Polymeric compounds comprising anti-tumor agents

Abstract

A water-soluble polymeric compound is disclosed. The compounds comprise: i) one or more antineoplastic agents; ii) a heteroalkylidene backbone comprising a group negatively charged at physiological pH; iii) one or more linear or branched linkers comprising a self-cleaving group and optionally a spacer group, the linkers linking each anti-tumor agent to the heteroalkylene backbone; and iv) a reactive group capable of forming a covalent bond with the targeting moiety. The water-soluble polymeric compounds have a variety of uses, including as therapeutic agents for various therapeutic methods.

Description

Background Technical Field

[0002] Embodiments of the present disclosure generally relate to polymeric bioactive compounds having anti-tumor agents, and methods for their preparation.

[0003] Description of Related Art

[0004] Unlike, for example, chemotherapy, targeted drug conjugates deliver drugs to target cells with little or no off-target activity. Generally, a targeted drug conjugate comprises a targeting molecule linked to a bioactive payload or drug. By combining unique targeting capabilities with the therapeutic efficacy of a bioactive drug (or moiety), the conjugate can deliver the drug only to the intended target and minimize potential side effects. Antibody-drug conjugates (ADCs, antibody-conjugated drugs) are a class of targeted drug conjugates of particular interest for treating various diseases. Although ADCs are becoming increasingly popular for cancer treatment, other uses of ADCs are still in their infancy.

[0005] Accordingly, there is a need in the art to develop effective, targeted drug conjugates with high therapeutic indices and methods for their preparation that are not limited to oncology. The present disclosure meets this need and provides further related advantages.

[0006] Brief Summary

[0007] In one embodiment, there is provided a water-soluble polymeric compound comprising: i) one or more anti-tumor agents; ii) a heteroalkyl backbone comprising groups that are negatively charged at physiological pH; iii) one or more linear or branched linkers comprising self-cleaving groups and optionally spacer groups, the linker connecting each anti-tumor agent to the heteroalkyl backbone; and iv) a reactive group capable of forming a covalent bond with a targeting moiety.

[0008] In another embodiment, there is provided a polymeric compound having the following structure (I):

[0009]

[0010] or a stereoisomer, tautomer, or salt thereof, wherein R 1 、R 2 、R 3 、R 4 、R 5 、L 1 、L 2 、L 3 、L 4 、L 5 、M 1 、M 2, p, m, and n are as defined herein. The compounds of structure (I) can be used in a variety of applications, including as therapeutic agents for various treatment methods.

[0011] In another embodiment, a conjugate is provided that comprises a compound of structure (I) covalently bound to a targeting moiety via a bond to a reactive group.

[0012] In another embodiment, a composition is provided that comprises a compound of structure (I) and a pharmaceutically acceptable carrier or a conjugate comprising a compound of structure (I) and a pharmaceutically acceptable carrier.

[0013] In yet another embodiment, a method of treating a disease is provided that comprises 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 2 is independently an anti-tumor agent effective in treating the disease.

[0014] These and other aspects of the disclosure will be apparent after reference to the following detailed description.

[0015] Detailed Description

[0016] In the following description, certain specific details are set forth to provide a thorough understanding of the various embodiments of the disclosure. However, one skilled in the art will understand that the disclosure may be practiced without these details.

[0017] Unless the context otherwise requires, throughout the specification and claims, the word "comprise" and its variants, such as "comprises" and "comprising," are to be construed in an open, inclusive sense, i.e., "including but not limited to."

[0018] References to "one embodiment" or "an embodiment" in this specification mean that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the disclosure. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" throughout the specification are not necessarily all referring to the same embodiment. Moreover, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0019] "Amino" means the -NH2 group.

[0020] "Carboxyl" means the -CO2H group.

[0021] "Cyano" means the -CN group.

[0022] "Formyl" means a -C(=O)H group.

[0023] "Hydroxy" or "hydroxyl" means a -OH group.

[0024] "Imino" means an =NH group.

[0025] "Nitro" means a -NO2 group.

[0026] "Oxo" means an =O substituent.

[0027] "Mercapto" means a -SH group.

[0028] "Thioxo" means an =S group.

[0029] "Alkyl" means a straight-chain or branched-chain hydrocarbon chain group consisting only of carbon and hydrogen atoms, containing no unsaturation, having 1 - 12 carbon atoms (C1 - C 12 alkyl), 1 - 8 carbon atoms (C1 - C8 alkyl) or 1 - 6 carbon atoms (C1 - C6 alkyl), and which is attached to the remainder of the molecule by a single bond, for example, methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (tert-butyl), 3-methylhexyl, 2-methylhexyl, etc. Unless specifically stated otherwise in the specification, the alkyl is optionally substituted.

[0030] "Alkylene" or "alkylene chain" means a straight-chain or branched-chain divalent hydrocarbon chain that connects the remainder of the molecule to a residue, which consists only of carbon and hydrogen, contains no unsaturation, and has 1 - 12 carbon atoms, for example, methylene, ethylene, propylene, n-butylene, etc. The alkylene chain is attached to the remainder of the molecule by a single bond and to the residue by a single bond. The points of attachment of the alkylene chain to the remainder of the molecule and to the residue can be through one carbon or any two carbons within the chain. Unless specifically stated otherwise in the specification, the alkylene is optionally substituted.

[0031] "Alkenylene" or "alkenylene chain" means a straight-chain or branched-chain divalent hydrocarbon chain that connects the remainder of the molecule to a residue, which consists only of carbon and hydrogen, contains at least one carbon-carbon double bond and has 2 - 12 carbon atoms, for example, vinylidene, propenylene, n-butenylene, etc. The alkenylene chain is attached to the remainder of the molecule by a single bond and to the residue by a double bond or a single bond. The points of attachment of the alkenylene chain to the remainder of the molecule and to the residue can be through one carbon or any two carbons within the chain. Unless specifically stated otherwise in the specification, the alkenylene is optionally substituted.

[0032] "Alkynylene" or "alkynylene chain" means a straight or branched divalent hydrocarbon chain that connects the remainder of the molecule to the residue, consisting only of carbon and hydrogen, containing at least one carbon-carbon triple bond and having 2-12 carbon atoms. For example, ethynylene, propynylene, n-butynylene, etc. The alkynylene chain is connected to the remainder of the molecule by a single bond and to the residue by a double bond or a single bond. The connection points of the alkynylene chain to the remainder of the molecule and to the residue can be through one carbon or any two carbons within the chain. Unless specifically stated otherwise in the specification, the alkynylene is optionally substituted.

[0033] "Alkyl ether" means any alkyl as defined above, in which at least one carbon-carbon bond is replaced by a carbon-oxygen bond. The carbon-oxygen bond can be at the end (as in an alkoxy group), or the carbon-oxygen bond can be internal (i.e., C-O-C). The alkyl ether includes at least one carbon-oxygen bond, but can include more than one. For example, polyethylene glycol (PEG) is included within the meaning of alkyl ether. Unless specifically stated otherwise in the specification, the alkyl ether group is optionally substituted. For example, in certain embodiments, the alkyl ether is substituted by an alcohol or -OP(=R a )(R b )R c , where each of R a , R b and R c is defined as for the compounds of structures (I)-(ID).

[0034] "Alkoxy" means a group of the formula -OR a , where R a is an alkyl as defined above containing 1-12 carbon atoms. Unless specifically stated otherwise in the specification, the alkoxy is optionally substituted.

[0035] "Alkoxyalkyl ether" means a group of the formula -OR a R b , where R a is an alkylene as defined above containing 1-12 carbon atoms, and R b is an alkyl ether group as defined herein. Unless specifically stated otherwise in the specification, the alkoxyalkyl ether group is optionally substituted, for example by an alcohol or -OP(=R a )(R b )R c , where each of R a , R b and R c is defined as for the compounds of structures (I)-(ID).

[0036] "Heteroalkyl" means an alkyl group as defined above that contains at least one heteroatom (e.g., N, O, P, or S) within the alkyl group or at the terminus of the alkyl group. In certain embodiments, the heteroatom is within the alkyl group (i.e., the heteroalkyl contains at least one carbon-[heteroatom] x -carbon bond, where x is 1, 2, or 3). In other embodiments, the heteroatom is at the terminus of the alkyl group and is thus used to attach the alkyl group to the remainder of the molecule (e.g., M1-H-A), where M1 is part of the molecule, H is the heteroatom, and A is the alkyl group). Unless specifically stated otherwise in the specification, heteroalkyl is optionally substituted. Exemplary heteroalkyls include oxyethylene (e.g., polyoxyethylene), optionally including a phosphorus-oxygen bond, such as a phosphodiester bond.

[0037] "Heteroalkoxy" means a group of the formula -OR a wherein R a is a heteroalkyl group as defined above containing 1 to 12 carbon atoms. Unless specifically stated otherwise in the specification, heteroalkoxy is optionally substituted.

[0038] "Heteroalkylene" means an alkylene group as defined above that contains at least one heteroatom (e.g., Si, N, O, P, or S) within the alkylene chain or at the terminus of the alkylene chain. In certain embodiments, the heteroatom is within the alkylene chain (i.e., the heteroalkylene contains at least one carbon-[heteroatom]-carbon bond, where x is 1, 2, or 3). In other embodiments, the heteroatom is at the terminus of the alkylene group and is thus used to attach the alkylene group to the remainder of the molecule (e.g., M 1 -H-A-M 2 where M 1 and M 2 are parts of the molecule, H is the heteroatom, and A is the alkylene group). Unless specifically stated otherwise in the specification, heteroalkylene is optionally substituted. Exemplary heteroalkylenes include oxyethylene (e.g., polyoxyethylene) and the "C", "HEG", and "PEG 1K" linker groups explained below:

[0039]

[0040] Polymers of the above C-linker, HEG linker, and / or PEG 1K linker are included in different embodiments of the heteroalkylene linker. In certain embodiments of the PEG 1K linker, n ranges from 19 to 25, for example, n is 19, 20, 21, 22, 23, 24, or 25. The polymer can comprise, for example, the following structures:

[0041]

[0042] where x is an integer of 0 or greater than 0. For example, x ranges from 0 to 100 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10).

[0043] "Sub-alkyl backbone" refers to the following structure:

[0044]

[0045] "Linker" refers to a continuous chain of at least one atom, such as carbon, oxygen, nitrogen, sulfur, phosphorus, and combinations thereof, which connects one part of a molecule to another part of the same molecule or to a different molecule, part, or solid support (e.g., microparticle). The linker can connect the molecules via covalent bonds or other means, such as ionic bond or hydrogen bond interaction. In certain embodiments, the linker is a heteroatom linker (e.g., containing 1 - 10 Si, N, O, P, or S atoms), sub-alkyl (e.g., containing 1 - 10 Si, N, O, P, or S atoms and an alkyl chain), or alkyl linker (e.g., containing 1 - 12 carbon atoms). In certain embodiments, the sub-alkyl linker contains the following structure:

[0046]

[0047] where:

[0048] x 9 and x 10 are each independently an integer greater than 0. In certain embodiments, the heteroatom linker is -O-, -S-, or -OP(=O)O - -O-. In certain embodiments, the sub-alkyl linker contains -OP(=O)O - -O-. In certain embodiments, the sub-alkyl linker contains at least one S - S bond.

[0049] "Physiologically cleavable linker" refers to a molecular connection that can be split or separated in a defined manner in the presence of an in vivo or in vitro environment of an organism or cell system, resulting in two or more separated molecules. Generally, the physiological conditions that induce such cleavage or breakage events can include a temperature in the range of about 20 to 40 °C, an atmospheric pressure of about 1 atm (101 kPa or 14.7 psi), a pH of about 4 to 8, a glucose concentration of about 1 to 20 mM, an atmospheric oxygen concentration, and earth gravity. In certain embodiments, the physiological conditions include enzymatic conditions (i.e., enzymatic cleavage). For example, cathepsin is a protease that cleaves proteins at low pH. Cathepsins include cathepsin A, cathepsin B, cathepsin D, cathepsin K, and cathepsin V. The bond cleavage or breakage can be homolytic or heterolytic.

[0050] "Physiological pH" refers to a temperature in the range of about 20 to 40 °C, an atmospheric pressure of about 1 atm (101 kPa or 14.7 psi), and a pH range of about 4 to 8.

[0051] "Heteroalkenyl" is a heteroalkyl group as defined above that contains at least one carbon-carbon double bond. Unless otherwise specifically stated in the specification, heteroalkenyl is optionally substituted.

[0052] "Heteroalkynyl" is a heteroalkyl group that contains at least one carbon-carbon triple bond. Unless otherwise specifically stated in the specification, heteroalkynyl is optionally substituted.

[0053] With respect to "heteroatom linker", "heteroatom" refers to a linker group composed of one or more heteroatoms. Exemplary heteroatom linkers include single atoms and multiple heteroatoms selected from O, N, P, and S, such as linkers and polymers having the formula -P(O-)(=O)O- or -OP(O-)(=O)O- and combinations thereof.

[0054] "Phosphate / ester" refers to the -OP(=O)(R a )R b group, where R a is OH, O-, or OR c ; and R b is OH, O-, OR c , a thiophosphate / ester group, or other phosphate / ester group, where R c is a counterion (e.g., Na + , etc.).

[0055] "Phosphoalkyl" refers to the -OP(=O)(R a )R b group, where R a is OH, O-, or OR c ; and R b is -Oalkyl, where R c is a counterion (e.g., Na + , etc.). Unless otherwise specifically stated in the specification, phosphoalkyl is optionally substituted. For example, in certain embodiments, the -Oalkyl portion of phosphoalkyl is optionally substituted with one or more of a hydroxyl group, an amino group, a mercapto group, a phosphate / ester, a thiophosphate / ester, a phosphoalkyl, a thiophosphoalkyl, a phosphoalkyl ether, a thiophosphoalkyl ether, or -OP(=R a )(R b )R c , where R a , R b and R cEach is defined as a compound regarding structures (I)-(ID).

[0056] "Phosphoryl alkyl ether" means -OP(=O)(R a )R b group, where R a is OH, O-, or OR c ; and R b is -O alkyl ether, where R c is a counterion (e.g., Na + etc.). Unless specifically stated otherwise in the specification, the phosphoryl alkyl ether group is optionally substituted. For example, in certain embodiments, the -O alkyl ether moiety in the phosphoryl alkyl ether group is optionally substituted by a hydroxyl group, an amino group, a mercapto group, a phosphate / ester, a thiophosphate / ester, a phosphoryl alkyl, a thiophosphoryl alkyl, a phosphoryl alkyl ether, a thiophosphoryl alkyl ether, or -OP(=R a )(R b )R c in which each of R a , R b and R c is defined as a compound regarding structures (I)-(ID).

[0057] "Thiophosphate / ester" means -OP(=R a )(R b )R c group, where R a is O or S, R b is OH, O-, S-, OR d or SR d ; and R c is OH, SH, O-, S-, OR d , SR d , a phosphate / ester group or another thiophosphate / ester group, where R d is a counterion (e.g., Na + etc.) and the conditions are: i) R a is S; ii) R b is S- or SR d ; iii) R c is SH, S- or SR d ; or iv) a combination of i), ii) and / or iii).

[0058] "Thiophosphoryl alkyl" means -OP(=R a )(R b )R c group, where R a is O or S, R bis OH, O−, S−, OR d or SR d ; and R c is -O alkyl, where R d is a counterion (e.g., Na + etc.) and the conditions are: i) R a is S; ii) R b is S− or SR d ; or iii) R a is S and R b is S− or SR d . Unless specifically stated otherwise in the specification, the thiophosphoryl alkyl group is optionally substituted. For example, in certain embodiments, the -O alkyl portion in the thiophosphoryl alkyl is optionally substituted by one or more of hydroxyl, amino, mercapto, phosphate / ester, thiophosphate / ester, phosphoryl alkyl, thiophosphoryl alkyl, phosphoryl alkyl ether, thiophosphoryl alkyl ether, or -OP(=R a )(R b )R c , where each of R a , R b and R c is defined as for the compounds of structures (I)-(ID).

[0059] "Thiophosphoryl alkyl ether" means the -OP(=R a )(R b )R c group, where R a is O or S, R b is OH, O−, S−, OR d or SR d ; and R c is -O alkyl ether, where R d is a counterion (e.g., Na + etc.) and the conditions are: i) R a is S; ii) R b is S− or SR d ; or iii) R a is S and R b is S− or SR d . Unless specifically stated otherwise in the specification, the thiophosphoryl alkyl ether group is optionally substituted. For example, in certain embodiments, the -O alkyl ether portion in the thiophosphoryl alkyl is optionally substituted by one or more of hydroxyl, amino, mercapto, phosphate / ester, thiophosphate / ester, phosphoryl alkyl, thiophosphoryl alkyl, phosphoryl alkyl ether, thiophosphoryl alkyl ether, or -OP(=R a )(R b )R c , where Ra , R b and R c each as defined for the compounds of structures (I)-(ID).

[0060] "Carbocyclic ring" means a stable 3- to 18-membered aromatic or non-aromatic ring containing 3 to 18 carbon atoms. Unless specifically stated otherwise in the specification, the carbocyclic ring can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which can include fused or bridged ring systems, and can be partially or fully saturated. Non-aromatic carbocyclic residues include cycloalkyl, while aromatic carbocyclic residues include aryl. Unless specifically stated otherwise in the specification, the carbocyclic group is optionally substituted.

[0061] "Cycloalkyl" means a stable non-aromatic monocyclic or polycyclic carbocyclic ring, which can include fused or bridged ring systems, having 3 to 15 carbon atoms, preferably 3 to 10 carbon atoms, and which is saturated or unsaturated and linked to the remainder of the molecule by a single bond. Monocyclic cycloalkyl includes, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. Polycyclic cycloalkyl includes, for example, adamantyl, norbornyl, decahydronaphthyl, 7,7-dimethyl-bicyclo-[2.2.1]heptanyl, etc. Unless specifically stated otherwise in the specification, the cycloalkyl is optionally substituted.

[0062] "Aryl" means a ring system containing at least one carbocyclic aromatic ring. In certain embodiments, aryl contains 6 to 18 carbon atoms. The aryl ring can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which can include fused or bridged ring systems. Aryl includes but is not limited to aryl derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, fluoranthene, fluorene, asymmetric indacene, symmetric indacene, indan, indene, naphthalene, phenalene, phenanthrene, heptacene, pyrene and triphenylene. Unless specifically stated otherwise in the specification, the aryl is optionally substituted.

[0063] "Heterocycle" means a stable 3- to 18-membered aromatic or non-aromatic ring containing 1 to 12 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specifically stated in the specification, the heterocycle 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 heterocycle can be optionally oxidized; the nitrogen atoms can be optionally quaternized; and the heterocycle can be partially or completely saturated. Examples of aromatic heterocycles are listed below in the definition of heteroaryl (i.e., heteroaryl is a subset of heterocycles). Examples of non-aromatic heterocycles include, but are not limited to, dioxolanyl, thienyl[1,3]dithiacyclohexyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidinonyl, pyrrolidinyl, pyrazolidinyl, pyrazolopyrimidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trioxanyl, trithiacyclohexyl, triazinyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless otherwise specifically stated in the specification, the heterocyclic group is optionally substituted.

[0064] "Heteroaryl" means a 5- to 14-membered ring system containing 1 to 13 carbon atoms, 1 to 6 heteroatoms selected from nitrogen, oxygen, and sulfur, and at least 1 aromatic ring. For the purposes of certain embodiments of the present disclosure, a heteroaryl residue 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 residue can be optionally oxidized; the nitrogen atoms can be optionally quaternized. Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxenyl, chromenyl, chromenone, benzofuranyl, benzofuranone, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, benzoxazolinonyl, benzimidazosulfenyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanone, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolinyl, indazinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1-oxido(pyridyl), 1-oxido(pyrimidinyl), 1-oxido(pyrazinyl), 1-oxido(pyridazinyl), 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, pteridinone, purinyl, pyrrolyl, pyrazolyl, pyridyl, pyridone, pyrazinyl, pyrimidinyl, pyrimidinonyl, pyridazinyl, pyrrolyl, pyrido[2,3-d]pyrimidinone, quinazolinyl, quinazolinone, quinoxalinyl, quinoxalinone, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, thieno[3,2-d]pyrimidin-4-one, thieno[2,3-d]pyrimidin-4-one, triazolyl, tetrazolyl, triazinyl, and thienyl (i.e., thiophenyl). Unless specifically stated otherwise in the specification, heteroaryl is optionally substituted.

[0065] The suffix "-ene" denotes a specific structural feature (e.g., alkyl, aryl, heteroalkyl, heteroaryl) that is connected to the remainder of the molecule by a single bond and to the residue by a single bond. In other words, the suffix "-ene" denotes a linker having the structural features of the moiety to which it is attached. The points of attachment of the "-ene" chain to the remainder of the molecule and to the residue can be through one atom or any two atoms within the chain. For example, heteroarylene denotes a linker containing a heteroaryl moiety as defined herein.

[0066] "Fused" means a ring system comprising at least two rings, wherein the two rings share at least one common ring atom, e.g., two common ring atoms. When the fused ring is a heterocyclyl ring or a heteroaryl ring, the common ring atoms may be carbon or nitrogen. Fused rings include bicyclic, tricyclic, tetracyclic, etc.

[0067] As used herein, the term "substituted" refers to any of the above groups (e.g., alkyl, alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene, alkoxy, alkyl ether, phosphate alkyl, phosphate alkyl ether, thiophosphoroalkyl, thiophosphoroalkyl ether, carbocyclic, cycloalkyl, aryl, heterocyclic and / or heteroaryl) wherein at least one hydrogen atom (e.g., 1, 2, 3 or all of the hydrogen atoms) is replaced by a bond to a non-hydrogen atom, such as, but not limited to, a halogen atom such as F, Cl, Br and I; oxygen atoms in groups such as hydroxyl, alkoxy 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, dialkylarylsilyl, alkyldiarylsilyl and triarylsilyl; and other heteroatoms in various other groups. "Substitution" also refers to any of the above groups in which one or more hydrogen atoms are replaced by a higher-order bond (e.g., a double bond or a triple bond) with a heteroatom, such as oxygen in oxo groups, carbonyl groups, carboxyl groups and ester groups; and nitrogen in groups such as imines, oximes, hydrazones and nitriles. For example, "substitution" includes any of the above groups in which one or more hydrogen atoms are replaced by -NR g R h 、-NR g C(=O)R h 、-NR g C(=O)NR g R h 、-NR g C(=O)OR h 、-NR g S02R 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 hSubstitution. "Substituted" also refers to any of the above groups in which one or more hydrogen atoms are replaced by -C(=O)R g 、-C(=O)OR g 、-C(=O)NR g R h 、-CH2SO2R g 、-CH2SO2NR g R h Substitution. In the foregoing, R g and R h are the same or different and are independently hydrogen, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclic group, N - heterocyclic group, heterocyclic group alkyl, 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 with the following groups: amino, cyano, hydroxy, imino, nitro, oxo, thio, halogen, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclic group, N - heterocyclic group, heterocyclic group alkyl, heteroaryl, N - heteroaryl and / or heteroarylalkyl. Additionally, each of the foregoing substituents may also optionally be substituted by one or more of the above substituents.

[0068] "Conjugated" means the overlap of one p - orbital with another p - orbital across an inserted σ - bond. Conjugation can occur in cyclic or acyclic compounds. "Degree of conjugation" means the overlap of at least one p - orbital with another p - orbital across an inserted σ - bond. For example, 1,3 - butadiene has one degree of conjugation, while benzene and other aromatic compounds typically have multiple degrees of conjugation. Fluorescent and colored compounds typically contain at least one degree of conjugation.

[0069] "Fluorescent" means a molecule that can absorb light of a specific frequency and emit light of a different frequency. Fluorescence is well known to those of ordinary skill in the art.

[0070] "Colored" means a molecule that absorbs light within the colored spectrum (i.e., red, yellow, blue, etc.).

[0071] The term "biomolecule" refers to any of a variety of biological materials, including nucleic acids, carbohydrates, amino acids, polypeptides, glycoproteins, hormones, aptamers, and mixtures thereof. More specifically, the term is intended to include, but is not limited to, RNA, DNA, oligonucleotides, modified or derivatized nucleotides, enzymes, receptors, prions, receptor ligands (including hormones), antibodies, antigens, and toxins, as well as bacteria, viruses, blood cells, and tissue cells. As further described herein, the visually detectable biomolecules of the present disclosure (e.g., compounds having structure (I) with a biomolecule attached thereto) are prepared by contacting a biomolecule with a compound having a reactive group capable of attaching the biomolecule to the compound via any available atom or functional group (such as an amino, hydroxyl, carboxyl, or thiol group on the biomolecule).

[0072] A "reactive group" is a moiety capable of reacting with a second reactive group (e.g., a "complementary reactive group") to form one or more covalent bonds, e.g., by substitution, oxidation, reduction, addition, or cycloaddition reactions. Exemplary reactive groups are provided in Table A and include, for example, nucleophiles, electrophiles, dienes, dienophiles, aldehydes, oximes, hydrazones, alkynes, amines, azides, acyl azides, acyl halides, nitriles, nitrones, thiols, disulfides, sulfonyl halides, isothiocyanates, imidates, activated esters, ketones, α,β-unsaturated carbonyls, alkenes, maleimides, α-haloimides, epoxides, aziridines, tetrazines, tetrazoles, phosphines, biotin, thietanes, etc.

[0073] "Bioconjugation" or "bioconjugate" and related variants denote a chemical reaction strategy for forming a stable covalent bond between two molecules. The term "bioconjugation" is typically used when one of the molecules is a biomolecule (e.g., an antibody), but can be used to describe the formation of a covalent bond with a non-biological molecule (e.g., a polymer resin). The product or compound resulting from such a reaction strategy is a "conjugate", "bioconjugate", or grammatical equivalent.

[0074] The terms "visible" and "visually detectable" are used herein to denote a substance that is observable by visual inspection without prior illumination or chemical or enzymatic activation. Such a visually detectable substance absorbs and emits light in a spectral region ranging from about 300 to about 900 nm. Preferably, such a substance has a strong color, preferably having a molar extinction coefficient of at least about 40,000 M -1 cm -1 , more preferably at least about 50,000 M -1 cm -1 , more preferably at least about 60,000 M -1 cm -1 , 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 compounds of the present disclosure can be detected by visual observation or with the aid of an optically based detection device, including but not limited to an absorption spectrophotometer, a transmission optical microscope, a digital camera, and a scanner. Visually detectable substances are not limited to those that emit and / or absorb light in the visible spectrum. Substances that emit and / or absorb light in the ultraviolet (UV) region (from about 10 nm to about 400 nm), the infrared (IR) region (from about 700 nm to about 1 mm), and substances that emit and / or absorb in other electromagnetic spectrum regions are also included within the scope of "visually detectable" substances.

[0075] For the purposes of the embodiments of the present disclosure, the term "light-stable visible dye" refers to a chemical moiety that is visually detectable as defined above and that does not undergo significant change or decomposition upon exposure to light. Preferably, after exposure to light for at least 1 hour, the light-stable visible dye does not exhibit significant decolorization or decomposition. More preferably, the visible dye is stable after exposure to light for at least 12 hours, more preferably at least 24 hours, more preferably at least 1 week, and most preferably at least 1 month. Non-limiting examples of light-stable visible dyes suitable for the compounds and methods of the present disclosure include azo dyes, thioindigo dyes, quinacridone pigments, dioxazines, phthalocyanines, perinones, diketopyrrolopyrroles, quinophthalones, and truarycarboniums.

[0076] As used herein, the term "perylene derivative" is intended to include any substituted perylene that is visually detectable. However, the term is not intended to include perylene itself. The terms "anthracene derivative", "naphthalene derivative", and "pyrene derivative" are used similarly. In certain preferred embodiments, the derivative (e.g., a perylene, pyrene, anthracene, or naphthalene derivative) is an imide, diimide, or hydrazamimide derivative of perylene, anthracene, naphthalene, or pyrene.

[0077] The visually detectable molecules of the various embodiments of the present disclosure can be used in a variety of analytical applications, such as biochemical and biomedical applications, where the presence, location, or amount of a specific analyte (e.g., a biomolecule) needs to be determined. Thus, in another aspect, the present disclosure provides a method for visually detecting a biomolecule, which includes: (a) providing a visually detectable biomolecule to a biological system, the biomolecule comprising a compound of structure (I) linked to the biomolecule; and (b) detecting the biomolecule by its visual properties. For the purposes of the present disclosure, the phrase "detecting the biomolecule by its visual properties" means observing the biomolecule with the naked eye or with the aid of an optical detection device, without illumination or chemical or enzymatic activation, the optical detection device including but not limited to an absorption spectrophotometer, a transmission optical microscope, a digital camera, and a scanner. A densitometer can be used to quantify the amount of the visually detectable biomolecule present. For example, by measuring the relative optical density, the relative amounts of biomolecules in two samples can be determined. If the stoichiometry of the dye molecule for each biomolecule is known and the extinction coefficient of the dye molecule is known, then the absolute concentration of the biomolecule can also be determined from the measured value of the optical density. The term "biological system" as used herein is used to denote any solution or mixture that contains one or more biomolecules in addition to the visually detectable biomolecule. Non-limiting examples of such biological systems include cells, cell extracts, tissue samples, electrophoretic gels, assay mixtures, and hybridization reaction mixtures.

[0078] "Antineoplastic agent" or "antineoplastic drug" refers to any molecule that inhibits the formation of neoplasms, where the growth of the neoplasm may become cancerous. Antineoplastic agents can include azacitidine, capecitabine, carmofur, cladribine, clofarabine, cytarabine, decitabine, floxuridine, fludarabine, fluorouracil, gemcitabine, mercaptopurine, nelarabine, pentostatin, tegafur, thioguanine, methotrexate, pemetrexed, raltitrexed, hydroxyurea, irinotecan, topotecan, daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone, valrubicin, etoposide, teniposide, cabazitaxel, docetaxel, paclitaxel, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, bendamustine, busulfan, carmustine, chlorambucil, mechlorethamine, cyclophosphamide, dacarbazine, fotemustine, ifosfamide, lomustine, melphalan, streptozocin, temozolomide, carboplatin, cisplatin, nedaplatin, oxaliplatin, hexamethylmelamine, bleomycin, bortezomib, actinomycin D (dacinomycin), estramustine, ixabepilone, mitomycin, or procarbazine. In certain embodiments, the antineoplastic agent includes monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), paclitaxel, camptothecin, doxorubicin, and PF-06380101.

[0079] "Self-cleaving group" means a group that undergoes an electron cascade that results in the release of the group to which it is attached. In certain embodiments, the self-cleaving group comprises one or more groups that can undergo 1,4-elimination, 1,6-elimination, 1,8-elimination, 1,6-cycloelimination, 1,5-cycloelimination, 1,3-cycloelimination, intramolecular 5-exo-trig cyclization, and / or 6-exo-trig cyclization. In certain embodiments, the self-cleaving group comprises valine (Val) and citrulline (Cit) or valine (Val) and alanine (Ala).

[0080] "Spacer group" means a linker covalently bonded between the self-cleaving group and the anti-tumor agent. In one embodiment, the spacer group is para-aminobenzyl carbamate (PBA).

[0081] "Solid support" or "solid resin" means any solid matrix known in the art for solid-phase support of molecules, e.g., "particulate" means any of a number of small particles that can be used to attach to the compounds of the present disclosure, including but not limited to glass beads, magnetic beads, polymer beads, non-polymer beads, etc. In certain embodiments, the particulate comprises polystyrene beads. In certain embodiments, the solid support or solid resin is controlled pore glass or macroporous polystyrene.

[0082] "Solid support residue" means a functional group that remains attached to the molecule when the molecule is cleaved from the solid support. Solid support residues are known in the art and can be readily deduced based on the structure of the solid support and the group to which the molecule is attached.

[0083] "Targeting moiety" is a moiety that selectively binds or associates with a specific target, such as an analyte molecule. "Selectively" binding or associating means that the targeting moiety preferentially associates or binds to the desired target relative to other targets. In certain embodiments, the compounds disclosed herein include a linkage to a targeting moiety for the purpose of selectively binding or associating the compound with a target analyte (i.e., the target of the targeting moiety), thereby allowing detection of the analyte. Exemplary targeting moieties include but are not limited to antibodies, antigens, nucleic acid sequences, enzymes, proteins, cell surface receptor antagonists, etc. A targeting moiety is a moiety (such as an antibody) that selectively binds or associates with a target feature on or in a cell, such as a target feature on the cell membrane or other cell structure, thereby allowing detection of the cell of interest. In certain embodiments, small molecules that selectively bind or associate with the desired analyte are also contemplated as targeting moieties. Those skilled in the art will appreciate other analytes and corresponding targeting moieties that will be useful in the various embodiments.

[0084] "Base pairing moiety" means a heterocyclic moiety capable of hybridizing to a complementary heterocyclic moiety by hydrogen bonding (e.g., Watson-Crick base pairing). Base pairing moieties include natural bases 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.

[0085] Embodiments of the disclosure herein are also intended to cover all compounds isotopically labeled by replacing one or more atoms with atoms having different atomic masses or mass numbers. Examples of isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, such as 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 I.

[0086] Isotopically labeled compounds of structure (I) can generally be prepared by conventional techniques known to those skilled in the art or by methods similar to those described below and in the methods of the following examples, using appropriate isotopically labeled reagents in place of the previously used unlabeled reagents.

[0087] "Stable compound" and "stable structure" are intended to indicate a compound that is sufficiently robust to survive isolation from the reaction mixture to a useful purity and formulation into an effective therapeutic agent.

[0088] "Optional" or "optionally" means that the subsequent described event or circumstance may or may not occur, and the description includes instances where the event or circumstance occurs and instances where it does not occur. For example, "optionally substituted alkyl" means that the alkyl may or may not be substituted, and the description includes substituted alkyls and unsubstituted alkyls.

[0089] "Salt" includes acid and base addition salts.

[0090] "Acid addition salts" refer to those salts formed with inorganic acids (such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.) and organic acids (such as, 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, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclohexanesulfamic acid, dodecyl sulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-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, undecylenic acid, etc.).

[0091] "Base addition salts" refer to those salts prepared by adding an inorganic base or an organic base to a free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, etc. Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, dimethylethanolamine (deanol), 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, phenethylbenzylamine, dibenzylethylenediamine, ethylenediamine, glucosamine, methylglucosamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine.

[0092] Crystallization can produce solvates of the compounds described herein. Embodiments of the present disclosure include all solvates of the described compounds. As used herein, the term "solvate" refers to an aggregate comprising one or more molecules of a compound of the present disclosure and one or more solvent molecules. The solvent can be water, in which case the solvate can be a hydrate. Alternatively, the solvent can be an organic solvent. Thus, the compounds of the present disclosure can exist as hydrates, including monohydrates, dihydrates, hemihydrates, sesquihydrates, trihydrates, tetrahydrates, etc., as well as the corresponding solvated forms. The compounds of the present disclosure can be true solvates, while in other cases, the compounds of the present disclosure can merely retain adventitious water or another solvent or a mixture of water plus some adventitious solvent.

[0093] Embodiments of the compounds of the present disclosure (e.g., compounds of Structure I) or their salts, tautomers, or solvates can contain one or more stereocenters and can thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that can be defined in terms of absolute stereochemistry as (R)- or (S)-, or for amino acids as (D)- or (L)-. Embodiments of the present disclosure are intended to include all such possible isomers, as well as their racemic and optically pure forms. The optically active (+) and (-), (R)- and (S)-, or (D)- and (L)-isomers can be prepared using chiral synthons or chiral reagents, or the optically active (+) and (-), (R)- and (S)-, or (D)- and (L)-isomers can be resolved using conventional techniques (e.g., chromatography and fractional crystallization). Conventional techniques for the preparation / separation of the 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 performance liquid chromatography (HPLC). When the compounds described herein contain alkenyl double bonds or other geometrically asymmetric centers, and unless otherwise specified, the compounds are intended to include E and Z geometric isomers. Likewise, all tautomeric forms are also intended to be included.

[0094] "Stereoisomers" refer to compounds that are composed of the same atoms bonded by the same bonds but have different three-dimensional structures that are not interchangeable. The present disclosure encompasses various stereoisomers and mixtures thereof, and includes "enantiomers", which refer to two stereoisomers whose molecules are non-superimposable mirror images of each other.

[0095] "Tautomers" refer to the transfer of a proton from one atom of a molecule to another atom of the same molecule. The present disclosure includes tautomers of any of the said compounds. Those of ordinary skill in the art can readily deduce the various tautomeric forms of a compound.

[0096] The chemical nomenclature and structural diagrams used herein are a modified form of the I.U.P.A.C. nomenclature system, using the ACD / Name version 9.07 software program and / or the ChemDraw Ultra version 11.0 software naming program (CambridgeSoft). Common names familiar to those of ordinary skill in the art are also used.

[0097] Compound

[0098] As noted above, in one embodiment of the present disclosure, a water-soluble polymeric compound is provided that can be used as a covalent linker between a bioactive moiety (such as an anti-tumor agent) and a targeting moiety. Thus, in certain embodiments, M 1 is independently, at each occurrence, a moiety comprising a fluorescent or colored dye; M 2 is independently, at each occurrence, a moiety comprising an anti-tumor agent. In certain embodiments, M 2 is monomethyl auristatin E and M 1 is a fluorescent dye (e.g., fluorescein, etc.).

[0099] The embodiments disclosed herein provide numerous advantages, including the ability to control: the number and type of anti-tumor agents M attached to the heteroalkyl main chain of the polymeric compound and any subsequent targeting moiety, the spacing between adjacent anti-tumor agents on the heteroalkyl main chain (e.g., the degree of proximity of the anti-tumor agent M), and the spacing between the heteroalkyl main chain and the anti-tumor agent (e.g., the length of the linker outside the polymer main chain). 2 The bioactive moiety can be attached to the heteroalkyl main chain via a straight-chain or branched linker. The procedures described in the present disclosure provide the ability to selectively install straight-chain or branched linkers. This allows the synthesis of compounds having both straight-chain and / or branched linkers, the linkers carrying one or more anti-tumor agents. In this regard, the anti-tumor agents can be sequentially cleaved according to physiological conditions. Additionally, compounds having both multiple anti-tumor agents and fluorescent moieties linked by a linker can be synthesized. 2 The embodiments of the present disclosure provide a combination of an anti-tumor agent, a targeting moiety, and a dye moiety (e.g., a chromophore or fluorophore) that can be used for simultaneous targeting, treatment, and detection. The ease of coupling a polymer-drug construct to a targeting reagent (such as an antibody, antibody fragment, protein, or other clinically interesting reagent) provides utility for a variety of applications of interest (such as surface chemistry, assay development, etc.). Thus, in certain embodiments, M

[0100] is a fluorophore (e.g., FITC, 5-FAM, 6-FAM, etc.) or a colored moiety.

[0101] The embodiments of the present disclosure provide a combination of an anti-tumor agent, a targeting moiety, and a dye moiety (e.g., a chromophore or fluorophore) that can be used for simultaneous targeting, treatment, and detection. The ease of coupling a polymer-drug construct to a targeting reagent (such as an antibody, antibody fragment, protein, or other clinically interesting reagent) provides utility for a variety of applications of interest (such as surface chemistry, assay development, etc.). Thus, in certain embodiments, M 1 is a fluorophore (e.g., FITC, 5-FAM, 6-FAM, etc.) or a colored moiety.

[0102] Compounds of certain embodiments also provide other desirable properties, including enhanced permeability and retention effects. In addition to providing the necessary solubility properties, the chemical characteristics of embodiments of the compounds of the present invention can be adjusted to modulate the ability of the compounds to penetrate diseased cells / tissues and remain inside them. These characteristics allow for the effective delivery of anti-tumor agents by increasing potency through increased penetration and enhanced retention.

[0103] Therefore, it should be understood that any embodiment of the compounds of structure (I) as described above can be independently combined with other embodiments to form embodiments of the present disclosure not specifically set forth above. It should be understood that in this specification, combinations of substituents and / or variables of the described formulas are permitted only if such combinations result in stable compounds.

[0104] Thus, one embodiment provides a water-soluble polymeric compound comprising: i) one or more anti-tumor agents; ii) a heteroalkyl backbone comprising groups that are negatively charged at physiological pH; iii) one or more linear or branched linkers comprising self-cleaving groups and optional spacer groups that link each anti-tumor agent to the heteroalkyl backbone; and iv) a reactive group capable of forming a covalent bond with a targeting moiety. In other embodiments, the polymeric compound further comprises i) at least one fluorescent or colored moiety; and ii) one or more linkers that link the fluorescent or colored moiety to the heteroalkyl backbone.

[0105] In one embodiment, the polymeric compound has the following structure (I):

[0106]

[0107] or a stereoisomer or tautomer thereof, wherein

[0108] M 1 is independently, each time it appears, a moiety comprising a fluorescent or colored moiety;

[0109] M 2 is independently, each time it appears, a moiety comprising an anti-tumor agent;

[0110] L 1 is independently, each time it appears, a linear or branched linker;

[0111] L 2 and L 3 is independently, each time it appears, an optional alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene or heteroatom linker;

[0112] L 4is independently a heteroalkyl linker each time it appears;

[0113] L 5 is independently a linker each time it appears;

[0114] R 1 is independently H, alkyl, or alkoxy each time it appears;

[0115] R 2 and R 3 are each independently H, OH, SH, -OP(=R a )(R b )R c or Q;

[0116] R 4 is independently OH, SH, O-, S-, OR d or SR d ;

[0117] R 5 is independently oxo, thioxo, or absent each time it appears;

[0118] R a is O or S;

[0119] R b is OH, SH, O - , S - , OR d or SR d ;

[0120] R c is OH, SH, O - , S - , OR d , OL′, SR d alkyl, alkoxy, heteroalkyl, heteroalkoxy, alkyl ether, alkoxyalkyl ether, phosphate / ester, thiophosphate / ester, phosphoalkyl, thiophosphoalkyl, phosphoalkyl ether, or thiophosphoalkyl ether;

[0121] R d is a counterion;

[0122] Q is independently a moiety containing a reactive group or a protected form thereof that is capable of forming a covalent bond with a complementary reactive group Q′ on a targeting moiety each time it appears;

[0123] L′ is, independently at each occurrence, a linker containing a covalent bond to Q, a targeting moiety, a linker containing a covalent bond to the targeting moiety, a linker containing a covalent bond to the solid support, a linker containing a covalent bond to a solid support residue, a linker containing a covalent bond to a nucleoside, or a linker containing a covalent bond to another compound of structure (I);

[0124] m is, independently at each occurrence, an integer from 1 to 5;

[0125] p is an integer of 0 or greater; and

[0126] n is an integer of 1 or greater.

[0127] The various linkers and substituents (e.g., R 1 , R 2 , R 3 , R 4 , R 5 , L 1 , L 2 , L 3 , L 4 , L 5 , M 1 , M 2 and Q) in the compounds of structure (I) are optionally substituted with one or more substituents. For example, in certain embodiments, the optional substituents are selected to optimize the water solubility or other properties of the compounds of structure (I). In certain embodiments, each chromophore, alkyl group, alkoxy group, alkyl ether, heteroarylene group, heteroalkyl group, alkylene group, alkenylene group, alkynylene group, heteroalkylene group, heteroalkenylene group, heteroalkynylene group, alkoxyalkyl ether, phosphoalkyl group, thiophosphoalkyl group, phosphoalkyl ether, and thiophosphoalkyl ether in the compounds of structure (I) is optionally substituted with one or more substituents selected from hydroxy, alkoxy, alkyl ether, alkoxyalkyl ether, mercapto, amino, alkylamino, carboxy, phosphate / ester, thiophosphate / ester, phosphoalkyl group, thiophosphoalkyl group, phosphoalkyl ether, and thiophosphoalkyl ether. In certain embodiments, the optional substituent is -OP(=R a )(R b )R c , where R a , R b and R c are as defined for the compounds of structure (I).

[0128] In certain embodiments, at least one fluorescent or colored moiety M 1 is a fluorescent moiety. In certain embodiments, the fluorescent or colored moiety M 1 is, independently at each occurrence, selected from pyrene, perylene, perylene monoimide, and 6-FAM moieties. In other embodiments, the fluorescent or colored moiety M1 independently has one of the following structures each time it appears:

[0129]

[0130]

[0131] wherein indicates the connection point to the fluorescent compound.

[0132] In certain specific embodiments, the fluorescent or colored moiety M 1 has one of the following structures:

[0133]

[0134] In certain other specific embodiments, wherein the fluorescent or colored moiety M 1 has the following structure:

[0135]

[0136] In certain embodiments, the polymeric compound comprises 1 - 10 fluorescent or colored moieties. In certain other embodiments, the polymeric compound comprises 1 - 5 fluorescent or colored moieties. In certain specific embodiments, the polymeric compound comprises 1 fluorescent or colored moiety.

[0137] In certain embodiments, the anti - tumor agent M 2 is independently selected from paclitaxel, camptothecin, doxorubicin, monomethyl auristatin E, monomethyl auristatin F, and PF - 06380101 each time it appears. In certain embodiments, the anti - tumor agent M 2 independently has one of the following structures each time it appears:

[0138]

[0139] In certain specific embodiments, the anti - tumor agent M 2 is monomethyl auristatin E. In certain more specific embodiments, the anti - tumor agent M 2 has the following structure:

[0140]

[0141] In certain embodiments, the polymeric compound comprises 1 to 20 anti-tumor agents. In certain other embodiments, the polymeric compound comprises 1 to 10 anti-tumor agents. In certain specific embodiments, the polymeric compound comprises 1 anti-tumor agent. In certain other specific embodiments, the polymeric compound comprises 3 anti-tumor agents. In certain other specific embodiments, the polymeric compound comprises 5 anti-tumor agents. In certain other specific embodiments, the polymeric compound comprises 7 anti-tumor agents. In certain other specific embodiments, the polymeric compound comprises 10 anti-tumor agents.

[0142] In certain embodiments, the heteroalkyl linker L 4 comprises one or more alkylene or oxyalkylene moieties. In certain more specific embodiments, L 4 has oxyethylene as the oxyalkylene. In certain more specific embodiments, the oxyethylene is polyoxyethylene. In certain embodiments, L 4 has the following structure each time it appears:

[0143]

[0144] wherein:

[0145] z is an integer from 1 to 100; and

[0146] * indicates a bond to an adjacent phosphorus atom.

[0147] In certain embodiments, z is an integer from 3 to 8. In certain embodiments, z is 3. In certain embodiments, z is 4. In certain embodiments, z is 5. In certain embodiments, z is 6. In certain embodiments, z is 7. In certain embodiments, z is 8.

[0148] In certain other embodiments, z is an integer from 22 to 26. In certain embodiments, z is 22. In certain embodiments, z is 23. In certain embodiments, z is 24. In certain embodiments, z is 25. In certain embodiments, z is 26.

[0149] In certain other embodiments, z ranges from 19 to 28. In certain embodiments, the average z is 23. In certain embodiments, the average z is 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28.

[0150] In certain embodiments, at least one occurrence of the compound of structure (I) comprises one of the following structures:

[0151]

[0152] where z is an integer from 1 to 100. In certain embodiments, z is an integer from 3 to 8. In certain other embodiments, z is an integer from 22 to 26. In certain other embodiments, the range of z is from 19 to 28.

[0153] In certain embodiments, each occurrence of in the compound of structure (I) comprises one of the following structures:

[0154]

[0155] In certain specific embodiments, the heteroalkyl linker L 4 comprises one of the following structures:

[0156]

[0157] In certain more specific embodiments, where the heteroalkyl linker L 4 comprises the following structure:

[0158]

[0159] In certain other specific embodiments, the heteroalkyl linker L 4 comprises the following structure:

[0160]

[0161] In certain embodiments, R 4 is O - or S - . In certain other embodiments, R 4 is O - . In certain embodiments, at least one occurrence of R 4 is O - . In certain embodiments, at least one occurrence of R 4 is S - . In certain embodiments, each occurrence of R 4 is O - .

[0162] In certain embodiments, R 5 is oxo or thioxo. In certain embodiments, at least one occurrence of R 5 is oxo (=O). In certain embodiments, at least one occurrence of R 5 is thioxo (=S). In certain embodiments, each occurrence of R 5 is oxo (=O).

[0163] Certain embodiments provide compounds having the following structure (IA) or (IB):

[0164]

[0165] In certain embodiments, L 2 and L 3 are each independently, upon each occurrence, i) absent or ii) C1-C6 alkylene, C2-C6 alkenylene or C2-C6 alkynylene.

[0166] Certain embodiments provide compounds having the following structure (IC) or (ID):

[0167]

[0168] wherein x 1 、x 2 、x 3 and x 4 are each independently, upon each occurrence, an integer from 0 to 6.

[0169] In certain embodiments, x 1 、x 2 、x 3 and x 4 are, upon each occurrence, the integer 0 (i.e., L 5 is a direct bond). In certain embodiments, x 1 、x 2 、x 3 and x 4 are, upon each occurrence, the integer 1. In certain embodiments, x 1 、x 2 、x 3 and x 4 are, upon each occurrence, the integer 2. In certain embodiments, x 1 、x 2 、x 3 and x 4 are, upon each occurrence, the integer 3. In certain embodiments, x 1 、x 2 、x 3 and x 4 are, upon each occurrence, the integer 4. In certain embodiments, x 1 、x 2 、x 3 and x 4 are, upon each occurrence, the integer 5. In certain embodiments, x 1 、x 2 、x 3 and x 4 are, upon each occurrence, the integer 6. In certain more specific embodiments, x1 , x 2 , x 3 and x 4 is each 0 or 1 upon each occurrence.

[0170] In certain embodiments, the self-cleaving group comprises two or more amino acids. In certain embodiments, the two amino acids are selected from valine, citrulline, glycine, and alanine.

[0171] In certain embodiments, the self-cleaving group comprises one of the following structures:

[0172]

[0173]

[0174] In certain embodiments, the self-cleaving group comprises one of the following structures (or a combination thereof):

[0175]

[0176]

[0177] In certain embodiments, the self-cleaving group has the following structure:

[0178]

[0179]

[0180] The optional spacer group can serve as the attachment point for M 2 and each anti-tumor agent. For example, in certain embodiments, synthetic precursors of compounds of structures (I)-(ID) are prepared, and the M 2 moiety is attached to the synthetic precursor using any number of coupling methods known in the art. In further embodiments, at least one occurrence of the optional spacer group comprises an amide bond, an ester bond, an ether bond, or a combination thereof. In other embodiments, the functional groups of the optional spacer group comprise amide, ester, ether, or a combination thereof.

[0181] In certain embodiments, the optional spacer group has one of the following structures:

[0182]

[0183] In certain specific embodiments, the optional spacer group has the following structure:

[0184]

[0185] Linker L 5 can serve as M1 The point of attachment of the moiety to the remainder of the compound (i.e., the heteroalkyl backbone). For example, in certain embodiments, a synthetic precursor of a compound of structure (I) is prepared and the moiety M is attached to the synthetic precursor using any number of coupling methods known in the art. 1 In more embodiments, the moiety L 5 contains, in at least one occurrence, an amide bond, an ester bond, a phosphodiester bond, a disulfide bond, a double bond, a triple bond, an ether bond, a hydrazone, an amino acid sequence containing one or more amino acid residues, a ketone, a diol, a cyano group, a nitro group, or a combination thereof. In other embodiments, the functional group contains an amide or an ester. In more specific embodiments, the moiety L 5 contains, in at least one occurrence, one of the following structures:

[0186]

[0187]

[0188] In more specific embodiments, each occurrence of the linker L 5 contains one of the following structures:

[0189]

[0190] In certain more specific embodiments, the linker L 5 has the following structure:

[0191]

[0192] In certain embodiments, at least one occurrence of R 1 is H. In certain embodiments, at least one occurrence of R 1 is alkyl. In certain embodiments, at least one occurrence of R 1 is alkoxy. In certain embodiments, each occurrence of R 1 is H.

[0193] In various other embodiments, R 2 and R 3 are each independently OH or -OP(=R a )(R b )R c . In certain different embodiments, R 2 or R 3 is OH or -OP(=R a )(R b )R c , and the other of R 2 or R 3 is Q or a linker containing a covalent bond to Q.

[0194] In more different embodiments of any of the compounds of the foregoing structures (I)-(ID), R 2 and R 3 are each independently -OP(=R a )(R b )R c . In some of these embodiments, R c is OL'.

[0195] In other embodiments, R 2 and R 3 are each independently -OP(=R a )(R b )OL', and L' is an alkylene or heteroalkylene linker to the following groups: Q, targeting moiety, analyte (e.g., analyte molecule), solid support, solid support residue, nucleoside, or other compound of structures (I)-(ID).

[0196] The linker L' can be any linker suitable for linking Q, targeting moiety, analyte (e.g., analyte molecule), solid support, solid support residue, nucleoside, or other compound of structures (I)-(ID) to the compound of structures (I)-(ID). Advantageously, certain embodiments include using a selected L' moiety to increase or optimize the water solubility of the compound. In certain embodiments, L' is a heteroalkylene moiety. In certain other specific embodiments, L' comprises an oxyalkylene or phosphodiester moiety or a combination thereof.

[0197] In certain other embodiments, R 2 or R 3 is L'. For example, in certain embodiments, L' is a linker to the targeting moiety. In another example, in certain embodiments, R 2 or R 3 is -NH2. In other embodiments, one of R 2 or R 3 is L' and L' is a linker comprising a covalent bond to the solid support. In certain embodiments, the solid support is a polymeric bead or a non-polymeric bead.

[0198] In certain specific embodiments, L' is a linker to the targeting moiety, the linker comprising an oxyalkylene or phosphodiester moiety or a combination thereof. In other embodiments, L' has one of the following structures:

[0199]

[0200] Wherein:

[0201] x 1 、x2 and x 3 and x 4 and x 5 and x 6 and x 7 and x 8 are independently integers from 1 to 10;

[0202] R b is H, an electron pair or a counterion; and

[0203] L″ is a targeting moiety or a linker to a targeting moiety.

[0204] In certain embodiments, x 1 and x 2 and x 3 and x 4 and x 5 and x 6 and x 7 and x 8 are independently the integer 1. In certain embodiments, x 1 and x 2 and x 3 and x 4 and x 5 and x 6 and x 7 and x 8 are independently the integer 2. In certain embodiments, x 1 and x 2 and x 3 and x 4 and x 5 and x 6 and x 7 and x 8 are independently the integer 3. In certain embodiments, x 1 and x 2 and x 3 and x 4 and x 5 and x 6 and x 7 and x 8 are independently the integer 4. In certain embodiments, x 1 and x 2 and x 3 and x 4 and x 5 and x 6 and x 7 and x 8 are independently the integer 5. In certain embodiments, x 1 and x 2 and x 3 and x 4 and x 5 and x6 , x 7 and x 8 is independently the integer 6. In certain embodiments, x 1 , x 2 , x 3 , x 4 , x 5 , x 6 , x 7 and x 8 is independently the integer 7. In certain embodiments, x 1 , x 2 , x 3 , x 4 , x 5 , x 6 , x 7 and x 8 is independently the integer 8. In certain embodiments, x 1 , x 2 , x 3 , x 4 , x 5 , x 6 , x 7 and x 8 is independently the integer 9. In certain embodiments, x 1 , x 2 , x 3 , x 4 , x 5 , x 6 , x 7 and x 8 is independently the integer 10.

[0205] In certain embodiments, R b is H. In certain embodiments, R b is a counterion. For example, in certain embodiments, R b is Na + . In certain embodiments, R b is K + .

[0206] In certain embodiments, L″ is a targeting moiety. In certain other embodiments, L″ is a linker to a targeting moiety. For example, the targeting moiety is an antibody. In another example, the targeting moiety is a cell surface receptor antagonist. In certain more specific 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 MET inhibitor. In other embodiments, the targeting moiety is a monoclonal antibody. For example, in certain embodiments, the monoclonal antibody is abciximab, adalimumab, alemtuzumab, alirocumab, avibactam, basiliximab, benralizumab, belotero solus, lintuzumab, brodalumab, brosimumab, canakinumab, caracizumab, pegsiticase, daclizumab, denosumab, dupilumab, eculizumab, emicizumab,erenumab, evolocumab, rimegepant, galsulizumab, golimumab, guselkumab, ibalizumab, idarucizumab, infliximab, ilotuzumab, icatibant, lanadelumab, logivumab, mepolizumab, natalizumab, otosiliximab, ocrelizumab, omalizumab, pertuzumab, ranibizumab, reslizumab, rezlizumab, Rmab, rovilizumab, rulizumab, sarilumab, secukinumab, telizumab, Thiomab, tocilizumab, ustekinumab, vedolizumab, avelumab, actoxumab, aducanumab, afacizumab, afimoxifene, anifrolumab, anrukinzumab (IMA-638), aselizumab, atorolimumab, bapineuzumab, BCD-100, bavituximab, besilesomab, bispecific monoclonal antibody, bimagrumab, bijinatumumab, bococizumab, braleukinumab, busocizumab, bococizumab, cabalizumab, carotuximab, cilizumab, clazakizumab, crizanlizumab, consortuzumab, cowesimab, CR6261, crizanlizumab, lirentelimab, crotedumab, ditoxizumab, mafodotin, delotuzumab biotin, dizamizumab, dilevalumab, domalizumab, dusigituzumab, emicizumab, epratuzumab, efalizumab, edralizumab, elenumab, enokizumab, eptinezumab, erlizumab, etrolizumab, evesacizumab, eweizumab, fasolizumab, faralimomab, faricimab, fasutinolizumab, fanolizumab, fazanolizumab, flolizumab, flusolizumab, flanotuzumab, gantenerumab, gavrilimomab, gevokizumab, gimsilumab, gonalizumab, ilesimab,Inalizumab, Inomomab, Iomab-B, Keliximab, Lampalizumab, Lanlotuzumab, Lavelizumab, Lirilumab, Lenvervimab, Lodamesumab, Lirilumab, Lirilumab, Ligoctant, Lodisatuximab, Perulizumab, Matuzumab, Mavilimumab, Metelimumab, Migalastat, Moxetumomab, Muromonab-CD3, Nebacumab, Nemolizumab, NEOD001, Nirsevimab, Odulimomab, Olokizumab, OMS721, OPN-305, Otilimomab, Ocrelizumab, Otilizumab, Oselumab, Ozanamab, Olaratumab, Paxalisib, Pabacizumab, Pacritinib, Patritumab, PDR001, Pexelizumab, Pexelizumab, Pralatrexate, Rozanolixizumab, Ponatinib, Puromycin, Puromycin, PRO 140, Quilizumab, Revilimumab, Repotrectinib, Renevimab, Ravagalimab, Reflizumab, Reflizumab, Regorafenib, Relatlimab, Linvoseltinib, Risankizumab, Roledumab, Romosozumab, Loncastuximab, SA237, Siltuximab, Cevimeline, SHP647, Cefazolin, Cimtozumab, Cilengitide, Cilengitide, Solanezumab, Sonelokimab, Spartalizumab, Stattic, Thiosulphate, Sutezolid, Sutilimab, Suvorexant, Sotrastaurin, Talizumab, Talizumab, Tanezumab, Tenecteplase, Teprotumumab, Tezepelumab, Tebipenem, Tocilizumab, Trodelvy, Trogituzumab, Tuvirumab, Ustekinumab, Ustekinumab, Ustekinumab, Varlilumab, Vapendavir, Vesenacumab, Vesiplimab, Vobarilizumab, Azoximer, Trastuzumab, Gemtuzumab, Brentuximab, Vorsetuzumab, Lovotuzumab, Cantuzumab, Bevacizumab or Icatibant or Vadastuximab.,

[0207] In still other more specific embodiments of any of the compounds of structures (I)-(ID) above, R 2 or R 3 has one of the following structures:

[0208]

[0209]

[0210] Certain embodiments of the compounds of structures (I)-(ID) can be prepared according to solid-phase synthesis methods similar to those known in the art for preparing oligonucleotides. Thus, in certain embodiments, L' is a linkage to a solid support, a solid support residue, or a nucleoside. Solid supports containing an activated deoxythymidine (dT) group are readily available and, in certain embodiments, can be used as starting materials for preparing the compounds of structures (I)-(ID). Thus, in certain specific embodiments, R 2 or R 3 has the following structure:

[0211]

[0212]

[0213] Those skilled in the art will understand that including the above dT group is merely for ease of synthesis and economic efficiency and is not essential. Other solid supports can be used and will result in the presence of different nucleosides or solid support residues on L', or the nucleoside or solid support residue can be removed or modified after synthesis.

[0214] In certain embodiments, the analyte molecule is a nucleic acid, an amino acid, or a polymer thereof. In certain embodiments, the analyte molecule is an enzyme, a receptor, a receptor ligand, an antibody, a glycoprotein, an aptamer, or a prion. In certain embodiments, the targeting moiety is an antibody or a cell surface receptor antagonist. In certain other embodiments, the solid support is a polymeric bead or a non-polymeric bead.

[0215] In still other embodiments, Q is independently, each time it appears, a moiety containing a reactive group that is capable of forming a covalent bond with the analyte molecule or the solid support. In other embodiments, Q is independently, each time it appears, a moiety containing a reactive group that is capable of forming a covalent bond with a complementary reactive group Q'. For example, in certain embodiments, Q' is present on another compound of structures (I)-(ID) (e.g., at the R 1 or R 2 position), and Q and Q' contain complementary reactive groups such that the reaction of the compounds of structures (I)-(ID) with other compounds of structures (I)-(ID) produces a dimer of the covalently bound compounds of structures (I)-(ID). Multimeric compounds of structures (I)-(ID) can also be prepared in a similar manner, and the multimeric compounds are included within the scope of the embodiments of the present disclosure.

[0216] The type of the Q group and the connectivity of the Q group to the remainder of the compounds of structures (I)-(ID) are not restricted, provided that Q contains a moiety having suitable reactivity for forming the desired bond.

[0217] In certain embodiments, Q is a moiety that is not readily hydrolyzable under aqueous conditions but has sufficient reactivity to form a bond with an analyte molecule or a corresponding group (e.g., amine, azide, or alkyne) on a solid support.

[0218] Certain embodiments of the compounds of structures (I)-(ID) contain Q groups commonly used in the field of bioconjugation. For example, in certain embodiments, Q contains a nucleophilic reactive group, an electrophilic reactive group, or a cycloaddition reactive group. In certain more specific embodiments, Q contains a thiol group, a disulfide group, an activated ester group, an isothiocyanate group, an azide group, an alkyne group, an alkene group, a diene group, a dienophile group, an acyl halide group, a sulfonyl halide group, a phosphine group, an α-haloamide group, a biotin group, an amino group, or a maleimide functional group. In certain embodiments, the activated ester is an N-succinimidyl ester, an imidate ester, or a polyfluorophenyl ester. In other embodiments, the alkyne is an alkyl azide or an acyl azide.

[0219] The Q group can be conveniently provided in a protected form to increase storage stability or other desired properties, and then the protecting group is removed at an appropriate time for conjugation with, for example, a targeting moiety or an analyte. Thus, the Q group includes "protected forms" of reactive groups, including any of the reactive groups described above and in Table A below. The "protected form" of Q refers to a moiety that has lower reactivity relative to Q under predetermined reaction conditions, but is convertible to Q under conditions that preferably do not degrade or react with other parts of the compounds of structures (I)-(ID). Those skilled in the art can deduce the appropriate protected form of Q based on the specific Q and the desired end use and storage conditions. For example, when Q is SH, the protected form of Q includes a disulfide, which can be reduced using known techniques and reagents to reveal the SH moiety.

[0220] Exemplary Q moieties are provided in Table A below.

[0221] Table A. Exemplary Q moieties

[0222]

[0223]

[0224]

[0225]

[0226] 1In certain embodiments, the electron-withdrawing group is a halogen (e.g., F, Cl, Br, or I), an oxygen-containing group (e.g., -OH or alkoxy), a nitrogen-containing group (e.g., -N(R)2, where R is H or alkyl), a haloalkyl, nitro, ammonium, cyano, carboxyl, -SO3H, etc.

[0227] It should be noted that in certain embodiments where Q is SH, the SH moiety will tend to form a disulfide bond with another thiol group (e.g., on another compound of structures (I)-(ID)). Thus, some embodiments include compounds of structures (I)-(ID) in the form of disulfide dimers, where the disulfide bond is derived from the SH Q group.

[0228] In other embodiments, the Q moiety is conveniently masked (e.g., protected) as a disulfide moiety, which can then be reduced to provide an activated Q moiety for binding to a desired analyte molecule or targeting moiety. For example, the Q moiety can be masked as a disulfide having the following structure:

[0229]

[0230] where R is an optionally substituted alkyl. For example, in certain embodiments, Q is provided as a disulfide moiety having the following structure:

[0231]

[0232] where n is an integer from 1 to 10.

[0233] In certain embodiments, -L 5 -M 1 has the following structure in at least one occurrence:

[0234]

[0235] In certain more specific embodiments, each occurrence of -L 5 -M 1 has the following structure:

[0236]

[0237] In certain embodiments, M 2 is the same in each occurrence; however, importantly, note that each occurrence of M 2 need not be the same M 2 , and some embodiments include compounds where M 2 is not the same in each occurrence. For example, in certain embodiments, each M 2 is not the same, and different Ms are selected 2The part has different anti-tumor agents.

[0238] In certain specific embodiments, the compound is a compound selected from Table 1. The compounds in Table 1 are prepared according to the procedures described in the Examples.

[0239]

[0240]

[0241]

[0242]

[0243]

[0244]

[0245]

[0246]

[0247]

[0248]

[0249]

[0250]

[0251]

[0252]

[0253]

[0254] Wherein FAM represents:

[0255]

[0256] The fluorescence intensity or the potency for treating diseases can also be adjusted by selecting different values of n. In certain embodiments, n is an integer of 1 or greater. In certain embodiments, n is an integer from 1 to 100. In other embodiments, n is an integer from 1 to 20. In other embodiments, n is an integer from 1 to 10. In certain embodiments, n is 1, 3, 5, 7, or 10. In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, n is 3. In certain embodiments, n is 4. In certain embodiments, n is 5. In certain embodiments, n is 6. In certain embodiments, n is 7. In certain embodiments, n is 8. In certain embodiments, n is 9. In certain embodiments, n is 10.

[0257] The fluorescence or the potency for treating diseases can also be adjusted by selecting the value of m. The value of m has the ability to control the spacing between adjacent M 1 and M 2 . In certain embodiments, m is an integer of 0 or greater. In certain embodiments, m is an integer from 0 to 100. In certain embodiments, m is an integer from 0 to 10. In certain embodiments, m is 0. In certain embodiments, m is 1. In certain embodiments, m is 2. In certain embodiments, m is 3. In certain embodiments, m is 4. In certain embodiments, m is 5. In certain embodiments, m is 6. In certain embodiments, m is 7. In certain embodiments, m is 8. In certain embodiments, m is 9. In certain embodiments, m is 10. In certain specific embodiments, m is 1, 2, 3, 4, or 5. The value of m depends on the hydrophobicity of M 1 or M 2 . Hydrophobic M 1 or M 2 may require more spacer units, which increases the value of m. In this regard, less hydrophobic M 1 or M 2 may require fewer spacer units, which decreases the value of m.

[0258] The fluorescence intensity can be adjusted by regulating the number of fluorescent dye moieties attached to the polymer backbone. The value of p has the ability to control the brightness of the compound. In certain embodiments, p is an integer of 0 or greater each time it appears. In certain more specific embodiments, p is from 0 to 10. In certain embodiments, p is from 0 to 5. For example, in certain embodiments, p is 0. In certain embodiments, p is 1. In certain embodiments, p is 2. In certain embodiments, p is 3. In certain embodiments, p is 4. In certain embodiments, p is 5. In certain other embodiments, p is 1.

[0259] The values of n, p, and m are closely related to provide the ability to control fluorescence and effectiveness against diseases. In certain specific embodiments, n is 1; p is 1; and m is 3. In certain specific embodiments, n is 3; p is 1; and m is 3. In certain specific embodiments, n is 5; p is 1; and m is 3. In certain specific embodiments, n is 7; p is 1; and m is 3. In certain specific embodiments, n is 10; p is 1; and m is 3.

[0260] In certain other specific embodiments, n is 1; p is 1; and m is 1. In certain other specific embodiments, n is 3; p is 1; and m is 1. In certain other specific embodiments, n is 5; p is 1; and m is 1. In certain other specific embodiments, n is 7; p is 1; and m is 1. In certain other specific embodiments, n is 10; p is 1; and m is 1.

[0261] Conjugate assembly

[0262] The conjugates of the present disclosure can comprise any antibody of the present disclosure and any linker of the present disclosure in combination therewith and any compound of the present disclosure in combination therewith. In certain embodiments, the conjugates of the present disclosure can be covalently bound to a targeting moiety via a bond to a reactive group. In certain embodiments, the targeting moiety has an affinity for a target analyte. In certain more specific embodiments, the targeting moiety comprises an antibody. In certain embodiments, the antibody comprises trastuzumab, gemtuzumab, brentuximab, vosaroxin, lomvotuzumab, canertinib, bevacizumab, or icatuximab, or vadastuximab. In certain embodiments, the antibody comprises brentuximab. In certain embodiments, the targeting moiety specifically binds brentuximab.

[0263] Pharmaceutical compositions

[0264] One embodiment provides a composition comprising a compound according to any one of the embodiments disclosed herein (e.g., a compound of structures (I)-(ID)) and a pharmaceutically acceptable carrier.

[0265] Other embodiments relate to pharmaceutical compositions. The pharmaceutical compositions comprise any one (or more) of the compounds of structures (I)-(ID) and a pharmaceutically acceptable carrier. In certain embodiments, the pharmaceutical composition is formulated for oral administration. In other embodiments, the pharmaceutical composition is formulated for injection. In further embodiments, the pharmaceutical composition comprises a compound of structures (I)-(ID) and an additional therapeutic agent (e.g., an anti-cancer agent). Non-limiting examples of such therapeutic agents are described below.

[0266] 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. Additionally, by way of example only, parenteral delivery includes intramuscular, subcutaneous, intravenous, intramedullary injections, as well as intrathecal, direct intraventricular, intraperitoneal, intralymphatic, and intranasal injections.

[0267] In certain embodiments, the compounds of structures (I)-(ID) are administered in a local rather than a systemic manner, for example, by direct injection of the compound into an organ, typically in a depot or sustained release formulation. In a specific embodiment, a long-acting formulation is administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Additionally, in other embodiments, in a targeted drug delivery system, such as in liposomes coated with an organ-specific antibody, the drug is delivered. In such embodiments, the liposomes target the organ and are selectively taken up by the organ. In other embodiments, the compounds of structures (I)-(ID) are provided in the form of an immediate release formulation, an extended release formulation, or an intermediate release formulation. In other embodiments, the compounds of structures (I)-(ID) are administered topically.

[0268] The compounds of structures (I)-(ID) are effective over a wide dosage range. For example, in the treatment of adults, doses of 0.01 - 1000 mg, 0.5 - 100 mg, 1 - 50 mg / day, and 5 - 40 mg / day are examples of doses used in certain embodiments. An exemplary dose is 10 - 30 mg / day. The exact dose will depend on the route of administration, the form of the compound being administered, the subject to be treated, the weight of the subject to be treated, and the preference and experience of the attending physician.

[0269] In certain embodiments, the compounds of structures (I)-(ID) are administered in a single dose. Typically, such administration will be by injection, for example, intravenous injection, to rapidly introduce the agent. However, other routes may also be used as appropriate. A single dose of the compounds of structures (I)-(ID) may also be used to treat acute conditions.

[0270] In certain embodiments, the compounds of structures (I)-(ID) are administered in multiple doses. In certain embodiments, the dosing is about once, twice, three times, four times, five times, six times, or more than six times per day. In other embodiments, the dosing is about once per month, once every two weeks, once per week, or once every other day. In another embodiment, the compounds of structures (I)-(ID) and another agent are co-administered about once a day to about six times a day. In another embodiment, the administration of the compounds of structures (I)-(ID) and the agent lasts less than about 7 days. In another embodiment, the administration lasts more than about 6, 10, 14, 28 days, two months, six months, or one year. In some cases, continuous administration can be achieved and maintained as long as needed.

[0271] The administration of the compounds of structures (I)-(ID) can continue as long as needed. In certain embodiments, the compounds of structure (I) are administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In certain embodiments, the compounds of structures (I)-(ID) are administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In certain embodiments, the compounds of structures (I)-(ID) are administered long-term on a continuous basis, such as for the treatment of chronic conditions.

[0272] In certain embodiments, the compounds of structures (I)-(ID) are administered in a dosage. It is known in the art that due to the inter-subject variability in compound pharmacokinetics, individualization of the dosing regimen is necessary for optimal treatment. Given the disclosure of the present invention, the dosage of the compounds of the present disclosure can be found by routine experimentation.

[0273] In certain embodiments, the compounds of structures (I)-(ID) are formulated into pharmaceutical compositions. In specific embodiments, one or more physiologically acceptable carriers, including excipients and adjuvants that facilitate the processing of the active compounds into pharmaceutically acceptable formulations, are used to formulate the pharmaceutical compositions in a conventional manner. Suitable formulations depend on the chosen route of administration. Any pharmaceutically acceptable techniques, carriers, and excipients are suitable for formulating the pharmaceutical compositions described herein: Remington: The Science and Practice of Pharmacy, Nineteenth Edition (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, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Edition (Lippincott Williams & Wilkins 1999).

[0274] The present disclosure provides pharmaceutical compositions that comprise a compound of structures (I)-(ID) and a pharmaceutically acceptable diluent, excipient, or carrier. In certain embodiments, the described compounds are administered as pharmaceutical compositions, wherein the compounds of structures (I)-(ID) are mixed with other active ingredients, such as in combination therapies. All combinations of active substances set forth in the following combination therapies section and throughout the present disclosure are encompassed herein. In specific embodiments, the pharmaceutical composition comprises one or more compounds of structures (I)-(ID).

[0275] The pharmaceutical compositions used herein represent mixtures of compounds of structures (I)-(ID) with other chemical components such as carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents, and / or excipients. In certain embodiments, the pharmaceutical compositions facilitate the administration of the compounds to an organism. In certain embodiments of practicing the therapeutic or use methods provided herein, a therapeutically effective amount of the compounds of structures (I)-(ID) provided herein is administered in a pharmaceutical composition to a mammal suffering from a disease, disorder, or medical condition to be treated. In a specific embodiment, the mammal is a human. In certain embodiments, the therapeutically effective amount varies 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 of structures (I)-(ID) are used alone or in combination with one or more therapeutic agents (as components of a mixture).

[0276] In one embodiment, one or more compounds of structures (I)-(ID) are formulated in an aqueous solution. In a specific embodiment, the aqueous solution is selected from, by way of example only, physiologically compatible buffers such as Hank's solution, Ringer's solution, or saline buffer. In other embodiments, one or more compounds of structures (I)-(ID) are formulated for transmucosal administration. In a specific embodiment, the transmucosal formulation includes a penetrant suitable for the barrier to be penetrated. In other embodiments in which 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.

[0277] In another embodiment, the compounds described herein are formulated for oral administration. The compounds described herein are formulated by combining the active compound with, for example, a pharmaceutically acceptable carrier or excipient. In different embodiments, the compounds described herein are formulated into oral dosage forms that include, by way of example only, tablets, powders, pills, dragees, capsules, liquids, gels, syrups, elixirs, slurries, suspensions, etc.

[0278] In certain embodiments, pharmaceutical compositions for oral use are obtained by mixing one or more solid excipients with one or more of the compounds described herein, optionally grinding the resulting mixture, and processing the granule mixture after adding suitable auxiliaries (if required) to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol or sorbitol; cellulose products such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose; or others such as polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. In specific embodiments, a disintegrant is optionally added. Disintegrants include, by way of example only, croscarmellose sodium, polyvinylpyrrolidone, agar or alginic acid or a salt thereof such as sodium alginate.

[0279] In one embodiment, the dosage forms (such as dragee cores and tablets) are provided with one or more suitable coatings. In specific embodiments, a concentrated sugar solution is used for coating the dosage forms. The sugar solution optionally contains additional components such as, by way of example only, gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol and / or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. Dyes and / or pigments are also optionally added to the coating for identification purposes. Additionally, dyes and / or pigments are optionally used to characterize different combinations of the active compound dosage.

[0280] 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 specific embodiments, the push-fit capsules contain the active ingredient admixed with one or more fillers. Fillers include, by way of example only, lactose, binders such as starch and / or lubricants such as talc or magnesium stearate and optionally 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 are optionally added.

[0281] 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 other embodiments, the compounds described herein are formulated for parenteral injection, including formulations suitable for bolus injection or continuous infusion. In a specific embodiment, the injectable formulation is present in a unit dosage form (e.g., in an ampoule) or a multi-dose container. A preservative is optionally added to the injection formulation. In other embodiments, the pharmaceutical composition is formulated in a form suitable for parenteral injection, such as a sterile suspension, solution or emulsion in an oily or aqueous vehicle. The parenteral injection formulation optionally contains formulating agents such as suspending agents, stabilizers and / or dispersing agents. In a specific embodiment, the pharmaceutical formulation for parenteral administration includes an aqueous solution of the active compound in a water-soluble form. In a further embodiment, a suspension of the active compound (e.g., a compound of structure (I)-(ID)) is prepared as a suitable oily injection suspension. Suitable lipophilic solvents or vehicles for the pharmaceutical compositions described herein include, by way of example only, fatty oils such as sesame oil or synthetic fatty acid esters such as ethyl oleate or triglycerides or liposomes. In certain specific embodiments, the aqueous injection suspension contains a substance that increases the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol or dextran. Optionally, the suspension contains a suitable stabilizer or a reagent that increases the solubility of the compound to allow the preparation of a highly concentrated solution. Alternatively, in other embodiments, the active ingredient is in powder form for formulation with a suitable vehicle (e.g., sterile pyrogen-free water) prior to use.

[0282] In other embodiments, the compounds of structure (I)-(ID) are administered topically. The compounds described herein are formulated into a variety of topical administration compositions, such as solutions, suspensions, lotions, gels, pastes, sticks, balms, creams or ointments. Such pharmaceutical compositions optionally contain solubilizing agents, stabilizers, tonicity enhancers, buffers and preservatives.

[0283] In other embodiments, the compounds of structures (I)-(ID) are formulated for transdermal administration. In specific embodiments, the transdermal formulations employ transdermal delivery devices and transdermal delivery patches and may be lipophilic emulsions or buffered aqueous solutions, dissolved and / or dispersed in a polymer or adhesive. In various embodiments, such patches are configured for continuous, pulsatile or on-demand delivery of a pharmaceutical agent. In additional embodiments, transdermal delivery of the compounds of structures (I)-(ID) is accomplished by means of iontophoresis patches and the like. In certain embodiments, the transdermal patch provides controlled delivery of the compounds of structures (I)-(ID). In specific embodiments, the absorption rate is slowed by using a rate controlling membrane or by trapping the compound within a polymeric matrix or gel. In alternative embodiments, absorption enhancers are used to increase absorption. Absorption enhancers or carriers include pharmaceutically acceptable solvents that are absorbable and assist in permeating the skin. For example, in one embodiment, the transdermal device is in the form of a bandage that includes a backing member, a reservoir containing the compound optionally with a carrier, an optional rate controlling barrier (to deliver the compound to the skin of the host at a controlled and predetermined rate over an extended period of time), and means for securing the device to the skin.

[0284] In other embodiments, the compounds of structures (I)-(ID) are formulated for administration by inhalation. Various forms suitable for administration by inhalation include, but are not limited to, aerosols, mists or powders. Using a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas), a pharmaceutical composition of any compound of structures (I)-(ID) is conveniently delivered in aerosol spray presentation form from a pressurized pack or nebulizer. In specific embodiments, the dosage unit of a pressurized aerosol is determined by providing a valve to deliver a metered amount. In certain embodiments, capsules and cartridges, such as of gelatin (by way of example only), formulated for use in an inhaler or insufflator, contain a powder mixture of the compound and a suitable powder base such as lactose or starch.

[0285] In still other embodiments, the compounds of structures (I)-(ID) are formulated as rectal compositions such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, gelatinous suppositories or retention enemas, which contain conventional suppository bases such as cocoa butter or other glycerides, and synthetic polymers such as polyvinylpyrrolidone, PEG, etc. In the suppository form of the composition, low melting point waxes such as, but not limited to, mixtures of fatty acid glycerides, are optionally combined with melted cocoa butter.

[0286] In certain embodiments, the pharmaceutical compositions are formulated in any conventional manner using one or more physiologically acceptable carriers, which carriers include excipients and auxiliaries that facilitate processing of the active compounds into pharmaceutically acceptable preparations. Suitable formulations depend on the chosen route of administration. Any pharmaceutically acceptable techniques, carriers, and excipients are optionally used as appropriate. Pharmaceutical compositions containing the compounds of structures (I)-(ID) are prepared in a conventional manner, such as, by way of example only, by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or compressing processes.

[0287] The pharmaceutical composition comprises at least one pharmaceutically acceptable carrier, diluent, or excipient and at least one compound of structures (I)-(ID) (described herein as the active ingredient). The active ingredient is in the form of the free acid or free base, or in the form of a pharmaceutically acceptable salt. Additionally, the methods and pharmaceutical compositions described herein include the use of N-oxides, crystal forms (also referred to as polymorphs), and active metabolites of these compounds having the same type of activity. All tautomeric forms of the compounds described herein are included within the scope of the compounds presented herein. Additionally, the compounds described herein encompass the unsolvated form as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, etc. The solvated forms of the compounds presented herein are also considered to be disclosed herein. Furthermore, the pharmaceutical composition optionally includes other drugs or pharmaceutical agents, carriers, adjuvants, such as preservatives, stabilizers, wetting agents or emulsifiers, solubilizing agents, salts for regulating osmotic pressure, buffers, and / or other therapeutically valuable substances.

[0288] 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 a solid, semi-solid, or liquid. Solid compositions include, but are not limited to, powders, tablets, dispersible granules, capsules, cachets, and suppositories. Liquid compositions include solutions in which the compound is dissolved, emulsions containing the compound, 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 forms of the pharmaceutical compositions described herein include liquid solutions or suspensions, solid forms suitable for dissolving or suspending in a liquid prior to use, or as an emulsion. These compositions also optionally contain small amounts of non-toxic auxiliary substances such as wetting agents or emulsifiers, pH buffers, etc.

[0289] In certain embodiments, a pharmaceutical composition comprising a compound of at least one of structures (I)-(ID) exemplary takes a liquid form, wherein the medicament is present in a solution, a suspension, or both. Generally, when the composition is administered as a solution or a suspension, a first portion of the medicament is present in the solution and a second portion of the medicament is suspended in the liquid matrix in particulate form. In certain embodiments, the liquid composition comprises a gel formulation. In other embodiments, the liquid composition is aqueous.

[0290] 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., hydroxypropyl methylcellulose) and water-insoluble polymers such as cross-linked carboxyl-containing polymers. Certain pharmaceutical compositions described herein contain mucoadhesive polymers, such as those selected from carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methyl methacrylate), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate, and dextran.

[0291] Useful pharmaceutical compositions also optionally include solubilizing agents that aid in the dissolution of the compounds of structures (I)-(ID). The term "solubilizing agent" generally includes agents that result in the formation of a micellar solution or a true solution of the reagent. Certain acceptable nonionic surfactants, such as polysorbate 80, can be used as solubilizing agents, and ophthalmically acceptable diols, polyglycols (e.g., polyethylene glycol 400), and diol ethers can also be used as solubilizing agents.

[0292] In addition, useful pharmaceutical compositions optionally include one or more pH regulators or buffers, 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(hydroxymethyl)aminomethane; and buffers such as citrate / dextrose, sodium bicarbonate, and ammonium chloride. Such acids, bases, and buffers are included in the amounts necessary to maintain the pH of the composition within an acceptable range.

[0293] In addition, useful compositions also optionally include one or more salts in the amounts necessary to bring the osmolality of the composition within an acceptable range. Such salts include salts 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.

[0294] Other useful pharmaceutical compositions optionally include one or more preservatives to inhibit microbial activity. Suitable preservatives include mercury-containing substances such as phenylmercuric borate (merfen) and thimerosal; stabilized chlorine dioxide; and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride.

[0295] Still other useful compositions include one or more surfactants to enhance physical stability or for other purposes. Suitable nonionic surfactants include polyoxyethylene fatty acid glycerides and vegetable oils, for example, polyoxyethylene (60) hydrogenated castor oil; and polyoxyethylene alkyl ethers and alkyl phenyl ethers, for example, octoxynol 10, octoxynol 40.

[0296] Still other useful compositions include one or more antioxidants to enhance chemical stability when needed. Suitable antioxidants include, by way of example only, ascorbic acid and sodium metabisulfite.

[0297] In certain embodiments, the aqueous suspension composition is packaged in a single-dose non-reclosable container. Alternatively, a multi-dose reclosable container is used, in which case a preservative is typically included in the composition.

[0298] 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, sustained release systems are used to deliver the compounds described herein, such as a semipermeable matrix of a solid hydrophobic polymer containing a therapeutic agent. A variety of sustained releases are useful herein. In certain embodiments, the sustained release capsules release the compound for weeks up to over 100 days. Depending on the chemical nature and biological stability of the therapeutic agent, additional strategies for protein stabilization are employed.

[0299] In certain embodiments, the formulations described herein contain one or more antioxidants, metal chelators, thiol-containing compounds, and / or other general stabilizers. Examples of such stabilizers include, but are not limited to: (a) about 0.5% to about 2% 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.

[0300] In certain embodiments, the concentration of one or more compounds provided in the pharmaceutical composition is less than 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%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002% or 0.0001% w / w, w / v or v / v.

[0301] In certain embodiments, the concentration of one or more compounds is greater than 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%, 1.25%, 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 0.0001% w / w, w / v or v / v.

[0302] In certain embodiments, the concentration of one or more compounds is in the range of 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 23%, from about 0.09% to about 22%, from about 0.1% to about 21%, from about 0.2% to about 20%, from about 0.3% to about 19%, from about 0.4% to about 18%, from about 0.5% to about 17%, from about 0.6% to about 16%, from about 0.7% to about 15%, from about 0.8% to about 14%, from about 0.9% to about 12%, from about 1% to about 10% w / w, w / v or v / v.

[0303] In certain embodiments, the concentration of one or more compounds is in the range of from about 0.001% to about 10%, from about 0.01% to about 5%, from about 0.02% to about 4.5%, from about 0.03% to about 4%, from about 0.04% to about 3.5%, from about 0.05% to about 3%, from about 0.06% to about 2.5%, from about 0.07% to about 2%, from about 0.08% to about 1.5%, from about 0.09% to about 1%, from about 0.1% to about 0.9% w / w, w / v or v / v.

[0304] In certain embodiments, the amount of one or more compounds is equal to or less than 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, 0.15 g, 0.1 g, 0.09 g, 0.08 g, 0.07 g, 0.06 g, 0.05 g, 0.04 g, 0.03 g, 0.02 g, 0.01 g, 0.009 g, 0.008 g, 0.007 g, 0.006 g, 0.005 g, 0.004 g, 0.003 g, 0.002 g, 0.001 g, 0.0009 g, 0.0008 g, 0.0007 g, 0.0006 g, 0.0005 g, 0.0004 g, 0.0003 g, 0.0002 g or 0.0001 g.

[0305] In certain embodiments, the amount of one or more compounds exceeds 0.0001 g, 0.0002 g, 0.0003 g, 0.0004 g, 0.0005 g, 0.0006 g, 0.0007 g, 0.0008 g, 0.0009 g, 0.001 g, 0.0015 g, 0.002 g, 0.0025 g, 0.003 g, 0.0035 g, 0.004 g, 0.0045 g, 0.005 g, 0.0055 g, 0.006 g, 0.0065 g, 0.007 g, 0.0075 g, 0.008 g, 0.0085 g, 0.009 g, 0.0095 g, 0.01 g, 0.015 g, 0.02 g, 0.025 g, 0.03 g, 0.035 g, 0.04 g, 0.045 g, 0.05 g, 0.055 g, 0.06 g, 0.065 g, 0.07 g, 0.075 g, 0.08 g, 0.085 g, 0.09 g, 0.095 g, 0.1 g, 0.15 g, 0.2 g, 0.25 g, 0.3 g, 0.35 g, 0.4 g, 0.45 g, 0.5 g, 0.55 g, 0.6 g, 0.65 g, 0.7 g, 0.75 g, 0.8 g, 0.85 g, 0.9 g, 0.95 g, 1 g, 1.5 g, 2 g, 2.5 g, 3 g, 3.5 g, 4 g, 4.5 g, 5 g, 5.5 g, 6 g, 6.5 g, 7 g, 7.5 g, 8 g, 8.5 g, 9 g, 9.5 g or 10 g.

[0306] In certain embodiments, the amount of one or more compounds ranges from 0.0001 - 10 g, 0.0005 - 9 g, 0.001 - 8 g, 0.005 - 7 g, 0.01 - 6 g, 0.05 - 5 g, 0.1 - 4 g, 0.5 - 4 g or 1 - 3 g.

[0307] Treatment method

[0308] Certain compounds of the present disclosure can be used to treat diseases (i.e., compounds of structures (I)-(ID)). Those compounds disclosed herein provide a targeting scheme for drug delivery strategies. Accordingly, one embodiment provides a method of treating a disease (or its symptoms), the method comprising administering to a mammal (e.g., a human) in need thereof a therapeutically effective amount of a compound of structures (I)-(ID).

[0309] For example, in certain embodiments, the present disclosure provides a method of treating an autoimmune disease such as rheumatoid arthritis, juvenile idiopathic arthritis, psoriatic arthritis, ankylosing spondylitis, adult Crohn's disease, pediatric Crohn's disease, ulcerative colitis, plaque psoriasis, hidradenitis suppurativa, uveitis, Behçet's disease, spondyloarthropathy, psoriasis or osteoarthritis.

[0310] Other therapeutic agents that can be combined with the compounds of the present disclosure can be found in Goodman and Gilman's “The Pharmacological Basis of Therapeutics” (Tenth Edition, edited by Hardman, Limbird, and Gilman) or the Physician’s Desk Reference, both of which are incorporated herein by reference in their entirety.

[0311] Depending on the disorder being treated, the compounds of Structures (I)-(ID) described herein can be used in combination with the agents disclosed herein or other suitable agents. Thus, in certain embodiments, one or more compounds of the present disclosure will be co-administered with other agents as described above. When used in combination therapy, the compounds described herein are administered simultaneously or separately from the second agent. Such co-administration can include simultaneous administration in the same dosage form, simultaneous administration in separate dosage forms, and separate administration of the two agents. That is, the compounds described herein and any of the above agents can be formulated together in the same dosage form and administered simultaneously. Alternatively, the compounds of the present disclosure and any of the above agents can be administered simultaneously, where the two agents are present in separate formulations. In another alternative, the compounds of the present disclosure can be administered immediately following any of the above agents, or vice versa. In certain embodiments of the separate administration scenario, the compounds of the present disclosure and any of the above agents are administered minutes, or hours, or days apart.

[0312] The examples and preparations provided below further illustrate and exemplify the compounds of the present disclosure and methods for preparing such compounds. It should be understood that the scope of the present disclosure is not limited in any way by the scope of the following examples and preparations. In the following examples and throughout the specification and claims, unless otherwise indicated, molecules and moieties having a single stereocenter are present as a racemic mixture. Unless otherwise indicated, those molecules and moieties having two or more stereocenters are present as a racemic mixture of diastereomers. The single enantiomers / diastereomers can be obtained by methods known to those skilled in the art.

[0313] Representative DNA synthesis cycle

[0314]

[0315] Oligomerization is typically initiated by removing a protecting group (e.g., dimethoxytrityl, DMTr) to expose the free -OH (hydroxyl) group (step 1, detritylation). In the subsequent coupling step, a phosphoramidite monomer is introduced, which reacts with the free OH group to form a new covalent bond to phosphorus, accompanied by the loss of a diisopropylamine group (step 2, coupling). The resulting phosphite triester is oxidized (e.g., with I2 and pyridine) to a more stable phosphate ester (step 3, oxidation), and a capping step renders any remaining free OH groups non - reactive (step 4, capping). The new product phosphate ester oligomer contains a DMTr - protected OH group, which can be deprotected to restart the synthesis cycle so that another phosphoramidite monomer can be attached to the oligomer.

[0316] Customization occurs in step 2 and is carried out by selecting the phosphoramidite monomer. The nature of L (i.e., the linker group) and M (i.e., the chemotherapeutic agent) in the above - described scheme is chosen so as to synthesize a compound of the desired structure (I)-(ID). M can optionally be absent to incorporate the desired spacing between M moieties. A person of ordinary skill in the art can select a variety of monomer types to obtain compounds of the present disclosure that contain a variety of therapeutic agents and / or other moieties (e.g., fluorophores or chromophores) and in which the linker group is variable simultaneously. Examples

[0317] General methods

[0318] Mass spectrometry was performed on a Waters / Micromass Quattro micro MS / MS system (in MS - only mode) using MassLynx 4.1 acquisition software. The mobile phase for dye LC / MS was 100 mM 1,1,1,3,3,3 - hexafluoro - 2 - propanol (HFIP), 8.6 mM triethylamine (TEA), pH 8. The phosphoramidites and precursor molecules were also analyzed using a Waters Acquity UHPLC system, which had a 2.1 mm×50 mm Acquity BEH - C 18 column maintained at 45 °C, with an acetonitrile / water mobile phase gradient. The molecular weights of the monomer intermediates were obtained using on - column infusion enhanced ionization on a Waters / Micromass Quattro micro MS / MS system (in MS - only mode). Excitation and emission curve experiments were recorded on a Cary Eclipse spectrophotometer.

[0319] All reactions were carried out in dried glassware under a nitrogen atmosphere unless otherwise specified. Commercially available DNA synthesis reagents were purchased from Glen Research (Sterling, VA). Anhydrous pyridine, toluene, dichloromethane, diisopropylethylamine, triethylamine, acetic acid, pyridine, and THF were purchased from Aldrich. All other chemicals were purchased from Aldrich or TCI and used as received without further purification.

[0320] Example 1

[0321] Synthesis of Compound I-1

[0322] Preparation of Stock Solutions

[0323] Prepare borate buffer at 250 mM, pH 10

[0324] Prepare fluorescein-NHS solution at 350 mM (300 mg in 1.35 mL of 25:75 DMSO:acetonitrile)

[0325] Solid-Phase Synthesis

[0326] Compound I-1 was prepared on a DNA synthesizer using solid support with standard DNA synthesis techniques (i.e., DMT-protected 2-cyanoethyl phosphoramidite). The polymer was removed from the solid support using ammonium hydroxide and lyophilized to a paste. A 250 mg aliquot was reconstituted in water. A small aliquot was taken and serial dilutions were prepared in 100 mM NaCO3 (pH 9) to determine the concentration (A 263ε = 10,000). The final stock concentration was found to be 14.5 mM.

[0327] Dye Conjugation Reaction

[0328] Place water (1.110 μL), borate buffer (1.800 μL), Compound I-1 polymer solution (466 μL), acetonitrile (137.5 μL), triethylamine (313 μL), and fluorescein-NHS solution (675 μL) in a 50 mL centrifuge tube equipped with a magnetic stir bar. Wrap the tube in aluminum foil and stir the mixture overnight at room temperature.

[0329] Size Exclusion Filtration

[0330] Add 1 mL of water to an Amicon Ultra-15 centrifugal filter (Millipore UFC900324, molecular weight cut-off = 3000). Add the crude product (4.5 mL) resulting from the dye coupling reaction to the filtration device. Rinse the reaction vessel twice with 4 mL of 100 mM NaOH and transfer the rinse to the filtration device. Centrifuge the filtration device at maximum speed (3220 g, swinging bucket, 30 minutes). Remove the filtrate and treat the retentate with an additional 10 mL of 100 mM NaOH. Centrifuge the filtration device as before. Remove the filtrate again and add a third 10 mL aliquot of 100 mM NaOH to the retentate. Centrifuge the device as before and remove the filtrate. Add a fourth 10 mL aliquot of 100 mM NaOH to the retentate and centrifuge as before. Remove the filtrate and add 10 mL of water to the filtration device. Centrifuge the mixture as before. Remove the retentate, wash the filtration container with water and combine the rinses to a final volume (3.5 mL). Confirm the desired product by LC-MS and determine the concentration using absorbance.

[0331] Synthetic scheme for compounds I-2 and I-7

[0332]

[0333] Example 2

[0334] Activation and antibody conjugation of compound I-2 or I-7

[0335]

[0336] Prepare maleimide-functionalized compound I-2 or I-7 according to the method described in Example 1. In parallel, treat the brentuximab antibody with bis-maleimidobutane (“BMOE”) to reduce disulfide bonds. React the reduced antibody with compound I-2 or I-7 at a polymer-to-antibody molar ratio of 5:1. By size exclusion chromatography detection, the reaction produces a final product with a polymer-to-antibody ratio of 1:1. In certain embodiments, anti-CD33, anti-CD70 or anti-CD123 can be used with bis-maleimidobutane (“BMOE”) to reduce disulfide bonds.

[0337] All U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications, and non-patent publications mentioned in this specification and / or listed in the Application Data Sheet, including U.S. Provisional Patent Application No. 63 / 384,712, filed Nov. 22, 2022, are hereby incorporated by reference in their entirety to the extent that they are not inconsistent with this specification. Aspects of the embodiments may be modified, if necessary, to employ concepts of different patents, applications, and publications to provide other embodiments.

[0338] It will be understood from the foregoing that, although particular embodiments of the disclosure have been described herein for purposes of illustration, various modifications may be made without departing from the spirit and scope of the disclosure. Accordingly, the disclosure is only limited by the appended claims.

Claims

1. A water-soluble polymeric compound, comprising: i) one or more anti-tumor agents; ii) a heteroalkyl main chain containing groups negatively charged at physiological pH; iii) one or more straight-chain or branched linkers containing a self-cleaving group and an optional spacer group, said linker connecting each anti-tumor agent to said heteroalkyl main chain; and iv) a reactive group capable of forming a covalent bond with a targeting moiety.

2. The polymeric compound according to claim 1, further comprising: i) at least one fluorescent or colored moiety; and ii) one or more linkers, each linker connecting the fluorescent or colored moiety to said heteroalkyl main chain.

3. The polymeric compound according to any one of claims 1-2, wherein the polymeric compound has the following structure (I): or a stereoisomer, salt or tautomer thereof, wherein: M 1 is, independently at each occurrence, a moiety comprising a fluorescent or colored moiety; M 2 independently at each occurrence is a moiety comprising an anti-tumor agent; L 1 is independently a straight-chain or branched linker each time it appears; L 2 and L 3 is independently, at each occurrence, an optional alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, heteroalkynylene or heteroatom linker; L 4 is independently a heteroalkyl linker at each occurrence; L 5 is independently a linker each time it appears; R 1 is independently H, alkyl or alkoxy each time it appears; R 2 and R 3 are each independently H, OH, SH, -OP(=R a )(R b )R c or Q; R 4 is independently OH, SH, O - , S - , OR d or SR d ; R 5 is independently oxo, thioxo or absent at each occurrence; R a is O or S; R b is OH, SH, O - , S - , OR d or SR d ; R c is OH, SH, O - , S - , OR d , OL′, SR d , alkyl, alkoxy, heteroalkyl, heteroalkoxy, alkyl ether, alkoxyalkyl ether, phosphate / ester, thiophosphate / ester, phosphoalkyl, thiophosphoalkyl, phosphoalkyl ether or thiophosphoalkyl ether; R d is a counter ion; Q is independently at each occurrence a moiety containing a reactive group or a protected form thereof, which is capable of forming a covalent bond with a complementary reactive group Q' on a targeting moiety; L' is independently at each occurrence a linker containing a covalent bond with Q, a targeting moiety, a linker containing a covalent bond with a targeting moiety, a linker containing a covalent bond with a solid support, a linker containing a covalent bond with a solid support residue, a linker containing a covalent bond with a nucleoside or a linker containing a covalent bond with another compound of structure (I); m is independently at each occurrence an integer from 1 to 5; p is an integer of 0 or greater; and n is an integer of 1 or greater.

4. The polymeric compound according to any one of claims 1-3, wherein the at least one fluorescent or colored moiety M1 is a fluorescent moiety.

5. The polymeric compound according to any one of claims 1-4, wherein the fluorescent or colored moiety M1 is independently at each occurrence selected from pyrene, perylene, perylene monoimide and 6-FAM moiety.

6. The polymeric compound according to any one of claims 1-5, wherein the fluorescent or colored moiety M 1 independently has one of the following structures each time it appears: Among them Indicates the connection point with the fluorescent compound.

7. The polymeric compound according to any one of claims 1-6, wherein the fluorescent or colored moiety M 1 has one of the following structures:

8. The polymeric compound according to any one of claims 1-7, wherein the fluorescent or colored moiety M 1 has the following structure:

9. The polymeric compound according to any one of claims 1-8, wherein the polymeric compound contains 1-10 fluorescent or colored moieties.

10. The polymeric compound according to any one of claims 1-9, wherein the polymeric compound contains 1-5 fluorescent or colored moieties.

11. The polymeric compound according to any one of claims 1-10, wherein the polymeric compound contains 1 fluorescent or colored moiety.

12. The polymeric compound according to any one of claims 1-11, wherein the anti-tumor agent M2 is independently at each occurrence selected from paclitaxel, camptothecin, doxorubicin, monomethyl auristatin E, monomethyl auristatin F and PF-06380101.

13. The polymeric compound according to any one of claims 1-12, wherein the anti-tumor agent M 2 independently has one of the following structures each time it appears:

14. The polymeric compound according to any one of claims 1-13, wherein the anti-tumor agent M 2 is monomethyl auristatin E.

15. The polymeric compound according to any one of claims 1-14, wherein the anti-tumor agent M 2 has the following structure:

16. The polymeric compound according to any one of claims 1-15, wherein the polymeric compound contains 1-20 anti-tumor agents.

17. The polymeric compound according to any one of claims 1-16, wherein the polymeric compound contains 1-10 anti-tumor agents.

18. The polymeric compound according to any one of claims 1-17, wherein the polymeric compound contains 1 anti-tumor agent.

19. The polymeric compound according to any one of claims 1-18, wherein the polymeric compound contains 3 anti-tumor agents.

20. The polymeric compound according to any one of claims 1-19, wherein the polymeric compound comprises 5 anti-tumor agents.

21. The polymeric compound according to any one of claims 1-20, wherein the polymeric compound comprises 7 anti-tumor agents.

22. The polymeric compound according to any one of claims 1-21, wherein the polymeric compound comprises 10 anti-tumor agents.

23. The polymeric compound according to any one of claims 1-22, wherein the heteroalkyl linker L4 comprises one or more alkylene or oxyalkylene moieties.

24. The polymeric compound according to any one of claims 1-23, wherein the oxyalkylene moiety comprises a polyoxyethylene moiety.

25. The polymeric compound according to any one of claims 1-24, wherein the negatively charged group of the heteroalkyl backbone is a phosphate / ester.

26. The polymeric compound according to any one of claims 1-25, wherein the polymeric compound comprises at least 4 negatively charged phosphate / ester groups.

27. The polymeric compound according to any one of claims 1-26, wherein the heteroalkyl linker L 4 comprises one of the following structures: where m is an integer of 1 or greater.

28. The polymeric compound according to any one of claims 1-27, wherein the heteroalkylene linker L 4 comprises one of the following structures:

29. The polymeric compound according to claim 28, wherein the heteroalkylene linker L 4 comprises the following structure:

30. The polymeric compound according to claim 28, wherein the heteroalkyl linker L 4 comprises the following structure:

31. The polymeric compound according to any one of claims 1-30, wherein R 4 is O - and R 5 is oxo.

32. The polymeric compound according to any one of claims 1-31, wherein the polymeric compound has one of the following structures (IA) or (IB):

33. The polymeric compound according to any one of claims 1-32, wherein L 2 and L 3 is independently, each time it appears, a C1-C6 alkylene group, a C2-C6 alkenylene group or a C2-C6 alkynylene group.

34. The polymeric compound according to any one of claims 1-33, wherein the compound has one of the following structures (IC) or (ID): wherein: x 1 、x 2 、x 3 and x 4 is independently an integer from 0 to 6 at each occurrence.

35. The polymeric compound according to claim 34, wherein x 1 , x 2 , x 3 and x 4 is each 0 or 1 at each occurrence.

36. The polymeric compound according to any one of claims 1-35, wherein the self-cleaving group has one of the following structures:

37. The polymeric compound according to any one of claims 1-36, wherein the optional spacer group has one of the following structures:

38. The polymeric compound according to any one of claims 1-37, wherein the optional spacer group has the following structure: The polymeric compound according to any one of claims 1 to 38, wherein the linker L 5 has one of the following structures: The polymeric compound according to any one of claims 1-39, wherein the linker L 5 has the following structure:

41. The polymeric compound according to any one of claims 1-40, wherein R 1 is H each time it appears.

42. The polymeric compound according to any one of claims 1-41, wherein Q comprises a thiol, disulfide, activated ester, isothiocyanate, azide, alkyne, alkene, diene, dienophile, acyl halide, sulfonyl halide, phosphine, α-haloamide, biotin, amino or maleimide functional group.

43. The polymeric compound according to claim 42, wherein the activated ester is an N-succinimidyl ester, imidate or polyfluorophenyl ester.

44. The polymeric compound according to any one of claims 1-43, wherein Q comprises one of the following structures: The polymeric compound according to any one of claims 1 to 44, wherein R 2 or R 3 has one of the following structures:

46. The polymeric compound according to any one of claims 1-45, wherein R 2 or R 3 has one of the following structures:

47. The polymeric compound according to any one of claims 1-46, wherein n is from 1 to 20.

48. The polymeric compound according to any one of claims 1-47, wherein n is from 1 to 10.

49. The polymeric compound according to any one of claims 1-48, wherein n is 1, 3, 5, 7 or 10.

50. The polymeric compound according to any one of claims 1-49, wherein n is 1.

51. The polymeric compound according to any one of claims 1-50, wherein n is 3.

52. The polymeric compound according to any one of claims 1-51, wherein n is 5.

53. The polymeric compound according to any one of claims 1-52, wherein n is 7.

54. The polymeric compound according to any one of claims 1-53, wherein n is 10.

55. The polymeric compound according to any one of claims 1-54, wherein m is 1, 2, 3, 4 or 5.

56. The polymeric compound according to any one of claims 1-55, wherein m is 1, 2 or 3.

57. The polymeric compound according to any one of claims 1-56, wherein m is 1.

58. The polymeric compound according to any one of claims 1-57, wherein m is 3.

59. The polymeric compound according to any one of claims 1-58, wherein p is 1, 2, 3 or 4.

60. The polymeric compound according to any one of claims 1-59, wherein p is 1.

61. The polymeric compound according to any one of claims 1-60 having one of the following structures: wherein FAM represents 62. A conjugate comprising a compound according to any one of claims 1-61 covalently bound to a targeting moiety via a bond to a reactive group.

63. The conjugate according to claim 62, wherein the targeting moiety has an affinity for a target analyte.

64. The conjugate according to claim 62, wherein the targeting moiety comprises an antibody.

65. The conjugate according to claim 62, wherein the antibody comprises trastuzumab, gemtuzumab, brentuximab, vosaroxin, lomustine, canertinib, bevacizumab or icatibant or vadastuximab.

66. The conjugate according to claim 65, wherein the antibody comprises brentuximab.

67. The conjugate according to any one of claims 62-65, wherein the targeting moiety specifically binds to brentuximab.

68. A pharmaceutical composition comprising a compound according to any one of claims 1-61 or a pharmaceutically acceptable salt, tautomer, prodrug or stereoisomer thereof, or a conjugate according to any one of claims 62-67 or a pharmaceutically acceptable salt, tautomer, prodrug or stereoisomer thereof, and a pharmaceutically acceptable excipient.

69. A method for forming a conjugate according to any one of claims 62-67, the method comprising contacting a compound according to any one of claims 1-61 with a targeting moiety.

70. A method for treating cancer in a subject in need thereof, the method comprising administering to the subject a compound according to any one of claims 1-61 or a pharmaceutically acceptable salt, tautomer, prodrug or stereoisomer thereof, a conjugate according to any one of claims 62-67 or a pharmaceutically acceptable salt, tautomer, prodrug or stereoisomer thereof, or a pharmaceutical composition according to claim 68.

71. Use of a compound according to any one of claims 1-61 or a pharmaceutically acceptable salt, tautomer, prodrug or stereoisomer thereof, a conjugate according to any one of claims 62-67 or a pharmaceutically acceptable salt, tautomer, prodrug or stereoisomer thereof, or a pharmaceutical composition according to claim 68 for the treatment of cancer. The method according to claim 70 or the use according to claim 71, wherein the cancer is breast cancer, gastric cancer, lung cancer, ovarian cancer, lymphoma and bladder cancer.

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