Process for preparing polymeric dyes with drug conjugates

Abstract

Methods for preparing polymer-drug conjugates containing a bioactive moiety and a fluorescent or colored dye are provided. The process comprises reacting a compound of structure (I): Formula (I) with an amine-containing bioactive compound of structure (II): wherein G comprises a group capable of forming a covalent bond with an amine group, and R1, R2, R3, R4, R5, R6, R7, R8, L1, L2, L3, L4, L5, L6, L7, M1, m, n, p and w are as defined herein. , (I)

Description

[0001] background

[0002] field

[0003] The embodiments of this disclosure generally relate to methods for preparing polymer-drug conjugates having a fluorescent or colored portion and a biologically active portion.

[0004] Description of related technologies

[0005] Unlike chemotherapy, targeted drug conjugates deliver drugs directly to target cells with little or no off-target activity. Typically, targeted drug conjugates contain a target molecule linked to a bioactive load or drug. By combining unique targeting capabilities with the therapeutic efficacy of a bioactive drug, conjugates can deliver drugs only to the intended target and minimize potential side effects. Polymer-drug conjugates (PDCs) are a class of targeted drug conjugates that have attracted considerable attention in the treatment of various diseases. Polymer conjugates have been found to improve drug pharmacokinetic parameters, enhance drug resistance to degradation, provide high loading capacity and sustained release modes, and avoid premature drug release. Polymer-drug conjugates integrated with imaging agents are attractive due to their ability to provide therapeutic and diagnostic or imaging functions. PDCs integrated with imaging agents facilitate the monitoring of drug biodistribution and localization, which contributes to the precise activation of therapeutic responses.

[0006] There remains a need in the art to develop methods for preparing polymer-drug conjugates that combine a therapeutic agent and an imaging agent on the same polymer backbone to allow for simultaneous diagnosis / imaging during drug therapy progression. This disclosure addresses this need and provides further related advantages.

[0007] Brief Overview

[0008] In short, embodiments of this disclosure generally relate to methods for preparing polymer-drug conjugates.

[0009] In some embodiments, a method is provided for preparing polymer-drug conjugates that allow for simultaneous diagnostic and therapeutic selection. The method comprises a compound that gives structure (I):

[0010]

[0011] Reaction with compounds of structure (II):

[0012]

[0013] Where R 1 R 2 R 3 R4 R 5 R 6 R 7 R 8 L 1 L 2 L 3 L 4 L 5 L 6 L 7 G, M 1 M 2 m, n, p and w are defined as in this article.

[0014] In some embodiments, a compound of structure (I) is provided. The compound of structure (I) has the following structure:

[0015]

[0016] Where R 1 R 2 R 3 R 4 R 5 R 6 R 7 L 1 L 2 L 3 L 4 L 5 L 6 L 7 G, M 1 m, n, p and w are defined as in this article.

[0017] In some embodiments, a compound of structure (III) is provided, which is formed by reacting a compound of structure (I) with a compound of structure (II). The compound of structure (III) has the following structure:

[0018]

[0019] Where R 1 R 2 R 3 R 4 R 5 R 6 R 7 L 1 L 2 L 3 L 4 L 5 L 6 L 7 M 1 M 2m, n, p and w are defined as in this article.

[0020] In some embodiments, a compound of structure (V) is provided, which can be used as an intermediate for the preparation of the compound of structure (I). The compound of structure (V) has the following structure:

[0021]

[0022] Where R 1 R 2 R 3 R 4 R 5 R 6 R 7 L 1 L 2 L 3 L 4 L 5 L 6 L 7 G, G 1 m, n, p and w are defined as in this article.

[0023] In some embodiments, a method for preparing a compound of structure (I) is provided, the method comprising mixing a compound of structure (V) with M. 1 -G 1’ The reaction of compounds, in which M 1 -G 1’ As defined in this article.

[0024] These and other aspects of this disclosure will become clear upon referring to the following detailed description.

[0025] Detailed description

[0026] In the following description, certain specific details are set forth to provide a thorough understanding of various embodiments of this disclosure. However, those skilled in the art will understand that this disclosure can be practiced without these details.

[0027] Unless the context otherwise requires, throughout this specification and the claims, the word “comprise” and its variations, such as “comprises” and “comprising”, shall be interpreted in an open and inclusive sense, meaning “including, but not limited to”.

[0028] In this specification, the reference to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with that embodiment is included in at least one embodiment of this disclosure. Therefore, the phrases "in one embodiment" or "in an embodiment" appearing throughout this specification do not necessarily refer to the same embodiment. Furthermore, the particular feature, structure, or characteristic may be combined in any suitable manner in one or more embodiments.

[0029] "Amino" NH2 group.

[0030] "Carboxyl group" CO2H group.

[0031] "Cyano" table CN group.

[0032] "Formyl group" C(=O)H group.

[0033] "Hydroxy" or "hydroxyl" indicates OH group.

[0034] "Imine" represents the =NH group.

[0035] "Nitro" table NO2 group.

[0036] "O" indicates an O substituent.

[0037] "Thiol" table SH group.

[0038] "Thioxo" means =S group.

[0039] "Alkyl" indicates a straight-chain or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, without unsaturation, and having 1-12 carbon atoms (C1-C2). 12 Alkyl groups (1-8 carbon atoms (C1-C8 alkyl) or 1-6 carbon atoms (C1-C6 alkyl) attached to the remainder of the molecule by single bonds, such as methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (tert-butyl), 3-methylhexyl, 2-methylhexyl, etc. Unless otherwise specifically stated in the specification, alkyl groups may optionally be substituted.

[0040] "alkylene" or "alkylene chain" refers to a straight-chain or branched divalent hydrocarbon chain in which the remainder of a molecule is attached to a residue group. It consists only of carbon and hydrogen, is unsaturated, and has 1-12 carbon atoms, such as methylene, ethylene, propylene, n-butylene, vinylene, propenylene, n-butynylene, propynylene, n-butynylene, etc. The alkylene chain is attached to the remainder of the molecule and to the residue group via single bonds. The connection point between the alkylene chain and the remainder of the molecule and to the residue group can be through one carbon or any two carbons within the chain. Unless otherwise specifically stated in the specification, the alkylene may optionally be substituted.

[0041] "Alkenyl" or "alkenyl chain" refers to a straight-chain or branched divalent hydrocarbon chain that links the remainder of a molecule to a residue group. It consists only of carbon and hydrogen, contains at least one carbon-carbon double bond, and has 2-12 carbon atoms, such as vinylene, propyleneene, n-butene, etc. The alkenyl chain is connected to the remainder of the molecule by a single bond and to the residue group by a double or single bond. The connection point between the alkenyl chain and the remainder of the molecule and the residue group can be through one carbon or any two carbons within the chain. Unless otherwise specifically stated in the specification, the alkenyl group may optionally be substituted.

[0042] "Imyynyl" or "Imyynyl chain" refers to a straight-chain or branched divalent hydrocarbon chain in which the remainder of a molecule is attached to a residue group. It consists only of carbon and hydrogen, contains at least one carbon-carbon triple bond, and has 2-12 carbon atoms; examples include ethynylene, propynylene, n-butynylene, etc. The ethynyl chain is attached to the remainder of the molecule by a single bond and to the residue group by a double or single bond. The connection point between the ethynyl chain and the remainder of the molecule, and between the ethynyl chain and the residue group, can be through one carbon or any two carbons within the chain. Unless otherwise specifically stated in the specification, the ethynyl group may optionally be substituted.

[0043] "Alkyl ether" means any alkyl group as defined above, wherein at least one carbon-carbon bond is replaced by a carbon-oxygen bond. The carbon-oxygen bond may be terminal (as in an alkoxy group) or internal (i.e., COC). Alkyl ethers include at least one carbon-oxygen bond, but may include more than one. For example, polyethylene glycol (PEG) is included within the meaning of alkyl ether. Unless otherwise specifically stated in the specification, the alkyl ether group may optionally be substituted. For example, in some embodiments, the alkyl ether is replaced by an alcohol or -OP (=R a (R) b )R c Replace, where R a R b and R c Each of them is defined as per the compounds with respect to structures (I), (III), and (V).

[0044] "alkoxy" indicates OR a The group, wherein R a It is an alkyl group as defined above, containing 1-12 carbon atoms. Unless otherwise specifically stated in the specification, the alkoxy group may optionally be substituted.

[0045] "Alkoxyalkyl ether" indicates OR a R b The group, wherein R a It is an alkylene group as defined above, containing 1-12 carbon atoms, and R b It is an alkyl ether group as defined herein. Unless otherwise specifically stated in the specification, the alkoxyalkyl ether group may optionally be substituted, for example, with an alcohol or -OP (=R a (R) b )R c Replace, where R a R b and R c Each of them is defined as per the compounds with respect to structures (I), (III), and (V).

[0046] "Heteroalkyl" refers to an alkyl group as defined above, which contains at least one heteroatom (e.g., N, O, P, or S) within or at the end of an alkyl group. In some embodiments, the heteroatom is within the alkyl group (i.e., the heteroalkyl group contains at least one carbon-[heteroatom]). x - A carbon bond, where x is 1, 2, or 3). In other embodiments, the heteroatom is at the alkyl terminus and thus serves to connect the alkyl group to the rest of the molecule (e.g., M1-HA), where M1 is part of the molecule, H is a heteroatom, and A is an alkyl group). Unless otherwise specifically stated in the specification, the heteroalkyl group is optionally substituted. Exemplary heteroalkyl groups include ethylene oxide (e.g., polyethylene oxide), optionally including phosphorus-oxygen bonds, such as phosphate diester bonds.

[0047] "Hexaalkoxy" indicates OR a The group, wherein R a It is a heteroalkyl group as defined above, containing 1-12 carbon atoms. Unless otherwise specifically stated in the specification, the heteroalkoxy group may optionally be substituted.

[0048] "Heteroalkylene" refers to an alkylene group as defined above, which contains at least one heteroatom (e.g., Si, N, O, P, or S) within or at the end of the alkylene chain. In some embodiments, the heteroatom is within the alkylene chain (i.e., the heteroalkylene contains at least one carbon-[heteroatom]-carbon bond, where x is 1, 2, or 3). In other embodiments, the heteroatom is at the end of the alkylene and thus serves to link the alkylene to the rest of the molecule (e.g., M1-HA-M2, where M1 and M2 are portions of the molecule, H is a heteroatom, and A is an alkylene). Unless otherwise specifically stated in the specification, the heteroalkylene may optionally be substituted. Exemplary heteroalkylenes include ethylene oxide (e.g., polyethylene oxide) and the "C", "HEG", and "PEG 1K" linking groups explained below:

[0049]

[0050] The polymers of the C-linker, HEG linker, and / or PEG 1K linker are included in different embodiments of the heteroalkylene linker. In some 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 may comprise, for example, the following structures:

[0051]

[0052] Where x is 0 or an integer greater than 0, for example, x ranges from 0 to 100 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10).

[0053] A "linking group" refers to a continuous chain of at least one atom, such as carbon, oxygen, nitrogen, sulfur, phosphorus, and combinations thereof, which links a portion of a molecule to another portion of the same molecule or to different molecules, portions, or solid supports (e.g., particles). Linking groups can connect molecules via covalent bonds or other means, such as ionic bonds or hydrogen bonds. In some embodiments, the linking group is a heteroatom linking group (e.g., containing 1-10 Si, N, O, P, or S atoms), a heteroalkylene linking group (e.g., containing 1-10 Si, N, O, P, or S atoms and an alkylene chain), or an alkylene linking group (e.g., containing 1-12 carbon atoms). In some embodiments, the heteroalkylene linking group comprises the following structure:

[0054]

[0055] in:

[0056] x 9 and x 10Each is an independent integer greater than 0. In some embodiments, the heteroatom linker is -O-, -S-, or -OP(=O)O. - -O-. In some embodiments, the heteroalkyl linker comprises -OP(=O)O. - -O-. In some embodiments, the heteroalkyl linker contains at least one SS bond.

[0057] A "physiologically cleavable linker" refers to a molecular linker that, in the presence of an in vivo or in vitro environment of an organism or cellular system, can be split or separated in a prescribed manner, resulting in two or more separate molecules. Typically, physiological conditions inducing such cleavage or breakage events can include a temperature in the range of about 20–40 °C, an atmospheric pressure of about 1 atm (101 kPa or 14.7 psi), a pH of about 6–8, a glucose concentration of about 1–20 mM, an atmospheric oxygen concentration, and gravity. In some embodiments, physiological conditions include enzymatic conditions (i.e., enzymatic cleavage). For example, cathepsins are proteases that cleave proteins at low pH. Cathepsins include cathepsin A, cathepsin B, cathepsin D, cathepsin K, and cathepsin V. Bond cleavage or breakage can be homolytic or heterolytic.

[0058] "Physiological pH" refers to a pH range of approximately 4 to 8 at a temperature of approximately 20 to 40°C and an atmospheric pressure of approximately 1 atm (101 kPa or 14.7 psi).

[0059] "Heteroalkylene" is a heteroalkylene group as defined above that contains at least one carbon-carbon double bond. Unless otherwise specifically stated in the specification, the heteroalkylene group may optionally be substituted.

[0060] "Hydynyl" is a heteroalkyl group containing at least one carbon-carbon triple bond. Unless otherwise specifically stated in the specification, the heteroalkyl group may optionally be substituted.

[0061] The term "heteroatomic linker" in the context of "heteroatomic linker" refers to a linker group consisting of one or more heteroatoms. Exemplary heteroatomic linkers include single atoms and multiple heteroatoms selected from O, N, P, and S, such as linkers having... P(O-)(=O) Or -OP(O-)(=O) and polymers and their combinations.

[0062] "Phosphate ester" represents -OP(=O)(R a )R b Group, wherein R a Is it OH, O- or OR? c And R bIt is OH, O-, OR c Thiophosphate group or other phosphate ester group, wherein R c It is a counter ion (e.g., Na+) + wait).

[0063] "Phosphoalkyl" indicates -OP (=O)(R a )R b Group, wherein R a Is it OH, O- or OR? c And R b O alkyl, wherein R c It is a counter ion (e.g., Na+) + (etc.). Unless otherwise specifically stated in the specification, the phosphoryl alkyl group may optionally be substituted. For example, in some embodiments, the phosphoryl alkyl group... The alkyl moiety is optionally replaced by a hydroxyl, amino, mercapto, phosphate ester, thiophosphate ester, phosphoryl alkyl, thiophosphoryl alkyl, phosphoryl alkyl ether, or -OP (=R) group. a (R) b )R c One or more substitutions in, where R a R b and R c Each of them is defined as per the compound with respect to structure (I).

[0064] "Phosphoalkyl ether" indicates -OP(=O)(R a )R b Group, wherein R a Is it OH, O- or OR? c And R b O alkyl ether, wherein R c It is a counter ion (e.g., Na+) + (etc.). Unless otherwise specifically stated in the specification, the phosphoryl alkyl ether group may optionally be substituted. For example, in some embodiments, the -O alkyl ether moiety in the phosphoryl alkyl ether group may optionally be replaced by a 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 One or more substitutions in, where R a R b and R c Each of them is defined as per the compounds with respect to structures (I), (III), and (V).

[0065] "Thiophosphate" indicates -OP (=R) a (R) b )R c Group, wherein R a Is it O or S, R? b It is OH, O-, S-, OR d or SR d And R c It is OH, SH, O-, S-, OR d SR d Phosphate ester group or other thiophosphate ester group, wherein R d It is a counter ion (e.g., Na+) + (etc.) and the condition is: i) R a It is S; ii) R b Is it S- or SR? d ;iii) R c Is it SH, S-, or SR? d ; or a combination of iv), i), ii) and / or iii).

[0066] "Thiophosphoryl alkyl" indicates -OP (=R) a (R) b )R c Group, wherein R a Is it O or S, R? b It is OH, O-, S-, OR d or SR d And R c O alkyl, wherein R d It is a counter ion (e.g., Na+) + (etc.) and the condition is: i) R a It is S; ii) R b Is it S- or SR? d ; or iii) R a It is S and R b Is it S- or SR? d Unless otherwise specifically stated in the specification, the thiophosphoryl alkyl group may optionally be substituted. For example, in some embodiments, the thiophosphoryl alkyl group contains... The alkyl moiety is optionally replaced by a hydroxyl, amino, mercapto, phosphate ester, thiophosphate ester, phosphoryl alkyl, thiophosphoryl alkyl, phosphoryl alkyl ether, or -OP (=R) group. a (R) b )R c One or more substitutions in, where R a R b and R c Each of them is defined as per the compounds with respect to structures (I), (III), and (V).

[0067] "Thiophosphoryl ether" indicates -OP (=R) a (R) b )R c Group, wherein R a Is it O or S, R? b It is OH, O-, S-, OR d or SR d And R c O alkyl ether, wherein R d It is a counter ion (e.g., Na+) + (etc.) and the condition is: i) R a It is S; ii) R b Is it S- or SR? d ; or iii) R a It is S and R b Is it S- or SR? d Unless otherwise specifically stated in the specification, the thiophosphoryl ether group may optionally be substituted. For example, in some embodiments, the -O alkyl ether moiety in the thiophosphoryl ether may optionally be replaced by a 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 One or more substitutions in, where R a R b and R c Each of them is defined as per the compounds with respect to structures (I), (III), and (V).

[0068] "Carbocyclic ring" refers to a stable 3-18 membered aromatic or non-aromatic ring containing 3-18 carbon atoms. Unless otherwise specifically stated in the specification, the carbocyclic ring can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems, and may be partially or completely saturated. Non-aromatic carbocyclic residues include cycloalkyl groups, while aromatic carbocyclic residues include aryl groups. Unless otherwise specifically stated in the specification, the carbocyclic group may optionally be substituted.

[0069] “Cycloalkyl” refers to a stable non-aromatic monocyclic or polycyclic carbon ring, which may include fused or bridged ring systems, having 3-15 carbon atoms, preferably 3-10 carbon atoms, and is saturated or unsaturated and connected to the remainder of the molecule by a single bond. Monocyclic cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl groups include, for example, adamantyl, norbornyl, decahydronaphthyl, 7,7-dimethyl-bicyclo-[2.2.1]heptyl, etc. Unless otherwise specifically stated in the specification, cycloalkyl groups may optionally be substituted.

[0070] "Aryl" indicates a ring system comprising at least one carbocyclic aromatic ring. In some embodiments, the aryl group comprises 6-18 carbon atoms. The aryl ring can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems. Aryl groups include, but are not limited to, those derived from acephenanthrene, acenaphthene, phenanthrene, anthracene, azulene, benzene, benzo[a], fluorene, asymmetric indacene, symmetric indacene, indene, indene, naphthalene, phenanthracene, pleiadene, pyrene, and benzo[a]phenanthrene. Unless otherwise specifically stated in the specification, the aryl group may optionally be substituted.

[0071] "Heterocycle" refers to a stable 3-18 membered aromatic or non-aromatic ring comprising 1-12 carbon atoms and 1-6 heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specifically stated in the specification, the heterocycle can be a monocyclic, bicyclic, tricyclic, or tetracyclic system, which may include fused or bridged ring systems; and the nitrogen, carbon, or sulfur atoms in the heterocycle may optionally be oxidized; the nitrogen atom may optionally be quaternized; and the heterocycle may 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, dioxacyclopentyl, thienyl[1,3]dithiohexacyclohexyl, decahydroisoquinolinyl, imidazolinyl, imidazoalkyl, isothiazolyl, isoxazolyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopiperylyl, oxazolyl, piperidinyl, piperazinyl, 4-piperidinoneyl, pyrrolylyl, pyrazolylyl, pyrazolopyrimidinyl, quininecycloyl, thiazoalkyl, tetrahydrofuranyl, trioxacyclohexyl, trithiohexyl, triazinyl, tetrahydropyranyl, thiomorpholinyl, thiomorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless otherwise specifically stated in the specification, heterocyclic groups may optionally be substituted.

[0072] "Heteroaryl" refers to a 5-14 membered ring system comprising 1-13 carbon atoms, 1-6 heteroatoms selected from nitrogen, oxygen, and sulfur, and at least one aromatic ring. For the purposes of certain embodiments of this disclosure, the heteroaryl residue can be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused or bridged ring systems; and the nitrogen, carbon, or sulfur atom in the heteroaryl residue may optionally be oxidized; the nitrogen atom may optionally be quaternized. Examples include, but are not limited to, nitrogen-containing heterocyclic heptadienyl, acridinel, benzimidazolyl, benzothiazolyl, benzoindolyl, benzom-dioxacyclopentenyl, benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[ b [1,4]dioxepinyl, 1,4-benzodioxepinyl, benzonaphthylfuranyl, benzoxazolyl, benzom-dioxepinyl, benzodioxepinyl, benzopyranyl, benzopyranoneyl, benzofuranyl, benzofuranoneyl, benzothiopheneyl, benzotriazolyl, benzo[4,6]imidazo[1,2- a Pyridyl, benzoxazolinone, benzimidazole thionyl, carbazole, cenyl, dibenzofuranyl, dibenzothiophene, furanyl, furanone, isothiazolyl, imidazolyl, indazole, indole, indazole, isoindole, indolinyl, isoindolelinyl, isoquinolinyl, inazinyl, isoxazolyl, naphridyl, oxadiazolyl, 2-oxoazacycloheptatrienyl, oxazolyl, oxacyclopropyl, 1-oxidopyridyl, 1-pyrimidinyl, 1-pyrazinyl, 1-pyridazinyl, 1-pyridazinyl, 1-phenyl-1 H -pyrrolyl, phenazinyl, phenothiazinyl, phenotoxazinyl, phthalazinyl, pteridinyl, pteridinoneyl, purine, pyrrolyl, pyrazolyl, pyridinyl, pyridinoneyl, pyrazinyl, pyrimidinoneyl, pyridazinyl, pyridine, pyridinoneyl, pyridazinyl, pyridino[2,3- d Pyrimidinone, quinazolinyl, quinazolinone, quinoxalinyl, quinoxalinone, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, thieno[3,2-] d ]Pyrimidine-4-keto, thiophene[2,3- d Pyrimidine-4-keto, triazolyl, tetrazolyl, triazine, and thiophene ( Right now Thiophene group). Unless otherwise specifically stated in the specification, heteroaryl groups may optionally be substituted.

[0073] The suffix "-ene" indicates a specific structural feature (e.g., alkyl, aryl, heteroalkyl, heteroaryl) that is linked to the rest of the molecule and to a free radical via a single bond. In other words, the suffix "-ene" indicates that the linking group has the structural feature of the portion to which it is connected. The connection point between the "-ene" chain and the rest of the molecule and the free radical can be through one atom or any two atoms in the chain. For example, arylene indicates a divalent linking group containing an aryl moiety as defined herein, and heteroarylene indicates a divalent linking group containing a heteroaryl moiety as defined herein.

[0074] "Fused" refers to a ring system containing at least two rings, wherein the two rings share at least one common ring atom, such as two common ring atoms. When the fused ring is a heterocyclic or heteroaryl ring, the common ring atom can be carbon or nitrogen. Fused rings include bicyclic, tricyclic, tetracyclic, etc.

[0075] As used herein, the term "substitution" refers to any of the above groups (e.g., alkyl, alkylene, alkenyl, ynylene, heteroalkyl, alkenylene, alkenylene, alkoxy, alkyl ether, phosphorylalkyl, phosphorylalkyl ether, thiophosphorylalkyl, thiophosphorylalkyl ether, carbocyclic, cycloalkyl, aryl, heterocyclic and / or heteroaryl) in which at least one hydrogen atom (e.g., 1, 2, 3 or all hydrogen atoms) is substituted by a bond with a non-hydrogen atom, such as, but not limited to, halogen atoms 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., double or triple bond) with a heteroatom, said heteroatom being oxygen in groups such as 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 NR g R h NR g C(=O)R h NR g C(=O)NR g R h NR g C(=O)OR h NR g SO2R h OC(=O)NR g R h OR g SR g SOR g SO2R g OSO2R g SO2OR g =NSO2R g SO2NR g R h Substitution. "Substitution" also refers to any of the above-mentioned groups, in which one or more hydrogen atoms... C(=O)R g C(=O)OR g C(=O)NR g R h CH2SO2R g CH2SO2NR g R h Alternative. In the foregoing, R... g and R h They are the same or different, and independently are hydrogen, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclic, N - Heterocyclic group, heterocyclic alkyl group, heteroaryl group N - Heteroaryl and / or heteroarylalkyl. "Substitution" further refers to any one of the above groups, wherein one or more hydrogen atoms are substituted with bonds to the following groups: amino, cyano, hydroxyl, imino, nitro, oxo, thio, halogen, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclic, N - Heterocyclic group, heterocyclic alkyl group, heteroaryl group N - Heteroaryl and / or heteroarylalkyl. Additionally, each of the foregoing substituents may optionally be substituted with one or more of the above substituents.

[0076] “Conjugation” refers to 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” refers to 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 usually contain at least one degree of conjugation.

[0077] "Fluorescent" refers to molecules that can absorb light of a specific frequency and emit light of different frequencies. Fluorescence is well known to those skilled in the art.

[0078] "Colored" refers to molecules that absorb light within the colored spectrum (i.e., red, yellow, blue, etc.).

[0079] 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 derived 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, visually detectable biomolecules of this disclosure (e.g., compounds having the structure (III) of the biomolecule to which they are linked) are prepared by contacting a biomolecule with a compound having a reactive group (which is capable of linking the biomolecule to a compound via any available atom or functional group, such as an amino, hydroxyl, carboxyl, or thiol group on the biomolecule).

[0080] A “reactive group” is a portion capable of reacting with a second reactive group (e.g., a “complementary reactive group”) to form one or more covalent bonds, for example, through substitution, oxidation, reduction, addition, or cycloaddition reactions. Exemplary reactive groups are provided in Table 1 and include, for example, nucleophiles, electrophiles, dienes, dienophiles, aldehydes, oximes, hydrazones, alkynes, amines, azides, acyl azides, acyl halides, nitriles, nitrile ketones, mercapto groups, disulfides, sulfonyl halides, isothiocyanates, imine esters, activated esters, ketones, α,β-unsaturated carbonyl groups, alkenes, maleimides, α-haloimides, epoxides, aziridines, tetrazides, tetrazolium, phosphine, biotin, thiapropylcyclohexane, etc.

[0081] "Bioconjugation" or "bioconjugate" and related variants refer to a chemical reaction strategy that forms a stable covalent bond between two molecules. The term "bioconjugation" is generally used when one of its molecules is a biological molecule (e.g., an antibody), but can also be used to describe the formation of a covalent bond with a non-biological molecule (e.g., a polymer resin). The product or compound produced by such a reaction strategy is a "conjugate," "bioconjugate," or grammatically equivalent term.

[0082] The terms "visually detectable" and "visually identifiable" are used herein to refer to substances that are observable by visual inspection without prior light, chemical, or enzymatic activation. Such visually detectable substances absorb and emit light in a spectral region ranging from about 300 to about 900 nm. Preferably, such substances have a strong color, and preferably have a wavelength of at least about 40,000 nm. -1 cm -1 The molar extinction coefficient, more preferably at least about 50,000 M -1 cm -1 More preferably at least about 60,000M -1 cm -1 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 this disclosure can be detected by visual observation or by means of optical detection devices, including but not limited to absorption spectrophotometers, transmission optical microscopes, digital cameras, and scanners. 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 (about 10 nm to about 400 nm), the infrared (IR) region (about 700 nm to about 1 mm), and other electromagnetic spectral regions are also included in the scope of "visually detectable" substances.

[0083] For the purposes of embodiments of this disclosure, the term "photostable visual dye" refers to a chemical component that is visually detectable as defined above and does not undergo significant alteration or decomposition upon exposure to light. Preferably, the photostable visual dye does not exhibit significant discoloration or decomposition after exposure to light for at least 1 hour. More preferably, the visual 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 photostable visual dyes suitable for the compounds and methods of this disclosure include azo dyes, thio-indigo dyes, quinophthalone pigments, dioxazine, phthalocyanine, perinone, diketopyrrolopyrrole, quinophthalone, and truarycarbonium.

[0084] As used herein, the term "dinaphthylbenzene derivative" is intended to include any visually detectable substituted dinaphthylbenzene. However, the term is not intended to include dinaphthylbenzene itself. The terms "anthracene derivative," "naphthalene derivative," and "pyrene derivative" are used similarly. In some preferred embodiments, the derivative (e.g., dinaphthylbenzene, pyrene, anthracene, or naphthalene derivative) is an imide, diimide, or hydrazamimide derivative of dinaphthylbenzene, anthracene, naphthalene, or pyrene.

[0085] Visually detectable molecules according to various embodiments of this disclosure can be used in a variety of analytical applications, such as biochemical and biomedical applications, where it is necessary to determine the presence, location, or amount of a specific analyte (e.g., a biomolecule). Therefore, in another aspect, this disclosure provides a method for visually detecting biomolecules, comprising: (a) providing a biological system with a visually detectable biomolecule comprising a compound of structure (III) linked to the biomolecule; and (b) detecting the biomolecule by its visual properties. For the purposes of this disclosure, the phrase “detecting a biomolecule by its visual properties” means observing the biomolecule with the naked eye or by means of an optical detection device, without illumination or chemical or enzymatic activation, 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 visually detectable biomolecules present. For example, the relative amount of biomolecules in two samples can be determined by measuring relative optical density. The absolute concentration of the biomolecule can also be determined from the measurement of optical density if the stoichiometry of the dye molecule for each biomolecule is known and the extinction coefficient of the dye molecule is known. As used herein, the term "biological system" refers to any solution or mixture containing one or more biomolecules in addition to those that are visually detectable. Non-limiting examples of such biological systems include cells, cell extracts, tissue samples, electrophoresis gels, assay mixtures, and hybridization reaction mixtures.

[0086] "Antitumor agents" or "tumor-resistant agents" refer to any molecule that inhibits the formation of growths that may develop into cancer. Antitumor agents can include azacitidine, capecitabine, carmoflu, cladribine, clofarabine, cytarabine, decitabine, fluorouridine, fludarabine, fluorouracil, gemcitabine, mercaptopurine, nerabine, pentostatin, tegafur, thioguanine, methotrexate, pemetrexed, raltitrexed, hydroxyurea, irinotecan, topotecan, daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone, pentoxorubicin, etoposide, teniposide, cabazitaxel, and docetaxel. The following are listed: paclitaxel, vincristine, vinblastine, vindesine, vinflunine, vinorelbine, bendamustine, busulfan, carmustine, chlorambucil, nitrogen mustard, cyclophosphamide, dacarbazine, formustine, ifosfamide, lomustine, melphalan, streptozotocin, temozolomide, carboplatin, cisplatin, nedaplatin, oxaliplatin, hexamethylmelamine, bleomycin, bortezomib, dacinomycin D, estradiol, ixaprolone, mitomycin, or procarbazine. In certain embodiments, the antitumor agents include monomethyloprestatin E (MMAE), monomethyloprestatin F (MMAF), paclitaxel, camptothecin, doxorubicin, and PF-06380101.

[0087] A "self-degrading group" refers to a group that undergoes an electronic cascade, resulting in the release of the group to which it is attached. In some embodiments, the self-degrading group comprises one or more groups that can undergo 1,4-elimination, 1,6-elimination, 1,8-elimination, 1,6-cyclization elimination, 1,5-cyclization elimination, 1,3-cyclization elimination, intramolecular 5-exo-trig cyclization, and / or 6-exo-trig cyclization. In some embodiments, the self-degrading group comprises valine (Val) and citrulline (Cit) or valine (Val) and alanine (Ala).

[0088] "Spacer group" refers to a linker covalently bonded between a self-degrading group and an antitumor agent. In one embodiment, the spacer group is para-aminobenzylcarbamate (PBA).

[0089] "Solid carrier" or "solid resin" refers to any solid matrix known in the art as a solid-phase carrier for molecules. For example, "microparticles" refers to any of a variety of small particles that can be used to attach to compounds of this disclosure, including but not limited to glass beads, magnetic beads, polymer beads, non-polymer beads, etc. In some embodiments, the microparticles comprise polystyrene beads. In some embodiments, the solid carrier or solid resin is controlled-porosity glass or macroporous polystyrene.

[0090] "Solid carrier residues" refer to functional groups that remain attached to the molecule when it is cleaved from the solid carrier. Solid carrier residues are known in the art and can be readily deduced based on the structure of the solid carrier and the groups of the molecule to which it is attached.

[0091] A “targeting moiety” is a portion that selectively binds to or associates with a specific target (such as an analyte molecule). “Selectively” binding or associating means that the targeting moiety preferentially associates with or binds to the desired target relative to other targets. In some embodiments, the compounds disclosed herein include a linker 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 the 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. In some embodiments, a targeting moiety is a portion (such as an antibody) that selectively binds to or associates with a target feature on or within a cell, such as a target feature on the cell membrane or other cellular structures, thereby allowing the detection of cells of interest. In some embodiments, small molecules that selectively bind to or associate with the desired analyte are also considered as targeting moieties. Other analytes and corresponding targeting moieties will be useful in different embodiments, as will be understood by those skilled in the art.

[0092] "Base-pairing moiety" refers to a heterocyclic moiety capable of hybridizing with a complementary heterocyclic moiety via hydrogen bonds (e.g., Watson-Crick base pairing). Base-pairing moieties include both native and non-native bases. Non-limiting examples of base-pairing moieties are RNA and DNA bases such as adenosine, guanosine, thymidine, cytosine, and uridine and their analogues.

[0093] The embodiments disclosed herein are also intended to cover all compounds that are 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.

[0094] Isotopically labeled compounds of structure (III) can generally be prepared using conventional techniques known to those skilled in the art, or by methods similar to those described below and the methods in the following examples, by using appropriate isotopically labeled reagents instead of previously used unlabeled reagents.

[0095] "Stable compound" and "stable structure" are intended to indicate a compound that is robust enough to withstand separation from the reaction mixture to a useful level of purity and formulation into an effective therapeutic agent.

[0096] "Optional" or "optionally" means that the event or situation described below may or may not occur, and the description includes instances where the event or situation occurs and instances where it does not occur. For example, "optionally substituted alkyl" means that the alkyl group may or may not be substituted, and the description includes substituted alkyl groups and unsubstituted alkyl groups.

[0097] "Salt" includes acid and base addition salts.

[0098] "Acid addition salt" refers to salts that react 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-acetaminobenzoic acid, camphoric acid, camphor-10-sulfonic acid, decanoic acid, hexanoic acid, octanoic acid, carbonic acid, cinnamic acid, citric acid, cyclohexanesulfonic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactopyric acid, gentianic acid, glucoheponic acid, etc.). Salts formed from glucuronic acid, glucuronic acid, glutamic acid, glutamate, 2-oxoglutamate, glycerophosphate, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucoic 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, pyruvic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanate, p-toluenesulfonic acid, trifluoroacetic acid, undecenoic acid, etc.

[0099] "Base addition salts" refer to salts prepared by adding an inorganic or organic base to a free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Salts derived from organic bases include, but are not limited to, the following: 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, diazonol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, phenethylbenzylamine, dibenzylethylenediamine, ethylenediamine, glucosamine, methyl-reduced glucosamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, etc. N -Ethylpiperidine, polyamine resins, etc. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0100] Crystallization can produce solvates of the compounds described herein. Embodiments of this disclosure include all solvates of the described compounds. As used herein, the term "solvate" refers to an aggregate of one or more molecules of a compound comprising this disclosure with one or more solvent molecules. The solvent may be water, in which case the solvate may be a hydrate. Alternatively, the solvent may be an organic solvent. Therefore, the compounds of this disclosure can exist as hydrates, including monohydrates, dihydrates, hemihydrates, sesquihydrates, trihydrates, tetrahydrates, etc., and corresponding solvated forms. The compounds of this disclosure may be true solvates, while in other cases, the compounds of this disclosure may retain only external water or another solvent, or a mixture of water and some external solvent.

[0101] Embodiments of the compounds of this disclosure (e.g., compounds of structure I) or their salts, tautomers, or solvates may contain one or more stereocenters, and thus may produce enantiomers, diastereomers, and other stereoisomers that can be defined (R)- or (S)-, or (D)- or (L)-, in absolute stereochemistry for amino acids. Embodiments of this disclosure are intended to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), ( R )-and( S (D)- and (L)- isomers, or optically active (+) and (-) isomers, can be separated using conventional techniques (e.g., chromatography and fractional crystallization). R )-and( S(D)- or (L)- isomers. Conventional techniques for the preparation / separation of each enantiomer include chiral synthesis from a suitable optically pure precursor, or resolution of the racemic mixture (or racemic mixture of the salt or derivative) using, for example, chiral high-performance liquid chromatography (HPLC). When the compounds described herein contain an alkene double bond or other geometrically asymmetric center, it is intended that the compounds include E and Z geometric isomers, unless otherwise stated. Similarly, it is intended to include all tautomer forms.

[0102] "Stereoisomers" refer to compounds consisting of identical atoms bonded by the same bonds but having different three-dimensional structures that are not interchangeable. This disclosure covers a variety of stereoisomers and mixtures thereof, and includes "enantiomers," which represent two stereoisomers whose molecules are non-overlapping mirror images of each other.

[0103] "Tautomer" refers to the transfer of a proton from one atom of a molecule to another atom within the same molecule. This disclosure includes tautomers of any of the compounds described. Various tautomer forms of the compounds can be readily deduced by those skilled in the art.

[0104] The chemical nomenclature schemes and structural diagrams used in this paper are improved versions of the IUPAC nomenclature system, using ACD / Name version 9.07 software and / or ChemDraw Ultra version 11.0 software (CambridgeSoft). Common names familiar to those skilled in the art are also used.

[0105] Methods for preparing compounds that can serve as covalent linkers between a bioactive moiety (such as an antitumor agent or an enediyne antitumor antibiotic) and a target moiety are provided. The methods of this disclosure allow the attachment of an amine-containing bioactive moiety to a polymer backbone via physiologically cleavable or non-cleavable linkers. The procedures described in this disclosure are capable of selectively attaching physiologically cleavable and / or non-cleavable linkers. This allows the synthesis of a compound having physiologically cleavable and non-cleavable linkers and one or more bioactive moieties. At this point, the bioactive moieties can be sequentially cleaved according to physiological conditions. Furthermore, compounds having multiple bioactive moieties and fluorescent or colored moieties linked by physiologically cleavable and / or non-cleavable linkers can be synthesized.

[0106] The method disclosed herein also allows control over the number and type of bioactive portions attached to the polymer backbone and any subsequent target portions, the spacing between adjacent bioactive portions on the polymer backbone (e.g., the distance or proximity between bioactive portions), and the spacing between the polymer backbone and the bioactive portions (e.g., the length of the linker of the polymer backbone).

[0107] The resulting compounds comprise combinations of therapeutic agents, targeting moieties, and dye moieties (e.g., chromophores or fluorophores), which can be used for simultaneous targeting, treatment, and detection. The ease of conjugating polymer-drug constructs with targeting agents (e.g., antibodies, antibody fragments, proteins, or other clinically interesting agents) offers practicality for a variety of applications of interest (e.g., surface chemistry, assay development, etc.).

[0108] The compounds formed by the methods of this disclosure also provide other desirable properties, including enhanced permeability and retention. In addition to providing the necessary solubility properties, the chemical characteristics of embodiments of the compounds of the present invention can be tuned to modulate the compounds' ability to penetrate diseased cells / tissues and remain within them. These characteristics allow for the efficient delivery of bioactive agents by increasing permeability and enhancing potency through enhanced retention.

[0109] In one embodiment, a method is provided for preparing a polymer-drug conjugate containing a bioactive moiety and a fluorescent or colored dye, the method comprising a compound having structure (I):

[0110]

[0111] Reaction with compounds of structure (II):

[0112]

[0113] in:

[0114] M 1 Each time it appears, it is independently a portion containing either fluorescent or colored dye;

[0115] M 2 It contains the bioactive portion or the fluorescent or colored dye portion;

[0116] G, each time it appears, is independently a part containing a reactive group, a group that can be converted into a reactive group, or a protected analogue thereof, which is capable of forming a covalent bond with an amino group;

[0117] L 1 and L 7 Each time it appears independently, it is an alkylene group, an alkenylene group, an ynylene group, a heteroalkylene group, an alkenylene group, an arylene group, or a heteroaryl linker or a combination thereof;

[0118] L 2 L 3 L 5 and L 6 It is either independent of any group that does not exist in each instance, or it is a alkylene group, alkenyl group, alynyl group, heteroalkyl group, alkenyl group, or alynyl group.

[0119] L 4 Each time it appears, it is independently an alkylene, alkenylene, ynylene, heteroalkylene, alkenylene, or ynylene linkage;

[0120] R 1 and R 2 Each of these can be independently H, OH, SH, alkyl, alkoxy, alkyl ether, heteroalkyl, -OP (=R) a (R) b )R c Or Q;

[0121] R 3 Each time it appears, it is independently H, alkyl, or alkoxy;

[0122] R 4 Each time it appears, it is independently OH, SH, or O. - S - OR d or SR d ;

[0123] R 5 Each occurrence is independently either oxidized, thiolated, or absent;

[0124] R 6 and R 7 Each time it appears, it is independently H, OH, or a halogen;

[0125] R 8 It is H or alkyl;

[0126] R a Is it O or S?

[0127] R b It is OH, SH, O - S - OR d or SR d ;

[0128] R c It is OH, SH, O - S - OR d OL' SR dAlkyl, alkoxy, heteroalkyl, heteroalkoxy, alkyl ether, alkoxyalkyl ether, phosphate ester, thiophosphate ester, phosphoryl alkyl, thiophosphoryl alkyl, phosphoryl alkyl ether or thiophosphoryl ether;

[0129] R d It is a counter ion;

[0130] Q is independently a portion containing a reactive group or its protected form each time it appears, which is capable of forming a covalent bond with the complementary reactive group Q′ on the target portion;

[0131] L' independently, each time it appears, is a linker containing a covalent bond with Q, a target portion, a linker containing a covalent bond with the target portion, a linker containing a covalent bond with the solid support, a linker containing a covalent bond with the solid support residue, a solid support residue, a linker containing a covalent bond with a nucleoside, or a linker containing a covalent bond with other compounds of structure (I).

[0132] m and p are independent integers equal to or greater than 0 each time they appear;

[0133] w is an integer equal to or greater than 1 each time it appears; and

[0134] n is an integer equal to or greater than 1.

[0135] Therefore: i) the G in the compound with structure (I) and the -NHR in the compound with structure (II) 8 Bonds are formed between them.

[0136] Various linking groups and substituents (e.g., M) in compounds with structure (I) 1 R 1 R 2 R 3 R 4 R 5 R 6 R 7 L 1 L 2 L 3 L 4 L 5 L 6 L 7(I) and (Q) are optionally substituted with one or more substituents. For example, in some embodiments, the optional substituents are selected to optimize the water solubility or other properties of the compound of structure (I). In some embodiments, each alkyl, alkoxy, alkyl ether, alkoxyalkyl ether, phosphorylalkyl, thiophosphorylalkyl, phosphorylalkyl ether, and thiophosphorylalkyl ether in the compound of structure (I) is optionally substituted with one or more substituents selected from hydroxyl, alkoxy, alkyl ether, alkoxyalkyl ether, mercapto, amino, alkylamino, carboxyl, phosphate ester, thiophosphate ester, phosphorylalkyl, thiophosphorylalkyl, phosphorylalkyl ether, and thiophosphorylalkyl ether. In some embodiments, the optional substituent is -OP (=R a (R) b )R c , where R a R b and R c As defined for compounds with structure (I).

[0137] In some embodiments, G is independently -C(=O)OH or -C(=O)H each time it appears. In some relevant embodiments, the compound of structure (I) has the following structure (IA) or (IB):

[0138] or

[0139]

[0140] Connecting base L 1 It can be used as a connection point between the G group and the rest of the compound.

[0141] In some implementations, L 1 At least once, L contains a linker that is not cleavable under physiological conditions. Therefore, in some embodiments, L 1 It includes amide bonds, ester bonds, disulfide bonds, hydrazones, triphosphates, diesters, β-glucuronides, double bonds, triple bonds, ether bonds, ketones, diols, or combinations thereof.

[0142] In some implementations, L 1 It contains tert-butyloxycarbonyl, p-methoxybenzyl, dialkyl or diaryldialkoxysilane, orthoester, acetal, β-thiopropionate, ketal, aminophosphate, hydrazone, vinyl ether, imine, aconitol, triphenylmethyl, polyketal, diarylhydrazone, diazobenzene, vivinal diol, pyrophosphate diester, or valine-citrulline.

[0143] In some implementations, L 1 Each time it appears, it is independently a linker that can be cleaved within a pH range of 6 to 8. For example, in some embodiments, L 1It is a linker that can be cleaved at pH 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9 or 8.0.

[0144] In some implementations, L 1 Each time L occurs, it is independently a linker that can cleave within a temperature range of 20°C to 40°C, 25°C to 35°C, 30°C to 35°C, 30°C to 37°C, 35°C to 37°C, 35°C to 40°C, or 32°C to 38°C. In some embodiments, L is independently a linker that can cleave within a temperature range of about 20°C, about 21°C, about 22°C, about 23°C, about 24°C, about 25°C, about 26°C, about 27°C, about 28°C, about 29°C, about 30°C, about 31°C, about 32°C, about 33°C, about 34°C, about 35°C, about 36°C, about 37°C, about 38°C, about 39°C, or about 40°C.

[0145] In some implementations, L 1 Each time it appears, it is independently a linker that can be cleaved by an enzyme. For example, in some embodiments, the enzyme is a hydrolase, an oxidoreductase, or a lyase. In some embodiments, the enzyme is an EC 4.1 (e.g., EC 4.1.1, EC 4.1.2, EC 4.1.3, or EC 4.1.99), EC 4.2, EC 4.3, EC 4.4, EC 4.5, EC 4.6, or EC 4.99 enzyme.

[0146] In some implementations, for L 1 L appears at least once. 1 -G has the following structure:

[0147] .

[0148] In some implementations, L 1a At least once, L appears as a optionally substituted 5-9 member heteroaryl linker. In some embodiments, L 1a It is the substituted 5-membered heteroaryl linker. In some embodiments, L 1a It is the substituted 6-membered heteroaryl linker. In some embodiments, L 1a It is the substituted 7-membered heteroaryl linker. In some embodiments, L 1a It is the substituted 8-membered heteroaryl linker. In some embodiments, L 1a It is the substituted 9-membered heteroaryl linker. In some relevant embodiments, L 1aIt is substituted with an oxo, an alkyl (e.g., methyl, ethyl, etc.) or a combination thereof. In some embodiments, L 1a It is not replaced each time it appears. In some more specific implementations, L 1a It is pyrimidine independently each time it appears. In some more specific embodiments, L 1a Each time it appears, it is independently either cytosine or thymine. In some embodiments, L 1a Each time it appears, it is independently selected from cytosine and thymine, such that the compound contains a sequence of cytosine and thymine bases capable of forming a triplet with the target DNA sequence.

[0149] In some implementations, L 1a It has one of the following structures:

[0150]

[0151] In some implementations, L 1b Each time it appears independently, it is optionally alkylene, alkenylene, alkyne, heteroalkylene, alkenylene, alkyne, alkyleneheteroarylalkylene, alkyleneheterocyclicalkylene, alkylenecarbocyclicalkylene, heteroalkyleneheteroarylalkylene, heteroalkyleneheterocyclicalkylene, heteroalkylenecarbocyclicalkylene, heteroalkyleneheteroarylalkylene, heteroalkyleneheterocyclicalkylene, heteroalkylenecarbocyclicalkylene Alkyl, alkyleneheteroarylheteroalkyl, alkyleneheterocyclic heteroalkyl, alkylenecarbamocyclic heteroalkyl, heteroaryl, heterocyclic, carbamocyclic, alkyleneheteroaryl, alkyleneheterocyclic, heteroarylalkylene, alkylenecarbamocyclic, carbamocyclic alkylene, heteroarylalkylene, heteroalkylheterocyclic, heteroarylalkylene, heteroalkylcarbamocyclic, carbamocyclic heteroalkyl or heteroatom-linked group. In some embodiments, L 1b It is an optional substituted hemienyl linker.

[0152] In some more specific implementations, L a and L b Together (i.e., It has one of the following structures:

[0153] or .

[0154] In some implementations, L 1 The presence of at least one of the amino acid residues is present. In some embodiments, the amino acid residue is valine. In some embodiments, L... 1 The at least one occurrence of contains one of the following structures:

[0155] ;

[0156] ;

[0157] ; ;

[0158] ; ;

[0159] ; ; ;

[0160] ;

[0161] ; ;

[0162] ; or

[0163] .

[0164] In some implementations, L 1 The at least one occurrence of contains one of the following structures:

[0165] ; ; ; ; ; ; ;

[0166] ; ; ; ; ;

[0167] ; ; ; ;

[0168] ; ;

[0169] ; ; ; or .

[0170] In some implementations, L 4At least once, L is a heteroalkylene group. In some embodiments, L 4 Each occurrence of L is a heteroalkylene group. In some embodiments, L 4 At least one occurrence of L contains an alkylene oxide. In some more specific embodiments, L 4 The alkyl oxide is ethylene oxide. In some more specific embodiments, the ethylene oxide is polyethylene oxide. In some embodiments, L 4 It has the following structure each time it appears:

[0171] ,

[0172] in:

[0173] z is an integer from 1 to 100; and

[0174] Indicates the bond with the adjacent phosphorus atom.

[0175] In some implementations, z is an integer from 3 to 8. In some implementations, z is 3. In some implementations, z is 4. In some implementations, z is 5. In some implementations, z is 6. In some implementations, z is 7. In some implementations, z is 8.

[0176] In some other embodiments, z is an integer from 22 to 26. In some embodiments, z is 22. In some embodiments, z is 23. In some embodiments, z is 24. In some embodiments, z is 25. In some embodiments, z is 26.

[0177] In some other embodiments, z ranges from 19 to 28. In some embodiments, the average z is 23. In some embodiments, the average z is 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28.

[0178] In some embodiments, in the compound of structure (I) The at least one occurrence of contains one of the following structures:

[0179] ; ; ;

[0180] ; ; or

[0181] ,

[0182] Where z is an integer from 1 to 100.

[0183] In some implementations, z is an integer from 3 to 8. In some other implementations, z is an integer from 22 to 26. In some other implementations, z ranges from 19 to 28.

[0184] In some embodiments, each time it appears, in the compound of structure (I) It contains one of the following structures:

[0185] ; ; ;

[0186] ; ; or

[0187] .

[0188] In some specific implementation schemes, It contains one of the following structures:

[0189] or .

[0190] In some more specific implementation schemes, It includes the following structure:

[0191] .

[0192] In some other specific implementations, It includes the following structure:

[0193] .

[0194] In some embodiments, the compound of structure (I) has the following structure (IC) or (ID):

[0195] or

[0196] .

[0197] In some implementations, L 2 L 3 L 5 and L 6 It is independently absent (i.e., a direct linker) or independently an alkylene, alkenylene, ynylene, heteroalkylene, alkenylene, or heteroynyl linker each time it appears. In some embodiments, L 2 L 3 L 5 and L 6It is an alkylene group each time it appears. In further embodiments, L 2 L 3 L 5 and L 6 Each time it appears, it is independently a C1-C6 alkylene, C2-C6 alkenylene, or C2-C6 ynylene. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C1 alkylene linker. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C2 alkylene linker. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C3 alkylene linker. In some more specific embodiments, L 3 and L 5 The alkylene linker is a C4 alkylene linker. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C5 alkylene linker. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C6 alkylene linker. In some other embodiments, L 2 L 3 L 5 and L 6 At least once, L appears as a direct-connect key. In some implementations, L 2 At least one occurrence of L is absent. In a more specific implementation, L 2 It does not exist each time it appears.

[0198] In some relevant embodiments, the compound of structure (I) has the following structure (IE) or (IF):

[0199] or

[0200] ,

[0201] Where y 1 y 2 y 3 and y4 Each occurrence is an integer from 0 to 6.

[0202] In some implementations, y 1 y 2 y 3 and y 4 It is an integer 0 each time it appears (i.e., L). 2 L 3 L 5 and L 6 Each of them is a direct-connect key. In some implementations, y 1 y 2 y 3 and y 4 It is an integer 1 each time it appears. In some implementations, y 1 y 2 y 3 and y 4 It is an integer of 2 each time it appears. In some implementations, y 1 y 2 y 3 and y 4 It is an integer 3 each time it appears. In some implementations, y 1 y 2 y 3 and y 4 It is an integer of 4 each time it appears. In some implementations, y 1 y 2 y 3 and y 4 It is an integer 5 each time it appears. In some implementations, y 1 y 2 y 3 and y 4 It is an integer 6 each time it appears. In some more specific implementations, y 1 , y 2 , y 3 , and y 4 Each occurrence is either 0 or 1.

[0203] Connecting base L 7 Can be used as M 1 The connection point between part and the rest of the compound (e.g., the heteroalkyl backbone). For example, in some embodiments, a synthetic precursor of the compound of structure (I) is prepared, and M is connected using any number of simple methods known in the art, such as methods known as "click chemistry". 1 Partially linked to the synthetic precursor. For this purpose, any rapid and substantially irreversible reaction can be used to link M... 1The compound is linked to a synthetic precursor to form structure (I). Exemplary reactions include copper-catalyzed reactions of azides and alkynes to form triazoles (Huisgen 1,3-dipolar cycloaddition), reactions of dienes and dienophiles (Diels-Alder), strain-promoted alkyne-nitroketone cycloaddition, strain-promoted cycloalkyne-azide cycloaddition (without Cu click), reactions of strained alkenes with azides, tetrazides, or tetrazoles, cycloadditions of alkenes and azides [3+2], reverse-demand Diels-Alder reactions of alkenes and tetrazides, photoreactions of alkenes and tetrazoles, and various substitution reactions, such as substitution of leaving groups by nucleophilic attack on electrophilic atoms. Exemplary substitution reactions include reactions of amines with activated esters; N-hydroxysuccinimide esters; isocyanates; isothiocyanates, etc. In some embodiments, L is formed. 7 The reaction can be carried out in an aqueous environment.

[0204] Therefore, in some implementations, L 7 Each time it appears, it is independently a linker group containing a functional group capable of being formed through a reaction of two complementary reactive groups, such as a functional group of the product of one of the aforementioned "click" reactions. In different embodiments, for L 7 The functional group can be formed by the reaction of an oxime, hydrazone, alkyne, amine, azide, acyl azide, acyl halide, nitrile, nitrone, mercapto, disulfide, sulfonyl halide, isothiocyanate, imine ester, activated ester (e.g., N-hydroxysuccinimide ester), ketone, α,β-unsaturated carbonyl, olefin, maleimide, α-haloimide, epoxide, aziridine, tetrazine, tetrazolium, phosphine, biotin, or thiamethoxamyl ring functional group with a complementary reactive group, for example, by the reaction of an amine with an N-hydroxysuccinimide ester or isothiocyanate.

[0205] In other implementations, for L 7 The functional group can be formed through the reaction of alkynes and azides, with at least one occurrence of [the specific component]. In other embodiments, for L [the specific component]... 7 The functional group can be formed by the reaction of an amine (e.g., a primary amine) with an N-hydroxysuccinimide ester or an isothiocyanate, with at least one occurrence of the functional group.

[0206] In more implementations, for L 7 The functional group includes at least one occurrence of olefin, ester, amide, thioester, disulfide, carbocyclic, heterocyclic, or heteroaryl group. In further embodiments, for L 7 The functional group comprises at least one olefin, ester, amide, thioester, thiourea, disulfide, carbocyclic, heterocyclic, or heteroaryl group. In other embodiments, the functional group comprises an amide or thiourea. In some more specific embodiments, for L... 7L appears at least once. 7 It is a linker group containing a triazole functional group. In some relevant embodiments, L 7 It independently contains a triazole functional group each time it appears. In other embodiments, however, for L... 7 L appears at least once. 7 It is a linker group containing amide or thiourea functional groups.

[0207] In some implementations, L 7 At least once, L is an alkylene or heteroalkylene group. In a more specific embodiment, L 7 The at least one occurrence of contains one of the following structures:

[0208] ; ; ; ; ;

[0209] ; ; ; ; ; ;

[0210] , ; ; ; ; ; or .

[0211] In a more specific implementation plan, L 7 The at least one occurrence of contains one of the following structures:

[0212] ; or .

[0213] In some more specific implementations, the connection base L 7 It has the following structure:

[0214] .

[0215] In other implementation schemes, for L 7 L appears at least once. 7 -M 1 It has one of the following structures:

[0216] or

[0217] Where L 7a and L 7b Each is an optional linker.

[0218] In some implementations, L 7a or L 7b Or neither of them exists. In other implementations, L 7a or L 7b Or both may exist.

[0219] In some implementations, L 7a and L 7b When present, each is independently an alkylene or heteroalkylene. For example, in some embodiments, L 7a and L 7b When it exists, it independently has one of the following structures:

[0220] ; ; ; ;

[0221] ; ;or .

[0222] In some implementations, M 1 -L 7 It has one of the following structures:

[0223] ; ;or ,

[0224] Where a, b, c, and d are each an independent integer in the range 1-6. In some embodiments, a, b, c, and d are each an independent integer 1. In some embodiments, a, b, c, and d are each an independent integer 2. In some embodiments, a, b, c, and d are each an independent integer 3. In some embodiments, a, b, c, and d are each an independent integer 4. In some embodiments, a, b, c, and d are each an independent integer 5. In some embodiments, a, b, c, and d are each an independent integer 6.

[0225] In some more specific implementations, L 7 The at least one occurrence of contains one of the following structures:

[0226] ; ;

[0227] ; or

[0228] or

[0229] .

[0230] In some implementations, L 7 The at least one occurrence of contains one of the following structures:

[0231] ; ; ; ; or .

[0232] In some implementations, R 3 The at least one occurrence of is H. In some implementations, R 1 At least once, R is an alkyl group. In some embodiments, R 1 At least once, it appears as an alkoxy group. In some embodiments, R 1 Each occurrence of is H.

[0233] In some implementations, R 4 At least one occurrence of is OH. In some embodiments, R 4 The at least one occurrence of is SH. In some implementations, R 4 The at least one occurrence of O is - In some implementations, R 4 The most frequent occurrence of S is - In some implementations, R 4 Each occurrence is O - .

[0234] In some implementations, R 5 At least once, it occurs as an oxygen (=O). In some implementations, R... 5 At least once, R is thiolated (=S). In some implementations, R 5 Each occurrence of is an oxygenation (=O).

[0235] In some implementations, R 1 and R 2 Each is independently OH or -OP (=R) a (R) b )R c In some different implementations, R 1 or R 2 Is it OH or -OP (=R) a (R) b )R c And R 2or R 3 The other one is Q or a linker containing a covalent bond with Q.

[0236] In further different embodiments of any of the aforementioned compounds of structure (I), R 2 and R 3 Each is independently -OP(=R) a (R) b )R c In some of these implementation schemes, R c She is an office lady (OL).

[0237] In other implementation schemes, R 1 and R 2 Each is independently -OP(=R) a (R) b )OL', where L' is an alkylene or heteroalkylene linker to: Q, the target moiety, the analyte (e.g., the analyte molecule), the solid support, the solid support residue, the nucleoside, or other compound of structure (I).

[0238] The linker L' can be any linker suitable for linking Q, the target moiety, the analyte (e.g., the analyte molecule), the solid support, solid support residues, the nucleoside, or other compounds of structure (I) to the compound of structure (I). Advantageously, some embodiments include the use of a selected L' moiety for increasing or optimizing the water solubility of the compound. In some embodiments, L' is a heteroalkylene moiety. In some other embodiments, L' comprises an alkylene oxide or a phosphate diester moiety, or a combination thereof.

[0239] In some implementations, L' has the following structure:

[0240] ,

[0241] in:

[0242] m'' and n'' are independent integers from 1 to 10;

[0243] R e It is H, an electron pair, or a counter ion;

[0244] L'' is R e Or a direct bond or connection with the following substances: Q, target moiety, analyte (e.g., analyte molecule), solid support, solid support residue, nucleoside or other compound of structure (I).

[0245] In some implementations, m'' is an integer from 4 to 10, such as 4, 6, or 10. In other implementations, n'' is an integer from 3 to 6, such as 3, 4, 5, or 6. In some implementations, n'' is an integer from 18 to 28, such as 21 to 23.

[0246] In some other embodiments, L'' is an alkylene, alkylene heterocyclic, alkylene heterocyclic alkylene, alkylene cyclocyclic, alkylene cyclocyclic alkylene, heteroalkyl, heteroalkylene heterocyclic, heteroalkylene heterocyclic, heteroalkylene heterocyclic, or heteroalkylene cyclocyclic heteroalkyl moiety. In some other embodiments, L'' comprises an alkylene oxide, a phosphate diester moiety, a mercapto, a disulfide, or a maleimide moiety, or a combination thereof.

[0247] In some of the foregoing embodiments, the targeting portion is an antibody or a cell surface receptor antagonist. In some 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 folic acid (folate) inhibitor, or a MET inhibitor. In other embodiments, the targeting portion is a monoclonal antibody. For example, in some embodiments, the monoclonal antibody is abciximab, adalimumab, alenzumab, alixiromumab, avibactam, baliximab, benaribizumab, belottosumab, lantomumab, brodamarab, ibuprofen, cananulumab, caralacilumab, pefozilumab, dacrolimus, denosumab, dupilumab, iculizumab, emecizumab, errenumab, evolocumab, remannetizumab, ganezumab, golimumab, gusekumab, ibaribizumab, idasarizumab, infliximab, itolizumab, or ixek. Izumab, Lanalimumab, Logivir, Meperizumab, Natalizumab, Otosaximab, Omarizumab, Omarzozab, Pallizumab, Rhabizumab, Recibakumab, Relizumab, Rmab, Roveizumab, Serlizumab, Sarilumab, Secukinumab, Titrajizumab, Thiomab, Tocilizumab, Ustenozumab, Vedolizumab, Aliruzumab, Actosumab, Adunazumab, Afasevikumab, Afimozumab, Aniluzumab, Amluzumab (IMA-638), Aserizumab, Atorumab Antibiotics, Bavizumab, BCD-100, Betemizumab, Besoxumab, Bisimab, Bimaluzumab, Bimecrolimus, Potoemizumab, Blelurumab, Butoxuzumab, Bercosizumab, Brazikumab, Brenuzumab, Bloxacillin, Karjuzumab, Carotuximab, Cializumab, Clarizumab, Klexixib, Constitución, Covirxixib, CR6261, Crenzumab, Lizanilizumab, Clotidumab, Depertuximab, Mofostatin, Dexamethasone, Biotin, Dizamizumab, Dilivoxib, Domalolimumab Antibiotics, Dustacizumab, Emexizumab, Ebazumab, Effluzumab, Efengumab, Edirumab, Elenumab, Enoxacizumab, Eptinezumab, Etrolizumab, Evezumab, Ivermab, Fanolesomab, Farimozumab, Faricimab, Fasnurumab, Fazolinumab, Fezanumab, Flanuzumab, Vorutuzumab, Avantuzumab, Flavoximumab, Flavoximumab, Flavoximumab, Gantinuzumab, Gavistumab, Gevozozabumab, Ginslurumab, GoliximabGosuranemab, Ilimumab, Inclacumab, Iomab-B, Keliximab, Lampalizumab, Lanroizumab, Raviriximab, Lejinizumab, Lenvervimab, Lelizumab, Rivirizumab, Ligligizumab, Lodicilizumab, Pego-Rullizumab, Matacizumab, Mafullimumab, Metilimumab, Mijizumab, Mavitizumab, Morozumab CD3, Nebacoozumab, Nemolizumab NEOD001, Nisevichimab, Odumozab, Olendalizumab, Olozumab, OMS721, Opinumab, Otetuzumab, Oxizumab, Otilimab, Oceluzumab, Ozanezumab, Olizumab, Paxizumab, Panocumumab, Pacozumab, Pallizumab, PDR001, Peregrinezumab, Pecrozumab, Pralucumab, Lozanezumab, Ponizumab, Porgaviximab, Prasinezumab, PRO 140. Quinolizumab, Raviolizumab, Ravigalimab, Ranevetmab, Ravagalimab, Refanelizumab, Regavir, Rilaribumab, Linusuzumab, Risankizumab, Roledozab, Lomoxolizumab, Rolizumab, SA237, Saturizumab, Sevelimumab, SHP647, Sifamumab, Simutuzumab, Silizumab, Sirukumab, Sulanzumab, Sonepizumab, Spartalizumab, Staphylocumab, Thioxamab, Sutavazolam, Sulizumab, Suvetazab, Sutosulamab, Taduzumab, Talizumab, Tantovelumab, Tanizumab, Tefizumab, Atemozumab, Tenexizumab, Tefizumab, Tetolimumab, Terzaluzumab, Teblizumab, Tolizumab, Traroluzumab, Tragoluzumab, Tovirazumab, Uroluzumab, Uruzumab, Varixuzumab, Verpamob, Vesenocurumab, Vexizumab, Wabalizumab, Azomob, Trastuzumab, Gelatuzumab, Brentuximab, Woseltuzumab, Lovotuzumab, Cantuzumab, Bivatuzumaborinotuzumab, or Vardatuzumab.

[0248] In some embodiments, the analyte molecule is a nucleic acid, an amino acid, or a polymer thereof. In other embodiments, the analyte molecule is an enzyme, a receptor, a receptor ligand, an antibody, a glycoprotein, an aptamer, or a prion.

[0249] In some embodiments, the solid carrier is a polymer bead or a non-polymer bead.

[0250] In other more specific embodiments of any of the aforementioned compounds of structure (I), R 1 Or R 2 It has one of the following structures:

[0251] ; ; ; ; ; ; or .

[0252] According to solid-phase synthesis methods similar to those known in the art for preparing oligonucleotides, certain embodiments of compounds of structure (I) can be prepared. Therefore, in some embodiments, L' is a link to a solid support, solid support residues, or a nucleoside. Solid supports containing activated deoxythymidine (dT) groups are readily available and, in some embodiments, can be used as starting materials for preparing compounds of structure (I) or (II). Therefore, in some embodiments, R... 1 Or R 2 It has the following structure:

[0253] or

[0254] .

[0255] Those skilled in the art will understand that the inclusion of the aforementioned dT group is merely for ease of synthesis and economic efficiency, and is not essential. Other solid supports may be used, resulting in the presence of different nucleoside or solid support residues at L', or the nucleoside or solid support residues may be removed or modified post-synthesis.

[0256] In some embodiments, the analyte molecule is a nucleic acid, an amino acid, or a polymer thereof. In some embodiments, the analyte molecule is an enzyme, a receptor, a receptor ligand, an antibody, a glycoprotein, an aptamer, or a prion. In some embodiments, the targeting portion is an antibody or a cell surface receptor antagonist. In other embodiments, the solid carrier is a polymer bead or a non-polymer bead.

[0257] In other embodiments, Q is an independently occurring portion containing a reactive group capable of forming a covalent bond with the analyte molecule or solid support. In other embodiments, Q is an independently occurring portion containing a reactive group capable of forming a covalent bond with a complementary reactive group Q′. For example, in some embodiments, Q′ is present on another compound of structure (I) (e.g., in R...). 2 Or R 3(Position), and Q and Q′ contain complementary reactive groups, such that the reaction of the compound of structure (I) with other compounds of structure (I) produces a dimer of the compound of structure (I) covalently bonded. Polymeric compounds of structure (I) can also be prepared in a similar manner, and said polymeric compounds are included within the scope of embodiments of the present invention.

[0258] The type of Q group and the connectivity of the Q group with the rest of the compound of structure (I) are not restricted, provided that Q contains a portion with appropriate reactivity for forming the desired bond.

[0259] In some embodiments, Q is the portion that is not easily hydrolyzed under aqueous conditions but is sufficiently reactive to form a bond with a corresponding group (e.g., amine, azide, or alkyne) on the analyte molecule or solid support.

[0260] Some embodiments of the compound of structure (I) include a Q group commonly used in the field of biological conjugation. For example, in some embodiments, Q comprises a nucleophilic reactive group, an electrophilic reactive group, or a cycloaddition reactive group. In some more specific embodiments, Q comprises a thiol, disulfide, activated ester, isothiocyanate, azide, alkyne, olefin, diene, dienophile, acyl halide, sulfonyl halide, phosphine, α-haloamide, biotin, amino, or maleimide functional group. In some embodiments, the activated ester is an N-succinimide ester, an imine ester, or a polyfluorophenyl ester. In other embodiments, the alkyne is an alkyl azide or an acyl azide.

[0261] The Q group can be conveniently provided in a protected form to increase storage stability or other desired properties, and then the protecting group can be removed at an appropriate time to conjugate with, for example, a target moiety or analyte. Therefore, the Q group includes the “protected form” of a reactive group, including any reactive group described in Table 1 above and below. The “protected form” of Q refers to a moiety that is less reactive than Q under predetermined reaction conditions, but can be converted to Q under conditions that preferably do not degrade other moieties of compounds with structure (I) or (II) or do not react with them. Depending on the specific Q and the desired end use and storage conditions, those skilled in the art can deduce the appropriate protected form of Q. 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.

[0262] An example Q section is provided in Table I below.

[0263] Table 1. Exemplary Q section

[0264]

[0265]

[0266]

[0267] It should be noted that in some embodiments, where Q is SH, the SH portion will tend to form a disulfide bond with another thiol group (e.g., on another compound of structure (I)). Therefore, some embodiments include compounds of structure (I) in the form of a disulfide dimer, with the disulfide bond derived from the SH Q group.

[0268] In other embodiments, the Q portion is conveniently masked (e.g., protected) as a disulfide portion, which can then be reduced to provide an activated Q portion for binding to a desired analyte molecule or targeting moiety. For example, the Q portion can be masked as a disulfide having the following structure:

[0269]

[0270] Wherein R is an alkyl group that is optionally substituted. For example, in some embodiments, Q is provided as a disulfide moiety having the following structure:

[0271]

[0272] Where n' is an integer from 1 to 10.

[0273] In some embodiments, the at least one fluorescent or colored portion M 1 It is the fluorescent portion. In some embodiments, M 1 Each time it appears, it independently contains two or more aryl or heteroaryl rings or combinations thereof, such as three or more or four or more aryl or heteroaryl rings or combinations thereof, or even five or more aryl or heteroaryl rings or combinations thereof. In some embodiments, M 1 Each occurrence independently comprises six aryl or heteroaryl rings or combinations thereof. In other embodiments, the rings are fused. For example, in some embodiments, M 1 Each occurrence independently contains two or more fused rings, three or more fused rings, four or more fused rings, five or more fused rings, or even six or more fused rings. In some more specific embodiments, M 1 Each occurrence independently contains a fused polycyclic aryl moiety, which comprises at least two fused rings.

[0274] In some specific implementations, M 1Each time it appears, it is independently selected from dimethylaminobis(2-)-styrene, quinacridone, fluorophenyl-dimethyl-BODIPY, difluorophenyl-BODIPY, acridine, trinaphthalene-diphenyl, hexaphenyl, porphyrin, benzo[a]pyrene, (fluorophenyl-dimethyl-difluoroborazine-diaza-indacene)phenyl, (difluorophenyl-difluoroborazine-diaza-indacene)phenyl, tetraphenyl, bibenzothiazole, tribenzothiazole, binaphthalene, bianthracene, squaric acid cyanine, squaric acid ontium, 9,10-ethynylanthracene, and trinaphthalene moiety.

[0275] In some implementations, M 1 Each time it appears, it is independently selected from p-terphenyl, dinaphthalene, azobenzene, phenazine, phenanthroline, acridine, thioxanthate, oxane, rubrene, benzoin, anthocyanin, dinaphthalene imide, dinaphthalene amide, and their derivatives. In some embodiments, M 1 Each time it appears, it is independently selected from coumarin dyes, halogen dyes, dipyrrole methylene boron difluoride dyes, ruthenium bipyridine dyes, thiazole orange dyes, polyacetylenes, and N-aryl-1,8-naphthalenediamide dyes. In some embodiments, M 1 M 2 Or M 3 Each time it appears, it is independently selected from coumarin dyes, boron-dipyrro methylene, rhodamine, anthocyanin, pyrene, tinamylene, tinamylene monoimide, 6-FAM, 5-FAM, 6-FITC, 5-FITC, and their derivatives.

[0276] In some other implementations, M 1 It independently possesses one of the following structures each time it appears:

[0277]

[0278]

[0279] In some implementations, M 1 It independently possesses one of the following structures each time it appears:

[0280] ; ; ; ; ;

[0281] ; or .

[0282] In some more specific implementations, M 1 The at least one occurrence of has the following structure:

[0283] .

[0284] In some more specific implementations, M 1 Each occurrence of has the following structure:

[0285] .

[0286] In some more specific implementations, -L 7 -M 1 The at least one occurrence of has the following structure:

[0287] .

[0288] In some more specific implementations, -L 7 -M 1 Each occurrence of has the following structure:

[0289] .

[0290] By selecting different values ​​of m, n, p, and w, the fluorescence intensity or the efficacy of treating diseases can be adjusted.

[0291] The value of m controls the neighboring M 1 Or M 2 The ability to specify the spacing between elements. In some embodiments, m is an integer equal to or greater than 0. In some embodiments, m is an integer from 0 to 10. In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, m is 3. In some embodiments, m is 4. In some embodiments, m is 5. In some embodiments, m is 6. In some embodiments, m is 7. In some embodiments, m is 8. In some embodiments, m is 9. In some embodiments, m is 10. In some more specific embodiments, m is 1 or 2.

[0292] The number of fluorescent dye moieties attached to the polymer backbone can adjust the fluorescence intensity. The value of p has the ability to control the brightness of the compound. In some embodiments, p is an integer equal to or greater than 0 each time it occurs. In some more specific embodiments, p is 0 to 10. In some embodiments, p is 0 to 5. For example, in some embodiments, p is 0. In some embodiments, p is 1. In some embodiments, p is 2. In some embodiments, p is 3. In some embodiments, p is 4. In some embodiments, p is 5.

[0293] The efficacy of fluorescence or treatment can also be adjusted by selecting the value of w. In some embodiments, w is an integer equal to or greater than 1. In some embodiments, w is an integer from 1 to 100. In other embodiments, w is an integer from 1 to 20. In other embodiments, w is an integer from 1 to 10. In some embodiments, w is 1, 3, 5, 7, or 10. In some embodiments, w is 1. In some embodiments, w is 2. In some embodiments, w is 3. In some embodiments, w is 4. In some embodiments, w is 5. In some embodiments, w is 6. In some embodiments, w is 7. In some embodiments, w is 8. In some embodiments, w is 9. In some embodiments, w is 10.

[0294] By selecting different values ​​of n, the fluorescence intensity or the efficacy of treating the disease can also be adjusted. In some embodiments, n is an integer equal to or greater than 1. In some embodiments, n is an integer from 1 to 100. In some embodiments, n is an integer from 1 to 20. In other embodiments, n is an integer from 1 to 10. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10.

[0295] In some specific implementations, m is 6, p is 1, w is 1, and n is 1. In some specific implementations, m is 6, p is 1, w is 2, and n is 1.

[0296] In some implementations, M 2 It is the biologically active portion. In some embodiments, M 2 It is an antitumor agent. In some embodiments, the antitumor agent M 2 Each time it appears, it is independently selected from paclitaxel, camptothecin, doxorubicin, monomethylolpropionate E (MMAE), Val-Cit-PAB-MMAE, and monomethylolpropionate F. In some embodiments, M 2 It has one of the following structures:

[0297] ;

[0298] ; ;

[0299] or

[0300] .

[0301] In some embodiments, the antitumor agent M 2 It is monomethylolpropionate E. In some more specific embodiments, the antitumor agent M... 2 It has the following structure:

[0302] .

[0303] In some implementations, M 2 It is an antitumor antibiotic. In some embodiments, the antitumor antibiotic M... 2 It is selected from pepromycin sulfate, pirarubicin hydrochloride, fentostatin ester, idarubicin hydrochloride, mitomycin C, bleomycin hydrochloride, doxorubicin hydrochloride, daunorubicin hydrochloride, and epirubicin hydrochloride.

[0304] In some implementations, M 2 It is an enediyne antitumor antibiotic. In some embodiments, the enediyne antitumor antibiotic M... 2 It is calichiomycin, or more specifically calichiomycin γ1.

[0305] In some implementations, M 2 It has one of the following structures:

[0306] ;

[0307] ;or

[0308] .

[0309] In some implementations, M 2 As mentioned in this article regarding M 1 The fluorescent or colored dye described.

[0310] In some specific embodiments, the compound of structure (I) is selected from the compounds in Table 2.

[0311] Table 2. Compounds with exemplary structure (I)

[0312]

[0313] In Table 2, FAM represents

[0314] or .

[0315] In some embodiments, the compounds of this disclosure formed by reacting the compound of structure (I) with the compound of structure (II) have the following structure (III):

[0316] ,

[0317] (III)

[0318] in:

[0319] M 1 Each time it appears, it is independently a portion containing either fluorescent or colored dye;

[0320] M 2 Each occurrence is independently a portion containing either a biologically active component or a fluorescent or colored dye, provided that M 2 At least once, it appears as the biologically active component;

[0321] L 1’ and L 7 It is either independent of or does not exist in each occurrence, or is an alkylene group, alkenylene group, ynylene group, heteroalkylene group, alkenylene group, arylene group, or a combination thereof;

[0322] L 2 L 3 L 5 and L 6 It is either independent of any group that does not exist in each instance, or it is a alkylene group, alkenyl group, alynyl group, heteroalkyl group, alkenyl group, or alynyl group.

[0323] L 4 Each time it appears, it is independently an alkylene, alkenylene, ynylene, heteroalkylene, alkenylene, or ynylene linkage;

[0324] R 1 and R 2 Each of these can be independently H, OH, SH, alkyl, alkoxy, alkyl ether, heteroalkyl, -OP (=R) a (R) b )R c Or Q;

[0325] R 3 Each time it appears, it is independently H, alkyl, or alkoxy;

[0326] R 4 Each time it appears, it is independently OH, SH, or O. - S - OR d or SR d ;

[0327] R 5 Each occurrence is independently either oxidized, thiolated, or absent;

[0328] R 6 and R 7Each time it appears, it is independently H, OH, or a halogen;

[0329] R a Is it O or S?

[0330] R b It is OH, SH, O - S - OR d or SR d ;

[0331] R c It is OH, SH, O - S - OR d OL' SR d Alkyl, alkoxy, heteroalkyl, heteroalkoxy, alkyl ether, alkoxyalkyl ether, phosphate ester, thiophosphate ester, phosphoryl alkyl, thiophosphoryl alkyl, phosphoryl alkyl ether or thiophosphoryl ether;

[0332] R d It is a counter ion;

[0333] Q is independently a portion containing a reactive group or its protected form each time it appears, which is capable of forming a covalent bond with the complementary reactive group Q′ on the target portion;

[0334] L' independently, each time it appears, is a linker containing a covalent bond with Q, a target portion, a linker containing a covalent bond with the target portion, a linker containing a covalent bond with the solid support, a linker containing a covalent bond with the solid support residue, a solid support residue, a linker containing a covalent bond with a nucleoside, or a linker containing a covalent bond with other compounds of structure (I).

[0335] m and p are independent integers equal to or greater than 0 each time they appear;

[0336] w is an integer equal to or greater than 1 each time it appears; and

[0337] n is an integer equal to or greater than 1.

[0338] In compounds with structure (III), M 1 M 2 R 1 R 2 R 3 R 4 R 5 R 6 R 7 L 2 L 3 L 4 L 5 L6 L 7 m, n, p, and w are as defined in any of the foregoing embodiments. Bioactive portion M 2 Through L 1’ Covalently linked to the polymer chain. In compounds of structure (III), in some embodiments, M 2 They are the same. In other implementations, each M 2 They are different. In more implementations, one or more M 2 They are the same, and one or more M 2 They are different.

[0339] In some implementations, L 1’ The at least one occurrence of contains one of the following structures:

[0340] or .

[0341] In some implementations, L 1’ The at least one occurrence of contains one of the following structures:

[0342] ; ; ;

[0343] ; ; ;

[0344] ;

[0345] ; ;

[0346] ;

[0347] ; ;

[0348] ; or

[0349] .

[0350] In some implementations, L 1’ The at least one occurrence of contains one of the following structures:

[0351] ; ; ; ; ; ;

[0352] ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; or .

[0353] In some embodiments, where G is -C(=O)OH, the resulting compound of structure (III) has the following structure (IIIA).

[0354]

[0355] (IIIA)

[0356] Where R 8 and L 1 As defined in any of the foregoing implementation schemes.

[0357] In some implementations, L 4 At least once, L is a heteroalkylene group. In some embodiments, L 4 Each occurrence of L is a heteroalkylene group. In some embodiments, L 4 At least one occurrence of L contains an alkylene oxide. In some more specific embodiments, L 4 The alkyl oxide is ethylene oxide. In some more specific embodiments, the ethylene oxide is polyethylene oxide. In some embodiments, L 4 It has the following structure each time it appears:

[0358] ,

[0359] in:

[0360] z is an integer from 1 to 100; and

[0361] Indicates the bond with the adjacent phosphorus atom.

[0362] In some embodiments, the compound of structure (III) has the following structure (IIIB):

[0363] .

[0364] In some implementations, L 2 L 3 L 5 and L 6 It is independently absent (i.e., a direct linker) or independently an alkylene, alkenylene, ynylene, heteroalkylene, alkenylene, or heteroynyl linker each time it appears. In some embodiments, L 2 L 3 L 5 and L 6 It is an alkylene group each time it appears. In further embodiments, L 2 L 3 L 5 and L 6 Each time it appears, it is independently a C1-C6 alkylene, C2-C6 alkenylene, or C2-C6 ynylene. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C1 alkylene linker. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C2 alkylene linker. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C3 alkylene linker. In some more specific embodiments, L 3 and L 5 The alkylene linker is a C4 alkylene linker. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C5 alkylene linker. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C6 alkylene linker. In some other embodiments, L 2 L 3 L 5 and L 6 At least once, L appears as a direct-connect key. In some implementations, L 2 At least one occurrence of L is absent. In a more specific implementation, L 2 It does not exist each time it appears.

[0365] In some relevant embodiments, the compound with structure (IIII) has the following structure (IIIC):

[0366] ,

[0367] Where y 1 y 2 y 3 and y 4 Each occurrence is an integer from 0 to 6.

[0368] In some implementations, y 1 y 2 y 3 and y 4 It is an integer 0 each time it appears (i.e., L). 2 L 3 L 5 and L 6 Each of them is a direct-connect key. In some implementations, y 1 y 2 y 3 and y 4 It is an integer 1 each time it appears. In some implementations, y 1 y 2 y 3 and y 4 It is an integer of 2 each time it appears. In some implementations, y 1 y 2 y 3 and y 4 It is an integer 3 each time it appears. In some implementations, y 1 y 2 y 3 and y 4 It is an integer of 4 each time it appears. In some implementations, y 1 y 2 y 3 and y 4 It is an integer 5 each time it appears. In some implementations, y 1 y 2 y 3 and y 4 It is an integer 6 each time it appears. In some more specific implementations, y 1 y 2 y 3 and y 4 Each occurrence is either 0 or 1.

[0369] In some specific embodiments, the compound of structure (III) is selected from the compounds in Table 3. The compounds in Table 3 are prepared according to the procedures described in the examples.

[0370] Table 3. Compounds with exemplary structure (III)

[0371]

[0372]

[0373]

[0374]

[0375]

[0376]

[0377] In Table 3, FAM represents

[0378] or .

[0379] The following reaction scheme illustrates an exemplary method for preparing a compound comprising one or more bioactive moieties and one or more fluorescent or colored moieties (e.g., a compound of structure (III)) by reacting a compound of structure (I) with a compound of structure (II).

[0380] Reaction scheme I illustrates that, in which G is a reactive group, G reacts directly with an amine-containing compound of structure (II) to form a compound of structure (III).

[0381] Reaction Scheme I

[0382]

[0383] Reaction Scheme II

[0384]

[0385] Reaction scheme II illustrates that, in which G is a group that can be converted into a reactive group, G reacts with an amine-containing compound of structure (II) to form a compound of structure (IIIA).

[0386] In the case where G is a carboxylic acid, the reaction of the compound of structure (IA) with the compound of structure (II) is carried out in the presence of an activator. The activator converts the carboxylic acid into a reactive group such as an activated ester or an anhydride. In some embodiments, the carboxylic acid is converted into an activated ester, and the compound of structure (I) reacts with the activator to generate a compound of structure (IA') containing an activated ester. In some embodiments, the activator comprises a guanidine salt reagent, a ureon reagent, preferably 2-(H-benzotriazol-1-yl)-1,1,3,3-tetramethylureon hexafluorophosphate (HBTU), O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethylureon tetrafluoroborate (TBTU), or 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethylureon hexafluorophosphate (HATU), a benzotriazole reagent, preferably 1-hydroxybenzotriazole reagent (HOBt), or an imine (immodium) N,N′-dicyclohexylcarbodiimide (DCC) or diisopropylcarbodiimide (DIPCDI), imidazolium reagent, organophosphorus reagent, acidic halogenating reagent, phosphonium reagent, preferably benzotriazol-1-yl-oxy-tris(dimethylamino)phosphonium hexafluorophosphate (BOP); also known as Castros reagent) or benzotriazol-1-yl-oxy-tripyrrolidinylphosphonium hexafluorophosphate (PyBOP), morpholine reagent, preferably N-methylmorpholine (NMM), chloroformate reagent and / or combinations thereof. In some embodiments, the coupling agent is HBTU. The activated ester then reacts with the amino group in the compound of structure (II) to form the compound of structure (IIIA). Bioactive part M 2 Therefore, it is covalently linked to the polymer chain through the reaction of activated esters and amines.

[0387] In compounds with structure (IA'), R 9 It is the leaving group that activates the ester. In some embodiments, R 9 It has one of the following structures:

[0388] ; ; ; ; ; ; ; ; ; ; ;

[0389] or .

[0390] In some embodiments, the reaction between the compound of structure (I) and the compound of structure (II) can be carried out in a solution containing an organic solvent and an activator. In some embodiments, the reaction can be carried out under alkaline conditions.

[0391] In some embodiments, the organic solvent may be one or more selected from the following: dimethyl sulfoxide (DMSO), N-methylformamide (NMP), N-methylpyrrolidone, dimethylacetamide, dimethylformamide (DMF), dichloromethane, dichloroethane, and chloroform. In some embodiments, the solvent is DMF.

[0392] In some embodiments, the base may be selected from one or more of the following: diisopropylethylamine (DIPEA), triethylamine, piperidine, pyrrolidine, pyridine, N-methylmorpholine (NMM), and trimethylpyridine. In some embodiments, the base is DIPEA.

[0393] In some more specific embodiments, the coupling of the compound of structure (I) with the compound of structure (II) is carried out in DMF in the presence of HBTU.

[0394] Compounds of structure (I) can be prepared by DNA synthesis methods. Monomers (e.g., phosphoramide monomers) can be commercially available (e.g., from ChemGenes Corporation, Wilmington Mass.) or synthesized using methods known in the art (see, for example, U.S. Publication No. 2019 / 0136065, which is hereby incorporated by reference). In the DNA synthesis step, the desired moiety can be introduced by including the desired moiety as part of the monomer (see, for example, L and M in the representative DNA synthesis cycle below). An exemplary DNA synthesis scheme is shown below.

[0395] Representative DNA synthesis cycle

[0396]

[0397] Oligopolymerization typically begins by removing a protecting group (e.g., dimethoxytriphenylmethyl, DMTr) to expose the free -OH (hydroxyl) group (step 1, detriphenylmethylation). In a subsequent coupling step, a phosphorus amide monomer is introduced, which reacts with the free OH group to form a new covalent bond with phosphorus, accompanied by the loss of the diisopropylamine group (step 2, coupling). The resulting phosphite trimer 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 unreactive (step 4, capping). The resulting phosphate ester oligomer contains the OH group protected by DMTr, which can be deprotected to restart the synthetic cycle, allowing another phosphorus amide monomer to be attached to the oligomer.

[0398] Customization occurs in step 2 by selecting a phosphorusamide monomer. The properties of L (i.e., the linker group) and M (i.e., the fluorescent or colored dye) in the above scheme are selected to synthesize a compound with the desired structure (I). M may optionally be absent to incorporate desired spacing between the M moieties. Those skilled in the art can select from a variety of monomer types to obtain compounds of this disclosure containing multiple therapeutic agents and / or other moieties (e.g., fluorophores or chromophores) and exhibiting differences in the linker groups.

[0399] In some embodiments, compounds with structure (I) can be prepared by oligomerization using well-known automated phosphorus amide chemistry with phosphorus amide compounds having the following structure (IV):

[0400]

[0401] in:

[0402] R 1 '' and R 2 Each time it appears, it is independently of H, a protecting group, or an activated phosphorus moiety;

[0403] R 4 '' is an alkoxy, haloalkyl, alkyl, optionally substituted aryl, or optionally substituted aralkyl.

[0404] In some embodiments of compounds with structure (IV), R 1 '' is H. In some embodiments, it is a protecting group, for example, a triphenylmethyl protecting group. In some embodiments, R 1 '' is triphenylmethyl. In some embodiments, R 1 '' is 4-methoxytriphenylmethyl. In a more specific embodiment, R 1 '' is 4,4'-dimethoxytriphenylmethyl.

[0405] In some implementations, R 2 '' is H. In some implementations, R 2 '' is the activated phosphorus moiety. For example, in some embodiments, R 2 '' contains the following structure:

[0406]

[0407] in:

[0408] R 5 '' is H or cyanoalkyl; and

[0409] R 6 Each time it appears, it is independently a C1-C6 alkyl group.

[0410] In some embodiments of compounds with structure (IV), R 5 '' is H. In other implementations, R 5 '' is 2-cyanoethyl.

[0411] In some implementations, R 6 The presence of '' at least once is isopropyl. In some embodiments, R 6 Each occurrence of '' represents isopropyl.

[0412] In some specific implementations, R 2 It has the following structure:

[0413] .

[0414] In some embodiments of compounds with structure (IV), R 4 '' is a C1-C4 haloalkyl group. In a more specific embodiment, R 4 '' is -CF3. In some implementations, R 4 '' is a C1-C4 alkoxy group. In a more specific embodiment, R 4 '' is a methoxy group.

[0415] In some specific embodiments, the compound with structure (IV) has the following structure:

[0416]

[0417] Alternatively, a compound with structure (V) can be synthesized first, and then a fluorescent or colored dye M can be applied. 1 and bioactive component M 2 The compound with structure (I) is prepared by coupling it to a compound with structure (V). Therefore, some embodiments provide intermediate compounds having structure (V):

[0418]

[0419] in:

[0420] G, each time it appears, is independently a part containing a reactive group, a group that can be converted into a reactive group, or a protected analogue thereof, which is capable of forming a covalent bond with an amino group;

[0421] G 1 Each time it appears, it is independently a portion containing a reactive group or its protected analogue, which is capable of reacting with M. 1 The complementary reactive groups in G form covalent bonds, provided that G 1 It does not react with amine groups.

[0422] L 1 and L 7’ Each time it appears independently, it is an alkylene group, an alkenylene group, an ynylene group, a heteroalkylene group, an alkenylene group, an arylene group, or a heteroaryl linker or a combination thereof;

[0423] L 2 L 3 L 5 and L 6 It is either independent of any group that does not exist in each instance, or it is a alkylene group, alkenyl group, alynyl group, heteroalkyl group, alkenyl group, or alynyl group.

[0424] L 4 Each time it appears, it is independently an alkylene, alkenylene, ynylene, heteroalkylene, alkenylene, or ynylene linkage;

[0425] R 1 and R 2 Each of these can be independently H, OH, SH, alkyl, alkoxy, alkyl ether, heteroalkyl, -OP (=R) a (R) b )R c Or Q;

[0426] R 3 Each time it appears, it is independently H, alkyl, or alkoxy;

[0427] R 4 Each time it appears, it is independently OH, SH, or O. - S - OR d or SR d ;

[0428] R 5 Each occurrence is independently either oxidized, thiolated, or absent;

[0429] R 6 and R 7Each time it appears, it is independently H, OH, or a halogen;

[0430] R a Is it O or S?

[0431] R b It is OH, SH, O - S - OR d or SR d ;

[0432] R c It is OH, SH, O - S - OR d OL' SR d Alkyl, alkoxy, heteroalkyl, heteroalkoxy, alkyl ether, alkoxyalkyl ether, phosphate ester, thiophosphate ester, phosphoryl alkyl, thiophosphoryl alkyl, phosphoryl alkyl ether or thiophosphoryl ether;

[0433] R d It is a counter ion;

[0434] Q is independently a portion containing a reactive group or its protected form each time it appears, which is capable of forming a covalent bond with the complementary reactive group Q′ on the target portion;

[0435] L' independently, each time it appears, is a linker containing a covalent bond with Q, a target portion, a linker containing a covalent bond with the target portion, a linker containing a covalent bond with the solid support, a linker containing a covalent bond with the solid support residue, a solid support residue, a linker containing a covalent bond with a nucleoside, or a linker containing a covalent bond with other compounds of structure (I).

[0436] m and p are independent integers equal to or greater than 0 each time they appear;

[0437] w is an integer equal to or greater than 1 each time it appears; and

[0438] n is an integer equal to or greater than 1.

[0439] G in compounds with structure (V) 1 The portion can be selected from any portion containing a group having a suitable reactive group for interaction with M. 1 Complementary groups on some parts form covalent bonds. In some embodiments, G 1Each occurrence independently includes a section applicable to reactions including: copper-catalyzed reactions of azides and alkynes to form triazoles (Huisgen 1, 3-dipolar cycloaddition), reactions of dienes and dienophiles (Diels-Alder), strain-promoted alkyne-nitroketone cycloaddition, reactions of strained alkenes with azides, tetrazides, or tetrazides, cycloaddition of alkenes with azides [3+2], reverse-demand Diels-Alder reactions of alkenes and tetrazides, photoreactions of alkenes and tetrazides, and various substitution reactions, such as nucleophilic attacks on electrophilic atoms to replace leaving groups.

[0440] In some implementations, G 1 Each time it appears, it is independently a part containing oxime, hydrazone, alkyne, amine, azide, acyl azide, acyl halide, nitrile, nitrile, mercapto, disulfide, sulfonyl halide, isothiocyanate, imine ester, activated ester, ketone, α,β-unsaturated carbonyl, olefin, maleimide, α-haloimide, epoxide, aziridine, tetrazine, tetrazolium, phosphine, biotin, or thiamethoxane functional group.

[0441] In some implementations, G 1 Each time it appears, it independently contains either an alkyne or an azide group. In other embodiments, G 1 Each time it appears, it independently contains an amino, isothiocyanate, or activated ester group. In different embodiments, G 1 Each time it appears, it independently contains a reactive group that, upon reaction with a complementary reactive group, forms a functional group comprising an olefin, ester, amide, thioester, disulfide, carbocyclic, heterocyclic, or heteroaryl group. For example, in some embodiments, the heteroaryl group is a triazole group.

[0442] In any of the other foregoing embodiments of compound (V), G 1 It is independent each time it appears. or In some implementations, G 1 The at least one occurrence of has one of the following structures:

[0443] ; or .

[0444] In some relevant implementation schemes, G 1 It independently possesses one of the following structures each time it appears:

[0445] ; or .

[0446] In some embodiments of compounds with structure (V), G1 At least one occurrence is In a more specific implementation plan, G 1 It is independent each time it appears. .

[0447] In some embodiments of compound (V), G 1 At least once, G appears as the protected form of the amine. In some embodiments, G 1 When it appears multiple times, it is independently the protected form of the amine. In some embodiments, G 1 It is an independent protected form of amine each time it appears.

[0448] In some of the foregoing embodiments, the amine is protected in the form of a trifluoroacetate-protected amine. In some embodiments, the amine is protected in the form of a BOC-protected amine. In some embodiments, the amine is protected in the form of an Fmoc-protected amine. For example, in some embodiments, G... 1 The at least one occurrence of has one of the following structures:

[0449] ; or .

[0450] In a more specific implementation plan, G 1 It independently possesses one of the following structures each time it appears:

[0451] ; or .

[0452] In some implementations, L 7’ -G 1 It has one of the following structures:

[0453] ; ; ; ; or .

[0454] In some implementations, R 1 R 2 R 3 R 4 R 5 R 6 R 7 L 2 L 3 L 4 L 5 or L 6As defined in any of the foregoing implementation schemes.

[0455] In some embodiments, G is independently -C(=O)OH or -C(=O)H each time it appears. In some relevant embodiments, the compound of structure (V) has the following structure (VA) or (VB):

[0456] or

[0457] (VA)

[0458] .

[0459] (VB)

[0460] Connecting base L 1 It can be used as a connection point between the G group and the rest of the compound.

[0461] In some implementations, L 1 At least once, L contains a linker that is not cleavable under physiological conditions. Therefore, in some embodiments, L 1 It includes amide bonds, ester bonds, disulfide bonds, hydrazones, triphosphates, diesters, β-glucuronides, double bonds, triple bonds, ether bonds, ketones, diols, or combinations thereof.

[0462] In some implementations, L 1 It contains tert-butyloxycarbonyl, p-methoxybenzyl, dialkyl or diaryldialkoxysilane, orthoester, acetal, β-thiopropionate, ketal, aminophosphate, hydrazone, vinyl ether, imine, aconitol, triphenylmethyl, polyketal, diarylhydrazone, diazobenzene, vivinal diol, pyrophosphate diester or valine citrulline.

[0463] In some implementations, L 1 Each time it appears, it is independently a linker that can be cleaved within a pH range of 6 to 8. For example, in some embodiments, L 1 It is a linker that can be cleaved at pH 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9 or 8.0.

[0464] In some implementations, L 1Each time L occurs, it is independently a linker that can cleave within a temperature range of 20°C to 40°C, 25°C to 35°C, 30°C to 35°C, 30°C to 37°C, 35°C to 37°C, 35°C to 40°C, or 32°C to 38°C. In some embodiments, L is independently a linker that can cleave within a temperature range of about 20°C, about 21°C, about 22°C, about 23°C, about 24°C, about 25°C, about 26°C, about 27°C, about 28°C, about 29°C, about 30°C, about 31°C, about 32°C, about 33°C, about 34°C, about 35°C, about 36°C, about 37°C, about 38°C, about 39°C, or about 40°C.

[0465] In some implementations, L 1 Each time it appears, it is independently a linker that can be cleaved by an enzyme. For example, in some embodiments, the enzyme is a hydrolase, an oxidoreductase, or a lyase. In some embodiments, the enzyme is an EC 4.1 (e.g., EC 4.1.1, EC 4.1.2, EC 4.1.3, or EC 4.1.99), EC 4.2, EC 4.3, EC 4.4, EC 4.5, EC 4.6, or EC 4.99 enzyme.

[0466] In some implementations, for L 1 L appears at least once. 1 -G has the following structure:

[0467] .

[0468] In some implementations, L 1a At least once, L appears as a optionally substituted 5-9 member heteroaryl linker. In some embodiments, L 1a It is the substituted 5-membered heteroaryl linker. In some embodiments, L 1a It is the substituted 6-membered heteroaryl linker. In some embodiments, L 1a It is the substituted 7-membered heteroaryl linker. In some embodiments, L 1a It is the substituted 8-membered heteroaryl linker. In some embodiments, L 1a It is the substituted 9-membered heteroaryl linker. In some relevant embodiments, L 1a It is substituted with an oxo, an alkyl (e.g., methyl, ethyl, etc.) or a combination thereof. In some embodiments, L 1a It is not replaced each time it appears. In some more specific implementations, L 1a It is pyrimidine independently each time it appears. In some more specific embodiments, L 1a Each time it appears, it is independently either cytosine or thymine. In some embodiments, L 1aEach time it appears, it is independently selected from cytosine and thymine, such that the compound contains a sequence of cytosine and thymine bases capable of forming a triplet with the target DNA sequence.

[0469] In some implementations, L 1a It has one of the following structures:

[0470]

[0471] In some implementations, L 1b Each time it appears independently, it is optionally alkylene, alkenylene, alkyne, heteroalkylene, alkenylene, alkyne, alkyleneheteroarylalkylene, alkyleneheterocyclicalkylene, alkylenecarbocyclicalkylene, heteroalkyleneheteroarylalkylene, heteroalkyleneheterocyclicalkylene, heteroalkylenecarbocyclicalkylene, heteroalkyleneheteroarylalkylene, heteroalkyleneheterocyclicalkylene, heteroalkylenecarbocyclicalkylene Alkyl, alkyleneheteroarylheteroalkyl, alkyleneheterocyclic heteroalkyl, alkylenecarbamocyclic heteroalkyl, heteroaryl, heterocyclic, carbamocyclic, alkyleneheteroaryl, alkyleneheterocyclic, heteroarylalkylene, alkylenecarbamocyclic, carbamocyclic alkylene, heteroarylalkylene, heteroalkylheterocyclic, heteroarylalkylene, heteroalkylcarbamocyclic, carbamocyclic heteroalkyl or heteroatom-linked group. In some embodiments, L 1b It is an optional substituted hemienyl linker.

[0472] In some more specific implementations, L a and L b Together (i.e., It has one of the following structures:

[0473] or .

[0474] In some implementations, L 1 The presence of at least one of the amino acid residues is present. In some embodiments, the amino acid residue is valine. In some embodiments, L... 1 The at least one occurrence of contains one of the following structures:

[0475] ;

[0476] ;

[0477] ; ;

[0478] ; ;

[0479] ; ; ; ;

[0480] ; ;

[0481] ; or

[0482] .

[0483] In some implementations, L 1 The at least one occurrence of contains one of the following structures:

[0484] ; ; ; ; ; ; ;

[0485] ; ; ; ; ;

[0486] ; ; ; ;

[0487] ; ;

[0488] ; ; ; or .

[0489] In some implementations, L 4 At least once, L is a heteroalkylene group. In some embodiments, L 4 Each occurrence of L is a heteroalkylene group. In some embodiments, L 4 At least one occurrence of L contains an alkylene oxide. In some more specific embodiments, L 4 The alkyl oxide is ethylene oxide. In some more specific embodiments, the ethylene oxide is polyethylene oxide. In some embodiments, L 4 It has the following structure each time it appears:

[0490] ,

[0491] in:

[0492] z is an integer from 1 to 100; and

[0493] Indicates the bond with the adjacent phosphorus atom.

[0494] In some embodiments, the compound of structure (V) has the following structure (VC) or (VD):

[0495] or

[0496] (VC)

[0497] .

[0498] (VD)

[0499] In some implementations, L 2 L 3 L 5 and L 6 It is independently absent (i.e., a direct linker) or independently an alkylene, alkenylene, ynylene, heteroalkylene, alkenylene, or heteroynyl linker each time it appears. In some embodiments, L 2 L 3 L 5 and L 6 It is an alkylene group each time it appears. In further embodiments, L 2 L 3 L 5 and L 6 Each time it appears, it is independently a C1-C6 alkylene, C2-C6 alkenylene, or C2-C6 ynylene. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C1 alkylene linker. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C2 alkylene linker. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C3 alkylene linker. In some more specific embodiments, L 3 and L 5The alkylene linker is a C4 alkylene linker. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C5 alkylene linker. In some more specific embodiments, L 2 L 3 L 5 and L 6 The alkylene linker is a C6 alkylene linker. In some other embodiments, L 2 L 3 L 5 and L 6 At least once, L appears as a direct-connect key. In some implementations, L 2 At least one occurrence of L is absent. In a more specific implementation, L 2 It does not exist each time it appears.

[0500] In some relevant embodiments, the compound with structure (V) has the following structure (VE) or (VF):

[0501] or

[0502] (VE)

[0503] ,

[0504] (VF)

[0505] Where y 1 y 2 y 3 and y 4 Each occurrence is an integer from 0 to 6.

[0506] In some implementations, y 1 y 2 y 3 and y 4 It is an integer 0 each time it appears (i.e., L). 2 L 3 L 5 and L 6 Each of them is a direct-connect key. In some implementations, y 1 y 2 y 3 and y 4 It is an integer 1 each time it appears. In some implementations, y 1 y 2 y 3 and y 4 It is an integer of 2 each time it appears. In some implementations, y1 y 2 y 3 and y 4 It is an integer 3 each time it appears. In some implementations, y 1 y 2 y 3 and y 4 It is an integer of 4 each time it appears. In some implementations, y 1 y 2 y 3 and y 4 It is an integer 5 each time it appears. In some implementations, y 1 y 2 y 3 and y 4 It is an integer 6 each time it appears. In some more specific implementations, y 1 y 2 y 3 and y 4 Each occurrence is either 0 or 1.

[0507] In some embodiments, the compound of structure (V) is selected from the compounds in Table 4.

[0508] Table 4. Exemplary compounds with structure V

[0509]

[0510] Reaction Scheme III

[0511]

[0512] Reaction scheme III illustrates an exemplary method for preparing compounds of structure (I), wherein M 1 G 1 G 1’ R 1 R 2 R 3 R 4 R 5 R 6 R 7 L 1 L 2 L 3 L 4 L 5 L 6 L 7 L 7’ m, n, p, and w are defined as above. Compounds with structure (V) are associated with M. 1 -G 1’ The reaction produces a compound with structure (I). Here, G1 and G 1’ This represents functional groups with complementary reactivity (i.e., functional groups that react to form covalent bonds). G 1’ It can be M 1 Accessories, or M 1 Part of the structural framework. G 1 and G 1’ It can be any number of functional groups described herein, such as alkynes and azides, amines and activated esters, or amines and isothiocyanates, etc. For example, in some embodiments, when G 1 When it is -NH2, G 1’ It is -C(=O)OH.

[0513] Compounds with structure (V) can be prepared using the DNA synthesis methods described above.

[0514] Those skilled in the art will understand that, in the methods described herein, the functional groups of the intermediate compounds may require protection by suitable protecting groups. Such functional groups include hydroxyl, amino, mercapto, and carboxylic acids. Suitable protecting groups for hydroxyl groups include trialkylsilyl or diarylalkylsilyl (e.g., tert-butyldimethylsilyl, tert-butyldiphenylsilyl, or trimethylsilyl), tetrahydropyranyl, benzyl, etc. Suitable protecting groups for amino, amidine, and guanidine groups include tert-butoxycarbonyl, benzyloxycarbonyl, etc. Suitable protecting groups for mercapto groups include -C(O)-R (where R is alkyl, aryl, or arylalkyl), p-methoxybenzyl, triphenylmethyl, etc. Suitable protecting groups for carboxylic acids include alkyl, aryl, or arylalkyl esters. Protecting groups may be added or removed according to techniques known to those skilled in the art and as described herein, as well as standard techniques. The application of protecting groups is described in detail in Green, TW, and PGM Wutz, Protective Groups in Organic Synthesis (1999), 3rd edition, Wiley. Those skilled in the art will understand that the protecting group can also be a polymeric resin such as Wang resin, Rink resin, or 2-chlorotriphenylmethyl-chloride resin.

[0515] Furthermore, all compounds of this disclosure existing in free base or acid form can be converted into their salts by treatment with suitable inorganic or organic bases or acids using methods known to those skilled in the art. Salts of the compounds of this disclosure can be converted into their free base or acid form using standard techniques.

[0516] Example

[0517] General methods

[0518] Mass spectrometry analysis was performed using MassLynx 4.1 acquisition software on a Waters / Micromass Quattro micro MS / MS system (MS mode only). The mobile phase used for dye-based LC / MS was 100 mM 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP), 8.6 mM triethylamine (TEA), pH 8, phosphorous acid. Precursor molecules were also analyzed using a Waters Acquity UHPLC system with a 2.1 mm × 50 mm Acquity BEH-C18 column maintained at 45 °C, employing an acetonitrile / water mobile phase gradient. The molecular weight of the monomer intermediate was obtained using tropylium cation infusion-enhanced ionization on the Waters / Micromass Quattro micro MS / MS system (MS mode only). Excitation and emission characteristics were recorded on a Cary Eclipse spectrophotometer.

[0519] Unless otherwise specified, all reactions were carried out in oven-dried glassware under a nitrogen atmosphere. 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 is without additional purification.

[0520] Example 1

[0521] Synthesis of Compound I-1 and Compound I-2

[0522] Compounds I-1 and I-2 were prepared by automated solid-phase synthesis (SPS) using standard phosphoramidite chemistry methods, similar to the synthetic chemistry methods used in oligonucleotide synthesis. The synthesis process included solid-phase synthesis, final deprotection and cleavage from the solid support, compound isolation, and analysis. The typical solid-phase synthesis process was a repeating four-step cycle with solvent washing between each step, as follows: Step 1 – detriphenylmethylation, Step 2 – monomer coupling, Step 3 – oxidation, and Step 4 – capping. The synthesis cycle was repeated until all monomers were sequentially incorporated into the polymer. The solid-phase synthesis of each of compounds I-1 and I-2 was performed using an ÄKTA oligopilot™ Plus synthesizer, UNICORN control software, and standard oligonucleotide synthesis programming steps. The ÄKTA synthesis column was loaded with a solid support of 0.13 g (40 μmol) Primer Support 5G T300 (purchased from Cytiva). The phosphoramidite monomers used in the synthesis (e.g., hexaethylene glycol phosphoramidite, 6-FAM phosphoramidite, 5'-thiol C-6 disulfide modifying agent CED phosphoramidite, carboxyl-dT phosphoramidite) were dissolved in anhydrous acetonitrile at a concentration of 0.2 M and loaded onto the ÄKTA system. In the monomer coupling step, 240 μmol (6 equivalents) of the phosphoramidite monomer was fed onto the solid support, allowing sufficient contact time to complete the coupling reaction. After solid-phase synthesis, compounds I-1 and I-2 were deprotected and cleaved from the solid support by holding a 20 mL solution of 0.4 M sodium hydroxide in methanol / water (80:20) at 37 °C for 18 hours. The solid support was then separated from the solution containing the compounds by filtration. The solid support was washed three times with 5 mL of ethanol / water (50:50), the filtrates were combined, and the filtrate solution was neutralized with 60 mL of 2 M triethylammonium acetate aqueous solution. The solution containing the compounds was desalted by ultrafiltration (using an Omega-modified polyethersulfone membrane and a Microsep centrifugal filter with a 1K MWCO, PALL Corporation). Samples of the desalted solution containing the compounds were then subjected to LC-UV / MS analysis and UV absorption spectroscopy, and subsequently concentrated to a dry residue by evaporation and low-pressure lyophilization. Compound I-1 yielded 74 mg (via UV spectroscopy at 266 nm), with a purity of approximately 85% (via LC-UV, diode array), and an observed mass of 2530.6 g (via mass spectrometry). Compound I-2 yielded 109 mg (via UV spectroscopy at 266 nm), with a purity of approximately 83% (via LC-UV, diode array), and an observed mass of 3923.6 g (via mass spectrometry).

[0523] 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 / 587,602, filed October 3, 2023, are incorporated herein by reference in their entirety to the extent that they do not conflict with this specification. Other embodiments may be provided by modifying aspects of the embodiments to adopt different patent, application and publication concepts, if necessary.

[0524] As can be understood from the foregoing, while specific embodiments of this disclosure have been described herein for illustrative purposes, various modifications may be made without departing from the spirit and scope of this disclosure. Therefore, this disclosure is limited only by the appended claims.

Claims

1. A method for preparing a polymer-drug conjugate, the method comprising a compound having structure (I): , (I) Reaction with compounds of structure (II): ， (II) in: M 1 Each time it appears, it is independently a portion containing either fluorescent or colored dye; M 2 It contains the bioactive portion or the fluorescent or colored dye portion; G, each time it appears, is independently a part containing a reactive group, a group that can be converted into a reactive group, or a protected analogue thereof, which is capable of forming a covalent bond with an amino group; L 1 and L 7 It is either independent of or does not exist in each occurrence, or is an alkylene group, alkenylene group, ynylene group, heteroalkylene group, alkenylene group, arylene group, or a combination thereof; L 2 L 3 L 5 and L 6 It is either independent of any group that does not exist in each instance, or it is a alkylene group, alkenyl group, alynyl group, heteroalkyl group, alkenyl group, or alynyl group. L 4 Each time it appears, it is independently an alkylene, alkenylene, ynylene, heteroalkylene, alkenylene, or ynylene linkage; R 1 and R 2 Each of these can be independently H, OH, SH, alkyl, alkoxy, alkyl ether, heteroalkyl, -OP (=R) a (R) b )R c Or Q; R 3 Each time it appears, it is independently H, alkyl, or alkoxy; R 4 Each time it appears, it is independently OH, SH, or O. - S - OR d or SR d ; R 5 Each occurrence is independently either oxidized, thiolated, or absent; R 6 and R 7 Each time it appears, it is independently H, OH, or a halogen; R 8 It is H or alkyl; R a Is it O or S? R b It is OH, SH, O - S - OR d or SR d ; R c It is OH, SH, O - S - OR d OL' SR d Alkyl, alkoxy, heteroalkyl, heteroalkoxy, alkyl ether, alkoxyalkyl ether, phosphate ester, thiophosphate ester, phosphoryl alkyl, thiophosphoryl alkyl, phosphoryl alkyl ether or thiophosphoryl ether; R d It is a counter ion; Q is independently a portion containing a reactive group or its protected form each time it appears, which is capable of forming a covalent bond with the complementary reactive group Q′ on the target portion; L' independently, each time it appears, is a linker containing a covalent bond with Q, a target portion, a linker containing a covalent bond with the target portion, a linker containing a covalent bond with the solid support, a linker containing a covalent bond with the solid support residue, a solid support residue, a linker containing a covalent bond with a nucleoside, or a linker containing a covalent bond with other compounds of structure (I). m and p are independent integers equal to or greater than 0 each time they appear; w is an integer equal to or greater than 1 each time it appears; and n is an integer equal to or greater than 1. Therefore: i) the G in the compound with structure (I) and the -NHR in the compound with structure (II) 8 The first bond is formed between them.

2. The method of claim 1, wherein G is -C(=O)OH or -C(=O)H.

3. The method of claim 2, wherein the compound of structure (I) has the following structure (IA) or (IB): or (IA) (IB).

4. The method according to any one of claims 1-3, wherein L 4 It has the following structure: in: z is an integer from 1 to 100; and Indicates the bond with the adjacent phosphorus atom.

5. The method of claim 4, wherein z is an integer from 3 to 8.

6. The method of claim 4, wherein z is an integer from 22 to 26.

7. The method according to any one of claims 3-6, wherein the compound of structure (I) has the following structure (IC) or (ID): or (IC) , (ID) Where m is an integer from 0 to 10.

8. The method of claim 7, wherein m is 1.

9. The method according to any one of claims 1-8, wherein L 2 L 3 L 5 and L 6 It is either absent independently each time it appears, or it is either a C1-C6 alkylene group, a C2-C6 alkenyl group, or a C2-C6 alkynyl group.

10. The method according to any one of claims 3-9, wherein the compound of structure (I) has the following structure (IE) or (IF): or (IE) , (IF) Where y 1 y 2 y 3 and y 4 Each occurrence is an integer from 0 to 6.

11. The method of claim 10, wherein y 1 y 2 y 3 and y 4 It is either 0 or 1 each time it appears.

12. The method according to any one of claims 1-11, wherein L 1 It includes amide bonds, ester bonds, disulfide bonds, hydrazones, triphosphates, diesters, β-glucuronides, double bonds, triple bonds, ether bonds, ketones, diols, or combinations thereof.

13. The method of claim 12, wherein L 1 It contains tert-butyloxycarbonyl, p-methoxybenzyl, dialkyldialkoxysilane, diaryldialkoxysilane, orthoester, acetal, β-thiopropionate, ketal, aminophosphate, hydrazone, vinyl ether, imine, aconitol, triphenylmethyl, polyketal, diarylhydrazone, diazobenzene, vivinal diol, pyrophosphate diester, or valine-citrulline.

14. The method according to any one of claims 1-11, wherein L 1 It contains one of the following structures: or .

15. The method according to any one of claims 1-11, wherein L 1 It contains one of the following structures: ； ； ； ； ； ； ； ； ； ； ； ； ; or .

16. The method according to any one of claims 1-11, wherein L 1 It contains one of the following structures: ； ; ； ； ； ； ； ； ； ； ； ； ； ； ； ； ； ； ; ; ; or .

17. The method according to any one of claims 1-16, wherein L 7 It has one of the following structures: ； ； ; ； ； ； ； ； ； ； ； , ; ; ; ; ; or .

18. The method of claim 17, wherein L 7 yes: 。 19. The method according to any one of claims 1-16, wherein L 7 It has one of the following structures: ； ； ; or 。 20. The method according to any one of claims 1-16, wherein L 7 It has one of the following structures: ； ； ; ; ; or 。 21. The method according to any one of claims 1-20, wherein R 3 It is H.

22. The method according to any one of claims 1-21, wherein R 4 It is O - OH or OR d .

23. The method according to any one of claims 1-22, wherein R 5 It is oxygenation.

24. The method according to any one of claims 1-23, wherein R 6 and R 7 It is H.

25. The method according to any one of claims 1-24, wherein R 1 Or R 2 One of them is -OP(=R) a (R) b )R c And R 1 Or R 2 The other one is Q.

26. The method of claim 25, wherein Q comprises a nucleophilic reactive group, an electrophilic reactive group, or a cycloaddition reactive group.

27. The method of claim 26, wherein Q comprises a thiol, disulfide, activated ester, isothiocyanate, azide, alkyne, olefin, diene, dienophile, acyl halide, sulfonyl halide, phosphine, α-haloamide, biotin, amino, or maleimide functional group.

28. The method of claim 27, wherein the activated ester is an N-succinimide ester, an imine ester, or a polyfluorophenyl ester.

29. The method of claim 27, wherein the azide is an alkyl azide or an acyl azide.

30. The method of any one of claims 25-29, wherein Q is selected from the portion of Table 1.

31. The method according to any one of claims 1-24, wherein R 1 and R 2 Each is independently -OP(=R) a (R) b )R c .

32. The method according to any one of claims 25-31, wherein R c She is an office lady (OL).

33. The method of claim 32, wherein L' is the target portion.

34. The method of claim 32, wherein L' is a linker to the target moiety, a linker to the analyte molecule, or a linker to the solid support, said linker comprising an alkylene oxide or a phosphate diester moiety or a combination thereof.

35. The method according to any one of claims 32-34, wherein L' has the following structure: ， in: m'' and n'' are independent integers from 1 to 10; R e It is H, an electron pair, or a counter ion; and L'' represents the target portion or a connection to the target portion.

36. The method of claim 35, wherein n'' is 6.

37. The method of any one of claims 33-36, wherein the targeting portion is an antibody or a cell surface receptor antagonist.

38. The method of claim 37, wherein the antibody or cell surface receptor antagonist is an epidermal growth factor receptor (EGFR) inhibitor, a hepatocyte growth factor receptor (HGFR) inhibitor, an insulin-like growth factor receptor (IGFR) inhibitor, a folic acid or MET inhibitor.

39. The method of claim 38, wherein the targeting portion is a monoclonal antibody.

40. The method of any one of claims 34-39, wherein the analyte molecule is a nucleic acid, an amino acid, or a polymer thereof.

41. The method of any one of claims 34-39, wherein the analyte molecule is an enzyme, receptor, receptor ligand, glycoprotein, aptamer, or prion.

42. The method of any one of claims 34-39, wherein the solid carrier is a polymer bead or a non-polymer bead.

43. The method according to any one of claims 25-42, wherein R 1 Or R 2 It has one of the following structures: ; ; ; ; ; ; ; ; or .

44. The method according to any one of claims 25-42, wherein R 1 Or R 2 It has the following structure: or .

45. The method according to any one of claims 1-44, wherein M 1 It has one of the following structures: ； ； ； ； ； ; or .

46. ​​The method of claim 45, wherein M 1 yes 。 47. The method of claim 6, wherein L 7 -M 1 It has the following structure: 。 48. The method according to any one of claims 1-44, wherein M 1 It has one of the following structures: 。 49. The method of any one of claims 1-48, wherein p is an integer from 0 to 4.

50. The method of claim 49, wherein p is 1.

51. The method of any one of claims 1-50, wherein w is an integer from 1 to 20.

52. The method of claim 51, wherein w is an integer from 1 to 10.

53. The method of claim 52, wherein w is 1.

54. The method of claim 52, wherein w is 2.

55. The method of any one of claims 1-54, wherein n is an integer from 1 to 20.

56. The method of claim 55, wherein n is 1.

57. The method of any one of claims 1-56, wherein the compound of structure (I) is selected from the compounds in Table 2.

58. The method according to any one of claims 1-57, wherein M 2 It is an anti-tumor agent.

59. The method of claim 58, wherein the antitumor agent is paclitaxel, camptothecin, doxorubicin, monomethylolpropionate E (MMAE), MMAE-Val-Cit, or monomethylolpropionate F.

60. The method of any one of claims 58-59, wherein the antitumor agent is monomethylolpropionate E.

61. The method according to any one of claims 58-59, wherein the antitumor agent is MMAE-Val-Cit.

62. The method of any one of claims 58-59, wherein the antitumor agent is doxorubicin.

63. The method according to any one of claims 58-59, wherein M 2 It has one of the following structures: ； ; ； or 。 64. The method according to any one of claims 1-58, wherein M 2 It is an anti-tumor antibiotic.

65. The method of claim 64, wherein M 2 It is selected from pepromycin sulfate, pirarubicin hydrochloride, fentostatin ester, idarubicin hydrochloride, mitomycin C, bleomycin hydrochloride, doxorubicin hydrochloride, daunorubicin hydrochloride, and epirubicin hydrochloride.

66. The method of claim 64, wherein M 2 It is an enediyne antitumor antibiotic.

67. The method of claim 66, wherein the enediyne antitumor antibiotic is calichiomycin.

68. The method according to any one of claims 1-67, wherein R 8 It is H or ethyl.

69. The method according to any one of claims 58-68, wherein the compound of structure (II) has one of the following structures: ； or 。 70. The method of any one of claims 1-69, wherein the reaction of the compound of structure (I) and the compound of structure (II) is carried out in a solution comprising the compound of structure (I) and the compound of structure (II).

71. The method of claim 70, wherein the solution further comprises a solvent selected from dimethyl sulfoxide (DMSO), N-methylformamide (NMP), N-methylpyrrolidone, dimethylacetamide, dimethylformamide (DMF), dichloromethane, dichloroethane, chloroform, and combinations thereof.

72. The method of claim 71, wherein the solvent is DMF.

73. The method of any one of claims 70-72, wherein the solution further comprises an activator.

74. The method of claim 73, wherein the activator is selected from 2-(H-benzotriazol-1-yl)-1,1,3,3-tetramethylureon hexafluorophosphate (HBTU), O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethylureon tetrafluoroborate (TBTU), 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethylureon hexafluorophosphate (HATU), 1-hydroxybenzotriazole reagent (HOBt), N,N′-dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIPCDI), benzotriazol-1-yl-oxy-tris(dimethylamino)phosphonium hexafluorophosphate (BOP), benzotriazol-1-yl-oxy-tripyrrolidinylphosphonium hexafluorophosphate (PyBOP), N-methylmorpholine (NMM), and combinations thereof.

75. The method of claim 74, wherein the activator is HBTU.

76. The method of any one of claims 70-75, wherein the solution further comprises a base selected from diisopropylethylamine (DIPEA), triethylamine, piperidine, pyrrolidine, pyridine, N-methylmorpholine (NMM), trimethylpyridine, and combinations thereof.

77. Compounds having the following structure (I): , (I) in: M 1 Each time it appears, it is independently a portion containing either fluorescent or colored dye; G, each time it appears, is independently a part containing a reactive group, a group that can be converted into a reactive group, or a protected analogue thereof, which is capable of forming a covalent bond with an amino group; L 1 and L 7 It is either independent of or does not exist in each occurrence, or is an alkylene group, alkenylene group, ynylene group, heteroalkylene group, alkenylene group, arylene group, or a combination thereof; L 2 L 3 L 5 and L 6 It is either independent of any group that does not exist in each instance, or it is a alkylene group, alkenyl group, alynyl group, heteroalkyl group, alkenyl group, or alynyl group. L 4 Each time it appears, it is independently an alkylene, alkenylene, ynylene, heteroalkylene, alkenylene, or ynylene linkage; R 1 and R 2 Each of these can be independently H, OH, SH, alkyl, alkoxy, alkyl ether, heteroalkyl, -OP (=R) a (R) b )R c Or Q; R 3 Each time it appears, it is independently H, alkyl, or alkoxy; R 4 Each time it appears, it is independently OH, SH, or O. - S - OR d or SR d ; R 5 Each occurrence is independently either oxidized, thiolated, or absent; R 6 and R 7 Each time it appears, it is independently H, OH, or a halogen; R a Is it O or S? R b It is OH, SH, O - S - OR d or SR d ; R c It is OH, SH, O - S - OR d OL' SR d Alkyl, alkoxy, heteroalkyl, heteroalkoxy, alkyl ether, alkoxyalkyl ether, phosphate ester, thiophosphate ester, phosphoryl alkyl, thiophosphoryl alkyl, phosphoryl alkyl ether or thiophosphoryl ether; R d It is a counter ion; Q is independently a portion containing a reactive group or its protected form each time it appears, which is capable of forming a covalent bond with the complementary reactive group Q′ on the target portion; L' independently, each time it appears, is a linker containing a covalent bond with Q, a target portion, a linker containing a covalent bond with the target portion, a linker containing a covalent bond with the solid support, a linker containing a covalent bond with the solid support residue, a solid support residue, a linker containing a covalent bond with a nucleoside, or a linker containing a covalent bond with other compounds of structure (I). m and p are independent integers equal to or greater than 0 each time they appear; w is an integer equal to or greater than 1 each time it appears; and n is an integer equal to or greater than 1.

78. The compound of claim 77, wherein G is -C(=O)OH or -C(=O)H.

79. The compound according to any one of claims 77-78, wherein the compound of structure (I) has the following structure (IA) or (IB): or (IA) (IB).

80. Compounds having the following structure (III): , (III) in: M 1 Each time it appears, it is independently a portion containing either fluorescent or colored dye; M 2 Each occurrence is independently a portion containing either a biologically active component or a fluorescent or colored dye, provided that M 2 At least once, it appears as the biologically active component; L 1’ and L 7 It is either independent of or does not exist in each occurrence, or is an alkylene group, alkenylene group, ynylene group, heteroalkylene group, alkenylene group, arylene group, or a combination thereof; L 2 L 3 L 5 and L 6 It is either independent of any group that does not exist in each instance, or it is a alkylene group, alkenyl group, alynyl group, heteroalkyl group, alkenyl group, or alynyl group. L 4 Each time it appears, it is independently an alkylene, alkenylene, ynylene, heteroalkylene, alkenylene, or ynylene linkage; R 1 and R 2 Each of these can be independently H, OH, SH, alkyl, alkoxy, alkyl ether, heteroalkyl, -OP (=R) a (R) b )R c Or Q; R 3 Each time it appears, it is independently H, alkyl, or alkoxy; R 4 Each time it appears, it is independently OH, SH, or O. - S - OR d or SR d ; R 5 Each occurrence is independently either oxidized, thiolated, or absent; R 6 and R 7 Each time it appears, it is independently H, OH, or a halogen; R a Is it O or S? R b It is OH, SH, O - S - OR d or SR d ; R c It is OH, SH, O - S - OR d OL' SR d Alkyl, alkoxy, heteroalkyl, heteroalkoxy, alkyl ether, alkoxyalkyl ether, phosphate ester, thiophosphate ester, phosphoryl alkyl, thiophosphoryl alkyl, phosphoryl alkyl ether or thiophosphoryl ether; R d It is a counter ion; Q is independently a portion containing a reactive group or its protected form each time it appears, which is capable of forming a covalent bond with the complementary reactive group Q′ on the target portion; L' independently, each time it appears, is a linker containing a covalent bond with Q, a target portion, a linker containing a covalent bond with the target portion, a linker containing a covalent bond with the solid support, a linker containing a covalent bond with the solid support residue, a solid support residue, a linker containing a covalent bond with a nucleoside, or a linker containing a covalent bond with other compounds of structure (III). m and p are independent integers equal to or greater than 0 each time they appear; w is an integer equal to or greater than 1 each time it appears; and n is an integer equal to or greater than 1.

81. The method of claim 80, wherein L 1’ It contains one of the following structures: or .

82. The method of claim 80, wherein L 1’ It contains one of the following structures: ； ; ； ； ； ； ； ; ； ； ； ； ； or 。 83. The method of claim 80, wherein L 1’ It contains one of the following structures: ； ； ； ； ； ； ; ； ； ； ; ； ； ; ; ; ; ; ; ; ; or 。 84. The compound according to any one of claims 80-83, wherein M 2 It has one of the following structures: ； ； ； or 。 85. The compound according to any one of claims 80-84, wherein the compound of structure (III) is selected from the compounds in Table 3.

86. The compound according to any one of claims 77-85, wherein L 7 It contains one of the following structures: ； ； ; ； ； ； ； ； ； ； ； , ; ; ; ; ; or .

87. The compound of claim 86, wherein L 7 yes: 。 88. The compound according to any one of claims 77-85, wherein L 7 It contains one of the following structures: ； ； ; or 。 89. The method according to any one of claims 77-86, wherein L 7 It contains one of the following structures: ； ； ; ; ; or 。 90. The compound according to any one of claims 77-89, wherein M 1 It has one of the following structures: ； ； ； ； ; ; or .

91. The compound according to any one of claims 77-90, wherein M 1 It has one of the following structures: 。 92. Compounds having the following structure (V): , (V) in: G, each time it appears, is independently a part containing a reactive group, a group that can be converted into a reactive group, or a protected analogue thereof, which is capable of forming a covalent bond with an amino group; G 1 Each time it appears, it is independently a portion containing a reactive group or its protected analogue, which is capable of reacting with M. 1 The complementary reactive groups in G form covalent bonds, provided that G 1 It does not react with amine groups; L 1 and L 7’ Each time it appears independently, it is an alkylene group, an alkenylene group, an ynylene group, a heteroalkylene group, an alkenylene group, an arylene group, or a heteroaryl linker or a combination thereof; L 2 L 3 L 5 and L 6 It is either independent of any group that does not exist in each instance, or it is a alkylene group, alkenyl group, alynyl group, heteroalkyl group, alkenyl group, or alynyl group. L 4 Each time it appears, it is independently an alkylene, alkenylene, ynylene, heteroalkylene, alkenylene, or ynylene linkage; R 1 and R 2 Each of these can be independently H, OH, SH, alkyl, alkoxy, alkyl ether, heteroalkyl, -OP (=R) a (R) b )R c Or Q; R 3 Each time it appears, it is independently H, alkyl, or alkoxy; R 4 Each time it appears, it is independently OH, SH, or O. - S - OR d or SR d ; R 5 Each occurrence is independently either oxidized, thiolated, or absent; R 6 and R 7 Each time it appears, it is independently H, OH, or a halogen; R 8 It is H or alkyl; R a Is it O or S? R b It is OH, SH, O - S - OR d or SR d ; R c It is OH, SH, O - S - OR d OL' SR d Alkyl, alkoxy, heteroalkyl, heteroalkoxy, alkyl ether, alkoxyalkyl ether, phosphate ester, thiophosphate ester, phosphoryl alkyl, thiophosphoryl alkyl, phosphoryl alkyl ether or thiophosphoryl ether; R d It is a counter ion; Q is independently a portion containing a reactive group or its protected form each time it appears, which is capable of forming a covalent bond with the complementary reactive group Q′ on the target portion; L' independently, each time it appears, is a linker containing a covalent bond with Q, a target portion, a linker containing a covalent bond with the target portion, a linker containing a covalent bond with the solid support, a linker containing a covalent bond with the solid support residue, a solid support residue, a linker containing a covalent bond with a nucleoside, or a linker containing a covalent bond with other compounds of structure (V). m and p are independent integers equal to or greater than 0 each time they appear; w is an integer equal to or greater than 1 each time it appears; and n is an integer equal to or greater than 1.

93. The compound of claim 92, wherein G 1 It has one of the following structures: ; or .

94. The compound of claim 92, wherein G 1 It has one of the following structures: ; or .

95. The compound of claim 92, wherein L7'-G 1 It has one of the following structures: ; ; ; ; or .

96. The compound according to any one of claims 92-95, wherein G is -C(=O)OH or -C(=O)H.

97. The compound of claim 96, wherein the compound of structure (V) has the following structure (VA) or (VB): or (VA) (VB).

98. The compound according to any one of claims 77-97, wherein L 4 It has the following structure: in: z is an integer from 1 to 100; and Indicates the bond with the adjacent phosphorus atom.

99. The compound of claim 97, wherein z is an integer from 3 to 8.

100. The compound of claim 97, wherein z is an integer from 22 to 26.

101. The compound according to any one of claims 77-100, wherein the compound of structure (I) has the following structure (IC) or (ID): or (IC) , (ID) The compounds with structure (III) have the following structure (IIIB): ;and (IIIB) Compounds with structure (V) have the following structures (VC) or V(D): or (IVC) , (IVD) Where m is an integer from 0 to 10.

102. The compound of claim 101, wherein m is 1.

103. The compound according to any one of claims 77-102, wherein L 2 L 3 L 5 and L 6 It is either absent independently each time it appears, or it is either a C1-C6 alkylene group, a C2-C6 alkenyl group, or a C2-C6 alkynyl group.

104. The compound of claim 103, wherein the compound of structure (I) has the following structure (IE) or (IF): or (IE) ； (IF) Compounds with structure (III) have the following structure (IIIC): ;and (IIIC) Compounds with structure (V) have the following structures (VE) or (VF): or (VE) , (VF) Where y 1 y 2 y 3 and y 4 Each occurrence is an integer from 0 to 6.

105. The compound of claim 104, wherein y 1 y 2 y 3 and y 4 It is either 0 or 1 each time it appears.

106. The compound according to any one of claims 77-79 and 92-105, wherein L 1 It has one of the following structures: or .

107. The compound according to any one of claims 77-79 and 92-106, wherein L 1 It has one of the following structures: ； ； ； ; ; ; ; ; ; ; ; ; ; or .

108. The compound according to any one of claims 77-79 and 92-107, wherein L 1 It has one of the following structures: ； ； ； ； ； ； ; ； ； ； ； ； ； ； ； ； ； ； ; ; ; or .

109. The compound according to any one of claims 77-108, wherein R 3 It is H.

110. The compound according to any one of claims 77-109, wherein R 4 It is O - OH or OR d .

111. The compound according to any one of claims 77-109, wherein R 5 It is oxygenation.

112. The compound according to any one of claims 77-111, wherein R 6 and R 7 It is H.

113. The compound according to any one of claims 77-112, wherein R 1 Or R 2 One of them is -OP(=R) a (R) b )R c And R 1 Or R 2 The other one is Q.

114. The compound of claim 113, wherein Q comprises a nucleophilic reactive group, an electrophilic reactive group, or a cycloaddition reactive group.

115. The compound of claim 114, wherein Q comprises a thiol group, a disulfide, an activated ester, an isothiocyanate, an azide, an alkyne, an olefin, a diene, a dienophile, an acyl halide, a sulfonyl halide, a phosphine, an α-haloamide, biotin, an amino group, or a maleimide functional group.

116. The compound of claim 115, wherein the activated ester is an N-succinimide ester, an imine ester, or a polyfluorophenyl ester.

117. The compound of claim 115, wherein the azide is an alkyl azide or an acyl azide.

118. The compound according to any one of claims 113-117, wherein Q is selected from the portion of Table 1.

119. The compound according to any one of claims 77-118, wherein R 1 and R 2 Each is independently -OP(=R) a (R) b )R c .

120. The compound according to any one of claims 113-119, wherein R c She is an office lady (OL).

121. The compound of claim 120, wherein L' is the target moiety.

122. The compound of claim 120, wherein L' is a linker to the target moiety, a linker to the analyte molecule, or a linker to the solid support, said linker comprising an alkylene oxide or a phosphate diester moiety or a combination thereof.

123. The compound of claim 120, wherein L' has the following structure: ， in: m'' and n'' are independent integers from 1 to 10; R e It is H, an electron pair, or a counter ion; and L'' represents the target portion or a connection to the target portion.

124. The compound of claim 123, wherein n'' is 6.

125. The compound of any one of claims 121-124, wherein the targeting portion is an antibody or a cell surface receptor antagonist.

126. The compound of claim 125, wherein the antibody or cell surface receptor antagonist is an epidermal growth factor receptor (EGFR) inhibitor, a hepatocyte growth factor receptor (HGFR) inhibitor, an insulin-like growth factor receptor (IGFR) inhibitor, a folic acid or MET inhibitor.

127. The compound of claim 125, wherein the targeting portion is a monoclonal antibody.

128. The compound of claim 122, wherein the analyte molecule is a nucleic acid, an amino acid, or a polymer thereof.

129. The compound of claim 122, wherein the analyte molecule is an enzyme, receptor, receptor ligand, glycoprotein, aptamer, or prion.

130. The compound of claim 122, wherein the solid support is a polymer bead or a non-polymer bead.

131. The compound according to any one of claims 113-130, wherein R 1 Or R 2 It has one of the following structures: ; ; ; ; ; ; ; ; or .

132. The compound according to any one of claims 113-130, wherein R 1 Or R 2 It has the following structure: or .

133. The compound according to any one of claims 92-132, wherein the compound of structure (V) is selected from the compounds in Table 4.

134. A method for preparing a compound of structure (I) of claim 1, comprising mixing a compound of structure (V) of claim 92 with M 1 G 1’ The reaction, in which G 1’ Includes G 1 Functional groups with complementary reactivity.

135. The method of claim 134, wherein G 1’ It is -C(=O)OH.

Images