Polymers with rigid spacing groups containing biologically active compounds - Patent Application 20070122997

Abstract

A compound useful as a biologically active compound is disclosed. The compound has the following structure (I): (I) (wherein A, R 1 , R 2 , R 3 , R 4 , R 5 , L, L 1 , L 2 , L 3 , L 4 , M, m, and n are as defined herein) or a stereoisomer, tautomer, or salt thereof. Related methods for the preparation and use of such compounds are also provided. (I)

Description

[Technical Field]

[0001] SUMMARY OF THE INVENTION Embodiments of the present invention are generally directed to biologically active polymeric compounds and methods for their preparation and use in various therapeutic methods. [Background technology]

[0002] 2. Description of Related Art Unlike chemotherapy, targeted drug conjugates are intended to target only diseased cells and not harm healthy cells. Typically, conjugates are composed of a targeting molecule linked to a biologically active payload or drug. By combining the inherent targeting ability and the therapeutic efficacy of biologically active drugs, 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 of particular interest for cancer treatment. ADCs combine the targeting characteristics of monoclonal antibodies with the cancer-killing potential of cytotoxic agents, resulting in a treatment with several advantages over other chemotherapeutic agents. However, the complexity of ADC construction, specifically the challenges associated with the chemical linker between the antibody and the drug, has significantly hindered 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. Currently, only Adcetris® and Kadcyla® are commercially available worldwide (Zevalin® is only approved in China). Pioneer Pfizer / Wyeth withdrew Mylotarg® in 2010 after safety issues were observed during controlled clinical trials. Therefore, there is a need in the art for potent targeted drug conjugates with large therapeutic indices. Ideally, such drug conjugates should achieve fine discrimination between healthy tissue and diseased tissue (e.g., tumor cells). The present invention addresses this need and achieves further related advantages. Summary of the Invention

[0003] Briefly, embodiments of the present invention are generally directed to compounds useful as targeted drug conjugates, which may include fluorescent and / or colored dyes that enable selective delivery to targets such as tumor cells, as well as reagents for their preparation. Methods for preparing such molecules and methods for using them to provide therapeutic treatment to patients in need thereof are also described. Embodiments of the presently disclosed compounds include one or more biologically active moieties covalently linked by a linker ("L"). Advantageously, embodiments of the present invention provide compounds that can be incorporated during polymer synthesis or attached post-synthetically. Furthermore, embodiments described herein allow for the incorporation of multiple biologically active moieties within the same compound, and the optional inclusion of a targeting moiety. In one embodiment, a compound having the following structure (I): [ka] (I) Alternatively, a stereoisomer, tautomer or salt thereof is provided (wherein A, R 1 , R 2 , R 3 , R 4 , R 5 , L, L 1 , L 2 , L 3 , L 4 , M, m, and n are as defined herein.) The compounds of structure (I) find utility in several applications, including use as therapeutic agents in a variety of treatment methods. In another embodiment, provided is a method of treating a disease 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 independently comprises a biologically active moiety effective to treat the disease. These and other aspects of the present invention will become evident upon reference to the following detailed description. DETAILED DESCRIPTION OF THE INVENTION

[0004] 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 their open and inclusive sense, i.e., "including but not limited to." References throughout this specification to "one embodiment" or "an embodiment" mean that the particular features, structures, or properties described in connection with these embodiments are included in at least one embodiment of the present invention. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification do not necessarily all refer to the same embodiment. Furthermore, the particular features, structures, or properties may be combined in any suitable manner in one or more embodiments. "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 an -SH group. "Thioxo" refers to the group ═S.

[0005] "Alkyl" means an alkyl group containing 1 to 12 carbon atoms (C1-C 12"C-C alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, having from 1 to 8 carbon atoms (C-C alkyl) or from 1 to 6 carbon atoms (C-C alkyl), attached to the rest of the molecule by a single bond, such as methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl, etc. Unless specifically stated otherwise in this specification, alkyl groups may be optionally substituted.

[0006] "Alkylene" or "alkylene chain" refers to a divalent straight or branched hydrocarbon chain, consisting solely of carbon and hydrogen, without unsaturation, having 1 to 12 carbon atoms, linking the rest 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 stated otherwise specifically in the specification, alkylene may be optionally substituted.

[0007] "Alkenylene" or "alkenylene chain" refers to a divalent straight or branched hydrocarbon chain, consisting solely of carbon and hydrogen, containing at least one carbon-carbon double bond and having 2 to 12 carbon atoms, e.g., ethenylene, propenylene, n-butenylene, etc., that links the rest of the molecule to a radical group. 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 stated otherwise specifically in the specification, alkenylene may be optionally substituted. "Alkynylene" or "alkynylene chain" refers to a divalent straight or branched hydrocarbon chain, consisting solely of carbon and hydrogen, containing at least one carbon-carbon triple bond and having 2 to 12 carbon atoms, e.g., ethenylene, propenylene, n-butenylene, etc., that links the rest of the molecule to a radical group. 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 stated otherwise specifically in the specification, alkynylene may be optionally substituted.

[0008] "Alkyl ether" refers to any alkyl group, as defined above, in which at least one carbon-carbon bond is 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 specifically stated herein, alkyl ether groups may be substituted. For example, in some embodiments, alkyl ethers are substituted with an alcohol or -OP(=R a )(R b )R c is substituted by R a , R b and R c are each as defined for compounds of structure (I). "Alkoxy" means a group of the formula -OR a R refers to the group a is an alkyl group, as defined above, containing 1 to 12 carbon atoms. Unless stated otherwise specifically in the specification, an alkoxy group may be optionally substituted.

[0009] "Alkoxyalkyl ether" means an alkoxyalkyl ether of the formula -OR a R b R refers to the groupa 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 stated herein, an alkoxyalkyl ether group may be optionally substituted, for example, by an alcohol or -OP(=Ra)(Rb)Rc, where Ra, Rb, and Rc are each as defined for 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 present within the alkyl group (i.e., a heteroalkyl contains at least one carbon-[heteroatom] x -carbon bond, and x is 1, 2, or 3). In other embodiments, the heteroatom is at the end of the alkyl group, thus serving to attach the alkyl group to the remainder of the molecule (e.g., M1-HA), where M1 is part of the molecule, H is the heteroatom, and A is the alkyl group. Unless otherwise specifically stated herein, heteroalkyl groups may be substituted. Exemplary heteroalkyl groups include ethylene oxide (e.g., polyethylene oxide), which may include a phosphorus-oxygen bond, such as a phosphodiester bond. "Heteroalkoxy" means a group of the formula -OR a R refers to the group a is a heteroalkyl group, as defined above, containing 1 to 12 carbon atoms. Unless stated otherwise specifically in the specification, a heteroalkoxy group may be optionally substituted.

[0010] "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). In other embodiments, the heteroatom is at the end of the alkylene and serves to attach the alkylene to the rest 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 specifically stated herein, heteroalkylene groups may be substituted. Exemplary heteroalkylene linking groups are illustrated below: [ka] "C linker" Various embodiments of heteroalkylene linkers include multimers of the C-linkers described above. "Heteroalkenylene" is a heteroalkylene, as defined above, containing at least one carbon-carbon double bond. Unless stated otherwise specifically in the specification, a heteroalkenylene group may be optionally substituted. A "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.

[0011] "Heteroatom" in reference 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- and multimers, and combinations thereof. "Phosphate" means -OP(=O)(R a )R b Group(R aOh, O - OR c and R b Oh, O - , OR c ), a thiophosphate group or a further phosphate group (R c is the counter ion (e.g., Na + It refers to something like that.

[0012] "Phosphoalkyl" means -OP(=O)(R a )R b R refers to the group a Oh, O - OR c and R b is -Oalkyl and R c is the counter ion (e.g., Na + Unless otherwise specifically stated herein, a phosphoalkyl group may be optionally 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 and R a , R b and R c are each as defined for compounds of structure (I). "Phosphoalkyl ether" means -OP(=O)(R a )R b R refers to the group a Oh, O - OR c and R b is an -O alkyl ether, and R c is the counter ion (e.g., Na +Unless otherwise specifically stated herein, a phosphoalkyl ether group may be substituted. For example, in certain embodiments, the -O alkyl ether moiety in a phosphoalkyl ether group is hydroxyl, amino, sulfhydryl, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkyl ether, thiophosphoalkyl ether, or -OP(=R a )(R b )R c and R a , R b and R c are each as defined for compounds of structure (I). "Thiophosphate" means -OP(=R a )(R b )R c Group(R a is O or S, and R b Oh, O - , S - , OR d or SR d and R c OH, SH, O - , S - , OR d , S.R. d , a phosphate group or a further thiophosphate group, and R d is the counter ion (e.g., Na + etc.), where i) R a is S, and ii) R b is S - or SR d and iii) R c , SH, S - or SR d or iv) a combination of i), ii) and / or iii).

[0013] "Thiophosphoalkyl" means -OP(=R a )(R b )R c R refers to the group a is O or S, and Rb Oh, O - , S - , OR d or SR d and R c is -Oalkyl and R d is the counter ion (e.g., Na + etc.), where i) R a is S, and ii) R b is S - or SR d or iii) R a is S and R b is S - or SR d Unless otherwise specifically stated herein, a thiophosphoalkyl group may be optionally substituted. For example, in certain embodiments, the -Oalkyl portion in a thiophosphoalkyl group may be hydroxyl, amino, sulfhydryl, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkylether, thiophosphoalkylether, or -OP(=R a )(R b )R c and R a , R b and R c are each as defined for compounds of structure (I).

[0014] "Thiophosphoalkyl ether" means -OP(=R a )(R b )R c R refers to the group a is O or S, and R b Oh, O - , S - , OR d or SR d and R c is an -O alkyl ether, and R d is the counter ion (e.g., Na + etc.), where i) R a is S, and ii) R b is S -or SR d or iii) R a is S and R b is S - or SR 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, phosphoalkylether, thiophosphoalkylether, or -OP(=R a )(R b )R c and R a , R b and R c are each as defined for compounds of structure (I).

[0015] "Carbocyclic" refers to a stable 3- to 18-membered aromatic or non-aromatic ring containing from 3 to 18 carbon atoms. Unless otherwise specifically stated herein, a carbocyclic ring can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems and may be partially saturated or fully saturated. Non-aromatic carbocyclyl radicals include cycloalkyl, while aromatic carbocyclyl radicals include aryl. Unless otherwise specifically stated herein, a carbocyclic group may be optionally substituted. "Cycloalkyl" refers to a stable non-aromatic monocyclic or polycarbocyclic ring, which may include saturated or unsaturated fused or bridged ring systems having 3 to 15 carbon atoms, preferably 3 to 10 carbon atoms, attached to the remainder of the molecule by a single bond. Monocyclic cyclocalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyls include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless specifically stated otherwise in the specification, cycloalkyl groups may be optionally substituted.

[0016] "Aryl" refers to a ring system containing at least one carbocyclic aromatic ring. In some embodiments, an 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, aryls 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 specifically stated herein, an aryl group may be optionally substituted.

[0017] "Heterocyclic" refers to a stable 3- to 18-membered aromatic or non-aromatic ring containing 1 to 12 carbon atoms and 1 to 6 heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur. Unless otherwise specifically stated herein, a heterocyclic ring may be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems, in which the nitrogen, carbon, or sulfur atoms in the heterocyclic ring may be oxidized, the nitrogen atoms may be quaternized, and the heterocyclic ring may be partially saturated 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, decahydroisoquinolyl, 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.

[0018] "Heteroaryl" refers to a 5- to 14-membered ring system containing 1 to 13 carbon atoms, 1 to 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 can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which can include fused or bridged ring systems, and the nitrogen, carbon, or sulfur atoms in the heteroaryl radical can be optionally oxidized, and the nitrogen atom can be optionally quaternized.Examples include, but are not limited to, 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, benz[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzo[b] ... Benzofuranonyl, benzothienyl (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, isoquinolyl, indolizinyl yl, isoxazolyl, 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

[0033] Unless stated otherwise specifically in the specification, heteroaryl groups may be optionally substituted.

[0019] "Fused" refers to a ring system containing at least two rings, where the two rings share at least one common ring atom, e.g., 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.

[0020] As used herein, the term "substituted" means 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) wherein at least one hydrogen atom (e.g., one, two, three, or all hydrogen atoms) has been replaced with, but is not limited to, F, Cl, Br, and I; oxygen atoms in groups such as hydroxyl groups, alkoxy groups, and ester groups; sulfur atoms in groups such as thiol groups, thioalkyl groups, sulfone groups, sulfonyl groups, and sulfoxide groups; nitrogen atoms in groups such as amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, imides, and enamines; silicon atoms in groups such as trialkylsilyl groups, dialkylarylsilyl groups, alkyldiarylsilyl groups, and triarylsilyl groups; and bonds to non-hydrogen atoms such as other heteroatoms in various other groups. "Substituted" also refers to any of the above groups in which one or more hydrogen atoms are replaced by a higher bond (e.g., a double bond or a triple bond) to a heteroatom, such as oxygen in oxo, carbonyl, carboxyl, and ester groups, and nitrogen in groups such as imines, oximes, hydrazones, and nitriles. For example, "substituted" refers to any of the above groups in which one or more hydrogen atoms are replaced by a higher bond (e.g., a double bond or a triple bond) to a heteroatom, such as oxygen in oxo, carbonyl, carboxyl, and ester groups, and nitrogen in imines, oximes, hydrazones, and nitriles. g R h , -NR g C(=O)Rh , -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" also includes any of the above groups replaced by -C(=O)R g , -C(=O)OR g , -C(=O)NR g R h , -CH2SO2R g , -CH2SO2NR g R h In the above, R g and R hare 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, an optional substituent is -OP(=R a )(R b )R c and R a , R b and R c are each as defined for compounds of structure (I). Additionally, any of the above substituents may also be substituted with one or more of the above substituents.

[0021] "Conjugation" or "bioconjugation" refers to a chemical strategy that forms 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, is conjugated, or a grammatical equivalent. "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 skilled in the art. "Colored" refers to molecules that absorb light within the color spectrum (ie, red, yellow, blue, etc.). "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). The linker can be attached to the molecule through covalent or other means, such as ionic or hydrogen-bonding interactions.

[0022] The term "biomolecule" refers to any of a variety of biological substances, 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 to them) are prepared by contacting a biomolecule with the above-described compounds having a reactive group that allows the binding of the biomolecule to the compound via any available atom or functional group, such as an amino, hydroxy, carboxyl, or sulfhydryl group, on the biomolecule, as further described herein.

[0023] 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 reaction. 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, biotin, thiirane, and the like. The terms "visible" and "visually detectable" are used herein to refer to substances observable by visual inspection without prior illumination or chemical or enzymatic activation. Such visually detectable substances absorb and emit light in the spectral region ranging from about 300 to about 900 nm. Preferably, such substances are intensely colored, preferably at least about 40,000 nm. -1 cm -1 , more preferably at least about 50,000 M -1 cm -1 , and even more preferably at least about 60,000 M -1 cm -1 , and even more preferably at least about 70,000 M -1 cm -1 , and most preferably at least about 80,000 M -1 cm -1 The compound has a molar extinction coefficient of 0.05. Embodiments of the compounds of the present disclosure may be detected by observation with the naked eye or with the aid of optically-based detection devices, including, but not limited to, absorption spectrophotometers, transmitted light microscopes, digital cameras, and scanners. Visually detectable substances are not limited to those that emit and / or absorb light in the visible spectrum. Also included within the scope of "visually detectable" substances are substances that emit and / or absorb light in the ultraviolet (UV) region (about 10 nm to about 400 nm), the infrared (IR) region (about 700 nm to about 1 mm), and substances that emit and / or absorb in other regions of the electromagnetic spectrum.

[0024] For purposes of the present invention, the term "photostable visible dye" refers to a chemical moiety that is visually detectable and does not significantly change or decompose upon exposure to light, as defined hereinabove. Preferably, the photostable visible dye does not exhibit significant discoloration or decomposition after exposure to light for at least 1 hour. More preferably, the visible dye is stable after exposure to light for at least 12 hours, even more preferably at least 24 hours, even more preferably at least 1 week, and most preferably at least 1 month. Non-limiting examples of photostable visible dyes suitable for use in the compounds and methods of the present invention include azo dyes, thioindigo dyes, quinacridone pigments, dioxazines, phthalocyanines, perinones, diketopyrrolopyrroles, quinophthalones, and truarycarboniums.

[0025] As used herein, the term "perylene derivative" is intended to include any visually detectable substituted perylene. However, the term is not intended to include perylene itself. The terms "anthracene derivative," "naphthalene derivative," and "pyrene derivative" are similarly used. In some preferred embodiments, the derivative (e.g., perylene, pyrene, anthracene, or naphthalene derivative) is an imide, bis-imide, or hydrazamimide derivative of perylene, anthracene, naphthalene, or pyrene. "Solid support" refers to any solid substrate known in the art for immobilized phase support of molecules, for example, "microparticle" refers to any of several small particles useful for binding to compounds of the invention, including, but not limited to, glass beads, magnetic beads, polymeric beads, non-polymeric beads, etc. In certain embodiments, microparticles comprise polystyrene beads. "Solid support residue" refers to a functional group that remains attached to a molecule once the molecule is cleaved from the solid support. Solid support residues are known in the art and can be easily derivatized based on the structure of the solid support and the group linking the molecule to the solid support.

[0026] A "targeting moiety" is a moiety that selectively binds to or associates with a particular target, such as a tumor cell antigen. "Selectively" binding or associating means that the targeting moiety preferentially associates with or binds to a desired target over other targets. In some embodiments, the compounds disclosed herein include a linking group to the 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 the 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, the targeting moiety is a moiety, such as an antibody, that selectively binds to or associates with a target feature on or within a cell, such as a target feature on the surface of a cell membrane or other cellular structure, thereby enabling delivery of the biologically active moiety to or within the cell of interest. Small molecules that selectively bind to or associate with a desired biological target are also contemplated as targeting moieties in certain embodiments. Those of skill in the art will recognize other biological targets and corresponding targeting moieties that are useful in various embodiments. "Base pairing moiety" refers to a heterocyclic moiety that can hybridize with a complementary heterocyclic moiety through hydrogen bonding (e.g., Watson-Crick base pairing). Base pairing moieties include natural and unnatural bases. Non-limiting examples of base pairing moieties are RNA and DNA bases such as adenosine, guanosine, thymidine, cytosine, and uridine, and their analogs. A "physiologically cleavable linker" refers to a molecular linking group that can be split or separated in a defined manner to yield two or more individual molecules while present in the in vivo or in vitro environment of an organism or cellular system. Generally, physiological conditions that encompass such cleavage or scission events include temperatures ranging from about 20 to 40°C, atmospheric pressure of about 1 atm (101 kPa or 14.7 psi), pH of about 6 to 8, glucose concentration of about 1 to 20 mM, atmospheric oxygen concentration, and Earth's gravity. In some embodiments, physiological conditions include enzymatic conditions (i.e., cleavage by an enzyme). Bond cleavage or scission can be homogeneous or heterogeneous.

[0027] 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 can be incorporated into the disclosed compounds include, respectively: 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) may generally be prepared by conventional techniques known to those skilled in the art, or by methods analogous to those described below and in the Examples below, substituting the appropriate isotopically labeled reagent for the previously used unlabeled reagent. "Stable compound" and "stable structure" are intended 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.

[0028] "Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances when the event or circumstance occurs as well as instances when it does not occur. For example, "optionally substituted alkyl" means that the alkyl group may be substituted or unsubstituted, and that the description includes both substituted and unsubstituted alkyl groups. "Salt" includes both acid and base addition salts.

[0029] "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 salts of, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, schizoic acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfonic acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, mucous acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutamine acid, glutamine It refers to salts formed with organic acids such as carboxylic acid, 2-oxo-glutaric 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 salts" refer to salts prepared from the addition of inorganic or organic bases to free acids. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. 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 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, purines, piperazine, piperidine, N-ethylpiperidine, and basic ion exchange resins such as polyamine resins. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine.

[0030] Crystallization may produce solvates of the compounds described herein. The embodiments of the present invention include all solvates of the compounds described herein. As used herein, the term "solvate" refers to an aggregate containing one or more molecules of the compound of the present invention and one or more solvent molecules. 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, while in other cases, the compounds of the present invention may simply retain incidental water or another solvent, or may be a mixture of water and an incidental solvent.

[0031] Embodiments of 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 may give rise to enantiomers, diastereomers, and other stereoisomers that may be defined, with respect to absolute stereochemistry, as (R)- or (S)-, or, in the case of amino acids, (D)- or (L)-. Embodiments of the present invention are intended 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 can be prepared using chiral synthons or chiral reagents or resolved using conventional techniques, such as 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 racemate (or racemate of a salt or derivative) using, for example, chiral high-pressure liquid chromatography (HPLC). When a compound described herein contains an olefinic double bond or other feature that results in geometric symmetry, unless otherwise specified, the compound is intended to include both E and Z geometric isomers, and all tautomeric forms are also intended to be included.

[0032] "Stereoisomers" refer to compounds composed of identical atoms joined by the same bonds, but having different three-dimensional structures, and are not interconvertible. The present invention contemplates various stereoisomers and mixtures thereof, and includes "enantiomers," which refer to two stereoisomers whose molecules are non-superimposable mirror images of one another. "Tautomer" refers to a proton shift from one atom of a molecule to another atom of the same molecule. The present invention includes tautomers of any of the compounds described above. Various tautomers of the compounds can be easily derived by one skilled in the art. The chemical naming protocols and structural diagrams used herein are a modification of the IUPAC nomenclature system using the ACD / Nomenclature version 9.07 software program and / or the ChemDraw Ultra version 11.0 software nomenclature program (CambridgeSoft). Common names familiar to those skilled in the art are also used.

[0033] As noted above, one embodiment of the present invention provides compounds useful as therapeutic molecules, and in certain embodiments, as colored dyes in various methods (e.g., treatments). In general terms, embodiments of the present invention are directed to dimeric or higher polymers of biologically active moieties. The biologically active moieties (including fragments and prodrugs thereof) and optional chromogenic moieties are linked by one or more rigid linkers (i.e., "A" as defined with respect to structure (I)). Without wishing to be bound by theory, it is believed that the linkers help maintain sufficient spatial distance between the biologically active moieties and each of the fluorescent and / or chromogenic moieties, thereby reducing or eliminating intramolecular interactions.

[0034] Thus, in some embodiments, the compound has the following structure (A): [ka] (A) (In the formula, L 5 is a linker that is sufficiently rigid to maintain spatial separation between one or more (e.g., each) M groups, resulting in reduced or no intramolecular interactions; R 1 , R 2 , R 3 , L, L 2 , M and n are as defined for structure (I).

[0035] In some embodiments, a compound having the following structure (I): [ka] (I) or a stereoisomer, pharmaceutically acceptable salt, or tautomer thereof, wherein 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, provided that at least one occurrence of M is not a fluorescent dye; L is a physiologically cleavable linker; L 1 , L 2 , L 3 and L 4 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, -OP(=R a )(R b )R c , Q, L', and R a is O or S, and R b OH, SH, O - , S - , OR d or SR d and R c OH, SH, O - , S - , OR d , S.R. d , alkyl, alkoxy, alkyl ether, alkoxyalkyl ether, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkyl ether or thiophosphoalkyl ether, and R d is the counterion, R 4 are independently assigned to each occurrence: OH, SH, O - , S -, OR d or SR d and R 5 is independently at each occurrence oxo, thioxo, or absent; Q is, independently at each occurrence, a moiety that contains a reactive group capable of forming a covalent bond with an analyte molecule, a solid support, or a complementary reactive group, Q'; L' is, independently at each occurrence, a linker that includes a covalent bond to Q, a targeting moiety, a linker that includes a covalent bond to a targeting moiety, a solid support or a solid support residue, a linker that includes a covalent bond to a solid support or a solid support residue, or a linker that includes a covalent bond to an additional compound of structure (I); m, independently at each occurrence, is 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 equal to or greater than 1) is provided.

[0036] The various linkers and substituents (e.g., M, A, Q, R) in the compounds of structure (I) 1 , R 2 , R 3 , R 4 , R 5 , L, L 1 , L 2 , L 3 , L 4and L') may be further substituted with one more substituent. For example, in some embodiments, optional substituents are selected to optimize the aqueous solubility, permeability, retention, or other properties of the compounds of structure (I). In certain embodiments, alkyl, alkoxy, alkyl ether, alkoxyalkyl ether, phosphoalkyl, thiophosphoalkyl, phosphoalkyl ether, and thiophosphoalkyl ether in the compounds of structure (I) may each be further substituted with one more 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.

[0037] In some embodiments, at least one occurrence of M comprises a biologically active moiety. In other embodiments, each occurrence of M independently comprises a biologically active moiety. In some embodiments, A is, independently at each occurrence, a moiety that includes one or more fused aryl or heteroaryl ring systems. In a different embodiment, A is, independently at each occurrence, a moiety that includes one or more bicyclic or tricyclic fused aryl or heteroaryl ring systems.

[0038] In other more specific embodiments, A is, independently at each occurrence, the following structure: [ka] (In the formula, a 1 , a 2 and a 3 is independently at each occurrence a 5-, 6-, or 7-membered carbocyclic or heterocyclic ring; L 6 is a direct bond or a linker) and a fused carbocyclic or heterocyclic ring system having one of the following ring structures:

[0039] In still other embodiments, A is, independently at each occurrence, the following structure: [ka] It has one of the following.

[0040] In some more specific embodiments, each occurrence of A has the following structure: [ka] It has.

[0041] The linker L can be used as the point of attachment of the M moiety to the remainder of the compound. For example, in some embodiments, a synthetic precursor to a compound of structure (I) is prepared, and the M moiety is attached to the synthetic precursor using any number of readily available methods known in the art, such as those referred to as "click chemistry." To this end, any of a number of rapid and substantially irreversible reactions can be used to attach M to the synthetic precursor for forming the compound of structure (I). Exemplary reactions include the copper-catalyzed reaction of an azide with an alkyne to form a triazole (Huisgen 1,3-dipolar cycloaddition), the reaction of a diene with a dienophile (Diels-Alder), strain-promoted alkyne-nitrone cycloaddition, the reaction of a strained alkene with an azide, tetrazine, or tetrazole, the [3 + 2] cycloaddition of an alkene with an azide, the reverse demand Diels-Alder reaction of an alkene with a tetrazine, the photoreaction of an alkene with a tetrazole, and various substitution reactions such as the replacement of a leaving group by nucleophilic attack on an electrophilic atom. Exemplary substitution reactions include reaction of amines with activated esters (e.g., N-hydroxysuccinimide esters), isocyanates, isothiocyanates, etc. In some embodiments, the reaction to form L may be carried out in an aqueous environment. Thus, in some embodiments, each occurrence of L is a linker that includes a functional group formable by the reaction of two complementary reactive groups, e.g., a functional group that is the product of one of the "click" reactions described above. In various embodiments, for at least one occurrence of L, the functional group can be formed by the 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., the reaction of an amine with an N-hydroxysuccinimide ester or an isothiocyanate.

[0042] In some embodiments, each occurrence of L is a linker that includes a functional group formable by reaction of two complementary reactive groups. In certain related embodiments, each occurrence of L 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, a heterocyclic, a heteroaryl, or a combination thereof. In other embodiments, for at least one occurrence of L, the functional group may be formed by the reaction of an alkyne and an azide. In other embodiments, for at least one occurrence of L, the functional group may be formed by the reaction of an amine (e.g., a primary amine) and an N-hydroxysuccinimide ester or an isothiocyanate.

[0043] In further embodiments, for at least one occurrence of L, the functional group comprises an alkene, ester, amide, thioester, disulfide, carbocyclic group, heterocyclic group, or heteroaryl group. In further embodiments, for at least one occurrence of L, the functional group comprises an alkene, ester, amide, thioester, thiourea, disulfide, carbocyclic group, heterocyclic group, or heteroaryl group. In other embodiments, the functional group comprises an amide or thiourea. In some further specific embodiments, for at least one occurrence of L, L is a linker comprising a triazolyl functional group. In other embodiments, for at least one occurrence of L, L is a linker comprising an amide or thiourea functional group. Some embodiments provide L that is cleavable 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 or oxo, 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, bivinaldiol, pyrophosphate diester, or valine citrulline.

[0044] In certain embodiments, L, independently at each occurrence, is a linker that is cleavable at a pH in the range of 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. In certain embodiments, L, independently at each occurrence, is a linker that is cleavable at a temperature in the range of 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, independently at each occurrence, is a linker that is cleavable at a temperature of 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.

[0045] In certain embodiments, each occurrence of L is independently a linker that is cleavable by an enzyme. For example, in some embodiments, the enzyme is a hydrolase, oxidoreductase, or lyase. In certain embodiments, the enzyme is an EC 4.1 (e.g., EC 4.1.1, EC 4.1.2, EC 4.1.3, or EC 4.1.99), EC 4.2, EC 4.3, EC 4.4, EC 4.5, EC 4.6, or EC 4.99 enzyme.

[0046] In certain embodiments, L is one of the following structures: [ka] (In the formula, R is H, methyl, ethyl, isopropyl, tert-butyl or phenyl; X is O or CH2; n is an integer greater than 0).

[0047] In still other embodiments, for at least one occurrence of L, LM has the following structure: [ka] (In the formula, L 1a and L 1b are each independently an optional linker. In a different embodiment, for at least one occurrence of L, LM has the following structure: [ka] (In the formula, L 1a and L 1b are each independently an optional linker. In the various embodiments described above, L 1a Or L 1b In other embodiments, L 1a Or L 1b , or both of them are present.

[0048] In some embodiments, L 1a or L 1b When present, each independently is alkylene or heteroalkylene. For example, in some embodiments, L 1a or L 1b is, if present, independently one of the following structures: [ka] It has.

[0049] In yet other different embodiments of structure (I), L is, independently at each occurrence, an optional alkylene or heteroalkylene linker. In certain embodiments, L is one of the following structures: [ka] It has. In a further embodiment, L 2 and L 3 is independently at each occurrence C1-C6 alkylene, C2-C6 alkenylene, or C2-C6 alkynylene.

[0050] In a further embodiment, L 2 , L 3 , L 4 and L 5is independently at each occurrence C-C alkylene, C-C alkenylene, or C-C alkynylene. For example, in some embodiments, the compound has the following structure (IA): [ka] (IA) (In the formula, x 1 , x 2 , x 3 , x 4 , x 5 and x 6 is independently an integer from 0 to 6 at each occurrence.

[0051] In certain embodiments of the compound of structure (IA), x 3 and x 4 In other embodiments, x 1 , x 2 , x 5 and x 6 is 1 for each occurrence. In some more specific embodiments of the compounds of structure (IA), L, independently at each occurrence, comprises a triazolyl functional group. In other embodiments of the compounds of structure (IA), L, independently at each occurrence, is an optional alkylene linker or heteroalkylene linker. In still other embodiments of either the compounds of structure (I) or (IA), R 4 are independently expressed as OH, O - OR d "OR d " and "SR d " is O bonded to a cation - and S - For example, the disodium salt of a phosphate group is:

[0052] [ka] (In the formula, R a is sodium (Na + ) is) It can be expressed as: In other embodiments of any of the compounds of structure (I) or (IA), R 5 is oxo at each occurrence. In some different embodiments of any of the compounds described above, R 1 Each occurrence of R is H. In some embodiments, R 1 is methyl at each occurrence.

[0053] In various other embodiments, R 2 and R 3 are each independently OH or -OP(=R a )(R b )R c In certain related 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 Q or a linker comprising a covalent bond to Q. In some such embodiments, Q comprises a nucleophilic reactive group, an electrophilic reactive group, or a cycloaddition reactive group. In more specific embodiments, Q is a sulfhydryl, a disulfide, an activated ester (e.g., an N-succinimide ester, an imide ester, or a polyphenylene ester). Ruoro Phenyl (polyfl u phenyl) ester), isothiocyanate, azide (e.g., alkyl azide or acyl azide), alkyne, alkene, diene, dienophile, acid halide, sulfonyl halide, phosphine, α-haloamide, biotin, amino, or maleimide functional groups. In certain embodiments, R 2 or R 3 one of is L', and L' is a linker comprising a covalent bond to the solid support. In some such embodiments, the solid support is a polymeric bead or a non-polymeric bead. In certain other embodiments, R 2 or R 3one of is L', where L' is a targeting moiety or a linker to a targeting moiety. In some of these embodiments, L' is a linker to a targeting moiety, and the linker comprises a heteroalkylene linker. 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 L'. In further different embodiments of any of the above compounds of structure (I), R 2 and R 3 are each independently -OP(=R a )(R b )R c In some of these embodiments, R c is OL'.

[0054] 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. In some embodiments, the linker is a peptidyl linker, or a linker comprising an alkylene oxide or 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 residue, an amino acid, a nucleoside, or an additional compound of structure (I). The linker L' can be any linker suitable for attaching 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 aqueous solubility of the compound. In certain embodiments, L' is a heteroalkylene moiety. In certain other embodiments, L' comprises an alkylene oxide or a phosphodiester moiety, or a combination thereof.

[0055] In certain embodiments, L' has the following structure: [ka] (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 linking group to a targeting moiety. In some embodiments, m" is an integer between 4 and 10, for example, 4, 6, or 10. In other embodiments, n" is an integer between 3 and 6, for example, 3, 4, 5, or 6.

[0056] 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, tovantinib (ARQ197), crizotinib, cabozantinib, tyrphostins (e.g., AG538, AG1024), pyrrolo(2,3-d)-pyrimidine derivatives (e.g., NVP-AEW541), monoclonal antibodies (e.g., Figitumab), or the like. The following are potential candidates for the treatment of rheumatoid arthritis: rivaroxaban, cetuximab, panitumumab, necitumumab, ganitumab, cixutumumab, dalotuzumab, lobatumumab, onartuzumab, K1, labetuzumab, milatuzumab, lorvotuzumab, inotuzumab), BMS-777607, PF-02341066, PF-04217903, AMG-458, MK-2461, JNJ-38877605, GSK1363089 (foretinib), XL880, XL184, ARQ197, E7050, or INCB28060.

[0057] In certain embodiments, the antibody or cell surface receptor antagonist targets EGFR (e.g., EGFRvIII), HER2, folate 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.

[0058] In some other embodiments, L" is an alkylene or heteroalkylene moiety. In certain other embodiments, L" comprises an alkylene oxide, a phosphodiester moiety, a peptidyl linker, a sulfhydryl, a disulfide, or a maleimide moiety, or a combination thereof.

[0059] In other further specific embodiments of any of the above-described compounds of structure (I), R 2 or R 3 is one of the following structures: [ka] It has TIFF0007813098000017.tif244170.

[0060] Certain embodiments of the compounds of structure (I) can be prepared according to solid-phase synthesis methods similar to those known in the art for preparing peptides or nucleic acids. Thus, in some embodiments, L' is a linking group to a solid support, a solid support residue, a peptide, or an amino acid. A variety of solid supports (e.g., polystyrene, polyamide, PEG-based), protecting groups (e.g., Fmoc, Boc, triphenylmethyl), activating groups (e.g., carbodiimide, triazole), and other materials are readily available and can, in some embodiments, be used to prepare compounds of structure (I). The solid support residue can be removed or modified after synthesis.

[0061] In still other embodiments, Q, independently at each occurrence, is a moiety that includes a reactive group capable of forming a covalent bond with an analyte molecule or a solid support. In other embodiments, Q, independently at each occurrence, is a moiety that includes a reactive group that is capable of forming a covalent bond with a complementary reactive group, Q'. For example, in some embodiments, Q' is present in an additional compound of structure (I) (e.g., R 2 or R 3In some embodiments, Q and Q' contain complementary reactive groups, such that reaction of a compound of structure (I) with an additional compound of structure (I) results in a covalent dimerization of the 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. The type of Q group and its attachability to the remainder of the compound of structure (I) are not particularly limited, provided that Q contains a moiety with appropriate reactivity to form the desired bond. In certain embodiments, Q is a moiety (e.g., an amine, azide, or alkyne) that is not susceptible to hydrolysis under aqueous conditions, yet is sufficiently reactive to form a bond with a corresponding group on the analyte molecule or solid support.

[0062] Certain embodiments of the compound of structure (I) include a Q group 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, disulfide, activated ester, isothiocyanate, azide, alkyne, alkene, diene, dienophile, acid halide, sulfonyl halide, phosphine, α-haloamide, biotin, amino, or maleimide functionality. In some embodiments, the activated ester is an N-succinimide ester, imidoester, or polyflourophenyl ester. In other embodiments, the alkyne is an alkyl azide or acyl azide. The Q group can be conveniently provided in a protected form that enhances storage stability or other desired properties, and the protecting group can then be removed at the appropriate time for coupling, for example, with a targeting moiety or analyte. Thus, the Q group includes "protected forms" of reactive groups, including any of the reactive groups listed above and in Table 1 below. A "protected form" of Q refers to a moiety that has lower reactivity under predetermined reaction conditions relative to Q, but that can be converted to Q under conditions that preferably do not decompose or react with other moieties of the compound of structure (I). One of skill in the art can derive an appropriate protected form of Q based on the particular Q and the desired end-use and storage conditions. For example, if Q is -SH, the protected form of Q includes a disulfide that can be reduced using common known techniques and reagents (e.g., TCEP) to reveal the -SH moiety.

[0063] Exemplary Q moieties are provided in Table 1 below. [Table 1-1] [Table 1-2] [Table 1-3]

[0064] It should be noted that in some embodiments, when Q is -SH, the -SH moiety tends to form a disulfide bond with another sulfhydryl group, for example, of another compound of structure (I). Thus, some embodiments include compounds of structure (I) in the form of a disulfide dimer, where the disulfide bond is derived from the Q group that is -SH. In other embodiments, the Q moiety is conveniently hidden (e.g., protected) as a disulfide moiety, which can be subsequently reduced to provide an activated Q moiety for conjugation to a desired targeting moiety. For example, the Q moiety may have the following structure: [ka] where R is an optionally substituted alkyl group. For example, in some embodiments, Q may be hidden as a disulfide having the following structure: [ka] (wherein n' is an integer from 1 to 10; for example, in some embodiments, n' is 4, 5, 6, or 7). The disulfide moiety is provided as

[0065] Similarly, compounds of structure (I) are included within the scope of certain embodiments, where R 2 and R 3 One or both of R 2 and R 3 One or both of -OP(=R a )(R b )R c and R c 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 having a "Q" suitable for reaction with a complementary Q' group on a second compound of structure (I). In this manner, compounds of structure (I) having any number of "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 the following structure (I'):

[0066] [ka] (I') (In the formula, A, R 1 , R 2 , R 3 , R 4 , R 5 , L, L 1 , L 2 , L 3 , L4 , M, m, and n at each occurrence are 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; α is an integer greater than 1, for example, 1 to 100, or 1 to 10. Other compounds of structure (I') can be derived by one skilled in the art, for example, by dimerizing or polymerizing the compounds of structure (I) provided herein.

[0067] 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. The value for m is another variable that can be selected based on the desired solubility, osmotic activity, or therapeutic use. In some embodiments, m, independently at each occurrence, is an integer from 1 to 20. In some embodiments, m, independently at each occurrence, is an integer from 1 to 10. In other embodiments, m, independently at each occurrence, is an integer from 1 to 5, e.g., 1, 2, 3, 4, or 5. Solubility, permeability, or retention can also be adjusted by selecting different values ​​of n. In certain embodiments, n is an integer between 1 and 100. In other embodiments, n is an integer between 1 and 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.

[0068] Tumor cell antigens include tumor-specific antigens and tumor-associated antigens, such as EGFR, HER2, folate receptor, CD20, CD33, oncofetal antigens (e.g., alphafetoprotein, carcinoembryonic antigen, immature laminin receptor, TAG-72), CA-125, MUC-1, epithelial tumor antigen, tyrosinase, melanoma-associated antigen (MAGE), and aberrant products of RAS or p53. Tumor cell antigens can also include antigens characterized as 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 testis (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 some embodiments, M is, independently at each occurrence, an NSAID, a kinase inhibitor, an anthracycline, an EGFR inhibitor, or an alkylating agent.

[0069] In some embodiments, at least one M is an anti-cancer drug. In certain particular embodiments, each occurrence of M is independently an anti-cancer drug and the targeting moiety is an antibody specific for a tumor cell antigen. In some embodiments, the tumor cell antigen is EGFR, HER2, folate receptor, CD20, or CD33. As used herein, anticancer drugs include derivatives, which are modified or derivatized anticancer drugs, such that the drug may be conjugated or attached to another molecule (e.g., to include a Q moiety). For example, maytansine is a cancer drug, and maytansinoids are derivatives of cancer drugs. 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. Anticancer drugs that may be modified and incorporated into 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, or methotrexate.

[0070] 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®)); alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenediaminetetraacetic acid (EDTA), and ethylenediaminetetraacetic acid (EDTA). melamine and methylmelamine (including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine); chlorambucil, chlornaphazine, clofosfamide, estramustine, ifosamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembicin, fenesterine, prednimustine, trofosfamide, nitrogen mustards such as uracil mustard; carmustine, chlorozotocin, fotemustine, loxacin Nitrosoureas such as mustine, nimustine, and ranimustine; aclacinomycins, actinomycin, ausramycin, azaserine, bleomycins, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, Casodex®, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, mycophenone Antibiotics such as benzodiazepine, nogalamycin, olivomycins, peplomycin, potfilomycin, puromycin, queramycin, rhodrubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, and zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, and trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, and thioguanine;Pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, and floxuridine; androgens such as calucelone, dromostanolone propionate, epitiostanol, mepitiostane, and testolactone; antiadrenal agents such as aminoglutethimide, mitotane, and trilostane; folic acid supplements such as florinic acid; aceglatone; aldophosphamide glycosides; aminolevulinic acid; amsacrine; Bestravcil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomitine; elliptinium acetate; etoglucide; nitrate Gallium; Hydroxyurea; Lentinan; Lonidamine; Mitoguazone; Mitoxantrone; Mopidamol; Nitracrine; Pentostatin; Fenameth; Pirarubicin; Podophyllic acid; 2-Ethylhydrazide; Procarbazine; PSK.RTM; Razoxane; Sizofiran; Spirogermanium; Tenuazonic acid; Triaziquone; 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) Squibb Oncology, Princeton, NJ), and docetaxel (TAXOTERE™, Rhone-Poulenc Rorer, Antony, France); retinoic acid; esperamicin or capecitabine. Similarly, for example, tamoxifen (Nolvadex™), raloxifene, aromatase-inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and toremifene (Fareston); and 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;Suitable cancer drugs include antihormonal agents that act to regulate or inhibit hormone action on tumors, such as mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; camptothecin-11 (CPT-11); the topoisomerase inhibitor RFS2000; and antiestrogens, including difluoromethylornithine (DMFO).

[0071] In certain embodiments, at least one M is selected from the group consisting of tyrosine kinase inhibitors (e.g., gefitinib, erlotinib), lapatinib, vandetanib, neratinib, osimertinib, tovantinib (ARQ197), crizotinib, cabozantinib, tyrphostins (e.g., AG538, AG1024), pyrrolo(2,3-d)-pyrimidine derivatives (e.g., NVP-AEW541), monoclonal antibodies (e.g., figitumumab, cetuximab, panitumumab), and the like. The following are potential candidates for the treatment of rheumatoid arthritis: rituzumab, necitumumab, ganitumab, cixutumumab, dalotuzumab, lobatuzumab, onartuzumab, K1, labetuzumab, milatuzumab, lorvotuzumab, inotuzumab), BMS-777607, PF-02341066, PF-04217903, AMG-458, MK-2461, JNJ-38877605, GSK1363089 (foretinib), XL880, XL184, ARQ197, E7050, or INCB28060.

[0072] Where desired, embodiments of the disclosed compounds or compositions may be used in combination with other compounds such as Herceptin®, Avastin®, Erbitux®, Rituxan®, Taxol®, Arimidex®, Taxotere®, ABVD, avicin, abagovomab, acridine carboxamide, adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, alfalazine, alvocidib, 3-allylamino-17-demethoxygeldanamycin ... Minopyridine-2-carboxaldehyde thiosemicarbazone, amonafide, anthracenedione, anti-CD22 immunotoxin, antineoplastic agents, antitumor herbs, apaziquone, atiprimod, azathioprine, belotecan, bendamustine, BIBW2992, biricodar, brostallicin, bryostatin, buthionine sulfoxime, CBV (chemotherapy), calyculin, cell cycle nonspecific antineoplastic agents, dichloroacetic acid, discodermolide, elsamitrucin, eno Citabine, epothilone, eribulin, everolimus, exatecan, exisulind, feldinol, holodesin, fosfestrol, ICE chemotherapy regimen, IT-101, imexon, imiquimod, indolocarbazole, irofulven, ranicuidar, larotaxel, lenalidomide, lucanton, lurtotecan, mefosfamide, mitozolomide, nafoxidine, nedaplatin, olaparib, ortataxel, PAC-1, pawpaw, pixantrone, prothrombin It may be used in combination with commonly prescribed anticancer drugs such as loteasome inhibitors, rebecamycin, resiquimod, rubitecan, SN-38, salinosporamide A, sapacitabine, Stanford V, swainsonine, talaprofin, taliquidar, tegafur-uracil, temodar, tesetaxel, triplatin tetranitrate, tris(2-chloroethyl)amine, troxacitabine, uramustine, vadimezan, vinflunine, ZD6126, or zosquidar.

[0073] M is selected based on desired therapeutic and / or optical properties, for example, treatment of a particular disease or condition (e.g., cancer), or generation of a particular color and / or fluorescent emission wavelength. In some embodiments, M is the same at each occurrence. However, it is important to note that each occurrence of M need not be the same M, and certain embodiments include compounds in which M is not the same at each occurrence. For example, in some embodiments, M is not each the same, and different M moieties are selected to have different therapeutic properties (e.g., cytotoxicity and anti-inflammatory). In some embodiments, M is not each the same, and different M moieties are selected to have the same or similar therapeutic properties (e.g., cytotoxicity). In certain embodiments, M is each of the following:

[0074] [ka] Selected independently from TIFF0007813098000025.tif64150.

[0075] For ease of illustration, specific points of attachment to the rest of the molecule (i.e., [ka] Although depicted with a 5'-point bond, the M moiety may be attached via any available point. One of ordinary skill in the art can determine the appropriate point of attachment.

[0076] In certain embodiments, LM has the following structure: [ka] I have one TIFF0007813098000028.tif217170. In further embodiments of any of the above, M are the same. In other embodiments, each M is different. In further embodiments, one or more M are the same and one or more M are different.

[0077] In some embodiments, the selected occurrences of M are not the same, and 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, different M moieties are selected so that absorbance of radiation at one wavelength causes emission of radiation at a different wavelength via a FRET mechanism. Exemplary M moieties can be appropriately selected by one 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 may be attached to the remainder of the molecule from any position (i.e., atom) on M. Those of skill in the art will recognize means by which M can be attached to the remainder of the molecule. Exemplary methods include the "click" reaction described herein.

[0078] In some embodiments, at least one M is a fluorescent or chromogenic moiety. Any fluorescent and / or chromogenic moiety may be used, such as those known in the art and commonly used in colorimetric, UV, and / or fluorescent assays. Examples of M moieties useful in various embodiments of the present invention include, but are not limited to, 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 squaraine dyes); naphthalene derivatives (e.g., dansyl and polodan derivatives); coumarin derivatives; oxadiazole derivatives (e.g., pyridyloxazole, nitro Examples of M moieties include benzoxadiazoles or benzoxadiazoles; anthracene derivatives (e.g., anthraquinones, including DRAQ5, DRAQ7, and CyTRAK 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 telapyrrole 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; Tamura; Texas Red, and Alexa Fluor® dyes.

[0079] In any of the above-mentioned further embodiments, at least one M comprises three or more aryl or heteroaryl rings, or a combination thereof, for example, four or more aryl or heteroaryl rings, or a combination thereof, or five or more aryl or heteroaryl rings, or even a combination thereof. In some embodiments, at least one M comprises six aryl or heteroaryl rings, or a combination thereof. In further embodiments, the rings are fused. For example, in some embodiments, at least one M comprises three or more fused rings, four or more fused rings, five or more fused rings, or even six or more fused rings.

[0080] In some embodiments, at least one M is cyclic. For example, in some embodiments, at least one M is carbocyclic. In other embodiments, at least one M is heterocyclic. In still other embodiments described above, at least one M, independently at each occurrence, comprises an aryl moiety. In some of these embodiments, the aryl moiety is polycyclic. In other further specific examples, the aryl moiety is a fused polycyclic aryl moiety, e.g., the fused polycyclic aryl moiety can comprise at least three, at least four, or even more than four aryl rings. In other embodiments of any of the above-described compounds of structure (I) or (I'), at least one 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 above, at least one M comprises at least one substituent, for example, in some embodiments, the substituent is 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.

[0081] In some more specific embodiments described above, at least one M is a dimethylaminostilbene, quinacridone, fluorophenyl-dimethyl-BODIPY, his-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, at least one M is p-terphenyl, perylene, azobenzene, phenazine, phenanthroline, acridine, thioxanthrene, chrysene, rubrene, coronene, cyanine, perylene imide, or perylene amide, or a derivative thereof. In further embodiments, at least one M is 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.

[0082] In some embodiments, at least one M is pyrene, perylene, perylene monoimide, or 6-FAM, or a derivative thereof. In some other embodiments, at least one M has the following structure: [ka] It has one of the following. The M moiety containing the carboxylic acid group is in the anionic form (CO2 - ), but one of skill in the art will understand that this will vary with pH and that the protonated form (COH) is included in various embodiments.

[0083] In some specific embodiments, the compound is a compound selected from Table 2. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] As used in Table 2 and throughout this application, R 2 , R 3 , L, M, and n have the definitions provided for the compounds of structure (I) unless otherwise indicated. In some embodiments, M is, independently at each occurrence, F, F', F", N', I', D', or D", provided that at least one M is N', I', D', or D". F, F', and F" are each selected from the following structures:

[0084] [ka] refers to a fluorescein moiety having the formula:

[0085] "N'" refers to the following structure: [ka] Refers to...

[0086] "I'" refers to the following structure: [ka] Refers to...

[0087] "D'" refers to the following structure: [ka] Refers to...

[0088] "D" means the following structure: [ka] Refers to...

[0089] "dT" has the following structure: [ka] (In the formula, R is H or a direct bond. In some embodiments, L′, R 2 or R 3 In more specific embodiments, R 2 In some embodiments, R 3 comprises dT. In some embodiments, L' comprises dT.

[0090] Some embodiments include any of the compounds presented in Table 2 and described above, including specific compounds conjugated to a targeting moiety, such as an antibody. In some embodiments, one of the compounds 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.

[0091] 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 useful for reacting with a complementary moiety, by any number of synthetic methodologies, to form a covalent bond between M and the reactive polymer (e.g., the "click" reaction described above), thus forming a compound of structure (I). Thus, in various embodiments, compounds of structure (I) have the following structure (II): [ka] (II) or a stereoisomer, salt or tautomer thereof, wherein G is, independently at each occurrence, a moiety containing a reactive group or a protected analogue thereof capable of forming a covalent bond with a complementary reactive group; L 1a , L 1 , L 2 , L 3 and L 4 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, -OP(=R a )(R b )R c , Q, L', and R a is O or S, and R b OH, SH, O - , S - , OR d or SR d and R c OH, SH, O - , S - , OR d , S.R. d , alkyl, alkoxy, alkyl ether, alkoxyalkyl ether, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkyl ether or thiophosphoalkyl ether, and R d is the counterion, R 4 are independently assigned to each occurrence: OH, SH, O - , S - , OR d or SR d and R 5 is independently at each occurrence oxo, thioxo, or absent; Q is, independently at each occurrence, a moiety that contains a reactive group capable of forming a covalent bond with an analyte molecule, a solid support, or a complementary reactive group, Q'; L' is, independently at each occurrence, a linker that includes a covalent bond to Q, a targeting moiety, a linker that includes a covalent bond to a targeting moiety, a linker that includes a covalent bond to a solid support or a solid support residue, a linker that includes a covalent bond to a solid support or a solid support residue, or a linker that includes a covalent bond to an additional compound of structure (I); m, independently at each occurrence, is 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 equal to or greater than 1) The polymer is formed using a reactive polymer having the formula:

[0092] In some embodiments, the reactive polymer is selected from Table 3 below. [Table 3]

[0093] In various embodiments, G in the compounds of Table 3 is alkynyl, such as ethynyl. In other embodiments, G in the compounds of Table 3 is azide. In other embodiments, G in the compounds of Table 3 is amino (NH). In other embodiments, G in the compounds of Table 3 is isothiocyanate. In other embodiments, G in the compounds of Table 3 is an activated ester, such as an ester of N-hydroxysuccinimide. Certain embodiments are directed 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 comprise 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 when excited by light, such as ultraviolet light.

[0094] composition Also provided is a composition comprising any one of the claimed compounds and a targeting moiety. The presently disclosed compound embodiments are "tunable," meaning that by appropriately selecting the variables in any of the compounds described above, one of skill in the art can arrive at a compound with a desired molar fluorescence and / or a predetermined molar fluorescence (molar brightness). The "tunability" of certain embodiments of the present compounds allows the user to easily arrive at a compound with a 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, A, M, L, and the like are particularly well-known. 1 , L 2 , L 4 It is believed that the proper selection of A, L, m, and n plays an important role in the molar fluorescence of embodiments of the disclosed compounds. Thus, in one embodiment, a method for obtaining a compound with a desired molar fluorescence comprises the steps of selecting an M moiety with known fluorescence, preparing a compound of structure (I) containing the M moiety, and adjusting A, L, and m to reach the desired molar fluorescence. 1 , L 2 , L 4 , m and n.

[0095] For ease of illustration, various compounds containing phosphorus moieties (e.g., phosphate, etc.) can be present in an anionic state (e.g., —OPO(OH)O - , -OPO3 2- ) Those skilled in the art will readily appreciate that the charge is pH dependent and that uncharged (e.g., protonated or salts such as sodium or other cations) forms are also included within the scope of embodiments of the present invention. Compositions comprising any of the above-described compounds and one or more targeting moieties (e.g., antibodies or cell surface receptor antagonists) are provided in various other embodiments. Also provided in some embodiments is the use of such compositions in a method for treating a disease, the method 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 to treat the disease.

[0096] Pharmaceutical Composition One embodiment provides a composition comprising a compound according to any one of the embodiments disclosed herein and a pharmaceutically acceptable carrier. Another embodiment provides a composition comprising a plurality of conjugates comprising the compound of claim 1 covalently attached to an antibody by a single linking group, wherein the plurality of conjugates have at least 90% structural homogeneity. "Structural homogeneity" refers to compounds having the same structure, including the point of attachment to the antibody. In more specific embodiments, the plurality of conjugates have at least 95% structural homogeneity. In related embodiments, the plurality of conjugates have greater than 99% structural homogeneity. In certain embodiments, R 2 and R 3 One of them is -OP(=R a )(R b ) OL' or L', where L' is an antibody or a linker that comprises a covalent bond to the antibody.

[0097] Other embodiments are directed to pharmaceutical compositions. The pharmaceutical compositions comprise any one (or more) of the compounds described above and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical compositions are formulated for oral administration. In other embodiments, the pharmaceutical compositions are formulated for injection. In further embodiments, the pharmaceutical compositions comprise 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, transmucosal, transdermal, vaginal, 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 directly injecting the compound into an organ, often in a depot preparation or sustained-release formulation. In specific embodiments, long-acting formulations are administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. In yet other embodiments, the drug is delivered in a targeted drug delivery system, for example, in a liposome coated with an organ-specific antibody. In such embodiments, the liposome is targeted to the organ and selectively taken up by the organ. In still other embodiments, the compounds described herein are provided in the form of an immediate-release formulation, a sustained-release formulation, or an intermediate-release formulation. In still other embodiments, the compounds described herein are administered locally.

[0098] The compounds according to embodiments of the present invention are effective over a wide dosage range. For example, in the treatment of adult humans, dosages of 0.01 to 1000 mg per day, 0.5 to 100 mg per day, 1 to 50 mg per day, and 5 to 40 mg per day are examples of dosages that may be used in certain embodiments. An exemplary dosage is 10 to 30 mg per day. The exact dosage will depend on the route of administration, the form in which the compound is administered, the subject being treated, the subject's weight, and the preference and experience of the attending physician. In some embodiments, the compound of the present invention is administered in a single dose.Usually, this administration will be by injection, for example, intravenous injection, in order to rapidly introduce the drug.However, if appropriate, other routes can be used.A single dose of the compound of the present invention can also be used to treat acute conditions.

[0099] 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 per day. In other embodiments, administration is about once a month, once every two weeks, 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 another agent continues for less than about seven 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 dosing is achieved and maintained for as long as necessary. Administration of an embodiment of a compound of the invention may continue for as long as necessary. In some embodiments, a compound of the invention is administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In some embodiments, a compound of the invention is administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, a compound of the invention is administered continuously over an extended period of time, for example, for long-term treatment. In some embodiments, the compound of the present invention is administered at a dosage.It is known in the art that due to the subject-to-subject variability of the pharmacokinetics of compounds, individualized administration regimens are necessary for optimal treatment.The dosage for the compound embodiment of the present invention can be found by routine experimentation in light of the present disclosure.

[0100] In some embodiments, the compound described herein is formulated into pharmaceutical composition.In specific embodiments, pharmaceutical composition is formulated by conventional method, using one or more physiologically acceptable carriers, including excipients and auxiliary agents, that facilitate the processing of active compound into the preparation that can be used pharmaceutically.Suitable formulation depends on the route of administration selected. Where appropriate, any pharmaceutically acceptable techniques, carriers, and excipients may be used to formulate 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, H.A. 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). Provided herein is a pharmaceutical composition comprising a compound of structure (I) and a pharmaceutically 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 active agents described in the combination therapy section below and throughout this disclosure are encompassed herein.In specific embodiments, the pharmaceutical composition comprises one or more compounds of structure (I).

[0101] As used herein, a pharmaceutical composition refers to a mixture of a compound of structure (I) with other chemical components, such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and / or excipients. In certain embodiments, the pharmaceutical composition facilitates administration of the compound to an organism. In some embodiments of practicing the treatment methods or methods of use provided herein, a therapeutically effective amount of a compound of structure (I) provided herein is administered in a pharmaceutical composition to a mammal having the 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 singly or as components of a mixture in combination with one or more therapeutic agents.

[0102] In one embodiment, one or more compounds of structure (I) are formulated in an aqueous solution. In a specific embodiment, the aqueous solution is selected from a physiologically compatible buffer, such as, by way of example only, Hank's solution, Ringer's solution, or physiological saline buffer. In other embodiments, one or more compounds of structure (I) are formulated for transmucosal administration. In a specific embodiment, the transmucosal formulation includes a penetrant appropriate for the barrier to be permeated. In yet other embodiments, when the compounds described herein are formulated for other parenteral injections, suitable formulations include aqueous or non-aqueous solutions. In a specific embodiment, such solutions include physiologically compatible buffers and / or excipients. In another embodiment, the compounds described herein are formulated for oral administration.The compounds described herein are formulated by combining active compounds with, for example, pharmaceutically acceptable carriers or excipients.In various embodiments, the compounds described herein are formulated into oral dosage forms, including, for example, tablets, powders, pills, dragees, capsules, liquids, gels, syrups, elixirs, slurries, suspensions, etc.

[0103] In certain embodiments, pharmaceutical preparations for oral use can be prepared by mixing one or more solid excipients with one or more compounds described herein, grinding the resulting mixture, and optionally adding suitable excipients, followed by processing the resulting granule mixture to obtain tablet or dragee cores. Suitable excipients include, in particular, sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations, such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, or fillers 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). In one embodiment, dosage forms such as dragee cores and tablets are coated with one or more suitable coatings. In a specific embodiment, a concentrated sugar solution is used to coat the dosage form. The sugar solution may contain additional components such as, by way of example only, gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol and / or titanium dioxide, a lacquer solution, and suitable organic solvents or solvent mixtures. Dyes and / or pigments may be added to the coating for identification purposes. Furthermore, dyes and / or pigments may be used to distinguish different combinations of active compound doses.

[0104] In certain embodiments, a therapeutically effective amount of at least one of the compounds described herein is formulated into other oral dosage forms. Oral dosage forms include push-fit capsules made of gelatin and soft, sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. In a specific embodiment, the push-fit capsules contain the active ingredient in a mixture 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 also 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. Additionally, stabilizers may be added.

[0105] In other embodiments, a therapeutically effective amount of at least one compound 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 still other embodiments, the compounds described herein are formulated for parenteral administration, including formulations suitable for bolus injection or continuous infusion. In specific embodiments, the injectable formulations are supplied in unit dosage form (e.g., in ampoules) or in multi-dose containers. Preservatives may be added to the injectable formulations. In still other embodiments, the pharmaceutical compositions are formulated in a form suitable for parenteral injection as sterile suspensions, solutions, or emulsions in oily or aqueous vehicles. Parenteral injection formulations may contain formulating agents such as suspending agents, stabilizers, and / or dispersing agents. In specific embodiments, pharmaceutical formulations for parenteral administration comprise aqueous solutions of the active compound in water-soluble form. In additional embodiments, suspensions of the active compound (e.g., a compound of structure (I)) are prepared as appropriate oily injection suspensions. The lipophilic solvent or vehicle suitable for use in the pharmaceutical compositions described herein includes, for example, fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes.In certain embodiments, aqueous injection suspensions 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 for 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.

[0106] In yet another embodiment, the compound of structure (I) is administered topically.The compounds described herein are formulated into various compositions that can be administered topically, such as solutions, suspensions, lotions, gels, pastes, medicated sticks, balms, creams or ointments.Such pharmaceutical compositions may contain solubilizers, stabilizers, tonicity enhancers, buffering agents and preservatives. In yet other embodiments, the compounds of structure (I) are formulated for transdermal administration. In specific embodiments, 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 pharmaceutical agents. In additional embodiments, transdermal delivery of compounds of structure (I) is achieved by iontophoretic patches, etc. In certain embodiments, transdermal patches provide controlled delivery of compounds of structure (I). In specific embodiments, the absorption rate is slowed by using rate-controlling membranes or by trapping the compound within a polymer matrix or gel. In alternative embodiments, absorption enhancers are used to increase absorption. The absorption enhancer or carrier includes absorbable pharmaceutically acceptable solvents that assist in passage through the skin. For example, in one embodiment, the transdermal device is in the form of a bandage comprising a backing member, a reservoir containing the compound (which may include a carrier), optionally a rate-controlling barrier for delivering the compound to the host's skin at a controlled and predetermined rate over an extended period of time, and a means for securing the device to the skin.

[0107] In other embodiments, the compound of structure (I) is formulated to be administered by inhalation.Various forms suitable for administration by inhalation 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 supply from a pressurized pack or nebulizer, together with the use of a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas).In specific embodiments, the dosage unit of the pressurized aerosol is determined by providing a valve to deliver a metered amount.In certain embodiments, capsules or cartridges, such as, by way of example only, gelatin, for use in an inhaler or insufflator, are formulated containing a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0108] In yet another embodiment, the compounds of structure (I) are formulated into rectal compositions such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, jelly suppositories, or retention enemas containing a conventional suppository base, such as cocoa butter or other glycerides, and a synthetic polymer, such as polyvinylpyrrolidone, PEG, etc. In suppository forms of the composition, a low melting wax, such as, but not limited to, a mixture of fatty acid glycerides, optionally in combination with cocoa butter, is first melted. In certain embodiments, pharmaceutical compositions are formulated in any conventional manner using one or more physiologically acceptable carriers, including excipients and auxiliary agents, which facilitate the processing of active compounds into pharmaceutically usable preparations.Suitable formulations depend on the selected route of administration.Any pharmaceutically acceptable technique, carrier and excipient may be used if appropriate.Pharmaceutical compositions comprising the compound of structure (I) are prepared in a conventional manner, for example, by conventional mixing, dissolving, granulating, making dragees, grinding, emulsifying, encapsulating, entrapping or compressing processes, etc.

[0109] Pharmaceutical compositions comprise at least one pharmaceutically acceptable carrier, diluent, or excipient and, as an active ingredient, at least one compound of structure (I) described herein. The active ingredient may be in free acid or free base form, or in pharmaceutically acceptable salt form. Furthermore, 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 provided herein. Furthermore, the compounds described herein encompass unsolvated forms and solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. Solvated forms of the compounds provided herein are also considered to be disclosed herein. Additionally, the pharmaceutical compositions may contain other medicinal or pharmaceutical agents, carriers, preservatives, stabilizers, wetting or emulsifying agents, solution promoters, adjuvants such as salts for regulating osmotic pressure, buffers, and / or therapeutically valuable substances.

[0110] Methods for preparing compositions containing the compounds described herein include formulating the compounds with one or more inert pharmaceutically acceptable excipients or carriers to form solid, semi-solid, or liquid forms. 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. The pharmaceutical compositions described herein may be in the form of liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid before use, or 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. In some embodiments, pharmaceutical compositions comprising at least one compound of structure (I) are illustratively in the form of a liquid in which the drug is present in solution, suspension, or both. Typically, when the composition is administered as a solution or suspension, a first portion of the drug is present in solution, and a second portion of the drug is present in particulate form in suspension in a liquid matrix. In some embodiments, the liquid composition comprises a gel formulation. In other embodiments, the liquid composition is aqueous.

[0111] In certain embodiments, useful aqueous suspensions contain one or more polymers as suspending agents. Useful polymers include water-soluble polymers such as cellulose polymers, e.g., hydroxypropylmethylcellulose, and water-insoluble polymers such as cross-linked carboxyl-containing polymers. Some pharmaceutical compositions described herein include a mucoadhesive polymer 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 to 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 nonionic surfactants, such as polysorbate 80, are useful as solubilizing agents, as are ophthalmically acceptable glycols, polyglycols, such as polyethylene glycol 400, and glycol ethers.

[0112] 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, sodium borate, 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 amounts necessary to maintain the pH of the composition within an acceptable range. Further useful compositions may also contain one or more salts in an amount necessary 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. 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.

[0113] Still other useful compositions contain one or more surfactants to enhance 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 alkylphenyl ethers, such as octoxynol 10 and octoxynol 40. Still other useful compositions optionally include one or more antioxidants to enhance chemical stability. Suitable antioxidants include, by way of example only, ascorbic acid and sodium metabisulfite. In certain embodiments, aqueous suspension compositions are packaged in single-dose non-reclosable containers. Alternatively, multi-dose reclosable containers are used, in which case it is customary to include a preservative in the composition. 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 semi-transparent matrices of solid hydrophobic polymers containing therapeutic agents. Various 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.

[0114] In certain embodiments, the formulations described herein include one or more antioxidants, metal chelators, thiol-containing compounds, and / or other general stabilizing agents. Examples of such stabilizers include, but are not limited to, (a) about 0.5% to about 2% w / v glycerol, (b) about 0.1% to about 1% w / v methionine, (c) about 0.1% to about 2% w / v monothioglycerol, (d) about 1 mM to about 10 mM EDTA, (e) about 0.01% to about 2% w / v ascorbic acid, (f) 0.003% to about 0.02% w / v polysorbate 80, (g) 0.001% to about 0.05% w / v polysorbate 20, (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.

[0115] In some embodiments, the concentration of one or more compounds provided in the pharmaceutical composition is 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%, 0.09%, 0.08%, 0.07%, 0.06%, 0.18%, 0.19%, 0.20%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.30%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39%, 0.40%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49 ...9%, 0.49%, 0.40%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48 %, 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 less than 0.0001% w / w, w / v or v / v. 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%, 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% w / w, w / v or v / v.

[0116] 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%, from about 0.07% to about 24%, from about 0.08% to about 100%, from about 0.09% to about 120%, from about 0.10% to about 130%, from about 0.11% to about 140%, from about 0.12% to about 150%, from about 0.13% to about 160%, from about 0.14% to about 170%, from about 0.15% to about 180%, from about 0.16% to about 190%, from about 0.17% to about 220%, from about 0.18% to about 230%, from about 0.19% to about 240%, from about 0.19% to about 250%, from about 0.19% to about 26 ...40%, from about 0.20% to about 250%, from about 0.21% to about 250%, from about 0.22% to about 250%, from about 0.23% to about 230%, from about 0.24% to about 240%, from about 0.25% to about 250%, from about 0.26% to The range is 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%, or about 1% to about 10% w / w, w / v, or v / v. In some embodiments, the concentration of the one or more compounds is in the range of about 0.001% to about 10%, about 0.01% to about 5%, about 0.02% to about 4.5%, about 0.03% to about 4%, about 0.04% to about 3.5%, about 0.05% to about 3%, about 0.06% to about 2.5%, about 0.07% to about 2%, about 0.08% to about 1.5%, about 0.09% to about 1%, or about 0.1% to about 0.9% w / w, w / v, or v / v.

[0117] 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.

[0118] 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, 0.003g, 0.004 ... g, 0.0035g, 0.004g, 0.0045g, 0.005g, 0.0055g, 0.006g, 0.0065g, 0.007g, 0.00 75g, 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 to 10 g, 0.0005 to 9 g, 0.001 to 8 g, 0.005 to 7 g, 0.01 to 6 g, 0.05 to 5 g, 0.1 to 4 g, 0.5 to 4 g, or 1 to 3 g.

[0119] Treatment method In still other embodiments, this compound is useful for various methods of treating disease or condition.Therefore, one embodiment provides a method for treating disease, comprising administering to a subject in need thereof a therapeutically effective amount of the compound according to any one of embodiments disclosed herein or the composition according to any one of embodiments disclosed herein, wherein each M is independently an effective biologically active moiety for treating the disease.In some embodiments, the disease is cancer, and each M is independently an anti-cancer drug.

[0120] For example, in certain embodiments, the present disclosure provides methods of treating solid tumors, multiple myeloma, glioma, clear cell renal 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, adenocarcinoma of the pancreas, ovary and / or lung, 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.

[0121] 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 pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate, or derivative thereof.In some embodiments, the method relates to the treatment of acute myeloid leukemia, adolescent cancer, adrenocortical carcinoma of 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 disorder, colon cancer. , colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, extrahepatic bile duct carcinoma in situ (DCIS), embryonal tumors, CNS cancer, endometrial cancer, ependymoma, esophageal cancer, nasal neuroblastoma, Ewing's sarcoma, extracranial germ cell tumors, extragonadal germ cell tumors, eye cancer, fibrous histiocytoma of bone, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors (GIST), germ cell tumors, gestational trophoblastic tumors, hairy cell leukemia, head and neck cancer, heart cancer, liver cancer, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, pancreatic islet cell tumors, pancreatic neuroendocrine tumors, kidney cancer, laryngeal cancer, lip and oral cavity cancer , liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma, metastatic squamous neck cancer of occult primary, midline duct 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 and osteosarcoma of bone, 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, pancreatic cancer, papillomatosis, paraganglioma for the treatment of cancers such as sarcoma, sinus and nasal cavity 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, stomach (gastric) cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, T-cell lymphoma, testicular cancer, laryngeal cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, chorioepithelial tumor, rare cancers of childhood, urethral cancer, uterine sarcoma, vaginal cancer, vulvar cancer or virus-induced cancers.In some embodiments, the methods relate to the treatment of non-cancerous hyperproliferative disorders such as benign hyperplasia of the skin (eg, psoriasis), restenosis, or prostate (eg, benign prostatic hyperplasia (BPH)).

[0122] 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-described compounds (or a pharmaceutical composition comprising the same). In certain embodiments, the lung cancer is non-small cell lung cancer (NSCLC), such as adenocarcinoma, squamous cell lung carcinoma, 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, adenoma, carcinoid tumor, and undifferentiated carcinoma. Thus, in some embodiments of the above-described method, R 2 is a linker that includes a covalent attachment group to a targeting moiety, such as an antibody or 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 folate, or a MET inhibitor. In a further embodiment, the method further comprises inducing apoptosis. Thus, embodiments of the present compounds find use in any number of methods, including but not limited to, drug delivery, quantification of apoptosis, qualifying the delivery of therapeutic agents, quantification of apoptosis, and the diagnosis and treatment of diseases such as hematological cancers.

[0123] In addition to the methods described above, embodiments of the compounds of structure (I) find utility in a variety of fields and methods, including, but not limited to, cancer treatment and imaging, and / or drug delivery, by including targeting moieties in the compounds of structure (I), such as antibodies or sugars or other moieties that preferentially bind to cancer cells. In some embodiments, the method of treatment includes treating a tumor having tumor cells with tumor cell receptors. In some embodiments, the tumor cells have receptors in the range of 1,000-100,000, 1,000-50,000, 1,000-25,000, or 1,000-10,000 receptors per cell. For example, in some embodiments, the tumor cells have about 1,000, about 10,000, or fewer than 100,000 receptors per cell.

[0124] Preparation method

[0023] Embodiments of the compounds of structure (I) above, as well as the variables A, R in the compounds of structure (I) above, 1 , R 2 , R 3 , R 4 , R 5 , L, L 1 , L 2 , L 3 , L 4 It is understood that any specific choices described herein for A, M, m, and / or n may 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. Furthermore, optimal listings may be combined with any particular A, R in a particular embodiment and / or claim. 1 , R 2 , R 3 , R 4 , R 5 , L, L 1 , L 2 , L 3 , L 4 In cases where there are lists of variables M, m and / or n, it is understood that each individual selection may be removed from the scope of a particular embodiment and / or claim, and that the most appropriate remaining list is considered to be within the scope of the present invention. It is understood that in this description, combinations of substituents and / or variables of the depicted formulae are permissible if such contributions result in stable compounds. It will also be appreciated by those skilled in the art that in the processes described herein, functional groups of intermediate compounds may need to be protected by 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" (where 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 Ed., Wiley. As will be appreciated by those skilled in the art, these protecting groups can also be polymeric resins such as Wang resin, Rink resin, or 2-chlorotrityl chloride resin. Additionally, all compounds of the present invention that exist in free base or free 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 present invention can be converted to their free base or acid form by standard techniques.

[0125] The following reaction scheme illustrates an exemplary method for preparing the compounds of the present invention.It will be understood by those skilled in the art that these compounds can be prepared by similar methods or by combining other methods known to those skilled in the art.Methods for preparing the embodiments of the compounds disclosed herein (e.g., structure (I) and (II)) can be found, for example, in PCT Publication No. WO2017 / 173348, the entire contents of which are incorporated herein by reference.It will also be understood by those skilled in the art that by using appropriate starting components and modifying synthetic parameters as necessary, other compounds of structure (I) that are not specifically exemplified below can be prepared by similar methods as described below. Generally, the starting components can be obtained from commercial sources such as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI, and Fluorochem USA, or can be synthesized according to sources 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 herein. The following examples are offered by way of illustration and not by way of limitation. [Example]

[0126] General method 1 H and NMR spectra are obtained on a JEOL 400 MHz spectrometer. 1H spectra are referenced to TMS. Reverse-phase HPLC dye analysis is performed using a Waters Acquity UHPLC system equipped with a 2.1 mm x 50 mm Acquity BEH-C18 column maintained at 45 °C. Mass spectral analysis is performed on a Waters / Micromass Quattro micro MS / system MS (MS mode only) using MassLynx 4.1 acquisition software. The mobile phase used for LC / MS for the dyes is 100 mM 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP), 8.6 mM triethylamine (TEA), pH 8. Phosphoramidite and precursor molecules are analyzed using an Agilent Infinity 1260 UHPLC system equipped with a diode array detector and high-performance autosampler, using an Aapptec © Spirit™ Peptide C18 column (4.6 mm x 100 mm, 5 μm particle size). Excitation and emission profile experiments are recorded on a Cary Eclipse spectrophotometer. All reactions were carried out in oven-dried glassware under a nitrogen atmosphere unless otherwise stated.

[0127] Example 1 Synthesis of ibuprofen-NHS and naproxen-NHS [ka] Ibuprofen-NHS and naproxen-NHS were synthesized using standard coupling conditions. Namely, ibuprofen and naproxen were dissolved separately in dichloromethane, and N,N'-dicyclohexylcarbodiimide (DCC) and N-hydroxysuccinimide (NHS) were added to this mixture. The product was then purified as needed and used in the next synthetic step. Ibuprofen-NHS is easily synthesized in one step on a multigram scale.

[0128] Example 2 Synthesis of ibuprofen polymers [ka] As shown in the reaction sequence above, an exemplary polymer (e.g., with amine functional side chains) is coupled with ibuprofen-NHS. The reaction is carried out using a borate-buffered HO / DMSO mixture containing magnesium chloride. This reaction successfully adds an ibuprofen moiety to each of the amine functional groups to give the desired product.

[0129] Example 3 Synthesis of naproxen polymers [ka] As shown in the reaction sequence above, an exemplary polymer (e.g., with amine functional side chains) is linked to naproxen-NHS. The reaction is carried out using a borate-buffered HO / DMSO mixture containing magnesium chloride. This reaction successfully adds an ibuprofen moiety to each of the amine functional groups to give the desired product.

[0130] Example 4 Synthesis of doxorubicin-NHS [ka] Doxorubicin is reacted with dihydrofuran-2,5-dione to give a carboxylic acid-containing intermediate, which is activated using TFA-NHS and pyridine to give doxorubicin-NHS.

[0131] Example 5 Synthesis of doxorubicin polymers [ka]

[0132] The exemplary polymer is linked to doxorubicin-NHS. The reaction conditions require experimentation using different solvent mixtures due to the limited solubility of the doxorubicin derivative. Alternative reaction conditions include increasing the content of organic solvent, increasing the reaction temperature, and adding sodium dodecyl sulfate (SDS) to support the reaction.

[0133] Example 6 Synthesis of doxorubicin-PEG-azide [ka]

[0134] Doxorubicin-PEG-azide was synthesized on a 50 mg scale according to the reaction sequence shown above. Azide 6-1 was reacted with dihydrofuran-2,5-dione to give intermediate 6-2, which was reacted with perfluorophenol to give intermediate 6-3. Intermediate 6-3 was coupled to doxorubicin to give the desired product, doxorubicin-PEG-azide, in good yield (74%). The presence of the desired product was confirmed by LC-MS.

[0135] Example 7 Synthesis of doxorubicin polymer (compound 18) [ka] An exemplary alkynyl-containing polymer is linked to doxorubicin-PEG-azide. The reaction conditions include CuSO4, tris(3-hydroxypropyltriazolylmethyl)amine (THPTA), and sodium ascorbate. The reaction is carried out in a phosphate buffered aqueous medium containing 60% DMS at pH 7.6. The reaction is carried out at room temperature, and the presence of the desired product is confirmed by LC-MS.

[0136] The embodiments disclosed herein offer numerous advantages, including the ability to control the number of biologically active or fluorescent dye moieties attached to the polymer and subsequently to any targeting moiety. The composition of the polymer backbone can also be selected to achieve desired solubility characteristics, for example, by controlling the incorporation of charged moieties (e.g., number, frequency, spacing, etc.). In addition to the properties provided by the backbone composition, the side chains can be selected to provide a basis for adjusting the solubility of the compounds disclosed herein.

[0137] The embodiments disclosed herein also provide compounds that can advantageously contain multiple therapeutic agents, for example, for complimentary or synergistic therapeutic strategies. Furthermore, 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 with which the polymer-drug constructs can be coupled to targeting agents such as antibodies, antibody fragments, proteins, or other drugs of clinical interest makes them useful for a wide range of interesting applications (e.g., surface chemistry, assay development, etc.).

[0138] The compounds of certain embodiments also achieve other desirable properties, including enhanced permeability and retention. In addition to achieving the necessary solubility, the chemical characteristics of the compound embodiments can be adjusted to modulate the compound's ability to penetrate and be retained within diseased cells / tissues. These characteristics allow for effective delivery of bioactive agents through enhanced permeability and increased efficacy through enhanced retention.

[0139] All U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications, and non-patent publications referred to in this specification, including U.S. Provisional Patent Application No. 62 / 616,636, filed January 12, 2018, are incorporated herein by reference in their entirety to the extent inconsistent with the present disclosure. From the foregoing, it will be appreciated that specific embodiments of the invention have been described herein for purposes of illustration, but that various modifications may be made without departing from the spirit and scope of the invention. Accordingly, the invention is not limited except as by the appended claims. Another aspect of the present invention may be as follows. 〔1〕 A compound having the following structure (I): [ka] (I) or a stereoisomer, pharmaceutically acceptable salt, or tautomer thereof, wherein 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, provided that at least one occurrence of M is not a fluorescent dye; L is a physiologically cleavable linker; L 1 、L 2 、L 3 and L 4 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, -OP(=R a )(R b )R c , Q, L', and R a is O or S, and R b OH, SH, O - 、S - , OR d or SR d and R c OH, SH, O - 、S - , OR d , S.R. d , alkyl, alkoxy, alkyl ether, alkoxyalkyl ether, phosphate, thiophosphate, phosphoalkyl, thiophosphoalkyl, phosphoalkyl ether or thiophosphoalkyl ether, and R d is the counterion, R 4 are independently assigned to each occurrence: OH, SH, O - 、S - , OR d or SR d and R 5 is independently at each occurrence oxo, thioxo, or absent; Q is, independently at each occurrence, a moiety that contains a reactive group capable of forming a covalent bond with an analyte molecule, a solid support, or a complementary reactive group, Q'; L' is, independently at each occurrence, a linker that includes a covalent bond to Q, a targeting moiety, a linker that includes a covalent bond to a targeting moiety, a solid support or a solid support residue, a linker that includes a covalent bond to a solid support or a solid support residue, or a linker that includes a covalent bond to an additional compound of structure (I); m, independently at each occurrence, is 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; where n is an integer greater than or equal to 1. [2] The compound according to [1] above, wherein A is, independently at each occurrence, a moiety containing one or more fused aryl or heteroaryl ring systems. [3] The compound according to [1] above, wherein A is, independently at each occurrence, a moiety containing one or more bicyclic or tricyclic fused aryl or heteroaryl ring systems. [4] A is, independently at each occurrence, the following structure:

change

change

change

[10] above, wherein each occurrence of 〔12〕R 4 However, each occurrence is independent, OH, O - OR d The compound according to any one of the above [1] to

[11] , 〔13〕R 5 is oxo at each occurrence. 〔14〕R 1 The compound according to any one of the above [1] to

[13] , wherein is H. 〔15〕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 compound according to any one of [1] to

[14] above, wherein the other is Q or a linker containing a covalent bond to Q. 〔16〕R 2 and R 3 are each independently OH or -OP(=R a )(R b )R c The compound according to any one of the above [1] to

[14] ,

[17] The compound according to any one of the above [1] to

[15] , wherein Q contains a nucleophilic reactive group, an electrophilic reactive group, or a cycloaddition reactive group.

[18] The compound according to

[17] , wherein Q comprises a sulfhydryl, disulfide, activated ester, isothiocyanate, azide, alkyne, alkene, diene, dienophile, acid halide, sulfonyl halide, phosphine, α-haloamide, biotin, amino, or maleimide functional group.

[19] The compound according to

[18] above, wherein the activated ester is an N-succinimide ester, an imido ester, or a polyfluorophenyl ester.

[20] The compound according to

[18] above, wherein the azide is an alkyl azide or an acyl azide.

[21] The compound according to any one of the above [1] to

[15] , wherein Q is a moiety selected from Table 1. 〔22〕R 2 or R 3 The compound according to any one of [1] to

[14] above, wherein one of the following is L', and L' is a linker comprising a covalent bond to a solid support.

[23] The compound according to

[22] above, wherein the solid support is a polymeric bead or a non-polymeric bead. 〔24〕R 2 or R 3 The compound according to any one of [1] to

[14] above, wherein one of the following is L', and L' is a targeting moiety or a linker to a targeting moiety.

[25] The compound according to

[24] above, wherein L' is a linker to a targeting moiety, and the linker comprises a heteroalkylene linker.

[26] The compound according to any one of [1] to

[25] above, wherein the targeting moiety is an antibody or a cell surface receptor antagonist.

[27] The compound according to

[26] above, 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, a folate or a MET inhibitor.

[28] The compound according to any one of the above [1] to

[27] , wherein n is an integer of 1 to 100.

[29] The compound according to any one of the above [1] to

[28] , wherein n is an integer of 1 to 10.

[30] The compound according to any one of the above [1] to

[29] , wherein m is independently an integer of 1 to 10 at each occurrence.

[31] The compound according to any one of the above [1] to

[30] , wherein m is independently an integer of 1 to 5 at each occurrence.

[32] The compound according to any one of [1] to

[31] above, wherein M is, independently at each occurrence, an NSAID, a kinase inhibitor, an anthracycline, an EGFR inhibitor, or an alkylating agent.

[33] The compound according to any one of [1] to

[32] above, wherein M is, independently at each occurrence, an anticancer drug, and the targeting moiety is an antibody specific to a tumor cell antigen.

[34] The compound according to

[33] above, wherein the tumor cell antigen is EGFR, HER2, folate receptor, CD20, or CD33.

[35] A is, independently at each occurrence, one of the following structures:

change

[36] The compound according to [1] above, selected from Table 2.

[37] A composition comprising the compound according to any one of the above [1] to

[36] and a pharmaceutically acceptable carrier.

[38] A composition comprising a plurality of conjugates comprising the compound of [1] covalently bound to an antibody by a single linking group, wherein the plurality of conjugates have at least 90% structural homogeneity.

[39] The composition described in

[38] above, wherein the multiple conjugates have at least 95% structural homogeneity.

[40] The composition described in

[38] above, wherein the multiple conjugates have a structural homogeneity of greater than 99%. 〔41〕R 2 and R 3 One of them is -OP(=R a )(R b ) OL' or L', wherein L' is an antibody or a linker comprising a covalent bond to the antibody.

[42] A method for treating a disease, comprising the step of administering to a subject in need thereof a therapeutically effective amount of the compound described in any one of [1] to

[36] or the composition described in any one of

[37] to

[41] , wherein each M is independently a biologically active moiety effective for treating the disease.

[43] The method according to

[42] above, wherein the disease is cancer and each M is independently an anticancer drug.

Claims

1. A compound having the following structure (IA): 【Chemistry 1】 (IA) or a stereoisomer, pharmaceutically acceptable salt, or tautomer thereof (In the formula, x 1 , x 2 , x 3 , x 4 , x 5 and x 6 is independently at each occurrence an integer from 0 to 6; A, independently at each occurrence, has the following structure: 【Chemistry 2】 (In the formula, a 1 , a 2 and a 3 is independently at each occurrence a 5-, 6-, or 7-membered carbocyclic or heterocyclic ring; L 6 is a direct bond or a linker as shown below 【Transformation 3】 ） or comprising one or more fused carbocyclic or fused heterocyclic ring systems having one of the following: A is, independently at each occurrence, one of the following structures: 【Chemistry 4】 and M, at each occurrence, is independently a biologically active agent selected from an NSAID and an anti-cancer drug. a moiety, or a fluorescent dye, with the proviso that at least one occurrence of M is not a fluorescent dye. provided that there is no L is cleavable at a pH in the range of 6 to 8 and cleavable at a temperature of 20°C to 40°C. a physiologically cleavable linker that can be cleaved by an enzyme, and wherein L is One of the structures below 【Transformation 5】 【Transformation 6】 【Transformation 7】 【Transformation 8】 or 【Chemistry 9】 wherein R is H, methyl, ethyl, isopropyl, tert-butyl, or phenyl; X is O or CH 2 and n is an integer greater than 0 Including, R 1 is independently at each occurrence H, alkyl, or alkoxy; R 2 and R 3 are each independently OH, —OP(═R a ) (R b ) R c , Q, L', and R a is O or S, and R b OH SH O - , S - , OR d or SR d and R c OH, OL', SH, O - , S - , OR d or SR d and R d is the counterion, R 4 are independently expressed as OH, SH, and O for each occurrence. - , S - , OR d or SR d and R d is the counterion, R 5 is independently at each occurrence oxo, thioxo, or absent; Q is, independently at each occurrence, a sulfhydryl, disulfide, N-succinimide ester, imidoester, polyfluorophenyl ester, isothiocyanate, azide, alkyne, alkene, diene, dienophile, acid halide, sulfonyl halide, phosphine, α-haloamide, biotin, amino, or maleimide functionality, or a moiety selected from Table 1; Table 1 Table 2 Table 3 L', independently at each occurrence, has the structure: 【Chemistry 10】 (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, said targeting moiety being an antibody or a cell surface receptor antagonist; m, independently at each occurrence, is 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; and n is an integer of 1 or greater.

2. x 3 and x 4 are both 2 for each occurrence, or x 1 , x 2 , x 5 and x 6 are 1 for each occurrence, The compound of claim 1.

3. R 4 However, each occurrence is independent, OH, O - or OR d 2. The compound of claim 1, wherein:

4. R 5 2. The compound of claim 1, wherein each occurrence of is oxo.

5. R 1 The compound of claim 1 , wherein is H.

6. 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 Q, or R 2 and R 3 are each independently OH or —OP(═R a ) (R b ) R c That is, The compound of claim 1.

7. R 2 or R 3 is L' and the targeting moiety is selected from the group consisting of an epidermal growth factor receptor (EGFR) inhibitor, a hepatocyte growth factor receptor (HGFR) inhibitor, an insulin-like growth factor receptor (IGFR) inhibitor, a folate, and a MET inhibitor; The compound of claim 1.

8. n is an integer from 1 to 100, n is an integer from 1 to 10, m is, independently at each occurrence, an integer from 1 to 10; or m is independently at each occurrence an integer from 1 to 5; The compound of claim 1.

9. the targeting moiety is an antibody specific for a tumor cell antigen, the tumor cell antigen being EGFR, HER2, folate receptor, CD20, or CD33; The compound of claim 1.

10. Each occurrence of A has the following structure: 【Chemistry 11】 2. The compound of claim 1 having the formula:

11. 2. The compound of claim 1, wherein the compound has one of the following structures: 【Chemistry 12】 【Chemistry 13】 【Chemistry 14】 【Chemistry 15】 【Chemistry 16】 【Chemistry 17】 [Chemistry 18] 【Chemistry 19】 【Chemistry 20】 【Chemistry 21】 【Chemistry 22】 【Chemistry 23】 【Chemistry 24】 【Chemistry 25】 【Chemistry 26】 【Chemistry 27】 【Chemistry 28】 【Chemistry 29】 【Transformation 30】 【Chemistry 31】 【Chemistry 32】 【Transformation 33】 (Wherein N' is 【Transformation 34】 and I′ is 【Chemistry 35】 and dT is 【Transformation 36】 (wherein R is H or a direct bond). It is.)

12. a plurality of molecules of the compound of claim 1 covalently attached to an antibody by a single linking group, said single linking group being the same in all said molecules; the plurality of molecules have at least 90% structural homogeneity; the plurality of molecules have at least 95% structural homogeneity; or the plurality of molecules have greater than 99% structural homogeneity; R 2 and R 3 One of them is -OP(=R a ) (R b ) OL' or L', wherein the targeting moiety of L', L", is an antibody as described above; composition.

13. 13. The composition of claim 12 for use in a method of treating cancer, the method comprising administering to a subject in need thereof a therapeutically effective amount of the composition, wherein each M is independently an anti-cancer drug.

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