tried, including increasing the organic solvent content, raising the reaction temperature, and adding sodium dodecyl sulfate (SDS) to accelerate the reaction.

[0177] The final reaction conditions were 100 mM borate buffer, pH 10, 500 mM magnesium chloride, 1% SDS, 75% DMSO. The reaction was kept at 80° C. overnight. After heating overnight, the reaction was concentrated to dryness and reconstituted in water. The reconstituted mixture was desalted using a microdesalting column and analyzed by PAGE and LC-MS.

[0178] PAGE analysis included visualization with a UV lamp at 365 nm wavelength (Figure 2). The desired product was detected at 5x higher and 10x lower than expected based on the calculated molecular weight of the desired product. The fluorescence detected is also consistent with the expected product. LC-MS analysis suggests a mixture of products.

[0179] [ka]

[0180] Example 11 Synthesis of doxorubicin-PEG-azide [ka]

[0181] Doxorubicin-PEG-azide was synthesized on a 50 mg scale following the reaction sequence shown above. Azide 9-1 was reacted with dihydrofuran-2,5-dione to give intermediate 9-2. The intermediate was reacted with perfluorophenol to give intermediate 9-3. Intermediate 9-3 was coupled to doxorubicin to give the desired product, doxorubicin-PEG-azide, in high yield (74%). The presence of the desired product was confirmed by LC-MS.

[0182] Example 12 Synthesis of doxorubicin polymer (compound 18)

[0183] [ka]

[0184] Polymer E (structure shown below), which has three pendant alkynyl functional groups, was coupled to doxorubicin-PEG-azide. Reaction conditions included CuSO4, tris(3-hydroxypropyltriazolylmethyl)amine (THPTA), and sodium ascorbate. The reaction was carried out in a phosphate-buffered aqueous solvent at pH 7.6 containing 60% DMS. The reaction was carried out at room temperature and the presence of the desired product was confirmed by LC-MS.

[0185] [ka]

[0186] The embodiments disclosed herein provide many advantages, including the ability to control the number of bioactive or fluorescent dye moieties that are coupled to the polymer and any subsequent targeting moieties.The composition of the polymer backbone can also be selected to obtain desired solubility properties, for example, by controlling the incorporation (e.g., number, frequency, spacing, etc.) of charged moieties.In addition to the properties provided by the backbone composition, side chains can be selected to provide a source for adjusting the solubility of the compounds disclosed herein.

[0187] The embodiments disclosed herein also provide compounds that can advantageously contain multiple therapeutic agents, for example, for complementary or synergistic therapeutic strategies.Furthermore, the embodiments of the present disclosure provide combinations of therapeutic agents, targeting moieties and dye moieties (e.g., fluorophores) that can be used for simultaneous targeting treatment and detection.The ease of coupling polymer-drug constructs to targeting agents such as antibodies, antibody fragments, proteins or other clinically interesting agents is useful for a wide variety of interesting applications (e.g., surface chemistry, assay development, etc.). Certain embodiment compounds also provide other desirable properties, including enhanced permeability and retention effects. In addition to providing the necessary solubility properties, the chemical features of the present compound embodiments can be tailored to modulate the ability of the compound to penetrate and be retained in diseased cells / tissues. These features allow for effective delivery of biologically active agents by increasing permeability, and increased efficacy by increasing retention.

[0188] (Example 13) Potency Assay of ADCs Made with Compound 19 and Herceptin The efficacy of a series of antibody-polymer drug conjugates (ADCs) was tested in a series of highly expressing Her2 positive cell lines and compared with the potency and selectivity of the antibody-polymer drug conjugate components.

[0189] Cytotoxicity Tumor cell cytotoxicity assays were performed using tumor cell lines including PC3 (CRL-1435), SKBR3 (HTB-30), SKOV3 (HTB-77), and HCC1954 (CRL-2338), all obtained from ATCC. Tumor cell lines were seeded at 1,000 cells per well in 96-well plates in RPMI medium (Life Technologies, Carlsbad CA) supplemented with 10% heat-inactivated FCS, pyruvate, non-essential amino acids, and gentamicin.

[0190] Compound 19 was used for the polymer drug moiety and Herceptin was used as the antibody moiety. Auristatin F was used as the drug moiety and Compound A (i.e., Compound III-32, where M is F") was also used in the assay.

[0191] For each assay, growth inhibition was assessed in duplicate samples by incubating the ADC with tumor cell lines for 72 hours followed by treatment with Alamar Blue (Life Technologies) for 1-4 hours. Fluorescence emission of reduced resazurin at 590 nm (excitation at 530-560 nm) was quantified using a molecular diagnostics plate reader. Each assay was performed in duplicate, data log transformed, and average values ​​were graphed on a semi-log scale. Prism software version 6 (GraphPad Software Inc, San Diego CA) was used to determine the half-maximal effective concentration (EC) of each treatment using a nonlinear least-squares method to fit a four-parameter logistic equation. 50 ) was calculated. EC 50 The means and standard deviations were generated, and the mean values ​​are shown in Figures 3 to 5. The results for HCC1954 and SKOV3 are shown in Figures 3 and 5, respectively. The results for HS578t ErbB2- are shown in Figure 4.

[0192] The results show that the ADC is more potent than the drug alone, which appears to be cell type dependent. Furthermore, the polymer-drug moiety is more potent than either the drug alone or the ADC. This may be due to concentration effects (e.g., 1 nM polymer drug = 5 nM drug), but is not explained by concentration alone. Without wishing to be bound by any particular theory, it is believed that auristatin F is added to the C-terminal position of the drug, which eliminates the carboxylate charge and increases potency. Furthermore, indeed the polymer alone appears to penetrate the cells.

[0193] All U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications, and non-patent publications cited herein, including U.S. Provisional Patent Application No. 62 / 568,706, filed October 5, 2017, are hereby incorporated by reference in their entirety to the extent not inconsistent herewith. From the foregoing, it will be appreciated that, although specific embodiments of the invention have been described herein for purposes of illustration, various modifications can be made without departing from the spirit and scope of the invention. Accordingly, the invention is not to be limited except as by the appended claims.

Claims

1. A compound having the structure (I): or a stereoisomer, pharma- ceutically acceptable salt, or tautomer thereof. 【Chemistry 1】 (I) (In the formula, M, at each occurrence, is independently a biologically active moiety or fragment thereof, a prodrug of a biologically active moiety or fragment thereof, a fluorescent dye, an imaging agent, or a radioisotope binding moiety, with the proviso that at least one occurrence of M is not a fluorescent dye; L is a physiologically cleavable linker; L 1 , L 2 and L 3 is independently at each occurrence an optional alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, or heteroatom linker; R 1 is independently at each occurrence H, alkyl or alkoxy; R 2 and R 3 are each independently H, OH, SH, alkyl, alkoxy, alkyl ether, heteroalkyl, -OP(=R a ) (R b ) R c , Q or a protected form thereof, or L′; R a is O or S, R b OH, SH, O - , S - , OR d or S.R. d and R c OH, SH, O - , S - , OR d , O.L.', S.R. d , alkyl, alkoxy, heteroalkyl, heteroalkoxy, alkyl ether, alkoxyalkyl ether, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkyl ether, or thiophosphoalkyl ether; R d is the counter ion, Q, in each occurrence, is independently a moiety that contains a reactive group or a protected form thereof that is capable of forming a covalent bond with a complementary reactive group Q′ of a targeting moiety; L′, in each occurrence, is independently a linker that includes a covalent bond to a targeting moiety, Q, a linker that includes a covalent bond to a targeting moiety, a linker that includes a covalent bond to a solid support, a linker that includes a covalent bond to a solid support residue, a linker that includes a covalent bond to a nucleoside, or a linker that includes a covalent bond to an additional compound of structure (I); n is an integer of 1 or more.

2. L 1 The compound of claim 1 , wherein is a heteroalkylene linker.

3. 3. The compound of claim 1 or 2 having the following structure (IA): 【Chemistry 2】 (IA) (In the formula, L 4 is independently at each occurrence an alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, or heteroalkynylene linker; R 4 is, in each occurrence, independently, OH, SH, O - , S - , OR d or S.R. d and R 5 is independently, at each occurrence, oxo, thioxo, or absent; m, in each occurrence, is independently an integer equal to or greater than 0.

4. L 4 The compound of claim 3 , wherein, in each occurrence, is independently a heteroalkylene linker.

5. L 4 The compound of claim 4 , wherein, in each occurrence, is independently an alkylene oxide linker.

6. L 4 4. The compound of claim 3, wherein: is ethylene oxide or polyethylene oxide, and the compound has the following structure (IB): 【Chemistry 3】 (IB) (wherein z is an integer from 1 to 100).

7. The compound according to claim 6, wherein z is an integer from 3 to 6.

8. L 2 and L 3 may, in each occurrence, independently: C 1 -C 6 Alkylene, C 2 -C 6 Alkenylene or C 2 -C 6 The compound according to any one of claims 1 to 7, which is an alkynylene.

9. The compound of any one of claims 1 to 8, having the following structure (IC): 【Chemistry 4】 (I C) (In the formula, x 1 , x 2 , x 3 and x 4 is independently, in each occurrence, an integer from 0 to 6; z is an integer from 1 to 100.

10. L 4 may, in each occurrence, independently: C 1 -C 6 Alkylene or C 2 -C 6 The compound according to any one of claims 1 to 3, which is an alkynylene.

11. 11. The compound of claim 10 having the following structure (ID): 【Chemistry 5】 (ID) (In the formula, x 1 , x 2 , x 3 and x 4 is independently, in each occurrence, an integer from 0 to 6; y is an integer from 1 to 6.

12. 12. The compound of claim 11, wherein y is 2.

13. x 1 , x 2 , x 3 or x 4 13. The compound according to any one of claims 9 to 12, wherein at least one occurrence of is 1.

14. x 1 , x 2 , x 3 and x 4 The compound according to any one of claims 9 to 12, wherein each occurrence of is 1.

15. R 4 may each occur independently: OH, O - OR d The compound according to any one of claims 3 to 14,

16. R 5 The compound according to any one of claims 3 to 15, wherein, at each occurrence, is oxo.

17. R 1 The compound according to any one of claims 1 to 16, wherein, in each occurrence, is H.

18. R 2 and R 3 are each independently OH or -OP(=R a ) (R b ) R c The compound according to any one of claims 1 to 17,

19. R 2 and R 3 One of the groups is OH or -OP (=R a ) (R b ) R c and R 2 and R 3 The compound according to any one of claims 1 to 18, wherein the other of is Q or a linker comprising a covalent bond to Q.

20. R 2 and R 3 One of the two is -OP(=R a ) (R b ) R c or L'.

21. R c The compound of claim 20, wherein is OL'.

22. 22. The compound of claim 20 or 21, wherein L' is a targeting moiety or a linker to a targeting moiety.

23. 23. The compound of claim 22, wherein L' is a linker to a targeting moiety, said linker comprising an alkylene oxide or a phosphodiester moiety, or a combination thereof.

24. 24. The compound of claim 23, wherein L' has the structure: 【Chemistry 6】 (In the formula, m″ and n″ are independently integers from 1 to 10; R e is H, an electron pair or a counterion, L" is a targeting moiety or a link to a targeting moiety.

25. The compound of any one of claims 1 to 24, wherein the targeting moiety is an antibody or a cell surface receptor antagonist.

26. 26. The compound of claim 25, wherein the antibody or cell surface receptor antagonist is an epidermal growth factor receptor (EGFR) inhibitor, a hepatocyte growth factor receptor (HGFR) inhibitor, an insulin-like growth factor receptor (IGFR) inhibitor, folate, or a MET inhibitor.

27. R 2 Or R 3 The compound of any one of claims 1 to 26, wherein: 【Chemistry 7】 【change】

28. R 2 Or R 3 The compound of any one of claims 1 to 26, wherein: 【Chemistry 8】

29. 29. The compound of any one of claims 1 to 28, wherein m, in each occurrence, is independently an integer from 1 to 10.

30. 29. The compound of any one of claims 1 to 28, wherein m in each occurrence is independently an integer from 1 to 5.

31. The compound according to any one of claims 1 to 30, wherein n is an integer from 1 to 100.

32. The compound according to any one of claims 1 to 30, wherein n is an integer from 1 to 10.

33. 33. The compound of any one of claims 1 to 32, wherein L, in each occurrence, is independently a linker that includes an amide bond, an ester bond, a disulfide bond, a double bond, a triple bond, an ether bond, a ketone, a diol, cyano, nitro, or a combination thereof.

34. 34. The compound of any one of claims 1 to 33, wherein M, at each occurrence, is independently an NSAID, a kinase inhibitor, an anthracycline, an EGFR inhibitor or an alkylating agent.

35. 35. The compound of any one of claims 1-34, wherein M, at each occurrence, is independently an anti-cancer drug and the targeting moiety is an antibody specific for a tumor cell antigen.

36. 36. The compound of claim 35, wherein the tumor cell antigen is EGFR, HER2, folate receptor, CD20 or CD33.

37. The compound of any one of claims 1 to 33, wherein at least one occurrence of M has one of the following structures: 【Chemistry 9】 【change】

38. 37. The compound of claim 36, wherein each occurrence of M has one of the following structures: 【Chemistry 10】 【change】

39. A compound selected from Tables 2, 3 or 4.

40. A composition comprising a compound according to any one of claims 1 to 39 and a pharma- ceutically acceptable carrier.

41. A composition comprising a plurality of conjugates, said conjugates comprising an antibody covalently bound to two or more biologically active moieties via a single linkage, said plurality of conjugates having at least 90% structural homogeneity.

42. 40. The composition of claim 39, wherein the single linkage is to a polymer backbone, the polymer backbone comprising two or more biologically active moieties covalently attached thereto.

43. 43. The composition of claim 41 or 42, wherein the plurality of conjugates have at least 95% structural homogeneity.

44. 42. The composition of claim 41 , wherein the plurality of conjugates has greater than 99% structural homogeneity.

45. Each conjugate is independently a compound according to claim 1, wherein R 2 and R 3 One of the two is -OP(=R a ) (R b 45. The composition of any one of claims 41 to 44, wherein: OL' or L', and L' is an antibody or a linker comprising a covalent bond to an antibody.

46. 11. A method of treating a disease, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1-39 or a composition of any one of claims 40-45, wherein each M is independently a biologically active moiety effective for treating the disease.

47. 45. The method of claim 44, wherein the disease is cancer and each M is independently an anti-cancer drug.

48. 48. The method of claim 46 or 47, wherein at least one occurrence of M has the structure: 【Chemistry 11】

49. 49. The method of any one of claims 46-48, wherein each occurrence of M has the structure: 【Chemistry 12】

50. 47. The method of claim 46, wherein the treatment comprises reducing pain or inflammation.

51. 51. The method of claim 46 or 50, wherein at least one occurrence of M has one of the following structures: 【Chemistry 13】

52. 52. The method of claim 46, 50, or 51, wherein each occurrence of M has one of the following structures: 【Chemistry 14】

53. 47. The method of claim 46, wherein the disease is an infectious disease.

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